To see the other types of publications on this topic, follow the link: Shona Names.
Journal articles on the topic 'Shona Names'
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the top 50 journal articles for your research on the topic 'Shona Names.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Browse journal articles on a wide variety of disciplines and organise your bibliography correctly.
1
Mheta, Gift, Esau Mangoya, and Livingstone Makondo. "Shona personal names of spiritual significance." Nomina Africana: Journal of African Onomastics 31, no. 1 (November 30, 2017): 01–09. http://dx.doi.org/10.2989/na.2017.31.1.1.1304.
Full text
2
Mhute, Isaac. "Are Names Really Empty: A Look into Shona Dog Names." European Scientific Journal, ESJ 12, no. 11 (April 27, 2016): 312. http://dx.doi.org/10.19044/esj.2016.v12n11p312.
Full textAbstract:
Following the popular Shakespearean saying that there is nothing in a name, the paper ventures into the linguistic area of onomastics focusing on uncovering the exact truth behind names in societies. It takes the Shona people’s dog names as a case study and reports on results from a qualitative research that used observations and open ended interviews as data collection techniques. Purposive sampling was employed and saw most of the data coming from districts in Masvingo province such as Zaka, Masvingo and Ndanga. Data were either recorded using a Samsung phone or recorded in the researcher’s notebook before being qualitatively analysed and interpreted. It came out that, though in certain situations names are just tags meant to enhance identification of certain dogs just like the Biblical names that were given to most African children following the coming of the former white masters, almost every Shona dog name has a story behind it.
3
Makondo, Livingstone. "Ethnicity and Matriarchal Protest: A Case of Dialoguing Shona Personal Names." Names 56, no. 1 (March 2008): 10–18. http://dx.doi.org/10.1179/175622708x282893.
Full text
4
Jenjekwa, Vincent. "Post-2000 revitalisation of Shona place names in Zimbabwe: recovering voices from the past." Nomina Africana: Journal of African Onomastics 35, no. 1 (January 2021): 1–14. http://dx.doi.org/10.2989/na.2021.35.1.1.1356.
Full text
5
Matiza, Vimbai Moreblessing, and Limukani T. Dube. "The Cultural and Historical Significance of Kalanga Place Names in Midlands Province of Zimbabwe." Journal of Law and Social Sciences 4, no. 2 (June 30, 2020): 21–28. http://dx.doi.org/10.53974/unza.jlss.4.2.470.
Full textAbstract:
The discipline of onomastics is still at its infancy yet it constitutes a very important aspect of the day to day survival of a people in the society. Naming is part of oral tradition in African societies, people were never used to write and record things but rather their names. This means that names are a historical record that would carry some aspects of a people's way of life which include their history, beliefs and customs among others. On the same note, Midlands Province constitute of people from different backgrounds mainly Shona and Ndebele. Of interest to this research is the presence of the Kalanga people through some toponyms that are found in the area. In light of this view, this study therefore seeks to identify and unlock the culture and history embedded in these names by looking at the significance of Kalanga place names in Midlands Province. The study argues that place names or toponyms of any people carry with them a history, meaning and significance to particular people that name the places, thus studying the place names in this community can be a valuable tool of unpacking the history surrounding the Kalanga people in Midlands Province in Zimbabwe. Guided by the Afrocentric paradigm, specifically nommoic creativity tenant, the study seeks to explore the cultural and historical significance of Kalanga toponyms in Midlands Province.
6
Chimhundu, Herbert. "Early Missionaries and the Ethnolinguistic Factor During the ‘Invention of Tribalism’ in Zimbabwe." Journal of African History 33, no. 1 (March 1992): 87–109. http://dx.doi.org/10.1017/s0021853700031868.
Full textAbstract:
There is evidence from across the disciplines that at least some of the contemporary regional names of African tribes, dialects and languages are fairly recent inventions in historical terms. This article offers some evidence from Zimbabwe to show that missionary linguistic politics were an important factor in this process. The South African linguist Clement Doke was brought in to resolve conflicts about the orthography of Shona. His Report on the Unification of the Shona Dialects (1931) shows how the language politics of the Christian denominations, which were also the factions within the umbrella organization the Southern Rhodesia Missionary Conference, contributed quite significantly to the creation and promotion of Zezuru, Karanga and Manyika as the main groupings of dialects in the central area which Doke later accommodated in a unified orthography of a unified language that was given the name Shona. While vocabulary from Ndau was to be incorporated, words from the Korekore group in the north were to be discouraged, and Kalanga in the West was allowed to be subsumed under Ndebele.Writing about sixty years later, Ranger focusses more closely on the Manyika and takes his discussion to the 1940s, but he also mentions that the Rhodesian Front government of the 1960s and 1970s deliberately incited tribalism between the Shona and the Ndebele, while at the same time magnifying the differences between the regional divisions of the Shona, which were, in turn, played against one another as constituent clans. It would appear then that, for the indigenous Africans, the price of Christianity, Western education and a new perception of language unity was the creation of regional ethnic identities that were at least potentially antagonistic and open to political manipulation.Through many decades of rather unnecessary intellectual justification, and as a result of the collective colonial experience through the churches, the schools and the workplaces, these imposed identities, and the myths and sentiments that are associated with them, have become fixed in the collective mind of Africa, and the modern nation states of the continent now seem to be stuck with them. Missionaries played a very significant role in creating this scenario because they were mainly responsible for fixing the ethnolinguistic maps of the African colonies during the early phase of European occupation. To a significant degree, these maps have remained intact and have continued to influence African research scholarship.
7
Mapara, Jacob, and Godwin Makaudze. "The interface between toponyms, hydronyms and geography: The case of selected Shona names from three provinces in Zimbabwe." South African Journal of African Languages 36, no. 2 (December 9, 2016): 243–49. http://dx.doi.org/10.1080/02572117.2016.1252028.
Full text
8
Beach, D. N. "An Innocent Woman, Unjustly Accused? Charwe, Medium of the Nehanda Mhondoro Spirit, and the 1896–97 Central Shona Rising in Zimbabwe." History in Africa 25 (1998): 27–54. http://dx.doi.org/10.2307/3172179.
Full textAbstract:
The rising of the Ndebele and southwestern and central Shona people against colonial rule in the 1890s has become one of the classic cases of such resistance. Yet, since the independence of Zimbabwe in 1980, very little fresh research has been carried out on the subject. This paper re-examines the role of Shona religious authorities in the rising, especially that of the medium of the Nehanda spirit of the Mazowe valley in the central Shona area. In just over a century, the figure of “Mbuya Nehanda” has become the best-known popular symbol of resistance to colonial rule in modern Zimbabwe. She has been commemorated since 1980 in statues, street names, a hospital, posters, songs, novels, and poems, and is soon to be the subject of a full-length feature film. This paper examines the historical basis behind the legend.This legend runs as follows: the historical “Nehanda” was supposed to have been the daughter of the founding ancestor of the Mutapa dynasty, who lived in the fifteenth century. Her ritual incest with her brother Matope gave supernatural sanction to the power of the Mutapa state. After her death, she became a mhondoro spirit, and this spirit possessed a number of mediums (masvikiro, singular svikiro). During periods of possession by the spirit, the svikiro was regarded as speaking with the voice and personality of the original Nehanda and not with her own. In the last part of the nineteenth century one medium, Charwe, was responsible for the organization of resistance to the government of the British South Africa Company and the settlers in the Mazowe valley, and in particular for the killing of H.H. Pollard, Kunyaira, the extremely oppressive Native Commissioner of the area. This resistance began in June 1896, and from then until her capture in late 1897 the Nehanda medium was a major factor in the war. Tried and sentenced to death in March 1898, she refused to convert to Christianity and struggled right up to the moment when she was hanged.
9
Makaudze, Godwin. "The significance of selected characters' personal and family names in the Shona novels, Pfumo Reropa and Mubairo." Nomina Africana: Journal of African Onomastics 34, no. 1 (January 2020): 13–20. http://dx.doi.org/10.2989/na.2020.34.1.2.1351.
Full text
10
Essenova, K. "RESEARCH OF ACADEMICIAN SHORA SARYBAYEVA ON PRESS LANGUAGE." BULLETIN Series of Philological Sciences 73, no. 3 (July 15, 2020): 32–35. http://dx.doi.org/10.51889/2020-3.1728-7804.05.
Full textAbstract:
The article analyzes the scientific achievements of academician Shora Sarybaev about the language of the press. It deals with opinions on violations of literary norms in the use of incorrect translations, and the use of many variant words and non-literary slang in the Kazakh media are presented. Scientist Sh. Sarybaev analyzes the conclusions about the dynamics of the penetration of new words into the literary language in 1920-30. A number of features of the language of newspapers and magazines of the period of independence are discussed in the article of the scientist Sh. Sarybaev "New applications of the language of the media", written in 1999. The language of the press of the society that supported the only ideology of the Soviet era was written in a literary language, with strict stylistic and spelling norms. However, the scientist Shora Sarybaev noted that there are difficulties with the translation of new foreign words in the media. Difficulties arose to give the correct translation of the neologisms. Given the proliferation of new use cases in periodicals, the scientist considered it prudent to use alternative names in newspapers and magazines along with the keyword before Terminсom approval.
11
Tyshchenko, K. "THE MEMORY ABOUT BYZANTINE JACOBITES AND MELEKITES: 1500 YEARS IN PLACE NAMES IN UKRAINE." Bulletin of Taras Shevchenko National University of Kyiv. History, no. 150 (2021): 68–77. http://dx.doi.org/10.17721/1728-2640.2021.150.11.
Full textAbstract:
The linguistic evidence of contacts of the Early Christian time between the Middle Dnieper region and Syria is highlighted. Found Aramaic Prototypes of such loanwords as Ukr. han'ba, shana, lylyk-nichka, norytsia, spyzharnia, saraka. It is shown that they are connected by means of realities designated by them with the wandering Syrian Monks-Sarakots (IV-VIth c.). These contacts get specific localization in persisted toponyms in Ukraine Sorokotychi, Rakhvalivka, Trypillia, Khalepya, Zoziv, Oskolonivka, in Ukrainian family names Sorokotiah, Rukhaylo, Zoza, Skolonets' etc. Now, the repeated spatial closeness to the already studied toponyms of a new "pattern" of names is revealed. Their bases are Sever(yn)-, Ladyh/zh-, Nastas/sh-, Melekh/sh-. The very fact of their relationship makes probable their prototypes: Syr. ܣܸܘܸܪܘܿܣ Severus 'Cyrus the Great' (from Antiochia), ܠܐܕܝܩܝܐ lâdîqîyâ, Λαοδίκεια, اللاذقية al-Lādhiq(iye) 'Laodicea in Syria', Ἀναστάσιος 'Anastasius I', ܡܠܟܝܐ malkoyo Μελχίτοι 'melekites'. All these concepts are connected not just by an epoch, but by a specific historical event, the Council of 512.
12
Tyshchenko, K. "THE MEMORY ABOUT BYZANTINE JACOBITES AND MELEKITES: 1500 YEARS IN PLACE NAMES IN UKRAINE." Bulletin of Taras Shevchenko National University of Kyiv. History, no. 150 (2021): 68–77. http://dx.doi.org/10.17721/1728-2640.2021.150.11.
Full textAbstract:
The linguistic evidence of contacts of the Early Christian time between the Middle Dnieper region and Syria is highlighted. Found Aramaic Prototypes of such loanwords as Ukr. han'ba, shana, lylyk-nichka, norytsia, spyzharnia, saraka. It is shown that they are connected by means of realities designated by them with the wandering Syrian Monks-Sarakots (IV-VIth c.). These contacts get specific localization in persisted toponyms in Ukraine Sorokotychi, Rakhvalivka, Trypillia, Khalepya, Zoziv, Oskolonivka, in Ukrainian family names Sorokotiah, Rukhaylo, Zoza, Skolonets' etc. Now, the repeated spatial closeness to the already studied toponyms of a new "pattern" of names is revealed. Their bases are Sever(yn)-, Ladyh/zh-, Nastas/sh-, Melekh/sh-. The very fact of their relationship makes probable their prototypes: Syr. ܣܸܘܸܪܘܿܣ Severus 'Cyrus the Great' (from Antiochia), ܠܐܕܝܩܝܐ lâdîqîyâ, Λαοδίκεια, اللاذقية al-Lādhiq(iye) 'Laodicea in Syria', Ἀναστάσιος 'Anastasius I', ܡܠܟܝܐ malkoyo Μελχίτοι 'melekites'. All these concepts are connected not just by an epoch, but by a specific historical event, the Council of 512.
13
Friday, Colleen, and John Derek Scasta. "Eastern Shoshone and Northern Arapaho Traditional Ecological Knowledge (TEK) and Ethnobotany for Wind River Reservation Rangelands." Ethnobiology Letters 11, no. 1 (May 11, 2020): 14–24. http://dx.doi.org/10.14237/ebl.11.1.2020.1654.
Full textAbstract:
The need to affirm and revitalize cultural knowledge of native plant communities is impera-tive for Indigenous people. This ethnobotanical study documents Traditional Ecological Knowledge (TEK) structured from an Indigenous paradigm by exploring the connection be-tween plants collected in two high-elevation basins and tribal members on the Wind River Indian Reservation (WRIR). We sought to qualitatively understand the plant resources by looking through the lens of Indigenous language and perspectives. Existing names of the ba-sin plants in both the Eastern Shoshone and Northern Arapaho languages were compiled through an ethnobotanical literature review, seven in-person interviews with Eastern Sho-shone and Northern Arapaho tribal members, and attendance at language workshops. We documented 53 Eastern Shoshone and 44 Northern Arapaho plant names, respectively. His-torical impacts of past Federal Indian policy eras have shaped TEK as it currently exists within tribal communities. Both tribes used and had Indigenous names for Northern sweetgrass (Hierochloe hirta ssp. hirta), bitterroot (Lewisia rediviva), junipers (Juniperus ssp.), and bear-berry or Kinnikinnick (Arctostaphylos uva-ursi). The resiliency of TEK is attributed to the perse-verance of Indigenous people continuing to practice and teach traditions. The historical con-text specific to both the Eastern Shoshone and Northern Arapaho tribes and their languages are important for enhancing our current understanding of the ethnobotanical TEK of plants on the WRIR. Recognizing the value of ethnobotanical TEK and incorporating it into natural resource management plans and decisions can bridge diverse perspectives on land use for meaningful collaboration with tribal communities.
14
TAKESI, OKADOME. "SIMPLE FLAT LANGUAGES: A LEARNABLE CLASS IN THE LIMIT FROM POSITIVE DATA." International Journal of Foundations of Computer Science 10, no. 04 (December 1999): 483–501. http://dx.doi.org/10.1142/s0129054199000344.
Full textAbstract:
The class of simple flat languages defined here is shonw to be learnable in the limit from positive data. In particular, its subclass named k-consecutive, which covers a part of the class of context-sensitive languages not belonging to the class of context-free languages, is polynomial-time learnable in the limit from positive data. The class of "disjunct" unions of simple flat languages is a nontrivial example which is learnable in the limit from positive data, but does not have Wright's finite elasticity. The learning algorithm presented here for identifying the subclasses of flat languages consists essentially of identifying an arithmetic progression in the limit from positive examples using Euclidean algorithm of mutual division.
15
Wang, Lijuan, Jianfei Qian, Yong Lu, Haiyan Li, Sungyoul Hong, Hanying Bao, Donghua He, et al. "Expression of B7-H1 in Mantle Cell Lymphoma Leads to Inhibition of T Cell Response to Tumor Cells." Blood 118, no. 21 (November 18, 2011): 2643. http://dx.doi.org/10.1182/blood.v118.21.2643.2643.
Full textAbstract:
Abstract Abstract 2643 Mantle cell lymphoma (MCL) is a unique subtype of incurable B-cell, non-Hodgkin lymphoma, and its overall survival currently remains only 4–5 years. Management of relapsed or refractory MCL patients is still challenging. Immunotherapy may provide an alternative treatment for patients with MCL. Recent studies demonstrated that PD-1/B7-H1 signaling plays a crucial role in T-cell regulation in various immune responses and is involved in peripheral tolerance, autoimmunity, infection, and antitumor immunity. In the present study, we examined whether B7-H1 plays an important role in immune evasion in MCL. We demonstrated that B7-H1 gene and protein were expressed in most MCL cell lines and primary MCL cells from all patients examined. CD3+ T cells were cultured with irradiated MCL cell lines and primary cells, which were pre-incubated with or without anti-B7-H1 monoclonal antibody or control antibody. The presence of anti-B7-H1 blocking antibody, but not control antibody, increased CD3+ T cell proliferation. We confirmed the effect of B7-H1 in suppression of T cell proliferation by knockdown of B7-H1 gene expression using B7-H1 specific and non-specific control shRNA lentiviral particles. Upon transfection, the B7-H1-specific shRNA reduced both B7-H1 gene and surface protein expression, while the control shRNA did not. The B7-H1 specific shRNA, but not control shRNA, augmented CD3+ T cell proliferation. To address whether B7-H1 contributed to the suppression of host antitumor immunity in MCL, allogeneic CD3+ T cells isolated from normal donors were cocultured with irradiated MCL cell line SP53, control shRNA SP53 (SP53-ctl), or B7-H1 targeted shRNA SP53 (SP53-kd), respectively. After 7 days of coculture, CD3+ T cells were harvested and restimulated with newly irradiated SP53, SP53-ctl or SP53-kd cells. After at least 3 repeated cycles of in vitro restimulation, three cytotoxic T lymphocyte (CTL) lines were generated, and named CTL-SP53, CTL-SP53-ctl and CTL-SP53-kd. CTL-SP53-kd showed increased killing of target cells as compared with CTL-SP53 (P <.01) or CTL-SP53-ctl (P <.01). We further showed that B7-H1 targeted shRNA MCL cell line (SP53-kd cells) displayed more specific lysis than SP53 (P <.01) or SP53-trl (P <.01). When SP53 cells were pre-incubated with a blocking anti-B7-H1 monoclonal antibody, it also showed more specific lysis as compared to the control antibody pre-treated cells. In these experiments, purified autologous blood B cells and PBMCs were used as target cells to demonstrate whether T cell lines were cytolytic to normal cells. The results showed that the three CTL cell lines did not kill B cells or PBMCs. Intracellular cytokine staining and ELISA assay demonstrated that T-cell lines express IFN-γ, but not IL-4, IL-6, IL-10 or IL-17, and were thus type I T cells. Moreover, T cell lines stimulated by SP53-kd cells express more IFN-γ than SP53 and SP53-ctl. The T cells also expressed CD45RO, CD28, CD44, but not CD45RA, CD27 or CD62L, indicating that they were memory effector cells. In conclusion, B7-H1 expression may be involved in immune evasion mechanism of MCL. Therefore, B7-H1 may be a promising target for immunotherapy in MCL. Disclosures: No relevant conflicts of interest to declare.
16
Sun, Jingnan, Wei Li, Yunpeng Sun, Dehai Yu, Xue Wen, Hong Wang, Jiuwei Cui, Guanjun Wang, Andrew R. Hoffman, and Ji-Fan Hu. "A novel antisense long noncoding RNA within the IGF1R gene locus is imprinted in hematopoietic malignancies." Nucleic Acids Research 42, no. 15 (August 4, 2014): 9588–601. http://dx.doi.org/10.1093/nar/gku549.
Full textAbstract:
AbstractDysregulation of the insulin-like growth factor type I receptor (IGF1R) has been implicated in the progression and therapeutic resistance of malignancies. In acute myeloid leukemia (AML) cells, IGF1R is one of the most abundantly phosphorylated receptor tyrosine kinases, promoting cell growth through the PI3K/Akt signaling pathway. However, little is known regarding the molecular mechanisms underlying IGF1R gene dysregulation in cancer. We discovered a novel intragenic long noncoding RNA (lncRNA) within the IGF1R locus, named IRAIN, which is transcribed in an antisense direction from an intronic promoter. The IRAIN lncRNA was expressed exclusively from the paternal allele, with the maternal counterpart being silenced. Using both reverse transcription-associated trap and chromatin conformation capture assays, we demonstrate that this lncRNA interacts with chromatin DNA and is involved in the formation of an intrachromosomal enhancer/promoter loop. Knockdown of IRAIN lncRNA with shRNA abolishes this intrachromosomal interaction. In addition, IRAIN was downregulated both in leukemia cell lines and in blood obtained from high-risk AML patients. These data identify IRAIN as a new imprinted lncRNA that is involved in long-range DNA interactions.
17
Иерей, Тимофей, Timofe Ierey, Екатерина Топалова, Ekaterina Topalova, Наталья Шафажинская, and Natalya Shafazhinskaya. "Continuity of Church Service and Orthodox Cultural Tradition at the Turn of the Soviet and Post-Soviet Periods." Scientific Research and Development. Socio-Humanitarian Research and Technology 8, no. 1 (March 27, 2019): 51–60. http://dx.doi.org/10.12737/article_5c8f500d7b2c32.53331275.
Full textAbstract:
The article is devoted to the disclosure of the content of the creative activities of representatives of the Russian Orthodox Church in a crucial period of spiritual service at the turn of the XX–XXI centuries. During this period, the continuity of religious enlightenment culture was interrupted and needed restoration in order to preserve church culture and art for future generations. In the twentieth century — the century of the New Martyrs and Confessors of Russia, when persecution and oppression in varying degrees affected the fate of every Orthodox Christian, preserved and survived examples of theological and spiritual educational work they represent a special value, which should be available for the study of modern citizens of Russia and, above all, young people. Spiritual mentors and elders of the past century, who have undergone persecution, but survived and managed to directly convey to their spiritual children their own inner experience of God communion and an example of asceticism — the great wealth of not only domestic, but also world spiritual culture. In the second half of the twentieth and early twenty-first centuries, the names of many representatives of the Russian Orthodox Church, true devotees of grace, shone with their spiritual experience, wise mentorship and literary works helping thousands of people of different classes to gain faith and the true meaning of earthly existence.
18
Song, Lei, Fei Zhang, Rui Zhou, Jun Xiao, Lei He, and Fei Dai. "hCTLA4-Gene-Modified Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMMSCs) Maintain POSTN Secretion to Enhance the Migration Capability of Allogeneic hBMMSCs through the Integrin αvβ3/FAK/ERK Signaling Pathway." Stem Cells International 2020 (March 24, 2020): 1–12. http://dx.doi.org/10.1155/2020/3608284.
Full textAbstract:
Cytotoxic T-lymphocyte-associated protein 4- (CTLA4-) modified human bone marrow-derived mesenchymal stem cells (hBMMSCs) might be promising seed cells for bone tissue engineering. However, the underlying mechanism is not clear. In the present study, we investigated whether CTLA4-modified hBMMSCs are involved in the migration of allogeneic hBMMSCs (allo-hBMMSCs) by maintaining POSTN secretion. hBMMSCs were isolated from different groups, named hBMMSCs and allo-hBMMSCs. hBMMSCs that were infected with the negative control (NC), empty adenovirus- or recombinant adenovirus-expressing CTLA4, POSTN, or CTLA4 plus the shRNA of POSTN were named NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs, respectively. They were then cocultured with PBMCs in a 1 : 5 ratio with 2.5 μg/mL phytohemagglutinin (PHA). The coculture supernatant was collected to treat allo-hBMMSCs with anti-integrin αvβ3 IgG, or negative control IgG, as a control. Following this, ELISA, Transwell assays, wound healing assays, and western blotting were performed. We found that the POSTN level was higher in the culture supernatant of CTLA4- and POSTN-modified hBMMSCs than in NC hBMMSCs cocultured with PBMCs treated with PHA. The migration capability of allo-hBMMSCs was enhanced, and the integrin αvβ3/FAK/ERK signaling pathway in allo-hBMMSCs was activated by the culture supernatant of CTLA4- and POSTN-modified hBMMSCs cocultured with PBMCs treated with PHA. Additionally, these induced effects can be weakened by POSTN knockdown, and the migration capability of allo-hBMMSCs was blocked by anti-integrin αvβ3 IgG. In conclusion, hCTLA4-gene-modified hBMMSCs maintain POSTN secretion to enhance the migration capability of allogeneic hBMMSCs through the integrin αvβ3/FAK/ERK signaling pathway in the T cell immune activation environment.
19
Kook, Hoon, Nakwon Choe, Hae-Jin Kee, Sung-Mi Kim, Ji-Young Kim, Dong-Kyun Han, Hee-Jo Baek, et al. "Identification and Characterization of Novel SET Domain-Containing Histone Lysine Methyltransferase and Implications of Histone Methylation in Acute Leukemia." Blood 108, no. 11 (November 1, 2006): 2227. http://dx.doi.org/10.1182/blood.v108.11.2227.2227.
Full textAbstract:
Abstract Evolutionally conserved SET domains, which methylate histone lysine residues, and thereby methylation of histones have been implicated in diverse malignancies including leukemias or multiple myelomas. Here we describe novel SET domain-containing proteins with histone methyltransferase (HMTase) activity and their characteristics. Using bioinformatics for homology screening, SET-domain containing proteins named WHISTLE, WHSC1-like 1 isoform 9 with methyltransferase activity to lysine, and RE-IIBP, interleukin-5 response element II binding protein, were identified. By mass spectrometric and immunoblot analysis, we demonstrated that WHISTLE dimethylates H9K4 and di-, and tri-methylates H3K27, while RE-IIBP methylates only H9K27. WHISTLE and RE-IIBP repressed transcription of the SV40 and IL-5 promoter activity and they recruited histone deacetylase. Chromatin immunoprecipitation analysis revealed that shRNA-mediated knockdown of RE-IIBP reduces histone methylation on the IL-5 promoter. Both proteins induce apoptosis in leukemic cells via caspase-3 activation. In acute lymphoblastic leukemia patients, the expression of RE-IIBP and WHISTLE was increased, which was accompanied with increase in the methylation histone 3. These data illustrate the important regulatory role of novel SET domain proteins with HMTase activity in transcriptional regulation and apoptosis, thereby pointing to the critical role in leukemogenesis.
20
Zhang, Li-qian, Su-qing Yang, Ying Wang, Qiao Fang, Xian-jun Chen, Hong-sheng Lu, and Ling-ping Zhao. "Long Noncoding RNA MIR4697HG Promotes Cell Growth and Metastasis in Human Ovarian Cancer." Analytical Cellular Pathology 2017 (2017): 1–9. http://dx.doi.org/10.1155/2017/8267863.
Full textAbstract:
Ovarian cancer is one of the three most common gynecological malignant tumors worldwide. The prognosis of patients suffering from this malignancy remains poor because of limited therapeutic strategies. Herein, we investigated the role of a long noncoding RNA named MIR4697 host gene (MIR4697HG) in the cell growth and metastasis of ovarian cancer. Results showed that the transcriptional level of MIR4697HG in cancerous tissues increased twofold compared with that in adjacent noncancerous tissues. MIR4697HG was differentially expressed in ovarian cancer cell lines, with the highest levels in OVCAR3 and SKOV3 cells. MIR4697HG knockdown by specific shRNA significantly inhibited cell proliferation and colony formation in both OVCAR3 and SKOC3 cells. Consistently, in a xenograft model of ovarian cancer, MIR4697HG depletion also significantly restricted tumor volumes and weights. Furthermore, MIR4697HG knockdown inhibited cell migration and invasion capacities. Invasion ability was inhibited by 58% in SKOV3 cells and 40% in OVCAR3 cells, and migration ability was inhibited by 73% in SKOV3 cells and 62% in OVCAR3 cells after MIR4697HG knockdown. MIR4697HG knockdown also caused a decrease in matrix metalloprotease-9, phosphorylated ERK, and phosphorylated AKT. These data suggested that MIR4697HG promoted ovarian cancer growth and metastasis. The aggressive role of MIR4697HG in ovarian cancer may be related to the ERK and AKT signaling pathways.
21
Ikeda, Marina, Akihiro Ito, Yuichi Sekine, and Masahiro Fujimuro. "UBE1a Suppresses Herpes Simplex Virus-1 Replication." Viruses 12, no. 12 (December 4, 2020): 1391. http://dx.doi.org/10.3390/v12121391.
Full textAbstract:
Herpes simplex virus-1 (HSV-1) is the causative agent of cold sores, keratitis, meningitis, and encephalitis. HSV-1-encoded ICP5, the major capsid protein, is essential for capsid assembly during viral replication. Ubiquitination is a post-translational modification that plays a critical role in the regulation of cellular events such as proteasomal degradation, protein trafficking, and the antiviral response and viral events such as the establishment of infection and viral replication. Ub-activating enzyme (E1, also named UBE1) is involved in the first step in the ubiquitination. However, it is still unknown whether UBE1 contributes to viral infection or the cellular antiviral response. Here, we found that UBE1a suppressed HSV-1 replication and contributed to the antiviral response. The UBE1a inhibitor PYR-41 increased HSV-1 production. Immunofluorescence analysis revealed that UBE1a highly expressing cells presented low ICP5 expression, and vice versa. UBE1a inhibition by PYR-41 and shRNA increased ICP5 expression in HSV-1-infected cells. UBE1a reduced and retarded ICP5 protein expression, without affecting transcription of ICP5 mRNA or degradation of ICP5 protein. Additionally, UBE1a interacted with ICP27, and both partially co-localized at the Hsc70 foci/virus-induced chaperone-enriched (VICE) domains. PYR-41 reduced the co-localization of UBE1a and ICP27. Thus, our findings provide insights into the mechanism of UBE1a in the cellular response to viral infection.
22
Lupu, Cristina, Hua Zhu, Narcis I. Popescu, Jonathan D. Wren, and Florea Lupu. "Novel protein ADTRP regulates TFPI expression and function in human endothelial cells in normal conditions and in response to androgen." Blood 118, no. 16 (October 20, 2011): 4463–71. http://dx.doi.org/10.1182/blood-2011-05-355370.
Full textAbstract:
Abstract Thrombosis and cardiovascular disease (CVD) represent major causes of morbidity and mortality. Low androgen correlates with higher incidence of CVD/thrombosis. Tissue Factor Pathway Inhibitor (TFPI) is the major inhibitor of tissue factor-factor VIIa (TF-FVIIa)–dependent FXa generation. Because endothelial cell (EC) dysfunction leading to vascular disease correlates with low EC-associated TFPI, we sought to identify mechanisms that regulate the natural expression of TFPI. Data mining of NCBI's GEO microarrays revealed strong coexpression between TFPI and the uncharacterized protein encoded by C6ORF105, which is predicted to be multispan, palmitoylated and androgen-responsive. We demonstrate that this protein regulates both the native and androgen-enhanced TFPI expression and activity in cultured ECs, and we named it androgen-dependent TFPI-regulating protein (ADTRP). We confirm ADTRP expression and colocalization with TFPI and caveolin-1 in ECs. ADTRP-shRNA reduces, while over-expression of ADTRP enhances, TFPI mRNA and activity and the colocalization of TF-FVIIa–FXa-TFPI with caveolin-1. Imaging and Triton X-114–extraction confirm TFPI and ADTRP association with lipid rafts/caveolae. Dihydrotestosterone up-regulates TFPI and ADTRP expression, and increases FXa inhibition by TFPI in an ADTRP- and caveolin-1-dependent manner. We conclude that the ADTRP-dependent up-regulation of TFPI expression and activity by androgen represents a novel mechanism of increasing the anticoagulant protection of the endothelium.
23
Ristic, Zorica, Simone M. R. Camargo, Elisa Romeo, Susana Bodoy, Joan Bertran, Manuel Palacin, Victoria Makrides, Esther M. Furrer, and François Verrey. "Neutral amino acid transport mediated by ortholog of imino acid transporter SIT1/SLC6A20 in opossum kidney cells." American Journal of Physiology-Renal Physiology 290, no. 4 (April 2006): F880—F887. http://dx.doi.org/10.1152/ajprenal.00319.2005.
Full textAbstract:
Most neutral l-amino acid acids are transported actively across the luminal brush-border membrane of small intestine and kidney proximal tubule epithelial cells by a Na+ cotransport system named B0 that has been recently molecularly identified (B0AT1, SLC6A19). We show here that the opossum kidney-derived cell line OK also displays a Na+-dependent B0-type neutral l-amino acid transport, although with a slightly differing substrate selectivity. We tested the hypothesis that one of the two B0AT1-related transporters, SLC6A18 (ortholog of orphan transporter XT2) or SLC6A20 (ortholog of the recently identified mammalian imino acid transporter SIT1), mediates this transport. Anti-sense RNA to OK SIT1 ( oSIT1) but not to OK XT2 ( oXT2) inhibited Na+-dependent neutral amino acid transport induced by OK mRNA injected in Xenopus laevis oocytes. Furthermore, inhibition of oSIT1 gene expression in OK cells by transfection of siRNA and expression of shRNA selectively reduced the Na+-dependent uptake of neutral l-amino acids. Finally, expression of OK cell oSIT1 cRNA in X. laevis oocytes induced besides the transport of the l-imino acid l-Pro also that of neutral l-amino acids. Taken together, the data indicate that in OK cells SIT1 (SLC6A20) is not only an apical imino acid transporter but also plays a major role as Na+-dependent neutral l-amino acid transporter. A similar double role could be envisaged for SIT1 in mammalian kidney proximal tubule and small intestine.
24
Serratto, Giulia M., Erika Pizzi, Luca Murru, Sara Mazzoleni, Silvia Pelucchi, Elena Marcello, Michele Mazzanti, Maria Passafaro, and Silvia Bassani. "The Epilepsy-Related Protein PCDH19 Regulates Tonic Inhibition, GABAAR Kinetics, and the Intrinsic Excitability of Hippocampal Neurons." Molecular Neurobiology 57, no. 12 (September 3, 2020): 5336–51. http://dx.doi.org/10.1007/s12035-020-02099-7.
Full textAbstract:
Abstract PCDH19 encodes for protocadherin-19 (PCDH19), a cell-adhesion molecule of the cadherin superfamily preferentially expressed in the brain. PCDH19 mutations cause a neurodevelopmental syndrome named epileptic encephalopathy, early infantile, 9 (EIEE9) characterized by seizures associated with cognitive and behavioral deficits. We recently reported that PCDH19 binds the alpha subunits of GABAA receptors (GABAARs), modulating their surface availability and miniature inhibitory postsynaptic currents (mIPSCs). Here, we investigated whether PCDH19 regulatory function on GABAARs extends to the extrasynaptic receptor pool that mediates tonic current. In fact, the latter shapes neuronal excitability and network properties at the base of information processing. By combining patch-clamp recordings in whole-cell and cell-attached configurations, we provided a functional characterization of primary hippocampal neurons from embryonic rats of either sex expressing a specific PCDH19 short hairpin (sh)RNA. We first demonstrated that PCDH19 downregulation reduces GABAAR-mediated tonic current, evaluated by current shift and baseline noise analysis. Next, by single-channel recordings, we showed that PCDH19 regulates GABAARs kinetics without altering their conductance. In particular, GABAARs of shRNA-expressing neurons preferentially exhibit brief openings at the expense of long ones, thus displaying a flickering behavior. Finally, we showed that PCDH19 downregulation reduces the rheobase and increases the frequency of action potential firing, thus indicating neuronal hyperexcitability. These findings establish PCDH19 as a critical determinant of GABAAR-mediated tonic transmission and GABAARs gating, and provide the first mechanistic insights into PCDH19-related hyperexcitability and comorbidities.
25
Schmidt, Eva, Jana Krosl, Jalila Chagraoui, Nadine Mayotte, Caroline Pabst, Tara McRae, and Guy Sauvageau. "Identification of Lats 1 As a Putative Tumor Suppressor in HoxA9/Meis Induced Leukemia." Blood 118, no. 21 (November 18, 2011): 2474. http://dx.doi.org/10.1182/blood.v118.21.2474.2474.
Full textAbstract:
Abstract Abstract 2474 Aberrant expression of Hox genes and their cofactors Pbx and Meis1 has been detected in approximately 50% of all human leukemias, and proteins interacting with these homeodomain factors could play a major role in leukemia development. Studies in drosophila showed that hth/MEIS directly interacts with YKI, a component of the Hippo signaling pathway (Peng HW et al., 2009). The core components of this pathway in the mammalian cells are the kinases MST 1 or 2 and LATS 1 or 2, and the downstream transcription cofactors WWTR1 and YAP (homologues of the drosophila Yki). The Hippo pathway has been proposed to play a tumor suppressive role in carcinoma development (Lu L et al. 2010), but little is known about its function in hematopoiesis and leukemia. To address this issue, we first determined the expression levels of the core Hippo pathway constituents in different subpopulations of primitive hematopoietic cells by quantitative RT-PCR. Hematopoietic stem cells (HSC) isolated from day 14.5 fetal liver (FL-HSC, phenotype: CD150+CD48-Lin-), or bone marrow from 3 and 4 week old mice (BM-HSC, phenotype: cKit+CD150+CD48-Lin-) express comparable levels of Lats 1/2 and Mst 1/2. FL-HSC, however, express approximately 3 fold higher levels of Wwtr1 and Yap than the BM-HSC. Expression of all core components of the Hippo pathway was also detected in the Hoxa9+Meis1-induced leukemia named FLA2 in which approximately 70% of cells represent leukemia stem cells (LSC). The role of this pathway in leukemia was assessed using the shRNA-mediated loss of function approach. For each core component, 5 different shRNAs were designed, and 2 achieving ≥40% decrease in the targeted transcript levels were selected for the in vivo experiments. Freshly isolated FLA2 leukemia cells were infected with recombinant retroviruses carrying the control shLuciferase or the targeting shRNA, and green fluorescent protein (GFP), and were transplanted into sub-lethally irradiated recipient mice. The proportions of shRNA transduced (GFP+) cells were determined at the time of transplantation (day 0), and at the time of sacrifice (day 18 ± 2). During this period, the proportions of shWwtr1(GFP+) cells to the leukemic cell populations decreased to 10–20% of the initial day 0 values. Conversely, the Lats1 knockdown leads to > 50% increase over the initial proportion of the GFP+ cells. The combined Lats1+Lats2 knockdown enhanced the competitiveness of the transduced cells compared shLuciferase controls. These significant results (p < 0.05, Mann-Whitney-Test) suggest that LATS kinases act as negative regulators of leukemic cell expansion. To exclude the possibility that this effect is limited to FLA2 leukemia we isolated the CD150+CD48-Lin- stem/progenitor cells from FL, co-infected them first with Hoxa9 and Meis1 cDNA carrying retroviruses, and then knocked down Wwtr1 or Lats1. Similar to observations in FLA2 leukemia model, Lats 1 depletion promoted ∼2-fold increase, and Wwtr 1 reduction >80% decrease in proportions of the transduced (GFP+) cells compared to their initial day 0 levels. Together, our observations suggest that LATS kinases act as negative modulators of Hox/Meis-induced leukemia and indicate a possibility for a specific targeting of the Hox/Meis-activated cellular pathways. Disclosures: No relevant conflicts of interest to declare.
26
Li, Shirong, Jing Fu, Xiaoming Xu, Shixian Deng, Markus Y. Mapara, Christophe Marcireau, and Suzanne Lentzsch. "Gck Kinase Activity Is Critical for RAS Mutated Myeloma - a Potential Treatment Approach for Targeting Specific Mutations." Blood 134, Supplement_1 (November 13, 2019): 1813. http://dx.doi.org/10.1182/blood-2019-129531.
Full textAbstract:
Introduction: Next-generation sequencing revealed frequent mutations of the RAS/mitogen-activated protein kinase (MAPK) pathway, with mutations in NRAS, KRAS or BRAF in up to 50% of newly diagnosed MM patients1. The majority of the NRAS, KRAS and BRAF mutations occur in hotspots causing constitutive activation of the corresponding pathways2. Given the upstream activator role of Germinal Center Kinase (GCK) in the MAPK pathway, GCK might be an attractive therapeutic target in MM. Indeed, we recently discovered the critical role of GCK, also named mitogen-activated protein kinase kinase kinase kinase 2 (MAP4K2), in MM cell survival and growth. Methods and Results: Our data show that GCK is higher expressed in RAS mutated MM cells compared to the wild type (WT) RAS MM cells. Silencing of GCK in RASmut MM cells (MM.1S and RPMI-8226) by using an inducible Tet-on-shGCK significantly decreased MM cell proliferation and induced cell death (Figure 1). In contrast, knockdown of GCK in RASwt MM cell lines (LP1 and U266) induced only modest inhibition of proliferation. The higher sensitivity to GCK knockdown in RASmut cells suggests that targeting GCK is especially effective in multiple myeloma which harbors RAS mutations. To exclude a potential off-target effect associated with GCK knockdown that led to the inhibition of MM proliferation, we set up an shGCK-resistant GCK allele (GCKshRNA-RES) by introducing mismatch mutations on the shRNA targeted sequences without changing the encoded amino acids. In the shGCK rescue experiment using GCKshRNA-RES we showed that all shRNA induced phenotypes (lack of growth, apoptosis and downstream effectors decrease) were corrected by the GCK resistant allele expression, ruling out the off-target hypothesis. Moreover, we expanded the in vivo studies of GCK knockdown on MM tumor progression. To monitor the tumor progression, we transduced MM.1S cells with firefly luciferase and established an inducible GCK knockdown system. Luciferase-expressing GCK inducible knockdown MM cells or non-targeting control shRNA (shCNTL) transduced MM cells were s.c. injected into SCID/Beige mice and the tumor progression was monitored by bioluminescence imaging. Doxycycline (for induction of shRNA) or vehicle treatment were started after the tumor was established on day 16 to induce shGCK and subsequently silence GCK expression. In contrast to the vehicle-treated MM.1S-Tet-on-shGCK or doxycycline-treated MM.1S-Tet-on-shCNTL tumors, doxycycline-treated animals bearing MM.1S-Tet-on-shGCK xenografts showed a significant inhibition (P<0.001) of tumor growth (Figure 2). Thus, GCK is also required for tumor growth. Lysine 45 is critical for GCK kinase activity. Point mutation of K45A will completely abolish its kinase activity. We introduced K45A mutation into GCKshRNA-RES (GCKshRNA-RESK45A→ shGCK resistant and kinase dead GCK). Tet-on-shGCK with GCKshRNA-RES or GCKshRNA-RESK45A were co-transduced in MM.1S cells. As expected, the GCK knockdown effects were rescued by GCKshRNA-RES but not by the kinase-dead mutant GCKshRNA-RESK45A. In contrast to GCKshRNA-RES, GCKshRNA-RESK45A failed to stimulate MM cell proliferation, to suppress MM cells apoptosis and to restore the downstream effectors expression. Our findings demonstrated that GCK kinase activity is required for its function in myeloma cell physiology. Conclusion: Taken together, our findings provide a rationale for the clinical evaluation of targeting GCK in MM patients and the role of GCK in MM tumorigenesis as well as drug resistance. The subsequent development of small molecules inhibiting this pathway, such as GCK kinase inhibitors, will address the unmet need of developing targeted treatments for RASmut myeloma and potentially for other RASmut malignancies. References 1. Walker, B.A., et al. Mutational Spectrum, Copy Number Changes, and Outcome: Results of a Sequencing Study of Patients With Newly Diagnosed Myeloma. J Clin Oncol33, 3911-3920 (2015). 2. Xu, J., et al. Molecular signaling in multiple myeloma: association of RAS/RAF mutations and MEK/ERK pathway activation. Oncogenesis6, e337 (2017). Disclosures Marcireau: Sanofi: Employment. Lentzsch:Caelum Biosciences: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy; Takeda: Consultancy; BMS: Consultancy; Proclara: Consultancy; Abbvie: Consultancy; Clinical Care Options: Speakers Bureau; Sanofi: Consultancy, Research Funding; Multiple Myeloma Research Foundation: Honoraria; International Myeloma Foundation: Honoraria; Karyopharm: Research Funding; Columbia University: Patents & Royalties: 11-1F4mAb as anti-amyloid strategy; Bayer: Consultancy.
27
Qu, Lin, Kosuke Murakami, James R. Broughman, Margarita K. Lay, Susana Guix, Victoria R. Tenge, Robert L. Atmar, and Mary K. Estes. "Replication of Human Norovirus RNA in Mammalian Cells Reveals Lack of Interferon Response." Journal of Virology 90, no. 19 (July 27, 2016): 8906–23. http://dx.doi.org/10.1128/jvi.01425-16.
Full textAbstract:
ABSTRACTHuman noroviruses (HuNoVs), named after the prototype strain Norwalk virus (NV), are a leading cause of acute gastroenteritis outbreaks worldwide. Studies on the related murine norovirus (MNV) have demonstrated the importance of an interferon (IFN) response in host control of virus replication, but this remains unclear for HuNoVs. Despite the lack of an efficient cell culture infection system, transfection of stool-isolated NV RNA into mammalian cells leads to viral RNA replication and virus production. Using this system, we show here that NV RNA replication is sensitive to type I (α/β) and III (interleukin-29 [IL-29]) IFN treatment. However, in cells capable of a strong IFN response to Sendai virus (SeV) and poly(I·C), NV RNA replicates efficiently and generates double-stranded RNA without inducing a detectable IFN response. Replication of HuNoV genogroup GII.3 strain U201 RNA, generated from a reverse genetics system, also does not induce an IFN response. Consistent with a lack of IFN induction, NV RNA replication is enhanced neither by neutralization of type I/III IFNs through neutralizing antibodies or the soluble IFN decoy receptor B18R nor by short hairpin RNA (shRNA) knockdown of mitochondrial antiviral signaling protein (MAVS) or interferon regulatory factor 3 (IRF3) in the IFN induction pathways. In contrast to other positive-strand RNA viruses that block IFN induction by targeting MAVS for degradation, MAVS is not degraded in NV RNA-replicating cells, and an SeV-induced IFN response is not blocked. Together, these results indicate that HuNoV RNA replication in mammalian cells does not induce an IFN response, suggesting that the epithelial IFN response may play a limited role in host restriction of HuNoV replication.IMPORTANCEHuman noroviruses (HuNoVs) are a leading cause of epidemic gastroenteritis worldwide. Due to lack of an efficient cell culture system and robust small-animal model, little is known about the innate host defense to these viruses. Studies on murine norovirus (MNV) have shown the importance of an interferon (IFN) response in host control of MNV replication, but this remains unclear for HuNoVs. Here, we investigated the IFN response to HuNoV RNA replication in mammalian cells using Norwalk virus stool RNA transfection, a reverse genetics system, IFN neutralization reagents, and shRNA knockdown methods. Our results show that HuNoV RNA replication in mammalian epithelial cells does not induce an IFN response, nor can it be enhanced by blocking the IFN response. These results suggest a limited role of the epithelial IFN response in host control of HuNoV RNA replication, providing important insights into our understanding of the host defense to HuNoVs that differs from that to MNV.
28
Tseliou, Melpomeni, Ahmed Al-Qahtani, Saud Alarifi, Saad H. Alkahtani, Christos Stournaras, and George Sourvinos. "The Role of RhoA, RhoB and RhoC GTPases in Cell Morphology, Proliferation and Migration in Human Cytomegalovirus (HCMV) Infected Glioblastoma Cells." Cellular Physiology and Biochemistry 38, no. 1 (2016): 94–109. http://dx.doi.org/10.1159/000438612.
Full textAbstract:
Background/Aims: Rho GTPases are crucial regulators of the actin cytoskeleton, membrane trafficking and cell signaling and their importance in cell migration and invasion is well- established. The human cytomegalovirus (HCMV) is a widespread pathogen responsible for generally asymptomatic and persistent infections in healthy people. Recent evidence indicates that HCMV gene products are expressed in over 90% of malignant type glioblastomas (GBM). In addition, the HCMV Immediate Early-1 protein (IE1) is expressed in >90% of tumors analyzed. Methods: RhoA, RhoB and RhoC were individually depleted in U373MG glioblastoma cells as well as U373MG cells stably expressing the HCMV IE1 protein (named U373MG-IE1 cells) shRNA lentivirus vectors. Cell proliferation assays, migration as well as wound-healing assays were performed in uninfected and HCMV-infected cells. Results: The depletion of RhoA, RhoB and RhoC protein resulted in significant alterations in the morphology of the uninfected cells, which were further enhanced by the cytopathic effect caused by HCMV. Furthermore, in the absence or presence of HCMV, the knockdown of RhoB and RhoC proteins decreased the proliferation rate of the parental and the IE1-expressing glioblastoma cells, whereas the knockdown of RhoA protein in the HCMV infected cell lines restored their proliferation rate. In addition, wound healing assays in U373MG cells revealed that depletion of RhoA, RhoB and RhoC differentially reduced their migration rate, even in the presence or the absence of HCMV. Conclusion: Collectively, these data show for the first time a differential implication of Rho GTPases in morphology, proliferation rate and motility of human glioblastoma cells during HCMV infection, further supporting an oncomodulatory role of HCMV depending on the Rho isoforms' state.
29
Lee, Y. Terry, Colleen Byrnes, Emily Riehm Meier, Antoinette Rabel, and Jeffery L. Miller. "Ineffective Erythropoiesis and Production of Normoblasts with a Beta Thalassemia Major Phenotype Using CD34+ Cells From Healthy Donors." Blood 118, no. 21 (November 18, 2011): 1085. http://dx.doi.org/10.1182/blood.v118.21.1085.1085.
Full textAbstract:
Abstract Abstract 1085 Reversal of anemia is the major target of thalassemia research, but studies of the molecular and cellular basis of the ineffective erythropoiesis of thalassemia are limited by access to donor progenitor cells. Here we demonstrate that thalassemic erythropoiesis may be recapitulated ex vivo by reducing the expression of hemoglobin in cultured CD34+ cells. Using lentiviral transduction of progenitor cells obtained from three healthy adult human donors, shRNA molecules were screened for their ability to reduce beta-globin gene and protein expression over 21 days in culture. Cells transduced with a scrambled vector served as donor-matched controls. Among the screened shRNA, one named HBB caused a consistent and significant reduction in beta-globin mRNA and protein. Beta-globin mRNA was reduced to levels <10% (p<0.001) compared to that of the controls (day14/21), while maintaining expression of gamma- and alpha-globin mRNA. HPLC was performed on an equivalent number of cells sampled on culture day 21 for hemoglobin type (HbA vs. HbF) and quantitation (area under each HPLC peak). The HbA peak was reduced by 96%, and there was a minor increase in the HbF peak (1.6 fold) after HBB transduction. Based upon these quantitative changes in hemoglobin, HbF represented 49.3±9.3% in the HBB transduced population compared with 2.9±0.7% (p<0.01) in controls. On culture day 14, there was no significant difference in glycophorin A (CD235), transferrin receptor (CD71), or cellular morphology despite the reduction in beta-globin mRNA. However, impaired terminal differentiation was detected by retainment of surface CD71 and a lack of enucleation during the third week of culture. Cell counts were lower in HBB transduced cells during the final stages of erythroid differentiation with a 61% (p=0.03) reduction in total cell counts by day 21 when compared to controls. Annexin V assay on day 21 also demonstrated increased phosphatidylserine expression in the HBB transduced cells [HBB=55.7±14.4% vs. Control=25.0±3.0%] in association with the decreased terminal differentiation. GDF15 quantitation demonstrated a significant (p=0.006) increase in the culture supernatants of HBB transduced cells. Sorted cytospin preparations revealed a distinct population of mature normoblasts containing a highly condensed nucleus surrounded by a thin ring of hypochromic cytoplasm. Reduction of erythroblast beta-globin gene and protein expression to levels associated with beta thalassemia major in humans causes ineffective erythropoiesis ex vivo by reducing cell production, increasing surface expression of phosphatidylserine, and impairing enucleation during terminal maturation. Efforts are now underway to use the culture system to explore mechanisms whereby reduced hemoglobin synthesis causes normoblast defects, and for screening of chemical and genetic rescue therapies for the thalassemic erythroid phenotype. Disclosures: No relevant conflicts of interest to declare.
30
Drakos, Elias, Konstantina Stathopoulou, Lingxiao Li, Daniel Bilbao, Vasiliki Leventaki, Pedro Farrajota Neves Da Silva, Vasileios Atsaves, et al. "NPM-ALK Upregulates Jab1/Csn5 through STAT3 Activation in Anaplastic Large Cell Lymphoma: A Novel Function of NPM-ALK That Contributes to PD1/PD-L1 Immune Checkpoint Regulation." Blood 134, Supplement_1 (November 13, 2019): 2796. http://dx.doi.org/10.1182/blood-2019-130457.
Full textAbstract:
Introduction: ALK+ anaplastic large cell lymphoma (ALCL) frequently carries the t(2;5) resulting in overexpression of NPM-ALK oncoprotein, which activates many oncogenic pathways. The Jab1 (c-Jun activation domain-binding protein-1), initially discovered as a c-Jun co-activator, represents the fifth component of an evolutionary highly conserved 8-subunit protein complex, named COP9 signalosome (CSN). The Jab1/Csn5 gene operates as an oncogene in cancer through multiple mechanisms including cell cycle control via the CDK inhibitor p27. Recent evidence suggests that Jab1/Csn5 is also involved in immune checkpoint regulation through PD-L1 stabilization by inhibition of PD-L1 proteasomal degradation. Recently, we reported that the protein levels of Jab1/Csn5 are the highest in ALCL as compared to other peripheral T-cell lymphomas (PTCL) and significantly correlate with PD-L1 levels in PTCLs (Drakos et al, HemaSphere 2019; (3):p596, EHA abstract). In this study, we hypothesized that NPM-ALK may regulate Jab1/Csn5 expression and thus contributes to cell proliferation as well as PD-L1/PD1 immune checkpoint regulation. Methods: The in vitro system included 4 ALK+ (Karpas 299, DEL, SUPM2, SUDHL1) and 2 ALK- (Mac1, Mac2a) ALCL cell lines as well as murine Ba/F3 parental and Ba/F3 clones stably transfected with NPM-ALK (Ba/F3-NPM-ALK), EEF1G (Ba/F3-EEF1G-ALK) or control plasmid (Ba/F3-MIG). Transient transfections with Jab1/CSN si-RNAs and STAT3 si-RNAs were also performed in ALCL cells. In addition, ALCL cells were treated with ALK (Crizotonib) or STAT3 (XIII) inhibitors. Two animal models were used in the study: 1) in the ex vivo model, Karpas-299 clones stably transfected with Jab1/CSN5 shRNA constructs were generated and injected in both thighs of SCID-beige immunocompromised mice. The Jab1/CSN5 shRNA and the control mice were followed for tumor development and their tumors were measured and analysed by immunohistochemistry. 2) in the patient derived xenograft (PDX) model of ALK+ ALCL, the mice were treated with the ALK inhibitor Ceritinib or control vehicle and were monitored for changes in tumor characteristics over two weeks. Tumor specimens were taken at early time points (24, 48, and 72 hrs) following treatment in order to obtain viable tumor cells for protein analysis. Results: Jab1/Csn5 was substantially upregulated in the Ba/F3-NPM-ALK and Ba/F3-EEF1G-ALK stable clones as compared to paternal or control Ba/F3-MIG clones. Jab1/Csn5 upregulation was associated with high STAT3 activation (Tyr705-phosphorylation) and increased PD-L1 gene expression in this system. Inversely, inhibition of ALK activity was associated with STAT3 de-activation and decreased protein levels of Jab1/Csn5 and PD-L1 in ALK+ ALCL cells. Knocking down STAT3 by siRNA or inhibition of its activity by the XIII inhibitor resulted in decreased levels of Jab1/Csn5 protein in both ALK+ and ALK- ALCL cell lines. The SCID-beige mice that received Jab1/CSN5-shRNA clones showed significant delay in tumor development and longer survival as compared to control mice, which was associated with significantly decreased PD-L1 protein levels and p27 upregulation in the tumor cells (xenografts). Treatment of the PDX mice with Ceritinib, a potent next generation ALK inhibitor, resulted in substantial tumor necrosis after 72 hrs and decreased tumor size at day 7 post-treatment. At earlier time points, de-activation (de-phosphorylation) of ALK kinase and STAT3 was observed in the Ceritinib-treated mice, which was linked to variably lower levels of Jab1/CSN5 and PD-L1 proteins as compared to control mice. Conclusion: Jab1/CSN5 is a novel downstream target of the NPM-ALK oncogenic kinase that regulates its expression, at least in part, through STAT3 activation. The Jab1/CSN5-mediated stabilization of PD-L1 can be efficiently inhibited by Ceritinib in preclinical animal models of ALK+ ALCL. Disclosures Österborg: BeiGene: Research Funding; Janssen: Research Funding; Abbvie: Research Funding; Kancera AB: Research Funding; Gilead: Research Funding. Vega:National Cancer Institute, national Institutes of Health: Other: Grant Funding-R01CA222918.
31
Shen, Q., Y. Liu, F. Yin, L. Yang, and G. Li. "86 EFFECT OF HDAC2 INTERFERENCES ON HISTONE MODIFICATIONS IN MOUSE EARLY EMBRYOS." Reproduction, Fertility and Development 25, no. 1 (2013): 191. http://dx.doi.org/10.1071/rdv25n1ab86.
Full textAbstract:
One of the histone deacetylase members, HDAC2 plays an important role in chromatin remodeling and transcriptional repression. The present study was designed to improve histone acetylations by knocking down the expression of histone deacetylase with RNA interferences. According to the published mouse HDAC2 mRNA sequence (NM_008229.2, GI:87162463), as well as shRNA design principles, 3 interference fragments of 107 to 126 bps, 879 to 898 bps, and 1240 to 1259 bps and a negative control sequence were designed and synthesized, respectively. Four interference vectors expressing a red fluorescent protein were successfully constructed, which were named pCDsRed2-shRNA107, pCDsRed2-shRNA879, pCDsRed2-shRNA1240, and pCDsRed2-shRNAcontrol, respectively. These vectors were then transfected respectively to mouse fetal fibroblast cells. Real-time, quantitative PCR of the transgenic cells showed that the vectors mentioned above resulted in 0.81, 0.73, 0.16, and 0.80 times knockdown of hdac2 expression when compared with the nontransfected cells, which suggested that the piece of 1240 to 1259 bp was the most effective RNAi region. Immunofluorescent staining of the transgenic cells showed that the histone acetylations and methylations of H4K5ac, H3K9ac, H3K9me2, H3K27me3, and H3K4me3 were significantly higher in all of the interference vectors than they were in the negative control. Vector pCDsRed2-shRNA1240 was the most effective RNAi. After injection of pCDsRed2-shRNA1240 into zygote pronuclei, embryos developed to 2-cell, 8-cell embryos and blastocysts decreased to 96.6, 85.3 (P < 0.05), and 35.6 (P < 0.01) of the control group, respectively. High levels of H3K9ac, H4K5ac, H3K4me3, and H3K27me3 were observed in the RNAi embryos when compared with the controls. These results indicated that the knockdown of hdac2 by RNAi decreased the expression of HDAC2 and induced high expression of histone acetylations in both somatic cells and early embryos. This work was supported by the national basic research program of china (No. 2012CB22306).
32
Cao, Yu, Deliang Cao, and Hongyan Ling. "The novel long non-coding RNA PANCR: A p53 activator and potential breast cancer biomarkers." Journal of Clinical Oncology 35, no. 15_suppl (May 20, 2017): e23016-e23016. http://dx.doi.org/10.1200/jco.2017.35.15_suppl.e23016.
Full textAbstract:
e23016 Background: Long non-coding RNAs (LncRNAs) may serve as a biomarker and potential therapeutic target of cancers. Chromosome 16q22.1 is frequently deleted in breast cancer and may contribute to breast carcinogenesis by inactivation of tumor suppressor genes. This study characterized a new LncRNA tumor suppressor in this region, named p53 activating non-coding RNA (PANCR). This LncRNA consists of 1.5kb in length. Methods: Quantitative real-time PCR was used for examine the PANCR expression in breast cancer tissues. RNA-pull down and RNA-Immunopreicitation were used to analyze PANCR targeted protein. Results: Our data showed that PANCR was downregulated in breast cancer cell lines and tissues. In the breast cancer cell lines, PANCR expression appeared reversely correlated with cell malignancy, and in breast cancer tissues, PANCR was downregulated over 2 times in 31(62.0%) of 50 cases compared to adjacent normal breast tissues. In breast cancer cells MCF7 and immortalized human mammary epithelial cells MCF10A, ectopic expression of PANCR induced marked apoptosis, suppressing cell proliferation in culture and tumor growth in xenografts, but in contrast, shRNA–mediated silencing of PANCR promoted cell growth and proliferation. Mechanistic approaches revealed that in both MCF7 and MCF10A cell, PANCR activated p53 and upregulated pro-apoptotic proteins bid and bim and cell cycle inhibitors p21waf/cip1 and p27Kip1. We further found that the PANCR binds to and activates p53 by dissociating the p53-MDM2 complex. We further characterized the functional domain of PANCR that interacts with p53. Conclusions: The LncRNA PANCR located in the deleted Chromosome 16q22.1 region is a novel intracellular p53 activator and tumor suppressor, which may be used as a target for cancer therapy through mimicking its binding domain and activation of p53.
33
Selliah, Nithianandan, Jiyi Yin, and Terri H. Finkel. "Silencing of HALP Decreases HIV-1 Infection of Primary CD4+ T-cells (45.4)." Journal of Immunology 178, no. 1_Supplement (April 1, 2007): S57—S58. http://dx.doi.org/10.4049/jimmunol.178.supp.45.4.
Full textAbstract:
Abstract HIV/AIDS is an ideal candidate for a novel gene therapy approach since it is an incurable and terminal disease. We have identified a set of candidate host cell genes, associated with survival of HIV-infected T-cells. We named one of these genes, “HALP”, for “HIV-Associated Life Preserver”. Recently, we used a lentiviral vector to stably express HALP shRNA in primary human CD4 T-cells. In the absence of HIV infection, silencing of HALP appeared to have no effect on viability of these cells. As predicted, silencing of HALP markedly reduced HIV infection of primary CD4+ T-cells, as measured by the percent of p24+ cells and by supernatant p24. Published data suggest that hypoxia induces HALP expression by increasing the activity of the transcription factor, Hif-1α. HIF-1 binding sites have been identified within the HALP promoter and HALP is responsive to HIF-1 in non-immune cells. We show here that HIV-1 infection of primary CD4+ T-cells increases Hif-1α mRNA expression, as measured by real time RT-PCR. Transfection of T-cells with wild-type Hif-1α or with a mutated form of Hif that is resistant to proteolysis (Hif-DPA) also increased expression of HALP mRNA. These data are consistent with a pathway of Hif-induced HALP expression in HIV-1 infected cells, leading to a hypoxia-like stress response and, ultimately, to cell survival. Our work represents one of the first attempts to search for novel cellular genes that contribute to viral inhibition of host cell apoptosis and, thus, to viral latency. HALP represents a candidate for a host cell anti-apoptotic protein, which is triggered by HIV infection, and may provide a novel molecular target for drug use and design.
34
Sun, Jingnan, Jifan Hu, Wei Li, and Yunpeng SUN. "A Novel Suppressive Long Noncoding RNA within the IGF1R Gene Locus Is Imprinted in Acute Myelocytic Leukemia." Blood 124, no. 21 (December 6, 2014): 3592. http://dx.doi.org/10.1182/blood.v124.21.3592.3592.
Full textAbstract:
Abstract Background：In acute myeloid leukemia (AML) cells, the insulin-like growth factor type I receptor (IGF1R) is one of the most abundantly phosphorylated receptor tyrosine kinases, promoting cell growth and therapeutic resistance through the PI3K/Akt signaling pathway. However, little is known regarding the molecular mechanisms underlying IGF1R gene dysregulation in AML. Long non-coding RNAs have been implicated in a variety of important biological processes by epigenetic mechanisms. Materials and methods：AML cell lines used in this study, KG-1, KG-1a were purchased from ATCC. The full length of IRAIN lncRNA was characterized by Marathon cDNA Amplification Kit. For qPCR, cDNA samples were amplified using CFX96TM real-time system (BIO-RAD) by SYBR PrimeScript™ RT-PCR Kit (TaKaRa). Both reverse transcription-associated trap (RAT) and chromatin conformation capture (3C) assays were used to examine the chromosome architecture of IGF1R and DNA-RNA interaction. The protocol of AMLpatients bone marrow and peripheral blood cell samples was approved by the Human Medical Ethical Review Committee from Jilin University First Hospital and informed consent was obtained from each AML patient and normal donor. Results： 1. we discovered a novel intragenic 5366bp long noncoding RNA (lncRNA) within the IGF1R locus, named IRAIN, which is transcribed in an antisense direction from an intronic promoter. 2. we demonstrated IRAIN was expressed exclusively from the paternal allele, with the maternal counterpart being silenced in normal donor and AML patients. 3. we confirmed that IRAIN interacts with IGF1R chromatin DNA and is involved in the formation of an intrachromosomal enhancer/promoter loop. Knockdown of IRAIN lncRNA with shRNA abolishes this intrachromosomal interaction. 4. We demonstrated IRAIN was downregulated both in AML cell lines and high risk AML patients. Furthermore，the expression of IRAIN was dramatical higher in APL patients achieved molecular remission. Conclusion：Our results identify IRAIN as a new imprinted suppressive lncRNA involved in long range IGF1R DNA interactions in AML. As a putative tumor suppressor and a novel treatment target， further studies are needed to delineate the specific role of this newly identified lncRNA in the uregulation of the IGF pathway in AML. Disclosures No relevant conflicts of interest to declare.
35
Li, Shirong, Jing Fu, Jun Yang, Markus Y. Mapara, Christophe Marcireau, and Suzanne Lentzsch. "Gck Inhibition Is a Novel Therapeutic Strategy for RAS Mutated Multiple Myeloma and Overcomes Resistance to IMiDs." Blood 136, Supplement 1 (November 5, 2020): 24. http://dx.doi.org/10.1182/blood-2020-143418.
Full textAbstract:
Introduction: RAS oncogenes are the most frequently mutated gene family in human cancers. 50% of newly diagnosed multiple myeloma patients carry a RAS/MAPK pathway mutation, with a rising percentage in the relapsed situation1. Thus, targeting RAS mutations in multiple myeloma will increase therapeutic efficiency and potentially overcome drug-resistance. Unfortunately, RAS mutations have been considered "undruggable" due to a lack of traditional small molecule binding pockets on the proteins. Therefore, key component in the RAS/MAPK pathway may represent an alternative therapeutic target for MM. Germinal center kinase (GCK), also named mitogen-activated protein kinase kinase kinase kinase 2 (MAP4K2), is an upstream activator in the MAPK pathway. Indeed, we recently discovered the critical role of GCK in RAS mutated (RASmut) MM cell survival and growth. GCK knockdown in RASmut MM cells induced MM cell growth inhibition both in vitro and in vivo. However, the detailed mechanism is yet to be defined. Methods and Results: Our previous data showed that GCK knockdown induces MM cell growth inhibition, associated with the blockage of MKK4/7-JNK phosphorylation and the downregulation of critical transcriptional factors (TFs) including IKZF1/3, BCL-6, and c-MYC proteins. To confirm that GCK knockdown downregulates IKZF1/3 etc at protein level but not mRNA level, we conducted real-time PCR on GCK knockdown MM cells and compared the expression of GCK and TFs to the empty vector (EV) infected MM cells. Results showed that shRNA induced GCK silencing only led to the significantly decreased GCK mRNA, however, did not affect IKZF1 and c-MYC expressions at mRNA level. Consistent with the effects of GCK knockdown, the GCK inhibitor TL4-12 dose-dependently downregulated IKZF1 and BCL-6 proteins, inhibited MM cell proliferation and induced cell apoptosis. IKZF1/3 are the key targets of the immunomodulatory drugs (IMiDs), which are the backbone of MM therapy. IMiDs bind to cereblon (CRBN) and induce IKZF1/3 protein degradation, which subsequently lead to MM cell growth inhibition. Importantly, our data showed that IMiDs-resistant RPMI-8226 MM cells have high expression of GCK. GCK knockdown and inhibition induced IKZF1 downregulation, triggered growth inhibition and cell apoptosis in RPMI-8226 cells, suggesting that GCK regulates IKZF1 degradation via a CRBN-independent mechanism. To confirm this hypothesis, we silenced CRBN in N-Rasmut H929 MM cells by shRNA lentiviral infection and examined the response to IMiDs and GCK inhibitor. CRBN knockdown was confirmed by western blotting. CRBN silencing in H929 cells resulted in lenalidomide (LEN) resistance, evidenced by the WTS proliferation assay. In contrast, CRBN silencing failed to rescue N-Rasmut H929 MM cells from TL4-12 induced proliferation inhibition and IKZF1 downregulation, confirming that GCK regulated IKZF1 and cell growth is independent of CRBN. Conclusion: Taken together, our data demonstrated that GCK inhibition induces cell growth inhibition and triggers apoptosis especially in RASmut MM cells. Importantly, GCK inhibitor downregulates IKZF1 via a CRBN-independent mechanism. Our findings thus provide a rationale for the clinical evaluation of targeting GCK in RASmut MM patients and further mechanistic insight into the role of GCK in MM tumorigenesis as well as drug resistance. GCK inhibitors may represent a novel therapy for the treatment of RASmut MM patients, especially those who are resistant to IMiDs as well as with refractory or relapsed MM. References Walker, B.A., et al. Mutational Spectrum, Copy Number Changes, and Outcome: Results of a Sequencing Study of Patients With Newly Diagnosed Myeloma. J Clin Oncol33, 3911-3920 (2015). Disclosures Marcireau: Sanofi: Current Employment. Lentzsch:Karyopharm: Research Funding; Mesoblast: Divested equity in a private or publicly-traded company in the past 24 months; Janssen: Consultancy; Sorrento: Consultancy; Caelum Biosciences: Current equity holder in private company, Membership on an entity's Board of Directors or advisory committees; Celularity: Consultancy; Magenta: Current equity holder in private company; Sanofi: Research Funding.
36
Roversi, Fernanda Marconi, Fernando Vieira Pericole, Adriana da Silva Santos Duarte, Karla Priscila Ferro, Flávia Adolfo Corrocher, Bruna Palodetto, Ana Leda Longhini, Maurizio Botta, and Sara T. Olalla Saad. "Knockdown of HCK Reduces Cell Death and Erythroid Differentiation in Human CD34+ Hematopoietic Progenitor Cells." Blood 126, no. 23 (December 3, 2015): 2860. http://dx.doi.org/10.1182/blood.v126.23.2860.2860.
Full textAbstract:
Abstract Myelodysplastic syndromes (MDS) are clonal disorders characterized by ineffective hematopoiesis and increased risk of transformation to acute myeloid leukemia (AML). The identification of genes and cellular pathways active in leukemia cells but not in normal hematopoietic stem/progenitor cells (HSC) may help to understand the key steps in the MDS and AML pathogenesis and lead to new approaches to further enhance the treatment of both diseases, considered incurable with non-transplantation therapy. Src kinase family (SFK) is a central mediator in multiple oncogenic signaling pathways and some SFK members (Hck, Lyn, Fgr, Fyn) had previously been described as overexpressed or activated in leukemic cells. However, to this moment, the role of hematopoietic cell kinase (HCK), the unique SFK member restricted expressed in hematopoietic cells, had not been characterized in MDS and AML pathogenesis as well as in HSC. In order to better understand the HCK importance in hematopoiesis, we used lentiviral shRNA vectors to knockdown the HCK expression in primary human CD34+ HSC. The HCK levels were reduced in approximately 70-80% (shHCK) compared to the control lentiviral shRNA (shControl-GFP). To promote erythroid differentiation, human CD34+ transduced cells were grown in methylcellulose for 7 days and in liquid media for another 6 days. During this experiment, shHCK cells showed decreased cell viability (fold change compared to shControl-GFP = 0.55, P<.0001, n=3) combined with an increase in CD71+ expression (fold change compared to shControl-GFP = 3, P<.01, n=3), indicating a delay in erythroid differentiation. As expected, shControl-GFP cells showed a decreased GATA1 expression during erythroid differentiation. Meanwhile, shHCK cells did not modulate GATA1 expression. Interestingly, without any stimulus, HCK knockdown in CD34+ cells significantly decreased apoptosis (AnnexinV+ cells) compared to shControl-GFP (fold change = 0.52, P<.01, n=4). Attempts have been made to overexpress HCK in CD34+ HSC, however more than 80% cells were apoptotic and further assays were not possible. Thus, in HSC, HCK participates of erythroid differentiation and apoptosis signaling. According to the HCK importance on HSC and that SFK inhibitors are undergoing early phase clinical testing, a specific inhibitory activity compound for HCK, named iHCK-37, had been developed by Dr Maurizzio Botta. We tested this compound on primary normal human CD34+ cells originated from healthy donors bone marrow samples and also from cord blood units. The iHCK-37 treatment did not change proliferation, survival and death of these normal CD34+ cells. Conversely, MDS and AML CD34+ cells treated with the same drug exhibited a dose-dependent growth inhibition. Likewise, following iHCK-37 treatment of MDS and AML total bone marrow mononuclear cells, the BFU-E and CFUs colony numbers were significantly decreased compared to untreated cells (vehicle). We also observed a potent in vitro antiproliferative activity of iHCK-37 against a panel of leukemia cell lines, with uM IC50 values in AML (5.0 - 5.8uM) and chronic myeloid leukemia (9.1 - 19.2uM). In addition, the combinatory in vitro treatment of iHCK-37 and 5-Azacitidine (Aza) also demonstrated additive effects relative to either drug alone. Interestingly, iHCK-37 or iHCK-37 plus Aza treatments of dysplastic and leukemia cells enhanced apoptosis and resulted in increased BAX and reduced BCL-XL protein levels. This result could be clinically relevant for MDS, as Aza is the only treatment available for higher-risk MDS, but with low response rates and frequent induced resistance and refractoriness over time. In summary, we herein have shown that HCK mRNA knockdown of normal CD34+ cells resulted in growth inhibition, decreased cell death and reduced erythroid differentiation, suggesting that HCK is essential for normal hematopoiesis. We presume that the deregulation of HCK pathway in leukemic cells might be crucial for MDS and AML pathogenesis. On the other hand, the inhibition of HCK protein activity with a specific inhibitor was able to restore the apoptotic pathways of leukemic cells, acting on cancer cells without alter any signaling of normal cells. Moreover, the specific inhibitor may have antineoplastic effect that can even be additive to current available drugs. Our study adds new insights to the role of HCK in MDS and AML as well as into potential new anticancer treatment strategies. Disclosures No relevant conflicts of interest to declare.
37
Bardhan, Kankana, Nikolaos Patsoukis, Alexandra Plessa, Niko Tsopoulidis, Duygu Sari, Lequn Li, and Vassiliki A. Boussiotis. "Rap1-GTP Augments TGF-b-Mediated Signaling in T Lymphocytes Via a Mechanism Dependent on the b Chain of LFA-1 Integrin." Blood 126, no. 23 (December 3, 2015): 3422. http://dx.doi.org/10.1182/blood.v126.23.3422.3422.
Full textAbstract:
Abstract Integrin-mediated adhesion of lymphocytes to antigen presenting cells (APCs) is a critical event linking innate and adaptive immunity. Integrins function as bidirectional receptors and can transmit signals from both sides of the plasma membrane, a property referred to as inside-out and outside-in signaling. Lymphocyte adhesion to APC is mainly accomplished through the principle adhesion molecule on the lymphocyte surface, the lymphocyte functional antigen 1 (LFA-1), which binds to intercellular adhesion molecule 1 (ICAM-1) on the surface of APCs. In order to mediate its function, LFA-1 must be activated via a process, which results in conformation changes of the receptor that extends the ectodomains of the α and β chains leading to a high affinity state. Among the few signaling molecules that have been implicated in integrin activation in hematopoietic cells are the small GTPase Rap1A (thereafter named Rap1) and its downstream effectors RapL and RIAM. In response to Rap1-GTP, RapL regulates LFA-1 activation by interacting with the integrin α chain, whereas RIAM mediates recruitment of talin to the cytoplasmic tail of the β chain leading to its conformational change to the high affinity state. To understand the role of Rap1 in T cell responses we generated transgenic (Tg) mice that selectively express the active, GTP-bound Rap1 mutant Rap1E63 in mature T cells. Rap1E63-Tg and littermate control mice had no statistically significant difference in absolute thymocyte numbers and differentiation profiles. In contrast, in peripheral lymphoid organs, Rap1E63-Tg mice had a significant reduction in total T cells but a 4-fold increase in the CD4+ CD103+ T cell fraction. CD103 (also known as αEβ7 integrin) defines a subset of peripherally generated Treg with potent suppressive function. TGF-β is the strongest stimulus for induction of CD103 (αEβ7). To examine whether Rap1-GTP can affect TGF-β-mediated signaling in T cells, we used stable Jurkat T cell lines expressing GTP-bound Rap1 mutant, Rap1E63, or Jurkat T cell lines in which the endogenous Rap1 was depleted by shRNA (Rap1-KD), and also primary T cells from Rap1E63-Tg mice and Rap1-KO mice. After interaction with two membrane-bound receptors, TGF-βRI and II, TGF-β propagates downstream signaling via the Smad family transcription factors. Incubation of Rap1E63 Jurkat T cells with TGF-β resulted in enhanced and sustained phosphorylation of Smad2 and Smad3, which was observed with very low concentrations of TGF-β that were incapable of inducing detectable phosphorylation of Smads in control Jurkat T cells. In contrast, diminished level and duration of Smad2 and Smad3 phosphorylation was observed in Rap1-KD Jurkat T cells. Similar patterns of responses to those observed in Rap1E63 Jurkat T cells and in Rap1-KD Jurkat T cells were observed in primary mouse T cells isolated from Rap1E63-Tg mice and Rap1-KO mice, respectively. To investigate whether the LFA-1 integrin α and/or β chain had an active role in the enhanced TGF-β-mediated signaling in the presence of Rap1-GTP, we used Rap1E63 Jurkat T cells in which RapL or RIAM were depleted by shRNA (Rap1E63/RapL-KD and Rap1E63/RIAM-KD) because these adaptors selectively regulate the LFA-1 α and the LFA-1 β chain, respectively, downstream of Rap1-GTP. Although in Rap1E63/RapL-KD cells the enhanced TGF-β-induced Smad3 phosphorylation remained unaffected, in Rap1E63/RIAM-KD cells the enhanced TGF-β-induced Smad3 phosphorylation was abrogated. To investigate the biological relevance of these observations, we used T cells from Rap1E63-Tg mice crossed with mice deficient for the LFA-1 α chain. TGF-β resulted in enhanced Smad3 phosphorylation in T cells from Rap1E63-Tg/LFA1-a KO mice similarly to T cells from Rap1E63-Tg mice, indicating that this effect was not dependent on the activation of LFA-1 α chain. In contrast, T cells from RIAMflox/flox -Lck-Cre mice, in which activation of the LFA-1 β chain is impaired, displayed abrogated activation of Smad3 in response to TGF-β. Our data reveal a novel mechanism by which Rap1 regulates T cell responses via outside-in integrin signaling and may have important implications on TGF-β-mediated T cell homeostasis, differentiation and immune quiescence. Disclosures No relevant conflicts of interest to declare.
38
Grosjean, Iris, Grégoire D’Andréa, Nathalie Yazbeck, Amine Belaid, Barnabé Roméo, Marie Angela Domdom, Nicolas Guillot, et al. "Abstract 1284: The tumor scaffold protein SQSTM1 at the crossroads of DNA damage and immunotherapy responses in solid tumors." Cancer Research 82, no. 12_Supplement (June 15, 2022): 1284. http://dx.doi.org/10.1158/1538-7445.am2022-1284.
Full textAbstract:
Abstract Introduction After the recent advent of Immune Checkpoint Inhibitors (ICIs), one of current challenge of clinical cancer trials is certainly to develop the optimal combinations of ICIs with DNA-Damaging Agents (chemotherapy and/or radiotherapy, named DDAs hereafter). Elucidating resistance mechanisms to these different treatments is pivotal to propose new predictive biomarkers and to develop therapeutic strategies to improve ICI efficiency. We hypothesized that resistance to DDA and ICIs is mediated in part by intrinsic tumor mechanisms, some of which may be shared. Methods To address this knowledge gap, we compared RNA expression signatures, antigen presentation, and PD-L1 expression from cohorts of cancer patients treated with radiotherapy, chemotherapy, and immunotherapy to identify shared molecular pathways that may mediate cross-resistance. Using a panel of lung cancer cell lines, we then confirmed that the tumor-cell-intrinsic expression of SQSTM1 is positively correlated with antigen presentation, and inversely with DNA damage repair and DDA/ICIs resistance gene signatures. This SQSTM1/HERV pathway was then supported in vitro using engineered silenced cell-lines (SQSTM1, MAVS, and STING ShRNA) treated with FDA-approved DDA (cisplatin, oxaliplatin, doxorubicin, and ionizing radiations) or synthetic double-stranded RNA (5’ppp dsRNAs and poly IC). Results Through three complementary approaches (in silico, ex vivo on patient cohorts, and in vitro), we identify the p62/SQSTM1 scaffold protein as a key molecular mediator able of predicting and controlling sensitivity for DDA and ICIs. Mechanistically, in response to DNA damage, we found that SQSTM1 is essential for the inhibition of DNA repair and the reactivation of human endogenous retroviruses (hERV). Analogous to a “viral alarm”, the hERV are sensed and activate IFN responses that drive the expression of MHC-I and PD-L1, leading to tumor immune evasion. Targeting the hERV pathways in SQSTM1-depleted tumor cells by poly(I:C)/Lyovec, or docetaxel treatment can rescue the hERV, IFN, and MHC pathway, providing a promising therapeutic avenue turning a “cold” into a “hot” tumor in non-responders cancer patients. Conclusion Depending on its levels, we thus propose SQSTM1 as a predictive biomarker for guiding treatment decisions from i) ICI alone, ii) ICI combined with cisplatin, or; iii) ICI combined with poly(I:C)/Lyovec or docetaxel, with the aim to increase immunotherapy efficacy. Citation Format: Iris Grosjean, Grégoire D’Andréa, Nathalie Yazbeck, Amine Belaid, Barnabé Roméo, Marie Angela Domdom, Nicolas Guillot, Eric Gilson, Emmanuel Chamorey, Gérard Milano, Véronique Hofman, Marius Ilié, Isabelle Benard-Thiery, Simon Heeke, Patrick Brest, François Ghiringhelli, Paul Hofman, Baharia Mograbi. The tumor scaffold protein SQSTM1 at the crossroads of DNA damage and immunotherapy responses in solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1284.
39
Tarnowski, Maciek, Magdalena Kucia, and Mariusz Z. Ratajczak. "Isolation and Functional Analysis of CXCR7 Promoter - a Novel Receptor for Stromal Derived Factor-1 (SDF-1): Different Regulation of Expression in Human Hematopoietic Cells Versus Pediatric Sarcomas." Blood 114, no. 22 (November 20, 2009): 4583. http://dx.doi.org/10.1182/blood.v114.22.4583.4583.
Full textAbstract:
Abstract Abstract 4583 Stromal derived factor-1 (SDF-1) binds to seven transmembrane-span G-protein coupled receptor CXCR4 and directs homing of CXCR4+ hematopoietic stem cells to bone marrow (BM) as well as metastasis of CXCR4+ cancer cells. Recently, a new SDF-1 binding receptor has been identified and named CXCR7. With identification of this receptor a role of SDF-1 in directing chemotaxis of CXCR7+ hematopoietic cells as well as metastasis of CXCR7+ tumors become more complex. While CXCR7 is expressed at very low level on normal hematopoietic stem cells, its expression becomes high on leukemic cells. Similarly we noticed high expression of CXCR7 on several pediatric sarcomas (e.g., rhabdomyosarcomas and neuroblastomas) that very often metastasize/infiltrate BM. The aim of our study was to evaluate 5′ fragment of CXCR7 gene for promoter activity and analyze how its expression is regulated in CXCR7+ human hematopoietic cells as well as in CXCR7+ human rhabdomyosarcoma cells (RMS). The putative CXCR7 promoter was cloned by employing specific primers for 5′ fragment of CXCR7 gene. We found that this 2.5 kb 5′ DNA fragment adjacent to CXCR7 gene contains three potential hypoxia responsive element (HRE)- (-100-104, -965-969, -1306-1310), five NF-kB- (-32-42, -308-318, -1019-1029, -1375-1379, -2145-2155), four NRF-1 binding sites (-1030-1040, -1468-1478, -1980-1990, -2085-2095), one c-myb binding site at -15-19 and at -702-706 a binding site for negative transcription regulatory factor YY1. We generated 8 constructs containing smaller CXCR7 promoter fragments and three constructs containing mutated distal NF-kB and HREs as well as c-myb that were subcloned into a pGL4.10 vector. The promoter activity of these fragments was tested in transfected human hematopoietic cells (THP-1) and RMS cell line (RD) by measuring luciferase activity. While the minimal promoter activity in human hematopoietic cells was retained in 80 bp short fragment containing c-myb binding site, similar activity in human rhabdomyosarcoma cells required longer 150 bp fragment containing proximal NF-kB binding element. We noticed that while mutation of c-myb binding site in CXCR7 promoter in THP-1 cells reduces promoter activity by ∼50%, mutation of proximal NF-kB-binding site in CXCR7 promoter completely inhibits promoter activity in RD cells. This was confirmed by knock-down of c-myb by shRNA and chemical inhibition of NF-kB respectively. Furthermore, we noticed that during hypoxia in contrast to CXCR4, CXCR7 expression does not change in hematopoietic cells, however is significantly downregulated in rhabdomyosarcoma cells. This could be explained by upregulation of negative transcripton factor YY1 during hypoxia, as evidenced by RQ-PCR and confirmed by CHIP assay. In conclusion we have demonstrated that 5′ fragment of CXCR7 possesses promoter activity and is differently regulated in hematopoietic versus sarcoma cells - in c-myb or NF-kB dependent manner respectively. Furthermore, we also found that in contrast to hematopoietic cells hypoxia inhibits CXCR7 promoter activity in RMS cells in YY1-dependent manner. Disclosures: No relevant conflicts of interest to declare.
40
Gong, Yuping, Ruiqing Zhoui, and Ting Niu. "The Role of Transcription Factor SCL/TAL-1 in the Hematopoiesis of Human Cord Blood Hematopoietic Stem Cell." Blood 118, no. 21 (November 18, 2011): 4795. http://dx.doi.org/10.1182/blood.v118.21.4795.4795.
Full textAbstract:
Abstract Abstract 4795 The role of transcription factor SCL/TAL-1 in the hematopoiesis was studied in the CD34+ hematopoietic stem cells purified from human cord blood. The sorted CD34+ cells were transfected by the plasmids pTRIPdU3-RNAiTALh-EF1a-GFP (SCL/TAL1 shRNA to reduce the expression of SCL/TAL-1), pTRIP-EF1a-TAL1 (SCL/TAL-1 cDNA to enhance the expression of SCL/TAL-1) and control plasmid pTRIP-dU3-RNAiluc-EF1-GFP expressing EGFP gene via lentiviral vector system, named the groups of SCL/TAL-1low, SCL/ TAL-1high and LUC control, and then cultured in methylcellulose medium. The clone forming unit (CFU) and the expression of hematopoietic related genes and surface markers of transfected CD34+ cells were examined by light microscope, RT-PCR and flow cytometry, and the role of SCL/TAL-1 in regulation of hematopoiesis was explored. After cultured in methylcellulose semi-solid medium with cell factors for 14 day, each 500 CD34+ cell purified from cord blood expanded to about 120 CFU/BFU. FCM detected the surface markers of hematopoietic differentiation including CD71/CD35a, CD33/CD13 and CD41a/CD42b in CD34+ cells cultured at d7. The results showed CD71 was 9.7±5.7%, CD235a 6.7±3.8%, CD33 49.4±12.6%, CD13 46.9±9.6%, CD41a 5.5±2.3% and CD42b 2.0±1.2%. During the hematopoietic differentiation of CD34+ cells, the overall tendency of SCL/TAL-1 mRNA increased significantly, especially after the 7 days. The levels of PU.1, LMO1 and LMO2 mRNA were almost similar with SCL/TAL-1's. The expression of GATA mRNA increased slightly during the differentiation process, and GATA2 showed a gradually increased expression in the first 10 days and then a decrease on the 14th day. However, there were no obvious alterations in RUNX1 mRNA level during the whole differentiation process. After interference of SCL/TAL-1, the CFU of SCL/TAL-1low significantly decreased in number and showed dysplasia with small sizes,especially in erythroid clones, there was no GEMM-BFU to be found. Hematopoiesis of SCL/ TAL-1high still developed successfully and was slightly better than LUC control. RT-PCR revealed that the level of PU.1 and GATA2 mRNA significantly decreased following a knockdown of SCL/TAL-1, the mRNA of LMO1 and LMO2 decreased slightly, while GATA1 varied reversely with SCL/TAL-1, and there was no obvious alteration in RUNX1 mRNA. The results of FCM showed that CD235a and CD71 decreased significantly in the group of SCL/TAL-1low and increased in the group of SCL/TAL-1high compared with LUC control. CD33 and CD13 expression in SCL/TAL-1low decreased significantly too, and CD33 and CD13 expressions in SCL/TAL-1high rose markedly. Expressions of CD41a and CD42b also decreased in SCL/TAL-1low and increased in SCL/TAL-1high, but there was no statistical significance. The results conclude SCL/TAL-1 plays a key role in the regulation of hematopoiesis by affecting the differentiation of erythroid and myeloid cells positively. Grant support: National Natural Science Foundation of China (No.30770912), Foundation of the Science & Technology Department of Sichuan Province (No.2008SZ0017). Disclosures: No relevant conflicts of interest to declare.
41
Bayanjargal, Ariunaa. "Abstract B018: Studying the role of FLI portion of EWS FLI in transcription regulation via modulation of chromatin 3D landscape in Ewing sarcoma." Cancer Research 82, no. 23_Supplement_2 (December 1, 2022): B018. http://dx.doi.org/10.1158/1538-7445.cancepi22-b018.
Full textAbstract:
Abstract Ewing sarcoma is an aggressive bone-associated tumor currently treated with dose-intense chemotherapy, radiation, and surgery. Ewing sarcoma affects adolescents and young adults with incidence rate of 3 per million. The hallmark of Ewing sarcoma is a translocated fusion transcription factor named EWS FLI that drives the oncogenic process. The lack of efficient and targeted treatment in Ewing sarcoma directly arises from the poor understanding of how EWS/FLI regulates expression of thousands of genes as well as the current lack of effective targeting of transcription factors. We hypothesize that FLI portion of EWS::FLI containing a crucial alpha-helix plays a novel role in transcription regulation of thousands of genes by modulating chromatin looping. We postulated this hypothesis based on recent evidence from our lab that established an alpha-helix immediately downstream of DNA binding domain as an important player in regulating transcriptional activity in A673 cell line. Structure-function mapping revealed that ETS domain alone was insufficient for full transcriptional regulation. The purpose of this study is to elucidate the mechanism underlying transcriptional regulation by FLI and it is likely to guide future studies on novel therapeutics for Ewing sarcoma patients as transcriptional factors remain elusive targets of therapy. We utilized knockdown/rescue experiments in which EWS FLI was depleted with shRNA and replaced with constructs containing the ETS DNA Binding Domain (DBD) alone or DBD+ (ETS DNA Binding Domain and 4th alpha-helix). The knockdown/rescue experiments were confirmed at mRNA level using qPCR, and at protein level using immunoblots. The binding pattern and transcriptional regulation of DBD and DBD+ were assessed with RNA-Seq respectively. The extent of roles each of these construct play in organization, structure, and function of chromatin are being assessed using Micro-C technique, a variation of Hi-C with improved resolution, higher signal-to-noise ratio and more information on chromatin domain boundaries and chromatin looping. The DNA binding and genomic localization of EWS FLI was unaltered by the deletion surrounding the DNA binding domain (which contains a 4th alpha-helix) in A673 cells. Despite this similarity in genomic localization and binding, the transcriptional output driven by EWS FLI was significantly diminished by the deletion. In addition, we observed this differential transcriptional output by the EWS FLI DBD and DBD+ in another Ewing sarcoma cell line called TTC466. Current work in our lab shows that EWS FLI has a substantial role in shaping the chromatin landscape of Ewing sarcoma cells. We observed this result with Hi-C technique upon the depleting EWS FLI from A673 cells. With this current study, we hope to understand if the flanking region neighboring the DNA binding domain contributes to the chromatin architecture remodeling function of EWS FLI and whether this function could be attributed to the transcriptional output differences in the DBD and DBD+ conditions. Citation Format: Ariunaa Bayanjargal. Studying the role of FLI portion of EWS FLI in transcription regulation via modulation of chromatin 3D landscape in Ewing sarcoma. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Epigenomics; 2022 Oct 6-8; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_2):Abstract nr B018.
42
Hidayatulloh, Taufik, Elindra Yetti, and Hapidin. "Movement and Song Idiom Traditional to Enhance Early Mathematical Skills: Gelantram Audio-visual Learning Media." JPUD - Jurnal Pendidikan Usia Dini 14, no. 2 (November 30, 2020): 215–30. http://dx.doi.org/10.21009/jpud.142.02.
Full textAbstract:
Many studies have shown a link between being competent in early mathematics and achievement in school. Early math skills have the potential to be the best predictors of later performance in reading and mathematics. Movement and songs are activities that children like, making it easier for teachers to apply mathematical concepts through this method. This study aims to develop audio-visual learning media in the form of songs with a mixture of western and traditional musical idioms, accompanied by movements that represent some of the teaching of early mathematics concepts. The stages of developing the ADDIE model are the basis for launching new learning media products related to math and art, and also planting the nation's cultural arts from an early age. These instructional media products were analyzed by experts and tested for their effectiveness through experiments on five children aged 3-4 years. The qualitative data were analyzed using transcripts of field notes and observations and interpreted in a descriptive narrative. The quantitative data were analyzed using gain score statistics. The results showed that there was a significant increase in value for early mathematical understanding of the concepts of geometry, numbers and measurement through this learning medium. The results of the effectiveness test become the final basis of reference for revision and complement the shortcomings of this learning medium. Further research can be carried out to develop other mathematical concepts through motion and song learning media, and to create experiments with a wider sample.
Keywords: Early Mathematical Skills, Movement and Song Idiom Traditional, Audio-visual Learning Media
References
An, S. A., & Tillman, D. A. (2015). Music activities as a meaningful context for teaching elementary students mathematics: a quasi-experiment time series design with random assigned control group. European Journal of Science and Mathematics Education, 3(1), 45–60. https://doi.org/10.1038/srep15999
An, S., Capraro, M. M., & Tillman, D. A. (2013). Elementary Teachers Integrate Music Activities into Regular Mathematics Lessons: Effects on Students’ Mathematical Abilities. Journal for Learning through the Arts: A Research Journal on Arts Integration in Schools and Communities, 9(1). https://doi.org/10.21977/d99112867
Austin, A. M. B., Blevins-Knabe, B., Ota, C., Rowe, T., & Lindauer, S. L. K. (2011). Mediators of preschoolers’ early mathematics concepts. Early Child Development and Care, 181(9), 1181–1198. https://doi.org/10.1080/03004430.2010.520711
Barrett, J. E., Cullen, C., Sarama, J., Miller, A. L., & Rumsey, C. (2011). Children ’ s unit concepts in measurement : a teaching experiment spanning grades 2 through 5. 637–650. https://doi.org/10.1007/s11858-011-0368-8
Basco, R. O. (2020). Effectiveness of Song, Drill and Game Strategy in Improving Mathematical Performance. International Educational Research, 3(2), p1. https://doi.org/10.30560/ier.v3n2p1
Bausela Herreras, E. (2017). Risk low math performance PISA 2012: Impact of assistance to Early Childhood Education and other possible cognitive variables. Acta de Investigación Psicológica, 7(1), 2606–2617. https://doi.org/10.1016/j.aipprr.2017.02.001
Buchoff, R. (2015). Childhood Education. January. https://doi.org/10.1080/00094056.1995.10521830
Clements, D. H. (2014). Geometric and Spatial Thinking in Young Children. In Science of Advanced Materials (Vol. 6, Issue 4). National Science Foundation. https://doi.org/10.1166/sam.2014.1766
Clements, D. H., Baroody, A. J., Joswick, C., & Wolfe, C. B. (2019). Evaluating the Efficacy of a Learning Trajectory for Early Shape Composition. XX(X), 1–22. https://doi.org/10.3102/0002831219842788
Clements, D. H., Swaminathan, S., Anne, M., & Hannibal, Z. (2016). Young Children ’ s Concepts of Shape. 30(2), 192–212.
Cross, C. T., Woods, T., & Schweingruber, H. (2009). Mathematics Learning in Early Chidhood Paths Toward Excellence and Equity. The National Academies Press.
Geary, D. C. (2011). Cognitive predictors of achievement growth in mathematics: A 5-year longitudinal study. Developmental Psychology, 47(6), 1539–1552. https://doi.org/10.1037/a0025510
Geary, D. C. (2012). Learning Disabilities and Persistent Low Achievement in Mathematics. J Dev Behav Pediatr., 32(3), 250–263. https://doi.org/10.1097/DBP.0b013e318209edef.Consequences
Gejard, G., & Melander, H. (2018). Mathematizing in preschool : children ’ s participation in geometrical discourse. 1807. https://doi.org/10.1080/1350293X.2018.1487143
Harususilo, Y. E. (2020). Skor PISA Terbaru Indonesia, Ini 5 PR Besar Pendidikan pada Era Nadiem Makarim. https://pusmenjar.kemdikbud.go.id/
Hsiao, T. (1999). Romanticism with Deep Affection: Selected Articles About the Music of Hsiao Tyzen (Hengzhe Lin (ed.)). Wang Chun Feng Wen Hua Fa Xing.
Kasuya-Ueba, Y., Zhao, S., & Toichi, M. (2020). The Effect of Music Intervention on Attention in Children: Experimental Evidence. Frontiers in Neuroscience, 14(July), 1–15. https://doi.org/10.3389/fnins.2020.00757
Kołodziejski, M., Králová, P. D. E., & Hudáková, P. D. J. (2014). Music and Movement Activities and Their Impact on Musicality and Healthy Development of a Child. Journal of Educational Revies, 7(4).
Kristanto, W. (2020). Javanese Traditional Songs for Early Childhood Character Education. 14(1), 169–184.
Litkowski, E. C., Duncan, R. J., Logan, J. A. R., & Purpura, D. J. (2020). When do preschoolers learn specific mathematics skills? Mapping the development of early numeracy knowledge. Journal of Experimental Child Psychology, 195, 104846. https://doi.org/10.1016/j.jecp.2020.104846
Logvinova, O. K. (2016). Socio-pedagogical approach to multicultural education at preschool. Procedia - Social and Behavioral Sciences, 233(May), 206–210. https://doi.org/10.1016/j.sbspro.2016.10.203
Lopintsova, O., Paloniemi, K., & Wahlroos, K. (2012). Multicultural Education through Expressive Methods in Early Childhood Education.
Ludwig, M. ., Marklein, M. ., & Song, M. (2016). Arts Integration: A Promising Approach to Improving Early Learning. American Institutes for Research.
Macdonald, A., & Lowrie, T. (2011). Developing measurement concepts within context : Children ’ s representations of length. 27–42. https://doi.org/10.1007/s13394-011-0002-7
Mans, M. (2002). Playing The Music- Comparing Perfomance of Children’s Song and dance in Traditional and Contemporary Namibian Education. In The Arts in Children’s Live (pp. 71–86). Kluwer Academic Publishers.
Maričić, S. M., & Stamatović, J. D. (2017). The Effect of Preschool Mathematics Education in Development of Geometry Concepts in Children. 8223(9), 6175–6187. https://doi.org/10.12973/eurasia.2017.01057a
Missall, K., Hojnoski, R. L., Caskie, G. I. L., & Repasky, P. (2015). Home Numeracy Environments of Preschoolers: Examining Relations Among Mathematical Activities, Parent Mathematical Beliefs, and Early Mathematical Skills. Early Education and Development, 26(3), 356–376. https://doi.org/10.1080/10409289.2015.968243
Moreno, S., Bialystok, E., Barac, R., Schellenberg, E. G., Cepeda, N. J., & Chau, T. (2011). Short-term music training enhances verbal intelligence and executive function. Psychological Science, 22(11), 1425–1433. https://doi.org/10.1177/0956797611416999
Nketia, J. H. K. (1982). Developing Contemporary Idioms out of Traditional Music. Studia Musicologica Academiae Scientiarum Hungaricae, 24, 81. https://doi.org/10.2307/902027
Nyota, S., & Mapara, J. (2008). Shona Traditional Children ’ s Games and Play : Songs as Indigenous Ways of Knowing. English, 2(4), 189–203.
Östergren, R., & Träff, U. (2013). Early number knowledge and cognitive ability affect early arithmetic ability. Journal of Experimental Child Psychology, 115(3), 405–421. https://doi.org/10.1016/j.jecp.2013.03.007
Pantoja, N., Schaeffer, M. W., Rozek, C. S., Beilock, S. L., & Levine, S. C. (2020). Children’s Math Anxiety Predicts Their Math Achievement Over and Above a Key Foundational Math Skill. Journal of Cognition and Development, 00(00), 1–20. https://doi.org/10.1080/15248372.2020.1832098
Papadakis, Stamatios, Kalogiannakis, M., & Zaranis, N. (2017). Improving Mathematics Teaching in Kindergarten with Realistic Mathematical Education. Early Childhood Education Journal, 45(3), 369–378. https://doi.org/10.1007/s10643-015-0768-4
Papadakis, Stamatios, Kalogiannakis, M., & Zaranis, N. (2018). The effectiveness of computer and tablet assisted intervention in early childhood students’ understanding of numbers. An empirical study conducted in Greece. Education and Information Technologies, 23(5), 1849–1871. https://doi.org/10.1007/s10639-018-9693-7
Papadakis, Stamatis, Kalogiannakis, M., & Zaranis, N. (2016). Comparing Tablets and PCs in teaching Mathematics: An attempt to improve Mathematics Competence in Early Childhood Education. Preschool and Primary Education, 4(2), 241. https://doi.org/10.12681/ppej.8779
Paul, T. (2019). Mathematics and music : loves and fights To cite this version.
PISA worldwide ranking; Indonesia’s PISA results show need to use education resources more efficiently, (2016).
Phyfferoen, D. (2019). The Dagbon Hiplife Zone in Northern Ghana Contemporary Idioms of Music Making in Tamale. 1(2), 81–104.
Purpura, D. J., Napoli, A. R., & King, Y. (2019). Development of Mathematical Language in Preschool and Its Role in Learning Numeracy Skills. In Cognitive Foundations for Improving Mathematical Learning (1st ed., Vol. 5). Elsevier Inc. https://doi.org/10.1016/b978-0-12-815952-1.00007-4
Ribeiro, F. S., & Santos, F. H. (2020). Persistent Effects of Musical Training on Mathematical Skills of Children With Developmental Dyscalculia. Frontiers in Psychology, 10(January), 1–15. https://doi.org/10.3389/fpsyg.2019.02888
Roa, R., & IA, C. (2020). Learning Music and Math, Together as One: Towards a Collaborative Approach for Practicing Math Skills with Music. In I. T. (eds) Nolte A., Alvarez C., Hishiyama R., Chounta IA., Rodríguez-Triana M. (Ed.), Collaboration Technologies and Social Computing. Col (Vol. 26, Issue 5, pp. 659–669). https://doi.org/https://doi.org/10.1007/978-3-030-58157-2_10
Sarama, J., & Clements, D. H. (2006a). Mathematics, Young Students, and Computers: Software, Teaching Strategies and Professional Development. The Mathematics Educato, 9(2), 112–134.
Sarama, J., & Clements, D. H. (2006b). Mathematics in early childhood. International Journal of Early Childhood, 38(1). https://doi.org/10.1007/bf03165980
Sarkar, J., & Biswas, U. (2015). The role of music and the brain development of children. 4(8), 107–111.
Sheridan, K. M., Banzer, D., Pradzinski, A., & Wen, X. (2020). Early Math Professional Development: Meeting the Challenge Through Online Learning. Early Childhood Education Journal, 48(2), 223–231. https://doi.org/10.1007/s10643-019-00992-y
Silver, A. M., Elliott, L., & Libertus, M. E. (2021). When beliefs matter most: Examining children’s math achievement in the context of parental math anxiety. Journal of Experimental Child Psychology, 201, 104992. https://doi.org/10.1016/j.jecp.2020.104992
Sterner, G., Wolff, U., & Helenius, O. (2020). Reasoning about Representations: Effects of an Early Math Intervention. Scandinavian Journal of Educational Research, 64(5), 782–800. https://doi.org/10.1080/00313831.2019.1600579
Temple, B. A., Bentley, K., Pugalee, D. K., Blundell, N., & Pereyra, C. M. (2020). Using dance & movement to enhance spatial awareness learning. Athens Journal of Education, 7(2), 153–167. https://doi.org/10.30958/aje.7-2-2
Thippana, J., Elliott, L., Gehman, S., Libertus, K., & Libertus, M. E. (2020). Parents’ use of number talk with young children: Comparing methods, family factors, activity contexts, and relations to math skills. Early Childhood Research Quarterly, 53, 249–259. https://doi.org/10.1016/j.ecresq.2020.05.002
Tsai, Y. (2017). Taiwanese Traditional Musical Idioms Meet Western Music Composition: An Analytical and Pedagogical Approach to Solo Piano Works by Tyzen Hsiao. http://aquila.usm.edu/dissertations/1398
Upadhyaya, D. (2017). Benefits of Music and Movement in young children. Furtados School of Music. https://www.linkedin.com/pulse/benefits-music-movement-young-children-dharini-upadhyaya
Vennberg, H., Norqvist, M., Bergqvist, E., Österholm, M., Granberg, C., & Sumpter, L. (2018). Counting on: Long Term Effects of an Early Intervention Programme. 4, 355–362. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-148101
Verdine, B. N., Lucca, K. R., Golinkoff, R. M., Hirsh-, K., & Newcombe, N. S. (2015). The Shape of Things : The Origin of Young Children ’ s Knowledge of the Names and Properties of Geometric Forms. 8372(October). https://doi.org/10.1080/15248372.2015.1016610
Wakabayashi, T., Andrade-Adaniya, F., Schweinhart, L. J., Xiang, Z., Marshall, B. A., & Markley, C. A. (2020). The impact of a supplementary preschool mathematics curriculum on children’s early mathematics learning. Early Childhood Research Quarterly, 53, 329–342. https://doi.org/10.1016/j.ecresq.2020.04.002
Wardani, I. K., Djohan, & Sittiprapaporn, P. (2018). The difference of brain activities of musical listeners. 1st International ECTI Northern Section Conference on Electrical, Electronics, Computer and Telecommunications Engineering, ECTI-NCON 2018, 181–184. https://doi.org/10.1109/ECTI-NCON.2018.8378307
Winter, E., & Seeger, P. (2015). The Important Role of Music in Early Childhood Learning. Independent School.
Zaranis, N., Kalogiannakis, M., & Papadakis, S. (2013). Using Mobile Devices for Teaching Realistic Mathematics in Kindergarten Education. Creative Education, 04(07), 1–10. https://doi.org/10.4236/ce.2013.47a1001
43
Malinge, Sebastien, Gina Kirsammer, Meghan Bliss-Moreau, Timothy M. Chlon, Lauren Diebold, Stella T. Chou, Mitchell Weiss, Sandeep Gurbuxani, and John Crispino. "Development of a Mouse Model of DS Megakaryoblastic Leukemia and Identification of the Hsa21 Gene DYRK1A as a Prominent Contributor to DS-AMKL." Blood 116, no. 21 (November 19, 2010): 469. http://dx.doi.org/10.1182/blood.v116.21.469.469.
Full textAbstract:
Abstract Abstract 469 Children with Down syndrome (DS) have a 500-fold increased risk of Acute Megakaryoblastic Leukemia (DS-AMKL). In addition to trisomy 21, DS-AMKL blasts harbor somatic mutations in the gene encoding GATA-1, an essential transcriptional regulator of erythroid and megakaryocytic differentiation. In every case, the GATA1 mutations lead to a block in expression of the full-length protein, but allow for the translation of a shortened isoform named GATA-1s, which lacks the N-terminal transcriptional activation domain. Recent studies have shown that mutations in of JAK2, JAK3 and MPL are another feature of AMKL. Whereas the impact on hematopoiesis of individual GATA-1s, JAK2 V617F and MPL W515L mutants is relatively well known, the specific effects of trisomy 21, the initiating event in DS leukemogenesis, remains unclear. Here, we used two different experimental models to identify specific genes on human chromosome 21 (Hsa21) that participate in leukemogenesis: human DS-AMKL cell lines and the Ts1Rhr mouse strain, which is trisomic for only 33 orthologs of Hsa21 genes. Comprehensive analysis of hematopoiesis in progressively more complex mouse models of DS disorders, including Ts1Rhr mice, Ts1Rhr/KI-Gata-1s compound mutant mice, and recipients of Ts1Rhr/KI-Gata-1s hematopoietic progenitors expressing MPL W515L, revealed that the presence of trisomy 21 impacts the phenotype of each genetic background. Ts1Rhr mice develop a progressive myeloproliferative disease with megakaryocytic features while Ts1Rhr/KI-Gata1s double transgenic mice develop thrombocytosis at an earlier age compared to Gata1s mutant or Ts1Rhr mice alone. However neither the Ts1Rhr nor the compound mutant mice develop AMKL. In stark contrast, expression of MPL W515L in Ts1Rhr/KI-Gata1s hematopoietic progenitors led to a rapid lethal megakaryoblastic disorder characterized by profound marrow and splenic fibrosis and the presence of immature megakaryoblasts in lethally irradiated recipients. Of note, transplantation of MPL W515L expressing singly mutant Ts1Rhr or KI-Gata-1s progenitors failed to cause a similar disease. These findings show that three oncogenic events are necessary and sufficient to induce a megakaryoblastic leukemia in recipient mice. In parallel, to identify specific genes on Hsa21 that contribute to AMKL, we performed shRNA screening in DS-AMKL cell lines by knocking down the human orthologs of the 33 genes trisomic in the Ts1Rhr mice. We then evaluated the consequence of knockdown on megakaryocyte proliferation, survival and/or differentiation. This study revealed that altered expression of ERG, DYRK1A, CHAF1B and HLCS affected megakaryocyte growth. Analysis of their expression in human samples suggest that these four genes likely play a role in etiology of DS-AMKL. Moreover expression evaluation of those genes in our in vivo models of 1, 2 or 3 genetic events revealed that ERG, a known megakaryoblastic oncogene, and DYRK1A are the most prominent genes implicated in these megakaryocytic disorders. DYRK1A (dual specificity tyrosine-regulated kinase 1A) has been functionally implicated in many developmental disorders associated with DS and shown to be constitutively phosphorylated/activated in human AMKL cell lines. We found that overexpression of DYRK1A cooperates with Gata1s to lead to a increase in megakaryocyte growth in liquid culture and in colony assays in vitro. In summary, we show that partially trisomic Ts1Rhr mice display progressive defects in the megakaryocytic compartment, that mutagenesis of Gata1 in the Ts1Rhr background exacerbates the Ts1Rhr phenotype, and that addition of an activating mutation of MPL leads to a fulminant megakaryocytic leukemia with myelofibrosis. Moreover, among the 33 genes present in 3 copies in the Ts1Rhr background, we identify DYRK1A as a likely megakaryoblastic oncogene in DS-AMKL. Disclosures: No relevant conflicts of interest to declare.
44
Chiabrando, Deborah, Carlotta Giorgi, Lorenzo Silengo, Fiorella Altruda, Paolo Pinton, and Emanuela Tolosano. "Mitochondrial Heme Export Through FLVCR1b Controls Erythroid Differentiation." Blood 120, no. 21 (November 16, 2012): 2090. http://dx.doi.org/10.1182/blood.v120.21.2090.2090.
Full textAbstract:
Abstract Abstract 2090 Feline Leukemia Virus subgroup C Receptor 1 (FLVCR1) is a cell membrane heme exporter that contributes to maintain the balance between heme level and globin synthesis in erythroid precursors. Consistently, FLVCR1-null mice died in utero due to a failure of erythropoiesis1. We previously reported the identification of a mitochondrial isoform of FLVCR1, named FLVCR1b, that was able to support fetal murine erythroid differentiation in the absence of the cell membrane isoform (herein called FLVCR1a)2. The aim of this work was to investigate the role of FLVCR1b during erythroid differentiation. To this end, overexpression and silencing experiments were performed. FLVCR1b overexpression promotes in vitro erythroid differentiation of K562 cells, as indicated by the increased transcription of globin mRNA and a higher percentage of benzidine positive cells, compared to control cells. On the contrary, FLVCR1a-overexpressing K562 cells were not able to differentiate. Interestingly, shRNA-mediated down-regulation of FLVCR1a led to the elevation of the percentage of benzidine positive cells compared to controls that further increased upon the stimulation of erythroid differentiation using sodium butyrate. A decrease of the percentage of benzidine positive cells was observed in K562 cells lacking both FLVCR1 isoforms compared to FLVCR1a-deficient cells. Moreover these cells were not able to differentiate upon stimulation with sodium butyrate. These results suggest a fundamental role of FLVCR1b during erythroid differentiation in vitro, supporting previous data obtained in the mouse model2. To understand the molecular mechanisms in which FLVCR1b is involved, we investigate whether FLVCR1b could affect heme export from mitochondria. The overexpression of FLVCR1b in HeLa cells led to increased intracellular heme content together with a strong induction of the heme degrading enzyme heme oxygenase 1 (HO-1) mRNA. Inhibition of the heme biosynthetic pathway using succinylacetone, completely prevented intracellular accumulation of heme observed when FLVCR1b was overexpressed. So, increased heme biosynthesis rate is responsible for the elevation of heme content. Accordingly, HeLa cells overexpressing FLVCR1b showed an alteration of heme biosynthesis enzymes and transporters. On the contrary, the specific loss of FLVCR1b using siRNA causes heme accumulation in mitochondria and a subsequent block of heme biosynthesis. These data are consistent with a role of FLVCR1b as a mitochondrial heme exporter. Similar results were also obtained in K562 cells thus suggesting that loss of FLVCR1b reduces the availability of heme for haemoglobin synthesis, a process essential during erythroid differentiation. To assess whether mitochondrial heme accumulation due to the loss of FLVCR1b affect mitochondrial functionality, the mitochondrial Ca2+ response after agonist stimulation was monitored as a highly sensitive readout of mitochondrial state. It is well known that mitochondrial alterations cause defects in Ca2+ uptake by the organelle. The silencing of FLVCR1a and FVLCR1b in HeLa cells caused a significant reduction of Ca2+spike in the mitochondrial matrix evoked by agonist stimulation. These data suggested that when FLVCR1b is lost heme accumulates in mitochondria resulting in the alterations of mitochondrial functionality. All together these data indicate that the impairment of erythroid differentiation observed in the absence of FLVCR1b is due to the block of heme export from mitochondria and a consequent impairment of mitochondrial functionality, which is essential for cell survival. These results, linking heme biosynthesis pathway to mitochondrial Ca2+signaling, will have broad implications in cellular metabolism. Disclosures: No relevant conflicts of interest to declare.
45
Nan, Li, Haiyan Chen, Jian Ruan, Rebecca S. Sollie, Cara Schafer, Francis T. Crittenden, Anurag Purushothaman, Ralph D. Sanderson, Amjad Javed, and Yang Yang. "Runx2 Transcription Factor Regulates Heparanase-Induced Bone Resorption in Multiple Myeloma." Blood 120, no. 21 (November 16, 2012): 567. http://dx.doi.org/10.1182/blood.v120.21.567.567.
Full textAbstract:
Abstract Abstract 567 Background. Despite advances in treatment strategies, myeloma remains incurable, and bone destruction is a major cause of morbidity in myeloma patients. We have documented in earlier studies that heparanase enzyme is preferentially expressed in myeloma cells and induces severe bone destruction in myeloma. We also discovered that heparanase increases the production of two major bone resorbing factors named Receptor Activator of NF-κB Ligand (RANKL) and Matrix Metalloproteinase 9 (MMP-9) by myeloma cells. Runx2, a member of the runt-related gene family, is a bone-specific transcription factor. Runx2 regulates osteoblast differentiation and is essential for bone tissue development. Interestingly, Runx2 also controls expression of RANKL and MMP-9 genes in osteoblasts. Recent evidence indicates that ectopic induction and overexpression of Runx2 in breast, uterine and prostate cancer cells is associated with bone-metastasis, and osteolytic bone disease in these cancers. However, very little is known about the function of Runx2 in myeloma cells. Here we report for the first time that heparanase engages the Runx2 pathway to promote expression of RANKL and MMP-9 in myeloma cells. Methods. Molecular, biochemical, cellular and in vivo approaches were used to assess the role of Runx2 in heparanase-induced expression of RANKL and MMP-9. These included: (1) Real-time PCR and Western blot analysis to monitor Runx2 levels in CAG myeloma cells expressing high level of heparanase (HPSE-high cells) or with knockdown of endogenous heparanase (HPSE k/d cells), and the corresponding control cells. (2) Chromatin Immunoprecipitation (ChIP) assay to determine in vivo occupancy of the RANKL and MMP-9 gene promoters in myeloma cells by Runx2. (3) Zymography to determine MMP-9 activity in both human and murine myeloma cells. (4) Real-time PCR to determine changes in RANKL and MMP-9 gene expression in Runx2 knockdown MM1.S and 5TGM1 myeloma cells. (5) Assessment of tumor growth/burden and bone resorption in the 5TGM1 syngenic model of murine myeloma. 5TGM1 cells with specific knockdown of Runx2 or non-target shRNA were injected into C57BL/KaLwRij mice through the tail vein and levels of IgG2b and TRAP5b were monitored in the sera of the mice by ELISA. Results. We find Runx2 expression is significantly increased in CAG myeloma cells expressing a high level of heparanase and dramatically reduced in HPSE k/d cells. Increased Runx2 in HPSE-high myeloma cells results in an increase in expression of RANKL and MMP-9 mRNA as well as MMP-9 activity. In sharp contrast, knockdown of Runx2 in human myeloma MM1.S and murine myeloma 5TGM1 cells results in a significant reduction of RANKL, MMP-9 gene expression and MMP-9 activity. Thus heparanase promotes RANKL and MMP-9 expression via Runx2. Analysis of initial 1-kb sequences of RANKL and MMP-9 gene promoter in human and mice, revealed presence of multiple high affinity Runx2-binding sites. ChIP assay confirmed that Runx2 mediates induction of RANKL and MMP-9 gene transcription in the both human and murine myeloma cells by direct association with the proximal promoter of these genes. Together these results demonstrate that Runx2 is a positive regulator of RANKL and MMP-9 gene expression in myeloma cells. Finally, in the syngenic model, knockdown of Runx2 in 5TGM1 murine myeloma cells remarkably inhibits the growth of these cells in vivo and bone resorption. Conclusions. Runx2 is a central regulator of heparanase induced myeloma bone disease. Our discoveries provide new insight into the mechanism of myeloma-induced bone disease and identify Runx2 as a novel target to block myeloma bone disease. Disclosures: No relevant conflicts of interest to declare.
46
Xu, Jing, Yu Liu, Chong Chen, and Ting Niu. "KMT2D Is a Haploinsufficient Tumor Suppressor in Acute Leukemia." Blood 132, Supplement 1 (November 29, 2018): 1511. http://dx.doi.org/10.1182/blood-2018-99-119038.
Full textAbstract:
Abstract Introduction: Epigenetic dysregulation plays a critical role in hematologic tumorigenesis and cancer progression. Histone-lysine N-methyltransferase 2D (KMT2D), also known as MLL4 (mixed-lineage leukemia 4), belongs to a family of mammalian histone H3 lysine 4 (H3K4) methyltransferases, which is amongst the most frequently mutated epigenetic genes in human cancers. It was reported that deletion of KMT2D inhibits MLL1-fusion-induced acute myeloid leukemia (Santos et al. 2014). However, in human cancers, the vast majority of the mutations in KMT2D is heterozygous deletion. Recent studies have revealed the role of KMT2D in lymphomagenesis, but the potential tumor suppressor function of KMT2D in leukemogenesis remains largely unclear. In addition, KMT2D was found to exist within a macromolecular complex named COMPASS (complex of proteins associated with Set1). KMT2C is another component of the complex, and was validated as another haploinsufficient tumor suppressor gene in acute myeloid leukemia (Chen et al. 2014). Mutations in KMT2D and KMT2C frequently co-occur in hematologic malignancies. Thus, we sought to first examine the role and molecular mechanism of KMT2D in acute leukemia, and then investigate whether co-mutated gene KMT2D and KMT2C have cooperative function in malignancies initiation and progression. Methods: Here we utilized RNAi approach to knockdown the expression of Kmt2d, and an approximate 50% reduction in gene dosage was validated by quantitative RT-PCR. We introduced two independent short-hairpin RNAs (shRNAs) targeting Kmt2d and a shRNA targeting Nf1 into p53 null hematopoietic stem and progenitor cells (hereafter referred to as shKmt2d;shNf1;p53-/- cells), and transplanted them into sublethally irradiated mice to model the impact of haploinsufficient tumor suppressors. The recipient mice were then monitored for the emergence of diseases by complete blood count as well as overall survival. Bone marrow was harvested and analyzed by flow cytometry after sacrifice. Histological analyses of blood smear, liver, spleen and bone marrow were conducted to accurately diagnose the disease. Besides, we established Kmt2d heterozygous and homozygous conditional knockout mouse models (Kmt2d fl/+; Mx1-Cre, Kmt2d fl/fl; Mx1-Cre) to further investigate the respective role of haploinsufficiency and deletion of KMT2D in acute leukemia. To study the cooperative function of KMT2D and KMT2C in leukemogenesis, two genes were co-suppressed by shRNAs in the same aforementioned genetic background (shNf1; p53-/-). Transplantation-based mouse modeling approach was used and the recipient mice were monitored for evidence of hematologic diseases. Results: A significant increase in peripheral white blood cell counts, anemia and thrombocytopenia were noticed in recipients of shKmt2d;shNf1;p53-/- cells over the same period after 4 weeks post-transplantation. Massive hepatomegaly and splenomegaly were displayed in those mice and a dramatic reduction in survival was observed (shKmt2d-1: median survival = 47 days, p = 0.0004 versus shRen; and shKmt2d-2: median survival = 74 days, p = 0.0127 versus shRen). Flow cytometry of cells from bone marrow of sacrificed mice showed a substantial enrichment of doule-positve cells (shRNAs targeting Kmt2d were linked to GFP and those targeting Nf1 were linked to mCherry) as compared to pre-injected, which were filled with leukemic cells that expressed myeloid/lymphoid lineage markers. Pathological analyses demonstrated the onset of acute myeloid/lymphoid leukemia. In Kmt2d and Kmt2c double-knockdown animal assay, the onset of acute myeloid leukemia was promoted compared with single-knockdown groups. (shKmt2d;shKmt2c: median survival = 48.5 days, p < 0.0001 versus shKmt2d; p = 0.0013 versus shKmt2c). Conclusions: KMT2D is a haploinsufficient tumor suppressor in acute leukemia. KMT2D deficiency cooperates with KMT2C to promote the development and progress of acute leukemia. Further understandings of the molecular mechanism of KMT2D as a haploinsufficient tumor suppressor in acute leukemia and the cooperative function of KMT2D and KMT2C in leukemogenesis are demanded, which can provide insights into developing novel therapeutic targets. Figure. Figure. Disclosures No relevant conflicts of interest to declare.
47
Godefroy, Emmanuelle, Patrice Chevallier, Thierry Guillaume, Pierre Peterlin, Alice Garnier, Amandine Le Bourgeois, Annabelle Pédron, Frédéric Altare, and Francine Jotereau. "Gut Microbiota-Induced Regulatory T Cells in Patients with Hematological Malignancies Receiving Allogeneic Hematopoietic Stem Cell Transplantation: Towards Deciphering a Role for These Tregs in aGVHD." Blood 136, Supplement 1 (November 5, 2020): 34–35. http://dx.doi.org/10.1182/blood-2020-136510.
Full textAbstract:
Prognosis for hematological malignancies treated with allogeneic stem cell transplantation (allo-HSCT) remains dismal and can often induce potentially deadly complications, such as graft versus host disease (GvHD). Steroids remain the first-line treatment for aGvHD. Nevertheless, new treatments are needed to improve allo-HSCT outcomes and reduce steroid complications. In this context, increasing evidences suggest that gut microbiota composition and the activity of regulatory T cells (Tregs) are involved in GvHD prevention (Elias and Rudensky, 2019, Br. J. Haematol. ; Shono and van den Brink, 2018, Nat. Rev. Cancer). We have identified a novel FoxP3-negative IL-10-secreting Treg subset, named DP8α, enriched in the colon mucosa and present in blood, characterized by its CD3+/CD4+/CD8αLOW/ CCR6+/CXCR6+ phenotype and its TCR-specificity for the gut ClostridiumIV bacterium Faecalibacterium prausnitzii (Sarrabayrouse et al., 2014, PLoS Biol ; Godefroy et al., 2018, Gastroenterology). Sizable fractions of these cells also expressed the membrane-bound ectonucleotidases CD39 and CD73 (gating strategy shown in Figure 1), which are directly involved in their suppressive activity in vitro (Godefroy et al., 2018, Gastroenterology). In vivo, these molecules play key regulatory roles both by degrading extracellular ATP and thus limiting its inflammasome-related proinflammatory role, and by inducing the production of immunosuppressive adenosine. Accordingly, an alteration in adenosine/purinergic signaling has been suggested in GvHD occurrence (Deaglio et al., 2007, J. Exp. Med. ; Wang et al., 2013, PLoS ONE). Thus, we postulated that F. prausnitzii-reactive DP8α Tregs, whose suppressive activity depends on purinergic signaling, could bridge microbiota dysbiosis and GvHD incidence in allo-HSCT patients. In support of this, decreased levels of Clostridium bacteria, especially Faecalibacteriumspp, in patient's stools, have been associated with aGvHD risk (Noor et al., 2019, J Innate Immun). Moreover, mice gavaged with butyrate-producing and Treg-inducing Clostridia displayed milder GvHD and improved survival (Mathewson et al., 2016, Nat. Immunol.). Therefore, we recently analyzed 63 patients with hematological malignancies, who received allo-HSCT, among which 21 developed an aGvHD (pathologies and treatments are listed Table I). We quantified circulating DP8α Tregs and analyzed their expression of CD39 and CD73 in donors, before and after mobilization, and in patients at one week before and 1, 2, and 3 months post allo-HSCT. Strikingly, results clearly showed that aGvHD development was strongly associated with a lack of expression of CD73 by DP8α Tregs (p<.0001) (Fig.2A), but not by any other T cell subsets analyzed. DP8α frequency and their CD39 expression were similar in GvHD+ versus GvHD- patients. Importantly, CD73 decrease did not result from their corticotherapy since it was equally observed before and after treatment (Fig.2B) and since in vitro treatment of PBMCs with methylprednisolone did not alter CD73 expression whatsoever. Therefore, the CD73 decrease observed in aGvHD patients, appears relevant and likely related to the aGvHD condition itself. It is noteworthy that this cohort was rather heterogeneous in terms of hematological diseases, conditioning regimens and prophylaxis treatments (Table I). Nevertheless, this drastic CD73 decrease in aGvHD patients was observed across all these conditions, strongly strengthening its relevancy. Altogether, these data strongly suggest that a CD73-dependent functional alteration of DP8α Tregs are, at least in part, involved in aGvHD occurrence. These results could therefore be used to predict GvHD risk and also give rise to the development of therapeutic strategies. Such innovative treatments could be based on the infusion of DP8α Tregs with appropriate features, since we have shown both the uniquely high proliferating potential and the stable immunosuppressive function of these cells in vitro. Moreover, administration of DP8α target antigens (F. prausnitzii-derived), in the form of peptides, proteins, or even bacterial lysates, such as probiotics, could also be foreseen to either directly in vivo expand these Tregs or indirectly through the induction of tolerogenic dendritic cells (Alameddine et al., 2019, Front Immunol) and subsequent Treg differentiation/priming to limit GvHD-related inflammation. Figure 1 Disclosures Chevallier: Incyte Corporation: Honoraria.
48
Amodio, Nicola, Marzia Leotta, Anna Maria Gullà, Lavinia Raimondi, Eugenio Morelli, Maria Angelica Stamato, Teresa Calimeri, et al. "MiR-29b Counteracts Aberrant HDAC4 Expression and Enhances Vorinostat Activity in Multiple Myeloma." Blood 124, no. 21 (December 6, 2014): 2060. http://dx.doi.org/10.1182/blood.v124.21.2060.2060.
Full textAbstract:
Abstract Gene transcription within the tumor cell and its microenvironment can be affected by epigenetic modulation in histones and inhibition of histone deacetylases is presently a promising therapeutic option in multiple myeloma (MM). MicroRNAs (miRNAs) are non-coding RNAs that control gene expression; a subclass of miRNAs, named epi-miRNAs, exerts anti-tumor activity by targeting effectors of the epigenetic machinery. We recently demonstrated a key role of tumor suppressor miR-29b as epi-miRNA able to target DNA-methyltransferases and to reduce aberrant hypermethylation in MM. In silico search of miR-29b targets further clarifying its epi-miRNA function, unveiled the histone deacetylase HDAC4. Here, we aimed at characterizing HDAC4 expression, function and its regulation by miR-29b in MM. HDAC4 mRNA, protein levels and activity were found increased in 11 out of 11 MM cell lines as compared to 3 healthy controls. The analysis of our microarray dataset consisting of plasmacells from 4 healthy donors, 55 MM and 29 plasma cell leukemia patients indicated HDAC4 overexpression in cancer samples, suggesting that high HDAC4 expression might be involved in MM pathogenesis. In the same dataset, HDAC4 mRNA expression inversely correlated with miR-29b levels. To investigate the mechanism by which miR-29b could affect HDAC4 expression, we transfected SKMM1 cells with miR-29b mimics, which resulted in down-modulation of HDAC4 at mRNA and protein level, along with increased acetylation at lysine 8 of histone H4 as well as of tubulin acetylation. This effect was isoform-specific, since levels of other HDACs (namely HDAC1, 2, 6) remained unchanged after miR-29b overexpression. Moreover, miR-29b mimics inhibited the 3’UTR of HDAC4 cloned in a luciferase vector whereas failed to regulate a mutant devoid of a predicted miR-29b target site. Through loss of function experiments, we assessed the functional significance of HDAC4 in MM cells. Stable shRNA-mediated silencing of HDAC4 phenocopied our previously reported miR-29b-triggered biological effects, since it led to growth inhibition, caspase 3/7-dependent apoptosis, inhibition of cell migration and induction of autophagy in KMS11 and NCI-H929 MM cells. HDAC4 depletion by shRNAs potentiated the in vitro anti-MM activity of dexamethasone, bortezomib and vorinostat. Notably, silencing of HDAC4 led to miR-29b upregulation, likely as consequence of histone H4 hyperacetylation at miR-29b promoter, as assessed by Chip experiments; conversely, HDAC4 overexpression strongly reduced miR-29b levels. These results underscore the presence of a negative feedback loop between miR-29b and its target. Adhesion to BM stromal cells reduced miR-29b levels while induced up-regulation of HDAC4 mRNA levels in NCI-H929 cells, thus suggesting that the huBMM might increase HDAC activity in MM cells via miR-29b down-modulation. Importantly, the pan-HDAC inhibitor vorinostat also triggered apoptosis, autophagy and anti-migratory effects in MM cells, along with the induction of miR-29b and the down-regulation of HDAC4 and of other miR-29b targets like Sp1, CDK6 and MCL-1; these phenotypical changes were potentiated by miR-29b mimics, whereas were strongly reduced by antagomiR-29b transduction, thus demonstrating a functional involvement of miR-29b in the anti-MM activity of vorinostat. Finally, in vivo studies using a KMS11 xenograft murine model of MM demonstrated a significant potentiation of vorinostat-induced tumor growth inhibition by subcutaneously delivered neutral lipid emulsion-formulated miR-29b synthetic mimics. Collectively, our results demonstrate that HDAC4 is aberrantly expressed and represents a novel therapeutic target in MM; moreover, our investigation provides the preclinical rationale for using miR-29b mimics to potentiate HDAC inhibitors activity in MM. Disclosures No relevant conflicts of interest to declare.
49
NİKABADZE, Nona, Lile TANDILAVA, and Maia BARAMIDZE. "Şota Rustaveli’nin “Kaplan Postlu Şövalye” Destanın Türkçe Çevirisinde Müzik Çalgıları." Rast Müzikoloji Dergisi, September 13, 2022. http://dx.doi.org/10.12975/rastmd.20221034.
Full textAbstract:
Shota Rustaveli's poem "The Knight in the Panther's Skin" is an epic poem containing medieval lyrical passages. The poem is distinguished by a variety of musical instruments. Our article deals with the issue of translating the most commonly used musical instruments in the Turkish translation of "The Knight in the Panther's Skin ". Using empirical material, source and target texts have been compared to understand how adequately the musical instruments are translated in the Turkish translation of "The Knight in the Panther's Skin " performed by Bilal Dindar.
Our goal is to investigate the principle of selection of equivalents and evaluate its adequacy. In this article, we will discuss the most commonly used musical instruments, which can be grouped as follows: String instruments - Chang, Eban, Barbuti, Chaghana; Wind instruments -Buki and Na; Percussion musical instrument -Tsintsilani; Drums - Noba and Nobati, Kosi, Tablaki. In many cases, these musical instruments have the corresponding words in the Turkish language, however, the translator does not translate them into the target language. Based on the comparison of the texts, we will empirically determine the proximity of the source text and the translation text.
The methodological basis of the study is a corpus-linguistic analysis of the original text and the translation of "The Knight in the Panther's Skin". In the process of work, the corpus-linguistic method is used, which is common in modern humanitarian studies. Using this method allows us to quickly and efficiently process large volumes of empirical material.
50
Guo, Qian. "Can Zimbabwe be saved from the Brink of Collapse?" Journal of Contemporary Educational Research 4, no. 1 (December 23, 2019). http://dx.doi.org/10.26689/jcer.v4i1.962.
Full textAbstract:
Zimbabwe, a country in the Southern Africa, is known as the “house of stones” which named the country with Shona language. In the past, Zimbabwe has been called as “the breadbasket of Africa”. However, Zimbabweans are living among no power, no water, and no money. What have caused for such situation and how can people save Zimbabwe from the brink of collapse?