HORMAD-specific TGF-beta resistant memory T cells for treatment of patients with Gastro-esophageal Cancer

项目摘要 这项建议解决了过继细胞治疗中的三个主要挑战：1)T细胞持久性；2)肿瘤 免疫抵抗和3)实体肿瘤的靶位识别，汇集了Dr。 Yee博士从事T细胞治疗，Rai博士从事表观遗传学研究，Morelli博士从事临床试验。我们已经制定了一项战略， 通过一种称为内源性T细胞的ACT方式从患者的外周血中产生记忆T细胞 (等)Yee实验室首创的治疗方法，使我们能够广泛地针对HORMAD1，一种癌症睾丸抗原 在胃癌和食道癌中表达，这是一个严重的未得到满足的需求。 通过应用合理设计的表观遗传调节器组合，我们计划优化复制 体外产生的HORMAD1特异性CTL(HMD-CTL)的容量和记忆特性 挑战有限的体内T细胞持久性(UG3目标1)。主导肿瘤的外在因素之一 胃癌、食道癌的免疫耐受是肿瘤微环境中转化生长因子-β的影响。 为了应对这一挑战，我们建议使用显性的负性转化生长因子-β受体通过以下方式隔离转化生长因子-β 逆转录病毒转导TGFBDNR2基因在HORMAD1特异性CTL中的工程表达 在细胞治疗制造中心(UG3目标2)确立的战略。最后，解决 针对胃癌和食道癌的挑战来自于正在进行的抗原发现流水线 经过5年的开发，研究了从MHC洗脱出来的串联质谱学定义的表位组合 超过30个肿瘤样本。从这条流水线上，我们已经经验性地验证了人类白细胞抗原A2和人类白细胞抗原A11 HORMAD1的表位与使用ETC工作流程产生的CTL具有很高的亲和力，并提出了 在这项试验中使用HORMAD1特异性CTL，使我们能够覆盖超过25%的胃和食道 给出HORMAD1在这些肿瘤中的表达(&gt；40%)和HLA等位基因表达(&gt； 65%)。在完成本提案的UG3部分后，我们将确定一个最佳的表观遗传方案(目标1) 和TGFBDNR2转导工作流(目标2)，满足进行/不进行决策的指标。在临床上 试验部分(UH3)这些策略将被组合在一起，使我们能够评估体内的安全性和持续时间 持久性，比较两种情况下HORMAD1特异性CTL和TGFBDNR2工程HORMAD-1特异性CTL 队列，同时评估扩张臂的初步疗效。 ETC平台在敏捷性方面提供了比其他ACT模式更大的灵活性，以将特定的 几乎任何想要的目标表位，证明了抗原传播的证据，最小的毒性，并具有 已建立的记忆潜力在几个临床试验中被证明是持续的。我们期待着，通过一个 增强的表观遗传记忆程序，并被赋予破坏至少一个特征的机制 肿瘤免疫耐药等治疗可为推进肿瘤过继细胞治疗提供载体 系统、循序渐进地治疗实体肿瘤。

Project Summary This proposal addresses three major challenges in adoptive cellular therapy: 1) T cell persistence; 2) Tumor immune resistance and 3) Target epitope identification for solid tumors, bringing together the expertise of Dr. Yee in T cell therapy, Dr. Rai in epigenetics and Dr. Morelli in clinical trials. We have established a strategy for generating memory T cells from peripheral blood of patients, via an ACT modality known as Endogenous T Cell (ETC) therapy pioneered in the Yee lab, allowing us to target HORMAD1, a cancer testis antigen broadly expressed in gastric and esophageal cancer, which in its advanced stages represents a significant unmet need. By applying rationally designed combinations of epigenetic modulators we plan to optimize the replicative capacity and memory properties of HORMAD1-specific CTL (HMD-CTL) generate ex vivo to address the challenge of limited in vivo T cell persistence (UG3 Aim 1). One of the dominant extrinsic factors mediating tumor immune resistance in gastric and esophageal cancer is the influence of TGF-β in the tumor microenvironment. In addressing this challenge, we propose the use of a dominant negative TGF-β receptor to sequester TGF-β by engineering expression of the TGFBDNR2 gene into HORMAD1-specific CTL using a retroviral transduction strategy that is well-established in the Cell Therapy Manufacturing Center (UG3 Aim 2). Finally, addressing the challenge of targeting gastric and esophageal cancers was borne out of an ongoing antigen discovery pipeline developed over 5 years examining the portfolio of tandem mass spectrometric defined epitopes eluted from MHC of over 30 tumor samples. From this pipeline we have empirically validated both an HLA-A2 and HLA-A11 epitopes of HORMAD1, demonstrated high affinity with CTL generated using the ETC workflow and propose the use of HORMAD1-specific CTL for this trial, allowing us to cover more than 25% of all gastric and esophageal patients given the prevalence of HORMAD1 expression in these tumors (> 40%) and HLA allele expression (> 65%). At completion of the UG3 portion of this proposal, we will identify an optimal epigenetic program (Aim 1) and TGFBDNR2 transduction workflow (Aim 2) that fulfills the metrics for a Go/ No-Go decision. In the clinical trial component (UH3) these strategies will be assembled to allow us to evaluate safety and duration of in vivo persistence, comparing HORMAD1-specific CTL and TGFBDNR2-engineered HORMAD-1-specitic CTL in two cohorts, while assessing for preliminary efficacy in an Expansion arm. The ETC platform provides greater flexibility than other ACT modalities in its agility to direct specificity to almost any desired target epitope, demonstrated evidence of antigen-spreading, minimal toxicities, and has an established memory potential proven to be sustained in several clinical trials. We anticipate, that with an enhanced epigenetic memory program and, endowed with a mechanism to undermine at least one feature of tumor immune resistance, ETC therapy can provide a vehicle for advancing adoptive cellular therapy for the treatment of solid tumors in a systematic and stepwise fashion.