Project-002

这项U19提案旨在通过混合嵌合来实现胰岛和肾移植耐受。 由三个核心支持的互补食蟹猴项目。这两个项目都有相关性 用于终末期肾病(ESRD)和1型糖尿病(T1D)的治疗。他们雇佣了相关人员 实现对已故和活体供体移植物的耐受性的方法。项目1旨在使用 扩增的多克隆受体Tregs开发一种可诱导耐受的压缩预适应方案 已故供者的骨髓、肾脏和胰岛同时移植。我们之前曾使用过6- 仅与活体供者相关的日间条件调节方案，以实现短暂嵌合体和肾脏 猴子和人类的同种异体移植耐受性。我们现在的目标是开发一种可以启动的条件化方案 在确认已故供者后，允许在24小时内进行移植(TX)。我们的方法建立在 我们的证据表明，扩大的多克隆受体Tregs可以实现更长的嵌合体和更多 健壮的耐受性比以前在食蟹猴模型中可能的耐受性更好。容忍消除了对 长期免疫抑制治疗及其对胰岛移植物的破坏性影响。此外，我们的研究在节点 小鼠已经表明，非清髓性诱导的持久混合嵌合体逆转了晚期抗胰岛抗体 自身免疫，同时避免同种免疫、自身免疫和药物毒性，这些目前限制了 胰岛TX治疗T1D的疗效观察项目2旨在开发一种耐受诱导策略，用于根治性治疗 应用活体亲属供体(LRD)复合胰岛肾(IK)TX治疗终末期糖尿病肾病。该项目 根据我们的观察，在大型动物身上移植预血管化胰岛作为复合iKs的一部分 与自由、无血管的胰岛相比，模型需要的胰岛要少得多，才能实现胰岛素独立。我们 最近通过一种新的低强度造血细胞移植实现了恒河猴对iKs的耐受性 一种“亲子结合”的协议。项目2旨在调整调理方案的组成部分 和/或可能对胰岛功能产生早期负面影响的供体细胞来源。利用这一能力 为了在LRD TX之前产生强大的供体特异性Tregs，我们将测试这些细胞促进耐受的能力 混合同种异体嵌合体，具有逆转T1D的潜力。这两个项目都将包括广泛的机械性 建立在高通量TCR测序方法基础上追踪同种异体反应性T细胞的分析 我们在人类身上开发的曲目，将适用于核心B中的食蟹猴。因此，我们的目标是 在两个项目中实现持久的混合嵌合体，以治愈自身免疫，同时防止 通过诱导耐受性的同种免疫攻击。A核将为这两个项目提供猴岛，B核将为这两个项目提供猴岛 开发高通量T细胞受体(TCR)测序平台，用于识别和跟踪 捐献者特异性同种异体反应T细胞克隆的命运，为这两个项目提供了一种独特的机械工具。 核心C将为所有项目和核心提供行政、生物库和数据管理支助。

This U19 proposal aims to achieve islet and kidney allograft tolerance through mixed chimerism in two complementary cynomolgus monkey projects that are supported by three cores. Both projects have relevance for end-stage renal disease (ESRD) and curative therapy for Type 1 diabetes (T1D). They employ related approaches to achieve tolerance for deceased and living donor grafts, respectively. Project 1 aims to use expanded polyclonal recipient Tregs to develop a compressed conditioning regimen that induces tolerance of simultaneous bone marrow, kidney and islet allografts from deceased donors. We have previously used a 6- day conditioning protocol that is relevant only for living donors, to achieve transient chimerism and kidney allograft tolerance in monkeys and humans. We now aim to develop a conditioning protocol that could be initiated after identification of a deceased donor, allowing transplantation (Tx) within 24 hours. Our approach builds on our demonstration that expanded polyclonal recipient Tregs can achieve far more prolonged chimerism and more robust tolerance than has previously been possible in the cynomolgus model. Tolerance obviates the need for long-term immunosuppressive therapy with its destructive effects on islet grafts. Moreover, our studies in NOD mice have shown that non-myeloablative induction of durable mixed chimerism reverses advanced anti-islet autoimmunity, simultaneously avoiding the alloimmunity, autoimmunity and drug toxicity that currently limit the efficacy of islet Tx in T1D. Project 2 aims to develop a tolerance induction strategy for curative treatment of end-stage diabetic nephropathy using living related donor (LRD) composite Islet-Kidney (IK) Tx. The project builds on our observation that transplanting pre-vascularized islets as part of composite IKs in large animal models requires far fewer islets to achieve insulin independence than Tx of free, non-vascularized islets. We recently achieved tolerance of IKs in rhesus monkeys using a novel, low intensity, hematopoietic cell transplant protocol in a “parent-to-offspring” combination. Project 2 aims to adjust components of the conditioning regimen and/or donor cell source that may have an early negative impact on islet function. Taking advantage of the ability to generate potent donor-specific Tregs prior to LRD Tx, we will test the ability of these cells to promote durable mixed allogeneic chimerism, with its potential to reverse T1D. Both projects will include extensive mechanistic analyses that build on a high-throughput TCR sequencing-based approach for tracking the alloreactive T cell repertoire that we have developed in humans and will apply to cynomolgus monkeys in Core B. Thus, we aim to achieve durable mixed chimerism in both projects to cure autoimmunity while simultaneously preventing alloimmune attack by inducing tolerance. Core A will provide monkey islets for both projects and Core B will develop a high throughput T cell receptor (TCR) sequencing platform that will be used to identify and track the fate of donor-specific alloreactive T cell clones, providing a unique mechanistic tool to be applied in both projects. Core C will provide administrative, biorepository and data management support for all of the projects and cores.