Immune response to allogeneic hESC-derived endothelial cells

对同种异体 hESC 衍生内皮细胞的免疫反应

• 批准号: 7826527
• 负责人: YONG-GUANG YANG
• 金额: $ 89.52万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-21 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7826527
• 关键词: Address; Allogenic; Allogenicity; Allografting; Antibody Formation; Antigens; Area; B-Lymphocytes; Biological Models; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cell Line; Cell Therapy; Cell surface; Cells; Chimerism; Chronic; Clinical; Dendritic Cells; Development; Embryo; Endothelial Cells; Exhibits; Goals; Graft Rejection; Harvest; Hematological Disease; Hematopoietic; Human; Human Characteristics; Immune; Immune response; Immune system; Immunization; Immunocompetent; Immunodeficient Mouse; Immunosuppression; In Vitro; Lead; Lung; Lymphoid; Measures; Mediating; Mus; Natural Killer Cells; Natural regeneration; Organ; Patients; Peripheral Blood Lymphocyte; Population; Protocols documentation; Regenerative Medicine; Replacement Therapy; Series; Staging; Stem cells; System; T-Lymphocyte; T-Lymphocyte Subsets; Time; Tissues; Translations; Transplantation; allograft rejection; base; clinically relevant; human embryonic stem cell; human embryonic stem cell line; human embryonic stem cell transplantation; immune function; immunogenicity; in vitro Assay; in vivo; in vivo Model; induced pluripotent stem cell; mouse model; novel; novel strategies; prevent; progenitor; public health relevance; reconstitution; response; self-renewal; somatic cell nuclear transfer; stem

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (11) Regenerative Medicine and specific Challenge Topic, 11-HL-101* Develop cell-based therapies for cardiovascular, lung, and blood diseases. Given their self-renewing and pluripotent nature, human embryonic stem (hES) and induced pluripotent stem (iPS) cells hold great promise as cell replacement therapy and regenerative medicine. We recently developed a 2D culture system for differentiating hES cells into the endothelial lineage, and demonstrated that hES cell-derived endothelial cells (hES-EC) can self-assemble into functional blood vessels that spontaneously integrate into the circulatory system after transplantation into immunodeficient mice. However, a key challenge for clinical translation of hES-EC-based therapies is the immunological rejection. The major goal of this project is to mechanistically understand the immunological responses to allogeneic hES-EC, and to develop effective approaches to preventing the rejection of hES cell-derived endothelial allografts. A key factor currently impeding the study of the immunogenicity and allogenicity of hES/iPS cells and their derivatives is the lack of a suitable in vivo model. We have developed a novel protocol for human immune reconstitution in immunodeficient mice. The humanized mice (hu-mice) exhibit sustained repopulation with multilineages of human lymphohematopoietic cells and formation of secondary lymphoid organs, develop strong antigen-specific human T cell and antibody responses upon immunization, and mediate transplant rejection. In this proposal, we will use this hu-mouse model to pursue the following two specific aims. In Aim 1, we will examine the allogenicity of hES-EC in hu-mice. The goal of this aim is to understand the allogenicity of hES-EC and determine whether hES-EC at different differentiation stages (e.g., early vs. late hES-EC) differ in allogenicity. First, we will determine the allogenicity of hES-EC using a series of in vitro assays. We will then assess in vivo immune responses to allogeneic hES-EC in hu-mice. We will also assess the survival/rejection of hES-EC allografts in hu-mice that are depleted of human T cells or T cell subset(s) for identifying the immune components that are essential to hES-EC allograft rejection. In Aim 2, we will explore the possibility of inducing tolerance to hES-EC allografts in hu-mice through establishment of mixed hematopoietic chimerism. We will develop protocols for in vitro differentiation of transplantable hematopoietic stem/progenitor cells from hES cells, and with these cells to induce chimerism and donor-specific tolerance in hu-mice. These studies will determine whether hES-EC derived from the same hES cell line that is used for establishing hematopoietic chimerism can be permanently accepted by the chimeric hu-mice without the need of immunosuppression. Successful completion of the proposed studies will help us to understand the mechanisms of alloimmune responses to hES-EC and develop effective strategies to overcome the immunological hurdle to hES cell-based regenerative medicine. PUBLIC HEALTH RELEVANCE: Immunological rejection is a key challenge for clinical translation of human embryonic stem (hES) and induced pluripotent stem (iPS) cell-based regenerative medicine. This proposal aims to use a humanized mouse model to assess the allogenicity of hES cell-derived endothelial cells and to develop strategies for controlling alloimmune responses to hES cell-derived endothelial allografts.

描述（由申请人提供）：本申请涉及广泛的挑战领域 (11) 再生医学和特定挑战主题 11-HL-101* 开发针对心血管、肺和血液疾病的细胞疗法。鉴于其自我更新和多能的性质，人类胚胎干（hES）和诱导多能干（iPS）细胞在细胞替代疗法和再生医学方面具有广阔的前景。我们最近开发了一种用于将 hES 细胞分化为内皮谱系的 2D 培养系统，并证明 hES 细胞衍生的内皮细胞 (hES-EC) 可以自组装成功能性血管，在移植到免疫缺陷小鼠体内后，这些血管会自发地融入循环系统。然而，基于 hES-EC 的疗法临床转化的一个关键挑战是免疫排斥。该项目的主要目标是从机制上了解对同种异体 hES-EC 的免疫反应，并开发有效的方法来防止 hES 细胞衍生的同种异体内皮移植物的排斥。目前阻碍hES/iPS细胞及其衍生物的免疫原性和同种异体性研究的一个关键因素是缺乏合适的体内模型。我们开发了一种新的免疫缺陷小鼠免疫重建方案。人源化小鼠（hu-mice）表现出人类淋巴造血细胞多谱系的持续再增殖和次级淋巴器官的形成，在免疫后产生强烈的抗原特异性人类T细胞和抗体反应，并介导移植排斥。在本提案中，我们将使用这种 hu-mouse 模型来实现以下两个具体目标。在目标 1 中，我们将检查 hu-mice 中 hES-EC 的同种异体性。该目的的目的是了解 hES-EC 的同种异体性，并确定不同分化阶段的 hES-EC（例如早期 hES-EC 与晚期 hES-EC）的同种异体性是否存在差异。首先，我们将使用一系列体外测定来确定 hES-EC 的同种异体性。然后我们将评估 hu-mice 对同种异体 hES-EC 的体内免疫反应。我们还将评估人类 T 细胞或 T 细胞亚群耗尽的 hu-mice 小鼠中 hES-EC 同种异体移植物的存活/排斥，以确定 hES-EC 同种异体移植物排斥所必需的免疫成分。在目标 2 中，我们将探索通过建立混合造血嵌合体诱导胡小鼠对 hES-EC 同种异体移植物耐受的可能性。我们将开发从 hES 细胞体外分化可移植造血干/祖细胞的方案，并用这些细胞在 hu-mice 中诱导嵌合和供体特异性耐受。这些研究将确定源自用于建立造血嵌合状态的同一 hES 细胞系的 hES-EC 是否可以被嵌合 hu-小鼠永久接受，而无需免疫抑制。成功完成拟议的研究将有助于我们了解 hES-EC 的同种免疫反应机制，并制定有效的策略来克服基于 hES 细胞的再生医学的免疫障碍。 公共健康相关性：免疫排斥是人类胚胎干 (hES) 和诱导多能干 (iPS) 细胞再生医学临床转化的关键挑战。该提案旨在使用人源化小鼠模型来评估 hES 细胞来源的内皮细胞的同种异体性，并制定控制 hES 细胞来源的内皮同种异体移植物的同种免疫反应的策略。