The Role of H3.3 histone variant in the pathogenesis of oral Kaposi's Sarcoma

H3.3组蛋白变异在口腔卡波西肉瘤发病机制中的作用

• 批准号: 10418661
• 负责人: ROLF F RENNE
• 金额: $ 35.53万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-04 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10418661
• 关键词: AIDS related cancer; ATRX gene; Acquired Immunodeficiency Syndrome; Address; Affect; B-Lymphocytes; Binding; Biology; Biopsy; CRISPR/Cas technology; Cell Line; Cells; Chromatin; Chromatin Structure; Clinical; Collaborations; Data; Dentistry; Deposition; Endothelial Cells; Epigenetic Process; Episome; Epithelial; Epithelial Cells; Equilibrium; Etiology; Formalin; Freezing; Gene Expression; Genes; Genetic Transcription; Genome; Gingiva; Goals; Growth; HIV; Head and neck structure; Herpesviridae; Histone H3; Histones; Horizontal Disease Transmission; Hour; Human; Human Herpesvirus 8; Incidence; Individual; Infection; Kaposi Sarcoma; Knock-in; Knock-out; Lymphoid; Lymphoid Cell; Lymphoma cell; Lymphoproliferative Disorders; Lytic; Lytic Phase; Maintenance; Malignant Neoplasms; Maps; Modification; Molecular Chaperones; Multicentric Angiofollicular Lymphoid Hyperplasia; Mutation; Names; Nucleosomes; Oral; Oral cavity; Palate Kaposi' s Sarcoma; Pathogenesis; Pathology; Pathway interactions; Patients; Pattern; Phenotype; Play; Post-Translational Protein Processing; Primary Infection; Process; Recombinants; Recurrence; Regulation; Role; Saliva; Sampling; Site; Small Interfering RNA; Specimen; Testing; Therapeutic; Transformed Cell Line; Translating; Variant; Viral; Viral Gene Expression Regulation; Viral Genes; Viral Genome; Viral Proteins; Virus; Work; antiretroviral therapy; base; cell type; centromere protein A; chromatin remodeling; college; comparative; epigenome; gene product; in vivo; knock-down; latency-associated nuclear antigen; latent infection; loss of function; lytic gene expression; lytic replication; malignant mouth neoplasm; neoplastic cell; new therapeutic target; novel; oral cavity epithelium; primary effusion lymphoma; transmission process; tumor; viral transmission

Abstract Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi's sarcoma (KS), and two lymphoproliferative diseases: primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). KS is the most common oral cancer in human immunodeficiency virus (HIV)-infected patients. The oral cavity is the major site for virus transmission via saliva. Although the incidence of AIDS-KS cases has been reduced in the US due to combination antiretroviral therapy (cART), recent studies suggest recurrence of oral KS in long-term treated AIDS patients. While KS tumor cells are mostly latently infected, and express a limited number of viral genes, gingival epithelial cells in the oral cavity actively replicate KSHV which is important for a) seeding the latency reservoir in B cells after primary infection and b) shedding in the oral cavity and transmission via saliva. Epigenetics and chromatin structure play a central role in the regulation of both transcription and replication. Deregulation, aberrant deposition, and mutations of histone variants such as H3.3, CENP-A, and H2A.Z and their associated chaperones have been implicated in multiple human cancers including head and neck. After de novo infection, KSHV genomes rapidly associate with nucleosomes which acquire specific epigenetic modifications that partition the viral episome into transcriptionally active and silenced domains. While multiple KSHV epigenetic marks have been mapped in lymphoid, epithelial and endothelial cells, the processes and potential host- viral interactions leading to the formation of latency permissive episomes are still poorly understood. Given that the KSHV latency-associated nuclear antigen (LANA) interacts with both chromatin remodelers and H3.3 histone chaperones Daxx, HIRA, and DEK, we hypothesize that H3.3 deposition plays an important role in the establishment and maintenance of KSHV latency. Our preliminary data demonstrate that H3.3 deposition on KSHV episomes can be detected early after de novo infection on episomes of long-term infected cells. Moreover, we demonstrated that genetically disrupting the H3.3 chaperone pathways HIRA and Daxx by CRISPR/Cas9 leads to marked changes in KSHV latency control, associated with alterations in the epigenetic status. We further hypothesize that latency-associated gene products including LANA modulate and or de-regulate histone chaperone pathways during de novo infection and latency. To directly address this hypothesis we propose to mechanistically study histone H3.3 deposition and the formation of histone post translational modifications (PTMs), with the goal of understanding their relationship with respect to viral gene expression and latent/lytic replication. A main aspect of these studies is to investigate how viral gene products interact with and modulate histone variant chaperone pathways. In addition, we use human oral primary epithelial cells which are more lytic than transformed epithelial cell lines to characterize epigenetic changes responsible for their lytic phenotype. Importantly, through a collaboration with Dr. Donald Cohen, College of Dentistry, we propose to establish in vivo epigenomes of oral KS in both formalin-fixed and snap frozen tumor samples. The overall goal is to investigate how histone variant deposition determines latent and lytic infection in a cell type-specific manner. The results may point to KSHV-specific novel therapeutic targets for oral KS.

摘要 卡波西肉瘤相关疱疹病毒（KSHV）是卡波西肉瘤（KS）的病原体， 和两种淋巴组织增生性疾病：原发性渗出性淋巴瘤（PEL）和多中心Castleman's 疾病（MCD）。KS是人类免疫缺陷病毒（HIV）感染者中最常见的口腔癌 患者口腔是病毒通过唾液传播的主要场所。虽然发病率 由于联合抗逆转录病毒治疗（cART），美国的艾滋病KS病例已经减少， 研究表明长期治疗的艾滋病患者的口服KS复发。虽然KS肿瘤细胞 大多数潜伏感染，并表达有限数量的病毒基因，牙龈上皮细胞中的 口腔主动复制KSHV，这对于a）在B细胞中接种潜伏储库是重要的 初次感染后和B）在口腔中脱落并通过唾液传播。表观遗传学和 染色质结构在转录和复制的调节中起中心作用。 组蛋白变体如H3.3、CENP-A和H3.3的失调、异常沉积和突变。 H2A.Z及其相关分子伴侣与多种人类癌症有关 包括头部和颈部。在从头感染后，KSHV基因组迅速与 核小体获得特定的表观遗传修饰，将病毒附加体分成 转录活性域和沉默域。虽然多个KSHV表观遗传标记 在淋巴、上皮和内皮细胞中，这些过程和潜在的宿主- 导致潜伏容许附加体形成的病毒相互作用仍然很差， 明白鉴于KSHV潜伏相关核抗原（拉娜）与 染色质重塑和H3.3组蛋白伴侣Daxx，HIRA和DEK，我们 假设H3.3沉积在建立和维持中起重要作用， KSHV潜伏期。我们的初步数据表明，H3.3沉积在KSHV游离体上， 在长期感染细胞的附加体上重新感染后早期检测到。而且我们 证明了通过基因干扰H3.3分子伴侣途径HIRA和Daxx， CRISPR/Cas9导致KSHV潜伏期控制的显着变化，与KSHV潜伏期控制的改变相关。 表观遗传状态我们进一步假设潜伏期相关基因产物，包括 拉娜在从头感染期间调节和/或去调节组蛋白伴侣途径， 延迟。为了直接解决这一假设，我们建议从机制上研究组蛋白H3.3 沉积和组蛋白翻译后修饰（PTM）的形成，目的是 了解它们与病毒基因表达和潜伏/裂解的关系 复制的这些研究的一个主要方面是研究病毒基因产物如何与 并调节组蛋白变体分子伴侣途径。此外，我们使用人口腔初级上皮细胞， 细胞比转化的上皮细胞系更具溶解性，以表征表观遗传 导致其裂解表型的变化。重要的是，通过与博士的合作。 唐纳德科恩，牙科学院，我们建议建立在体内表观基因组的口腔KS在这两个 福尔马林固定和快速冷冻的肿瘤样品。总的目标是研究组蛋白 变体沉积以细胞类型特异性方式决定潜伏性和溶解性感染。结果 可能指向KSHV特异性口服KS的新治疗靶点。

项目成果

Coordination of transcription, processing, and export of highly expressed RNAs by distinct biomolecular condensates.

• DOI: 10.1042/etls20190160
• 发表时间: 2020-12-11
• 期刊: Emerging topics in life sciences
• 影响因子: 3.8
• 作者: Ishov AM;Gurumurthy A;Bungert J
• 通讯作者: Bungert J

Inhibition of Mps1 kinase enhances taxanes efficacy in castration resistant prostate cancer.

MPS1激酶的抑制作用增强了抗castratiation抗性前列腺癌中紫杉烷的功效。

• DOI: 10.1038/s41419-022-05312-8
• 发表时间: 2022-10-13
• 期刊: CELL DEATH & DISEASE
• 影响因子: 9
• 作者: Sarwar, Sadia;Morozov, Viacheslav M.;Purayil, Hamsa;Daaka, Yehia;Ishov, Alexander M.
• 通讯作者: Ishov, Alexander M.

Sex-biased expression is associated with chromatin state in D. melanogaster and D. simulans.

性别偏向表达与黑腹果蝇和拟果蝇中的染色质状态相关。

• DOI: 10.1101/2023.01.13.523946
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Nanni,AdalenaV;Martinez,Natalie;Graze,Rita;Morse,Alison;Newman,JeremyRB;Jain,Vaibhav;Vlaho,Srna;Signor,Sarah;Nuzhdin,SergeyV;Renne,Rolf;McIntyre,LaurenM
• 通讯作者: McIntyre,LaurenM

Super-enhancer mediated regulation of adult β-globin gene expression: the role of eRNA and Integrator.

• DOI: 10.1093/nar/gkab002
• 发表时间: 2021-02-22
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Gurumurthy A;Yu DT;Stees JR;Chamales P;Gavrilova E;Wassel P;Li L;Stribling D;Chen J;Brackett M;Ishov AM;Xie M;Bungert J
• 通讯作者: Bungert J