Engineering T cells to Promote Islet Transplant

A.具体目标 1型糖尿病(T1D)是一种进行性自身免疫性疾病，由于免疫介导的β细胞破坏，使个体无法调节血糖水平，导致胰岛素产生损失和许多严重的健康并发症，如果不治疗，将导致死亡。仔细监测血糖，再加上胰岛素注射，使T1D成为一种慢性疾病，T1D患者的寿命比健康人少十年左右。因此，治疗T1D是非常可取的。细胞基因治疗已被证明是治疗顽固性疾病的有效方法，如儿童白血病，表达嵌合抗原受体(CAR)的T细胞获得90%的完全应答，使许多人处于非常长时间的缓解1，但到目前为止，还没有工程T细胞疗法试图治愈人类的T1D。 我们建议检验这样一种假设，即工程T细胞可以在最少或没有额外免疫抑制的情况下实现胰岛移植。胰岛移植代表了测试工程T细胞保护胰岛细胞免受免疫攻击的能力的最佳方案，并可能为开发治疗新发的T1D的策略奠定基础。在胰岛移植中，供者和受者的MHC匹配很少发生，产生高表达的、独特的胰岛特异性HLA抗原，这些抗原可以被CAR工程的T调节细胞(CAR树)或表达抑制免疫系统的分子(T抑制细胞或TSUPS)的T细胞靶向。此外，通过全面的筛选过程，我们已经确定了两个靶点，成纤维细胞激活蛋白(FAP)和二肽基肽酶样蛋白6(DPP6)，它们在α和β细胞上高表达，但在其他地方表达有限，可用于治疗最近诊断的个体和所有移植受者，而不存在所需的MHC错配。在这项提案中，我们将进一步开发这个工具箱，以开发更好的体内T1D临床前模型，以及将预防、延缓或逆转T1D的新细胞和基因疗法。 在过去的十年里，基于T细胞的治疗取得了快速的进步，使得考虑对T1D进行这样的治疗成为可能。在卡尔·朱恩的领导下，宾夕法尼亚大学细胞免疫治疗中心开发的CD19特异性CARS治疗3名癌症患者的长期缓解后，CAR T细胞革命启动。这一早期的成功推动了相当大的投资，使许多机构和公司能够开发方法来提高安全性和有效性，并降低生产工程T细胞的成本。这些创新中的许多还将有助于实现T1D的细胞和基因疗法，这是本RFA的首要目标。这项应用的最终目标是在胰岛移植后成功地使用工程T细胞治疗三种非人类灵长类动物(NHP)，我们预测这将引发对T1D细胞和基因治疗的类似热情，就像前三位CD19-CAR接受者在癌症汽车治疗中所做的那样。 为了实现这一目标，一支经验丰富、专业知识互补的调查小组将领导这项提议。该团队包括帮助开创胰岛移植作为T1D疗法的先驱(NAJI)的移植外科医生、具有开发NHP T调节细胞疗法模型(Duran-Struuck)的专业知识的兽医、以及拥有设计T细胞和开发首个人类临床试验(Riley)记录的细胞和基因治疗师，他们一直通过赫尔姆斯利基金会的资助密切合作，以获得本申请中提供的初步数据。该团队现在准备立即进行体内研究，以测试工程T细胞疗法预防、延缓或逆转T1D的能力。 目的1.在人源化小鼠模型中设计T抑制细胞(TSUPS)促进胰岛移植。虽然T调节细胞是强大的免疫抑制细胞，但还有其他有希望的方法来诱导耐受，这些方法可能作为独立治疗或与CAR树协同作用以保护β细胞功能。我们将探索使用针对胰岛的PD-L1、转化生长因子-β1和/或γ-氨基丁酸表达细胞单独或结合我们利用人源化小鼠开发的CAR-Treg方法来调节胰岛的接受。 目的2.优化T细胞途径，促进非霍奇金淋巴瘤胰岛移植。到目前为止，我们的努力主要集中在开发NHP汽车Tregs以实现胰岛移植。我们已经优化了NHP MHC特异性CAR T调节细胞的分离、转导和扩增，并准备立即开始胰岛移植实验。在我们使用CAR Tregs进行原理实验证明之后，我们将根据在目标1中获得的数据来确定未来实验的优先顺序。我们研究的一个特别令人兴奋的方面是测试由Garcia Lab3开发的Ortho-IL-2系统是否可以在非人类灵长类动物中工作，并支持工程化Treg或其他工程化T细胞的功能和扩增，同时对内源性效应细胞没有影响。如果成功，这些研究将形成人类第一阶段临床试验的基础和理论基础。

A. Specific Aims Type 1 diabetes (T1D) is a progressive autoimmune disease which renders individuals incapable of regulating their blood glucose levels due to immune-mediated β cell destruction, resulting in loss of insulin production and many severe health complications that, if untreated, lead to death. Careful monitoring of blood glucose coupled with insulin injections have made T1D a chronic disease in which T1D individuals live ~ a decade less than their healthy counterparts. Thus, a cure for T1D is highly desirable. Cell gene therapy has proven to be an effective way to treat recalcitrant diseases such as pediatric leukemia, where chimeric antigen receptor (CAR) expressing T cells achieve 90% complete response, putting many individuals into very long remissions1, but to date no engineered T cell therapies have been attempted to cure T1D in humans. We propose to test the hypothesis that engineered T cells can enable islet transplant with minimal or no additional immunosuppression. Islet transplantation represents the best-case scenario to test the ability of engineered T cells to protect islet cells from immune attack and will likely lay the foundation by which strategies are developed to treat new onset T1D. In islet transplant, MHC matching of donor and recipient rarely occurs, generating highly expressed, unique islet-specific HLA antigens that can be targeted by CAR engineered T regulatory cells (CAR Tregs) or T cells engineered to express molecules that suppress the immune system (T suppressor cells or Tsups). Additionally, through a comprehensive screening process, we have identified two targets, fibroblast activation protein (FAP) and dipeptidyl peptidase like 6 (DPP6), that are highly expressed on α and β cells and have limited expression elsewhere that could be used to treat recently diagnosed individuals, and all transplant recipients without the desired MHC mismatch. In this proposal, we will further develop this toolbox to both develop better in vivo, preclinical models of T1D and new cell and gene therapies that will prevent, stall or reverse T1D. Within the last decade, rapid progress made in T cell-based therapies makes it possible to consider such therapies for T1D. Following long-term remission of 3 cancer patients treated by CD19-specific CARs developed by the Center for Cellular Immunotherapies at Penn under the leadership of Carl June2, the CAR T cell revolution was launched. This early success drove considerable investment, empowering many institutions and companies to develop ways to improve both the safety and efficacy of, and reduce cost to produce engineered T cells. Many of these innovations will also help enable cell and gene therapies for T1D, which is the overarching goal of this RFA. The ultimate goal of this application is to successfully treat three non- human primates (NHP) with engineered T cells after an islet transplant, which we predict will launch similar enthusiasm for T1D cell and gene therapy as the first three CD19-CAR recipients did for cancer CAR therapy. To achieve this goal, an experienced team of investigators with complementary expertise will lead this proposal. This team, a transplant surgeon who helped pioneer islet transplantation as a T1D therapy (Naji), a veterinary physician with expertise developing NHP T regulatory cell therapy models (Duran-Struuck), and a cell and gene therapist with a track record of engineering T cells and developing first-in-human clinical trials (Riley), have been working closely together via Helmsley Foundation funding to obtain the preliminary data presented in this application. This team is now is poised to immediately perform in vivo studies to test the ability of engineered T cell therapies to prevent, stall, or reverse T1D. Aim 1. Engineer T Suppressor Cells (Tsups) to Facilitate Islet Transplant in Humanized Mouse Models. While T regulatory cells are potent immune suppressors, there are other promising ways to induce tolerance that may work as stand-alone therapies or synergize with CAR Tregs to protect β cell function. We will explore using PD-L1, TGF-β1, and/or γ-aminobutiric acid (GABA) expressing cells targeted to the islet to mediate islet acceptance alone or in combination with CAR Treg approaches we developed using humanized mice. Aim 2. Optimize T cell Approaches to Promote Islet Transplant in NHPs. To date our efforts have focused on developing NHP CAR Tregs to enable islet transplant. We have optimized isolation, transduction, and expansion of NHP MHC-specific CAR T regulatory cells and are poised to start islet transplant experiments immediately. After we perform proof of principle experiments using CAR Tregs, we will prioritize future experiments based on the data obtained in Aim 1. A particularly exciting aspect of our studies is testing whether the ortho-IL-2 system developed by the Garcia Lab3 can work in non-human primates and support engineered Treg or other engineered T cell function and expansion while having no effect on endogenous effector cells. If successful, these studies will form the basis and rationale for Phase I clinical trials in humans.