Gene-Editing of Human Pluripotent Stem Cell-Derived Cardiovascular Therapy Grafts to Improve Transplant Outcomes

对人类多能干细胞衍生的心血管治疗移植物进行基因编辑以改善移植结果

基本信息

批准号：
10683804
负责人：
Matthew E Brown
金额：
$ 38.21万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-13 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10683804
关键词：
Ablation Adherence Allogenic American Anatomy Animal Experiments Anti-Inflammatory Agents Antiinflammatory Effect Apoptosis Binding Biological Assay Biology Blood Vessels CCL2 gene CCL22 gene CRISPR/Cas technology Cardiac Cardiac Myocytes Cardiovascular Diseases Cardiovascular system Cell Adhesion Cell Adhesion Molecules Cell Communication Cell Therapy Cell-Mediated Cytolysis Cells Cellular immunotherapy Clinical Clinical Trials Cues Cytolysis Data Discipline Disease Elements Endothelial Cells Extravasation Female Fibroblasts Free Radical Scavenging Functional disorder Future Gene Expression Genes Goals HLA Antigens Health Histocompatibility Antigens Class I Human Immune Immune Targeting Immune Tolerance Immune response Immunocompetent Immunology Immunosuppression Immunotherapy In Vitro Individual Infarction Infiltration Inflammatory Inflammatory Response Intercellular adhesion molecule 1 Interleukin-10 Intervention Knock-out Laboratories Leukocytes Malignant Neoplasms Mediating Mission Modeling Myocardial Infarction Natural Killer Cells Outcome Oxidative Stress Pathology Pathway interactions Patients Pharmacologic Substance Phenotype Pluripotent Stem Cells Public Health Quality of life Research Research Project Grants Resistance Risk Source T-Lymphocyte TNF gene Testing Therapeutic Intervention Thick Transplantation Transplantation Tolerance United States National Institutes of Health Validation Vascular Graft Ventricular Work cell type chemokine clinical translation cytokine cytotoxic design disability donor-specific antibody heart function human pluripotent stem cell immunogenicity immunological synapse formation improved improved functioning in vivo in vivo Model innovation insight male mouse model next generation nonhuman primate novel novel strategies post-transplant preclinical study prevent repair function reparative capacity stem cell therapy therapeutic genome editing translational study transplantation therapy

项目摘要

ABSTRACT After a first myocardial infarction (MI), 36% of male and 47% of female patients die within 5 years. This illustrates the inadequacy of current therapeutic interventions. The long-term goal of our laboratory is to develop reparative pluripotent stem cell (PSC)-based therapies that are immune-tolerated and meaningfully improve patient health and quality of life. The overall objectives of this R01 application are to: 1) use CRISPR/Cas9 gene-editing approaches to target adhesion molecules (AMs) on human PSC-derived cardiovascular therapies (PSC-CVTs) to disrupt the adherence, infiltration, and destruction of vascularized grafts by allogeneic immune cells; and 2) define optimal cellular composition and immunogenicity profiles of next-generation hypoimmune PSC-CVT grafts to maximize their reparative capacity in the inflammatory setting of MI. Our central hypothesis is that targeted deletion of AM genes will facilitate immune tolerance of PSC- CVTs via two mechanisms: 1) diminished immune cell contact-mediated destruction; and 2) anti-inflammatory effects (e.g., secreted factor and gene expression changes) directly associated with genetically disrupting AM function. The rationale for this project is that hypoimmune PSCs will be key clinical platforms in the coming years and improved gene-editing approaches are needed to achieve effective immune tolerance of PSC grafts. Additionally, successful validation of AM gene-editing in this project will provide a new avenue for advancing future transplantation therapies for other diseases. To attain our objectives, we will pursue the following specific aims (SAs): SA1) Define the effects of AM ablation on immune cell contact-mediated PSC-CVT graft destruction; SA2) Define the inflammatory responses initiated by immune cell:PSC-CVT graft interactions; and SA3) Determine the in vivo reparative capacity and immune-tolerance potential of AM knockout PSC-CVT grafts in the inflammatory MI setting. This research is significant because it validates a new graft strategy and testing platform for hypoimmune PSC therapies, with great potential to save lives and improve quality of life for many patients with MI and other pathologies characterized by cellular dysfunction in immune-competent anatomical sites. It is innovative because: 1) of the new approach of targeting immune cell adhesion in a manner anticipated to impede both adaptive and innate immune cell-mediated graft destruction; 2) it uses a tri- cellular PSC-CVT graft optimized for superior reparative function and hypoimmunogenicity; and 3) we rigorously interrogate the human immune response using advanced assays and models developed in our lab. Ultimately, this work will develop a breakthrough cardiac therapy well-suited for clinical trials, with the potential to save lives and improve the quality-of-life for millions of patients.

抽象的第一次心肌梗塞（MI）之后，男性中有36％和47％的女性患者在5年内死亡。这说明当前治疗干预措施不足。我们实验室的长期目标是发展为免疫耐受性且有意义的基于修复的多能干细胞（PSC）的疗法改善患者的健康和生活质量。此R01应用程序的总体目标是：1）使用 CRISPR/CAS9基因编辑方法，用于人类PSC衍生的靶粘附分子（AMS）心血管疗法（PSC-CVT）破坏了血管化的粘附，浸润和破坏同种异体免疫细胞的移植物； 2）定义最佳细胞组成和免疫原样下一代低免疫PSC-CVT移植物在炎症环境中最大化其恢复能力 Mi。我们的中心假设是，AM基因的靶向缺失将促进PSC-的免疫耐受性 CVT通过两种机制：1）免疫细胞接触介导的破坏减少； 2）抗炎效果（例如，分泌因子和基因表达变化）与遗传破坏AM直接相关功能。该项目的基本原理是低免疫PSC将是即将到来的关键临床平台需要使用年和改进的基因编辑方法来实现PSC移植物的有效免疫耐受性。此外，该项目中成功验证AM基因编辑将为前进的新途径提供新的途径其他疾病的未来移植疗法。为了实现我们的目标，我们将追求以下具体目的（SAS）：SA1）定义了AM消融对免疫细胞接触介导的PSC-CVT移植物的影响破坏； SA2）定义免疫细胞引发的炎症反应：PSC-CVT移植物相互作用；和 SA3）确定AM基因敲除PSC-CVT的体内修复能力和免疫耐受性的潜力炎症MI环境中的移植物。这项研究很重要，因为它验证了一种新的移植策略和低免疫PSC疗法的测试平台，具有挽救生命并改善生活质量的巨大潜力许多具有MI和其他病理的患者，其特征是免疫能力的细胞功能障碍解剖站点。它具有创新性，因为：1）靶向免疫细胞粘附的新方法预计会阻碍适应性和先天免疫细胞介导的移植物破坏的方式； 2）使用三细胞PSC-CVT移植物优化用于优质的修复功能和低肿瘤生成性； 3）我们严格使用我们实验室中开发的高级测定法和模型对人的免疫反应进行询问。最终，这项工作将开发出非常适合临床试验的突破性心脏疗法，潜力挽救生命并改善数百万患者的生活质量。