Nano-Therapeutic Approaches for Oncogenic Herpesvirus-Mediated Malignancies

一些治疗癌症的化疗药物会因为药物诱导的细胞压力而触发重新激活。 病毒重新激活后，肿瘤表达的新抗原成为细胞毒细胞的新靶点。这些近地天体- 抗原包括病毒裂解抗原、病原体相关分子模式和替代肿瘤相关抗原。 在病毒重新激活时表达的分子。病毒在肿瘤中的重新激活也会导致肿瘤细胞死亡和 将肿瘤相关免疫原抗原释放到肿瘤微环境或循环中。后者可以 从系统上增强抗肿瘤免疫反应。最后，肿瘤产生的外切体进入循环，这 可能携带病毒基因组和抗原，可以是另一种形式的肿瘤相关免疫触发。因此， 事实上，如果我们只能控制和限制，病毒的重新激活可能有助于恢复患者的免疫细胞毒性 病毒重新激活的程度和位置。我们的体外初步数据表明，这种情况是 BTZ和OTX015联合治疗KSHV阳性的腹腔积液淋巴瘤(PEL)是可行的。 也就是说，用BTZ和OTX015治疗KSHV阳性的PEL可诱导KSHV和 在阻断终末病毒颗粒形成的同时表达裂解抗原，导致PEL细胞有限死亡 脱落成熟的病毒颗粒。此外，OTX015对炎性细胞因子也有很强的抑制作用 由PEL生产。在这个补充项目中，候选人下田博士将介绍一只人性化的小鼠 模型检验BTZ/OTX015治疗在小鼠体内创造免疫原性肿瘤微环境的假设 一个人性化的PEL小鼠模型。具体目标是： 目的：评价BTZ/OTX015治疗人源化PEL小鼠模型的免疫表型。 我们将验证BTZ/OTX015在HLA-A0201限制性BC-3肿瘤中重新激活KSHV的假设 在肿瘤浸润性免疫细胞增加的情况下，总体细胞毒性增强。免疫细胞表型与 采用流式细胞术检测脾和肿瘤组织中CD107a+和IFNG+细胞的表达情况。我们的 目的是确定该药物组合在抗肿瘤免疫反应中具有全面的有益作用。 目的2：对肿瘤微环境进行单细胞转录组分析。 BTZ/OTX015治疗。我们将检验这样一种假设，即在肿瘤中诱导的转录变化是微小的 用单细胞RNA测序技术可以鉴定HuPBL-NSG PEL小鼠体内的药物环境。我们 将在单细胞水平上评估药物对PEL肿瘤以及肿瘤浸润性免疫细胞的作用。我们的 目的是确定药物和细胞特有的转录组变化在肿瘤和免疫细胞亚群中的作用。

Some chemotherapeutic drugs for cancers trigger reactivation because of cellular stress induced by the drugs. Upon viral reactivation, neo-antigens expressed by tumors become new targets for cytotoxic cells. These neo- antigens include virus lytic antigens, pathogen-associated molecular patterns and surrogate tumor-associated molecules expressed upon viral reactivation. Viral reactivation in tumors also induces tumor cell death and releases tumor-associated immunogenic antigens into the tumor micro-environment or circulation. The latter may boost anti-tumor immune response systemically. Finally, exosomes produced by tumors into circulation, which potentially carry virus genomes and antigens, can be another form of tumor associated immune triggers. Thus, viral reactivation may, in fact, help revive a patients' immune cytotoxicity if we can only control and limit the level and location of viral reactivation. Our in vitro preliminary data suggests that this scenario is achievable in KSHV-positive peritoneal effusion lymphoma (PEL) by the BTZ and OTX015 combination therapy. That is, the treatment of KSHV-positive PELs with BTZ and OTX015 induced reactivation of KSHV and expression of lytic antigens while blocking terminal viral particle formation, resulting in PEL cell death with limited shedding of mature viral particles. In addition, OTX015 showed strong suppression of inflammatory cytokine production by PEL. In this supplement project, the candidate, Dr. Shimoda will introduce a humanized mouse model to test the hypothesis that BTZ/OTX015 therapy creates immunogenic tumor micro-environment in a humanized mouse model of PEL. Specific Aims are: Aim 1: To evaluate immune phenotype in a humanized mouse model of PEL under BTZ/OTX015 therapy. We will test the hypothesis that KSHV reactivation by BTZ/OTX015 in HLA-A0201-restricted BC-3 tumors results in increased tumor-infiltrating immune cells with overall enhanced cytotoxicity. Immune cell phenotype and the frequency of cytotoxic CD107a+ and IFNg+ cells in spleen and tumors will be evaluated by flow cytometry. Our goal is to establish that the drug combination has overall beneficial effect on anti-tumor immune response. Aim 2: To conduct single cell transcriptome analysis of the tumor micro-environment during BTZ/OTX015 therapy. We will test the hypothesis that transcriptional changes induced in a tumor micro- environment by drugs in huPBL-NSG PEL mice can be identified by single cell RNA sequencing technology. We will evaluate the drug effects on PEL tumors as well as tumor infiltrating immune cells at single cell level. Our goal is to identify the drug and cell-specific unique transcriptome changes in tumor and immune cell subsets.