CRISPR-Modified Cardiac Xenograft Transplantation

CRISPR 改良心脏异种移植

• 批准号: 10403525
• 负责人: Richard N Pierson
• 金额: $ 79.57万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-10 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10403525
• 关键词: Address; Adenosine; Adhesions; Adhesives; Antibodies; Binding; Biochemical; Biological Assay; Blood; Blood Coagulation Disorders; Blood Platelets; Cells; Clinic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coagulation Process; Complement; DNA cassette; Data; Development; Drug usage; Endothelium; Erythrocytes; Exhibits; Family suidae; Flow Cytometry; Functional disorder; Gene Expression; Genes; Genetic; Genetic Engineering; Genotype; Health; Heart; Heart Injuries; Histology; Hour; Human; Immunosuppression; In Vitro; Inflammation; Injury; Integrins; Intervention; Ischemia; Kidney; Knock-out; Learning; Leukocytes; Life; Liver; Lung; Measures; Mediating; Modeling; Modification; Monoclonal Antibodies; Organ Transplantation; Organ failure; Papio; Pathway interactions; Pharmaceutical Preparations; Phenotype; Physiology; Positioning Attribute; Publishing; Refractory; Regimen; Regulator Genes; Reperfusion Injury; Reporting; Residual state; Resources; Selectins; Steroids; Structure; TNFRSF5 gene; Testing; Thrombomodulin; Transferase; Translating; Translations; Transplantation; Treatment Protocols; United States National Institutes of Health; Work; Xenograft procedure; base; clinical application; clinically relevant; complement pathway; design; ex vivo perfusion; experience; experimental study; genetic regulatory protein; heart function; heart xenograft; immunomodulatory therapies; immunoregulation; improved; in vivo; innovation; insight; novel; pre-clinical; prevent; protein expression; targeted treatment; thrombotic; tool; trial design

SUMMARY ABSTRACT: CRISPR-Modified Cardiac Xenograft Transplantation The past three years have witnessed two major breakthroughs in heart xenotransplantation. First, the previously intractable barrier of delayed xenograft rejection (DXR) has been overcome in the pig-to-baboon heart xeno model. In this model, DXR is manifest primarily as consumptive coagulopathy (CC) in the recipient and thrombotic microangiopathy (TM) in the graft. Working in the heterotopic model in baboons, our `Mohiuddin/NIH' group used genetically modified GTKO.hCPRP.hTBM hearts and a treatment regimen including “induction” T and B depletion, MMF, and steroids – all clinically used drugs – with an experimental monoclonal antibody directed against CD40. DXR was prevented for as long as CD40 treatment was continued, and in one instance xenograft survival was extended beyond two years. The second major advance came from the Munich group, who reported consistent survival of orthotopic pig heart xenograft recipients beyond 180 days using our immunomodulatory regimen and GTKO.hCPRP.hTBM hearts. Accomplishing consistent survival of orthotopic heart xenografts had previously been prevented by initial xenograft dysfunction (IXD), a phenomenon refractory to concerted efforts by multiple experienced clinical transplant teams. Importantly, the Munich group found that minimizing graft ischemia was necessary and sufficient to consistently prevent IXD. Discovering that ischemia minimization was sufficient to overcome the IXD barrier is a `breakthrough' advance, even as the mechanism of improved graft protection by ischemia minimization is not yet well understood. In this Project, we hypothesize that IXD mechanisms that injure GTKO.hCPRP.hTBM hearts may be addressed by the additional xeno-focused genetic modifications expressed in one or more versions of Pig 2.0, created by our collaborators at eGenesis using innovative CRISPR-driven gene editing tools. Pig 2.0 is designed to address multiple known xeno-specific injury mechanisms through a combination of 12 xeno- targeted genetic modifications. This Project takes advantage of availability of several well-defined versions of Pig 2.0, our deep experience with multiple in vitro, ex vivo, and in vivo heart xeno models, and a robust mechanistic assay capability to determine whether Pig 2.0 is protected from heart IXD, with or without ischemia minimization (Aim 1). Specifically, we will use these unique resources to learn a) whether versions of Pig 2.0 containing a coagulation cascade regulatory gene cassette are protected from IXD; b) whether ischemia minimization is necessary and/or sufficient to prevent IXD in susceptible Pig 2.0 hearts; and c) to define a clinically acceptable immunosuppression regimen for Pig 2.0 hearts in baboons that is consistently safe and effective (Aim 2). Results from this Project will yield insights likely to catalyze progress for other cell and organ xenografts, and inform clinical heart xenotransplantation trial design.

摘要摘要：CRISPR修饰的心脏异种移植移植 在过去的三年中，心脏异种移植的两个重大突破。首先， 以前在猪到巴伯州已经克服了延迟Xenographographic拒绝（DXR）的顽固性障碍（DXR） 心脏Xeno模型。在此模型中，DXR在接受者中的紧凑型凝血病（CC）表现出来 和移植物中的血栓性微血管病（TM）。在狒狒的异位模型中工作，我们 “ Mohiuddin/NIH”组使用了一般修改的GTKO.hcprp.htbm心脏和治疗方案 包括“诱导” T和B耗竭，MMF和类固醇 - 所有临床使用的药物 - 针对CD40的单克隆抗体。只要CD40治疗是 继续，在一个实例中，Xenographographic生存延长了两年。 第二个重大进步来自慕尼黑集团，他们报告了原位的始终如一的生存 使用我们的免疫调节方案和 gtko.hcprp.htbm心。以前有一致的原位心脏异种移植物的生存率 最初的异种移植功能障碍（IXD）阻止 经验丰富的临床移植团队。重要的是，慕尼黑集团发现，最大程度地减少了移植缺血是 必要且足够以始终防止IXD。发现缺血最小化足以 克服IXD障碍是“突破性的”进步，即使通过通过 缺血最小化尚未得到很好的理解。 在这个项目中，我们假设损害gtko.hcprp.htbm心脏的IXD机制可能是 以一种或多种版本的Pig 2.0，以Xeno的其他遗传修饰来解决 由我们的e基因合作者使用创新的CRIS驱动基因编辑工具创建。猪2.0是 旨在通过12种Xeno-的组合解决多种已知的Xeno特异性损伤机制 靶向遗传修饰。该项目利用了几个定义明确的版本的可用性 Pig 2.0，我们对多个体外，体内和体内心脏Xeno模型的深厚经验，以及强大的 机械测定能力，可以确定Pig 2.0是否受到心脏IXD的保护，有或没有缺血 最小化（AIM 1）。特别是，我们将使用这些独特的资源来学习a）版本的猪2.0是否版本 含有凝血的级联调节基因盒受到IXD的保护； b）是否缺血 最小化是必要的和/或足以防止易感猪2.0心中的IXD的； c）定义 ba狒狒的猪2.0心的临床上可接受的免疫抑制方案，始终是安全且 有效（目标2）。该项目的结果将产生可能催化其他细胞和器官进展的见解 异种移植物，并为临床心脏异种移植试验设计提供信息。