TCR Gene Transfer for Treating HCV-Associated Malignancies

项目总结/摘要 丙型肝炎病毒（HCV）感染了大约3%的世界人口，通常引起相关疾病 包括肝硬化和肝细胞癌（HCC）。有效的治疗方法有限， 疫苗开发、治疗和预防感染的替代方法及其相关 疾病是必要的。一种基于免疫的靶向治疗方法利用过继性T细胞转移， 通过基因工程改造一个人的T细胞，使其对选择的抗原产生反应。我们的实验室以前 证明了其成功重定向外周血淋巴细胞（PBL）衍生的T细胞的能力， 逆转录病毒载体以识别靶抗原。研究重定向T细胞识别黑色素瘤- 相关抗原导致了正在进行的治疗转移性疾病的I/II期临床试验。我们相信这 该方法可应用于HCV感染和HCV相关的恶性肿瘤。我们的实验室克隆了 从能够识别HCV的人PBL衍生的T细胞克隆表达两种功能性TCR 肽NS 3：1406-1415和NS 3：1073-1081。此外，我们已经证明这些是高亲和力TCR， 因为它们具有CD 8非依赖性靶细胞识别，且可以识别天然发生突变逃逸 变体。我们相信，重新引导T细胞识别突变性HCV抗原的方法可能有好处 在理解TCR生物学以及产生HCV感染和相关疾病的临床治疗方面， 疾病基于这些观察结果，我们假设TCR具有广泛反应性的能力并不依赖于TCR的表达。 这种分离的克隆所特有的，并且在具有消退的HCV感染的患者中，可能存在多种 具有识别病毒蛋白的许多表位的能力的TCR。我们还假设HCV TCR 转导的T细胞具有识别和杀死表达野生型和突变型HCV的肿瘤细胞的能力 在体外可以抑制HCV抗原，并且在体内可以介导HCV+肿瘤细胞的消退。三个具体目标是 来检验这些假设。目的1将验证我们的假设，即来自患者的HCV反应性T细胞 那些治愈了HCV感染，但没有HCV慢性感染的患者表达TCR， 野生型和广谱突变型HCV抗原。目标2将检验以下假设： 来自解决其HCV感染的患者的HCV反应性T细胞可以将这种反应性转移到针对突变型HCV的T细胞。 HCV抗原转移到其他T细胞。目的3将验证HCVTCR转导的T细胞可以介导HCV-TCR的表达的假设。 小鼠异种移植肿瘤模型中表达野生型和突变型HCV抗原的HCC肿瘤的消退， vivo.总的来说，这项建议将产生新的信息，可能会导致更好地了解能力 的TCR识别多个突变的HCV表位，并将有助于开发新的治疗方法， HCV相关HCC患者。

Project Summary/Abstract Hepatitis C virus (HCV) infects approximately 3% of the world's population, often causing associated diseases including cirrhosis of the liver and hepatocellular carcinoma (HCC). With limited effective therapies and slow vaccine development, alternative approaches for treatment and prevention of infection and its associated diseases is imperative. An immune-based approach for targeted therapy utilizes adoptive T cell transfer to genetically engineer an individual's T cells to become reactive to an antigen of choice. Our lab has previously demonstrated its ability to successfully redirect peripheral blood lymphocyte (PBL)-derived T cells with retroviral vectors in order to recognize target antigens. Studies redirecting T cells to recognize melanoma- associated antigens have led to ongoing Phase I/II clinical trials for treating metastatic disease. We believe this approach can be applied to HCV infections and HCV-associated malignancies. Our lab has cloned and expressed two functional TCRs from PBL-derived T cell clones from humans capable of recognizing HCV peptides NS3:1406-1415 and NS3:1073-1081. Additionally, we have shown that these are high affinity TCRs since they have CD8-independent target cell recognition and can recognize naturally occurring mutant escape variants. We believe the approach to redirect T cells to recognize mutagenic HCV antigens may have benefits in understanding TCR biology as well as producing a clinical therapy for HCV infection and associated diseases. Based on these observations, we hypothesize that the ability of a TCR to have broad reactivity is not unique to such isolated clones and that in patients with resolved HCV infection there may exist a variety of TCRs with the capability of recognizing many epitopes of viral proteins. We also hypothesize that HCV TCR transduced T cells have the ability to recognize and kill tumor cells expressing wild type and mutant HCV antigens in vitro and can mediate regression of HCV+ tumor cells in vivo. Three specific aims have been developed to examine these hypotheses. Aim 1 will test our hypothesis that HCV reactive T cells from patients who resolved their HCV infections but not patients with HCV chronic infections express TCRs that recognize wild type and a broad spectrum of mutant HCV antigens. Aim 2 will test the hypothesis that the TCRs from HCV reactive T cells from patients who resolved their HCV infections can transfer this reactivity against mutant HCV antigens to other T cells. Aim 3 will test the hypothesis that HCV TCR transduced T cells can mediate regression of HCC tumors expressing wild type and mutant HCV antigens in mouse xenograft tumor models in vivo. Overall, this proposal will yield novel information that may lead to a better understanding of the capability of TCRs to recognize multiple mutant HCV epitopes and will aid in the development of novel therapeutics for patients with HCV-associated HCC.