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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

抽象的卡波西肉瘤相关疱疹病毒（KSHV）是卡波西肉瘤（KS）的病原体，和两种淋巴增殖性疾病：原发性渗出性淋巴瘤 (PEL) 和多中心卡斯尔曼氏病疾病（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。虽然发病率最近，由于联合抗逆转录病毒疗法 (cART)，美国 AIDS-KS 病例有所减少研究表明，长期接受治疗的艾滋病患者中口腔 KS 会复发。虽然 KS 肿瘤细胞是牙龈上皮细胞大多是潜伏感染，表达有限数量的病毒基因口腔主动复制 KSHV，这对于 a) 在 B 细胞中播种潜伏储存库非常重要原发感染后和 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着核心作用。组蛋白变体（例如 H3.3、CENP-A 和）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。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 表观遗传标记已已在淋巴样、上皮细胞和内皮细胞中绘制了图谱，其过程和潜在宿主导致潜伏允许附加体形成的病毒相互作用仍然很差明白了。鉴于 KSHV 潜伏相关核抗原 (LANA) 与染色质重塑剂和 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，KSHV 附加体上的 H3.3 沉积可以在长期感染细胞的附加体上从头感染后尽早检测到。此外，我们证明通过基因破坏 H3.3 伴侣通路 HIRA 和 Daxx CRISPR/Cas9 导致 KSHV 潜伏期控制发生显着变化，与表观遗传状态。我们进一步假设潜伏期相关基因产物包括 LANA 在从头感染期间调节和/或解除组蛋白伴侣通路的调节延迟。为了直接解决这个假设，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。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异伴侣通路。此外，我们使用人类口腔原代上皮细胞比转化的上皮细胞系更具裂解性的细胞来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。 Donald Cohen，牙科学院，我们建议在两种情况下建立口腔 KS 的体内表观基因组福尔马林固定并速冻的肿瘤样本。总体目标是研究组蛋白如何变异沉积以细胞类型特异性的方式决定潜伏性和裂解性感染。结果可能为口服 KS 指明 KSHV 特异性新治疗靶点。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

超级增强子介导的成人β珠蛋白基因表达的调节：eRNA 和整合器的作用。

DOI：
发表时间：
2021
期刊：
影响因子：
14.9
作者：
Gurumurthy, Aishwarya;Yu, David T;Stees, Jared R;Chamales, Pamela;Gavrilova, Ekaterina;Wassel, Paul;Li, Lu;Stribling, Daniel;Chen, Jinyang;Brackett, Marissa;Ishov, Alexander M;Xie, Mingyi;Bungert, Jörg
通讯作者：
Bungert, Jörg

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

通过不同的生物分子凝聚体协调高表达 RNA 的转录、加工和输出。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Ishov, Alexander M;Gurumurthy, Aishwarya;Bungert, Jörg
通讯作者：
Bungert, Jörg

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

抑制 Mps1 激酶可增强紫杉烷类药物在去势抵抗性前列腺癌中的疗效。

DOI：
发表时间：
2022-10-13
期刊：
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：
发表时间：
2023-01-13
期刊：
影响因子：
0
作者：
Nanni, Adalena V;Martinez, Natalie;Graze, Rita;Morse, Alison;Newman, Jeremy R B;Jain, Vaibhav;Vlaho, Srna;Signor, Sarah;Nuzhdin, Sergey V;Renne, Rolf;McIntyre, Lauren M
通讯作者：
McIntyre, Lauren M