Project 2-Tso

项目2-曹

• 批准号: 10664033
• 负责人: For Yue Tso
• 金额: $ 25.35万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10664033
• 关键词: AIDS related cancer; AIDS with Kaposi' s sarcoma; Adherence; Affect; Africa South of the Sahara; Antigen Presentation; Area; Attenuated; Base Sequence; Bioinformatics; Burkitt Lymphoma; CRISPR/Cas technology; Cell Culture Techniques; Cell model; Cells; Chronic; Country; Coupled; Development; Etiology; Gene Expression; Genes; Geography; Growth; HIV; HIV-1; Herpesviridae Infections; Human Herpesvirus 4; Human Herpesvirus 8; Immune; Immune Evasion; Immunologic Surveillance; Incidence; Individual; Kaposi Sarcoma; Louisiana; Maintenance; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of nasopharynx; Mentored Clinical Scientist Development Program; MicroRNAs; Modeling; Monitor; Multicentric Angiofollicular Lymphoid Hyperplasia; Neoplasms; Oncogenic; Oncology; Pathway interactions; Persons; Play; Property; Reporting; Research; Resistance; Risk; Role; Seroprevalences; System; Testing; Therapeutic; United States; Viral; Viral Cancer; Viral Genes; Viral Load result; Virus; Virus Diseases; Virus Latency; antiretroviral therapy; design; experience; gene interaction; health disparity; immunoregulation; infection rate; insight; latency-associated nuclear antigen; malignant phenotype; neoplastic cell; new technology; novel; people of color; primary effusion lymphoma; programs; protein expression; transcriptome; transcriptome sequencing; tumorigenesis; vector

PROJECT SUMMARY/ABSTRACT Kaposi’s sarcoma (KS) is an HIV/AIDS-associated malignancy, that remains prevalent, despite antiretroviral therapy, in areas where there is high incidence of KS-associated herpesvirus (KSHV) infection. KSHV seroprevalence is geographically uneven but is high in sub-Saharan Africa (SSA) and in parts of the United States, such as Southern Louisiana, where HIV-1 infection rates are high. In this region, both HIV and KS disproportionately affect people of color who also experience other health disparities. Because HIV- and KSHV- infected individuals remain at risk to develop KS and other KSHV-associated malignances despite HIV viral load suppression, novel research strategies are needed for people living with HIV (PLWH). Chronic oncogenic viral infections possess mechanisms that engender immune avoidance and viral persistence that leads to neoplastic growth. The KSHV latency locus encodes a small number of viral genes that are involved in viral persistence. The latency-associated nuclear antigen (LANA) plays a key role in the maintenance of latency and help the virus to evade host immune surveillance by inhibiting MHC-1 antigen presentation, while other latency genes, such as kaposin (K12) was reported to have oncogenic properties, as well as microRNAs that could be essential for tumorigenesis. Although the individual function of KSHV latency-associated genes have been studied, there remain unresolved questions regarding their specific contribution to latency, immune evasion and neoplasia. It is also critical to know, which host genes and pathways interact with the viral genes to elicit the latency program, and to answer how this virus-host interplay contributes to malignant transformation. We recently demonstrated that our unique CRISPR/Cas9 vector, specifically designed to target a single KSHV LANA nucleotide sequence, reduced ~70-80% of KSHV episomal copies, and attenuated the growth of KSHV-positive cells. Our major objective here is to utilize this new technology to disrupt/eliminate multiple genes of the KSHV latency locus, and to investigate their individual and combined effects on KSHV latency. We will seek to identify cellular genes and pathways affected by, and possibly contributing to the development and maintenance of the malignant phenotype. We hypothesize that the components of the KSHV latency locus individually dysregulate expression of specific sets of cellular genes to promote KSHV persistence and synergistically to promote growth dysregulation and tumorigenesis. We will test this concept by editing multiple viral gene sequences within the latency locus with CRISPR/Cas9 using KS and PEL cell culture models. Through bioinformatics analyses of transcriptome profiles of the CRISPR/Cas9 edited KSHV infected cells, we will identify unique host genes/cellular pathway and their roles/interaction with viral latency genes in viral latency, growth dysregulation and tumorigenesis.

项目总结/摘要 卡波西肉瘤（KS）是一种HIV/AIDS相关的恶性肿瘤，尽管抗逆转录病毒治疗， 在KS相关疱疹病毒（KSHV）感染高发地区进行治疗。KSHV 血清阳性率在地理上是不均衡的，但在撒哈拉以南非洲（SSA）和美国部分地区很高。 美国路易斯安那州南部等HIV-1感染率较高的州。在该地区，艾滋病毒和KS 不成比例地影响有色人种，他们也经历其他健康差异。因为艾滋病毒和KSHV- 尽管有HIV病毒载量，感染者仍有发生KS和其他KSHV相关恶性肿瘤的风险 抑制，新的研究策略需要与艾滋病毒感染者（PLWH）。慢性致癌病毒 感染具有产生免疫回避和病毒持久性的机制，导致肿瘤性 增长KSHV潜伏基因座编码参与病毒持久性的少量病毒基因。 潜伏相关核抗原（拉娜）在维持潜伏期和帮助病毒 通过抑制MHC-1抗原呈递来逃避宿主免疫监视，而其他潜伏基因，如 由于Kaposin（K12）被报道具有致癌特性，以及可能是必不可少的microRNA， 肿瘤发生虽然KSHV潜伏相关基因的个体功能已经研究过， 关于它们对潜伏期、免疫逃避和肿瘤形成的具体作用，仍然存在未解决的问题。它 同样重要的是要知道，哪些宿主基因和途径与病毒基因相互作用，引发潜伏期程序， 并回答这种病毒与宿主的相互作用如何导致恶性转化。我们最近展示了 我们独特的CRISPR/Cas9载体，专门设计用于靶向单个KSHV拉娜核苷酸序列， 减少~70-80%的KSHV附加型拷贝，并减弱KSHV阳性细胞的生长。我们的主要 目的是利用这种新技术破坏/消除KSHV潜伏基因座的多个基因， 研究它们对KSHV潜伏期的单独和组合影响。我们将寻求识别细胞基因， 受影响的途径，并可能有助于恶性肿瘤的发展和维持， 表型我们假设KSHV潜伏期基因座的组分单独失调表达， 特定的细胞基因组可促进KSHV的持续存在并协同促进生长 失调和肿瘤发生。我们将通过编辑多个病毒基因序列来测试这一概念。 使用KS和PEL细胞培养模型，使用CRISPR/Cas9与潜伏基因座进行比较。通过生物信息学分析， 通过CRISPR/Cas9编辑的KSHV感染细胞的转录组谱，我们将鉴定独特的宿主基因/细胞 途径及其在病毒潜伏期、生长失调和 肿瘤发生