Combining repetitive element-specific T cells with epigenetic therapy to treat ovarian cancer

20多年来，卵巢癌患者的五年生存率一直保持在47%。 OC的特点是高度抑制肿瘤微环境，目前的研究工作集中在 逆转这种免疫抑制。激活针对OC的免疫反应的一种方法是使用表观遗传学 治疗学，通过诱导重复转录来刺激肿瘤细胞中的干扰素反应 元素(RES)--类似于某些病毒遗传物质的基因组区域。因为Res通常是 在终末分化的细胞中没有表达，它们在肿瘤细胞中的上调表明这些基因组元件 可作为诱导性治疗靶点。针对一种RE来源的抗原的T细胞，即无功能的 内源性逆转录病毒K的包膜基因(ERV-K-Env)，能识别其同源抗原并杀伤OC细胞 同时保留健康的细胞。这些T细胞在与免疫原性表观遗传学结合时是否会更具裂解性 治疗方法尚不清楚。除ERV-K-Env外，其他表观遗传学上调的RE是一个未被勘探的矿藏 肿瘤相关抗原可能成为新的治疗靶点。我假设结合表观遗传学 用RE特异性T细胞治疗OC将具有强大的免疫原性和靶向性抗肿瘤效果。 在目标1中，我将确定联合表观遗传疗法和ERK-K-Env特异性T细胞在小鼠体内的作用。 OC。初步数据表明，该候选RE，ERV-K-Env，是OC中的靶向肿瘤相关抗原。我 假设表观遗传疗法与体外扩增的ERV-K-Env特异性T细胞相结合将 导致有针对性的免疫原性OC细胞杀伤。ERV-K-Env特异性T细胞将从体外扩增 并在体外进行抗原特异性评估。我会将扩增的T细胞与 对OC细胞进行表观遗传学处理，并在体外和体内评估T细胞的激活和细胞毒性。 在目标2中，我将鉴定和靶向更多与肿瘤相关的RE多肽作为新的OC肿瘤抗原。 来自我的实验室的初步数据表明，表观遗传治疗导致RES和他们的 在OC细胞表面呈现给免疫系统。我假设不同的肿瘤相关 表观遗传疗法上调的RES可作为OC的治疗相关T细胞靶点。生物信息学 工具将被用来确定表观遗传疗法上调的哪些RE有可能成为T细胞抗原。 我将通过扩增来自健康捐赠者的RE特异性T细胞和 使用OC患者T细胞评估实验验证的RES作为治疗靶点的临床相关性。 拟议项目的完成将为表观遗传疗法的组合提供新的知识 用RE特异性T细胞作为一种新的免疫原性和潜在的治疗OC的策略。总而言之， 这些创新的跨学科实验将阐明RES作为治疗药物的作用尚未得到充分研究。 靶点，同时进一步阐明激活抗肿瘤免疫反应的机制，该免疫反应可能 适用于其他实体瘤。

The five year survival rate for ovarian cancer (OC) patients has remained at 47% for over two decades. OC is characterized by a highly suppressive tumor microenvironment and current research efforts focus on reversing this immune suppression. One way to activate the immune response against OC is using epigenetic therapeutics, which stimulate an interferon response in cancer cells by inducing the transcription of repetitive elements (REs)—genomic regions that resemble the genetic material of some viruses. As REs are normally silent in terminally differentiated cells, their upregulation in tumor cells suggests that these genomic elements can be used as inducible treatment targets. T cells specific for an RE-derived antigen, the non-functional envelope gene of endogenous retrovirus K (ERV-K-Env), can recognize their cognate antigen and kill OC cells while sparing healthy cells. Whether these T cells will be more lytic when combined with immunogenic epigenetic therapy is unknown. Aside from ERV-K-Env, other epigenetically upregulated REs serve as an unexplored pool of tumor-associated antigens that may be novel treatment targets. I hypothesize that combining epigenetic therapy with RE-specific T cells will have potent immunogenic and targeted anti-tumor efficacy in OC. In Aim 1, I will determine the effect of combination epigenetic therapy and ERK-K-Env-specific T cells in OC. Preliminary data suggest this candidate RE, ERV-K-Env, is a targetable tumor-associated antigen in OC. I hypothesize that combining epigenetic therapy with ex vivo expanded ERV-K-Env-specific T cells will result in targeted immunogenic OC cell killing. ERV-K-Env-specific T cells will be expanded ex vivo from whole blood donors and assessed in vitro for antigen specificity. I will co-culture the expanded T cells with epigenetically treated OC cells and assess T cell activation and cytotoxicity in vitro and in vivo. In Aim 2, I will identify and target additional tumor-associated RE peptides as novel OC tumor antigens. Preliminary data from my lab suggest that epigenetic treatment results in the upregulation of REs and their presentation on the surface of OC cells to the immune system. I hypothesize that diverse tumor-associated REs upregulated by epigenetic therapy can be therapeutically relevant T cell targets in OC. Bioinformatic tools will be used to identify which REs upregulated by epigenetic therapy have the potential to be T cell antigens. I will validate the immunogenicity of these antigens by expanding RE-specific T cells from healthy donors and assess the clinical relevance of the experimentally validated REs as treatment targets using OC patient T cells. Completion of the proposed project will provide new knowledge on the combination of epigenetic therapy with RE-specific T cells as a novel immunogenic and potentially curative treatment strategy for OC. Collectively, these innovative interdisciplinary experiments will shed light on the understudied role of REs as therapeutic targets while further elucidating the mechanisms involved in activating an anti-tumor immune response that may be applicable to other solid tumors.