Stem Cell Therapies for Primary Immune Deficiency

原发性免疫缺陷的干细胞疗法

• 批准号: 7347251
• 负责人: Gay M Crooks
• 金额: $ 127.56万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-18 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7347251
• 关键词: Stem; Cell; Therapies; Primary; Immune

The overall goal of this Program is to develop novel stem cell transplantation and gene therapy approaches to produce an intact immune system in patients with Severe Combined Immune Deficiency (SCID) and other Primary Immune Deficiencies. The manipulation of stem and progenitor cells in the profoundly immune deficient patient with SCID provides a unique biological and clinical setting to unravel mechanisms of stem cell engraftment, lymphopoiesis and immune reconstitution. The central hypothesis of the Program is that the age of the hematopoietic graft and the host environment influences the ultimate reconstitution of the immune system in the patient with SCID. Each of the Projects will test this hypothesis from a different and complementary perspective. The goal of Project 1 (Crooks) is to delineate the role of the thymic vascular niche in the regulation of homing to the neonatal thymus after bone marrow transplantation (BMT). Project 2 (Dorshkind) will provide new information regarding fetal B cell development and the development of the humoral immune response, with a specific focus on the ontogeny of B-1 B cells. This information will be highly relevant to understanding the restoration of humoral immunity following BMT and gene therapy. The Program brings together an experienced and collaborative team of investigators from UCLA, with interactions facilitated through an Administrative Core. Two scientific cores will support the Projects by providing specialized expertise in cell and tissue isolation and analysis, and in animal models of transplantation. These Projects and Cores will provide synergy and focus for basic and clinical studies in Stem Cell therapies for SCID, the most lethal form of Primary Immune Deficiency. Project 1: Thymic Reconstitution and the Vascular Niche (Gay Crooks, M.D.) Revised Abstract Section (No Change) Studies of age-related changes in thymic function and reconstitution after bone marrow transplant (BMT) have focused on the epithelial component of the thymic microenvironment; little attention has been paid to the influence of the endothelial compartment of the thymus in this process. The goal of this proposal is to delineate the role of the thymic vascular niche in the regulation of homing of bone marrow cells to thymus after transplantation. Clinical data from patients with Severe Combined Immune Deficiency (SCID) and our own data in experimental immune deficient murine models, show that when non-conditioned BMT is performed specifically in the neonatal period, rapid thymic engraftment results in donor lymphopoiesis, with little or no contribution of donor cells to non-lymphoid lineages. These findings demonstrate that although microenvironmental signals for T cell differentiation exist throughout postnatal life, different host mechanisms for thymic engraftment may exist in neonatal and adult hosts. We propose that in the neonatal setting, bone marrow can home to the thymus directly without the need for an intermediate stage of marrow engraftment. We will use the striking dichotomy between neonatal and adult thymic engraftment patterns to define the endothelial mechanisms regulating these differences. Data from murine allogeneic models will be translated to the human setting by testing our findings in a similar xenogeneic BMT model and by analysis of human thymus samples at different stages of postnatal development. Project 2: Identification of Human Cord Blood B Lineage Cells (Kenneth Dorshkind, Ph.D.) Revised Abstract Section Studies of murine B cell development have demonstrated the existence of two distinct waves of cell production. The first initiates during embryogenesis and results in the production of progenitors destined to generate B-1 B cells that are part of the innate immune system and which are particularly efficient at recognizing encapsulated bacteria such as S. pneumoniae. The second major wave generates B-2 B cells that are produced in the bone marrow throughout life and function in adaptive immunity. The central hypothesis of this application continues to be that successful reconstitution of humoral immunity in humans following bone marrow transplantation is dependent upon the recapitulation of these two distinct ontogenic programs. However, other than the fact that B cells emerge in the embryo, nothing is known about human fetal B cell development. Studies to characterize novel human cord blood progenitors that include an intermediate that co-expresses B and myeloid lineage determinants will be performed. The information that will be obtained is relevant to restoration of humoral immunity following bone marrow transplantation, the treatment of immunodeficiency diseases, and the recovery of the immune response following various myeloablative treatments. Administrative Core A Gay Crooks Revised Abstract Section Dr Gay Crooks is the Principal Investigator of the Program Project Grant and Director of the Administrative Core. The Administrative Core will be an essential mechanism to enable all the scientific components of the Program to interact by providing the structure and financial support for regular planning and evaluation of the research. Regular communication between investigators will be critical for the success of the Program. The Core will be responsible for ensuring that the investigators act as a cohesive and interactive group by providing the planning and leadership required for regular and frequent interactions between projects and cores. The Administrative Core will also be responsible for supervising budgetary and regulatory aspects of the Program, and providing the logistical support for transporting samples between institutions. Program interactions and oversight will be facilitated by both an Internal Advisory Board and an External Advisory Board composed of experts in the fields of Primary Immune Deficiency, Bone marrow Transplantation and basic immuno-biology. Core B: Cell Isolation and Analysis Core Director: Linda Baum, MD, PhD Revised Abstract Section The Cell Isolation and Analysis Core (Core B) will consist of two components, flow cytometry and histology, and will provide technical and advisory support for both Projects. The Core will aid investigators in experimental design for cell isolation and in analysis of flow cytometry data from murine and human samples. Core B will be administered by Dr Linda Baum. The core will provide FACS analysis and sorting and preparation of tissue sections and interpretation of specific tissue samples. FACSAria, and BD LSRII flow cytometry machines will be available for cell isolation and analysis. The histology component will support Projects 1 and 2 by preparing whole mounts and tissue sections by cryostat and microtome, and performing staining with standard or immunofluorescent techniques. Core C: Animal Core Director: Donald B. Kohn, MD Revised Abstract Section The focus of the Animal Core Laboratory (ACL) is to provide immunodeficient mice for the Program projects. Chimeric mouse human hybrids have become an important research tool for the evaluation of reconstitution and differentiation potential of hematopoietic and immunologic progenitor populations. The ACL will provide the following services for the projects in the Program: 1) establishment and maintenance of breeding colonies of Immunodeficient mice 3) provision of pups and adult for experiments 3) assistance with transplantation of immune deficient mice with human cells 4) health monitoring and day-to-day care of immune deficient mice, and 5) training of research personnel on the care and techniques used with immunodeficient mice. The Core will be housed within the animal care facility at UCLA under the direction of Dr. Donald Kohn.

该计划的总体目标是开发新型的干细胞移植和基因治疗方法，以在严重的免疫缺乏症（SCID）和其他原发性免疫缺陷患者中产生完整的免疫系统。严重免疫缺陷患者SCID的茎和祖细胞的操纵为解开干细胞植入，淋巴细胞和免疫重建的机制提供了独特的生物学和临床环境。该计划的中心假设是造血移植的年龄和宿主环境会影响SCID患者免疫系统的最终重构。每个项目都将从不同和互补的角度检验这一假设。项目1（骗子）的目的是描述胸骨髓移植（BMT）后胸腺血管生态裂市场在调节新生儿胸腺的作用。项目2（Dorshkind）将提供有关胎儿B细胞发育和体液免疫反应的发展的新信息，并特别关注B-1 B细胞的个体发育。该信息将与理解BMT和基因治疗后的体液免疫的恢复高度相关。该计划汇集了来自加州大学洛杉矶分校的经验丰富且合作的调查人员团队，并通过行政核心促进了互动。两个科学核心将通过提供细胞和组织隔离和分析以及移植动物模型方面的专业专业知识来支持这些项目。这些项目和核心将为SCID干细胞疗法的基础和临床研究提供协同和重点，这是原发性免疫缺陷的最致命形式。 项目1：胸腺重构和血管生态位（同性恋Crooks，M.D。） 修订的摘要部分 （没有更改） 骨髓移植后胸腺功能和重建年龄变化的研究集中在胸腺微环境的上皮成分上。在此过程中，对胸腺内皮室的影响很少。该提议的目的是描述 胸腺血管生态位在移植后骨髓细胞向胸腺的归巢中的作用。来自实验性免疫缺陷鼠模型中严重合并免疫缺陷（SCID）患者的临床数据，我们自己的数据表明，当在新生儿期间特别进行非条件的BMT时，快速的胸腺植入会导致供体淋巴结疫苗，几乎没有供体细胞对非淋巴机的供体细胞的贡献。这些发现表明，尽管在整个产后生活中都存在用于T细胞分化的微环境信号，但新生儿和成人宿主中可能存在不同的胸腺植入宿主机制。我们建议在新生儿环境中，骨髓可以直接回到胸腺，而无需骨髓植入的中间阶段。我们将使用新生儿和成人胸腺植入模式之间的引人注目的二分法来定义 调节这些差异的内皮机制。来自鼠类同种异体模型的数据将通过在类似的异构BMT模型中测试我们的发现以及通过分析产后发育的不同阶段的人类胸腺样品来转化为人类环境。 项目2：鉴定人脐带血谱系细胞（Kenneth Dorshkind，Ph.D。） 修订的摘要部分 对鼠B细胞发育的研究表明，存在两个不同的细胞产生波。第一个在胚胎发生过程中的启动，并导致产生的祖细胞的产生，以生成是先天免疫系统的一部分的B-1 B细胞，并且特别有效地识别封装 细菌，例如肺炎链球菌。第二个主要波会产生B-2 B细胞，这些B-2 B细胞在自适应免疫中在骨髓中产生。该应用的核心假设继续是，骨髓移植后人类中的体液免疫的成功重建取决于对这两个不同的化体基础计划的概括。但是，除了B细胞出现在胚胎中的事实外，对人类胎儿B细胞的发育尚无任何了解。将表征包括中间体的新型人脐带血祖细胞的研究，该中间体将进行表达B和髓样谱系决定因素。将获得的信息与骨髓移植后体液免疫的恢复，免疫缺陷疾病的治疗以及各种骨髓性治疗后免疫反应的恢复有关。 行政核心a 同性恋骗子 修订的摘要部分 盖伊·克鲁克斯（Gay Crooks）博士是计划项目赠款的主要研究员，也是行政核心的主任。行政核心将是使程序的所有科学组成部分通过为研究的定期计划和评估提供结构和财务支持，以使该计划的所有科学组成部分进行互动。调查人员之间的定期沟通对于该计划的成功至关重要。核心将负责确保调查人员通过提供项目与核心之间的常规和频繁互动所需的计划和领导来确保成为一个凝聚力和互动群体。行政核心还将负责监督该计划的预算和监管方面，并为机构之间运输样本提供后勤支持。计划的互动和监督将由内部顾问委员会和外部顾问委员会促进，该咨询委员会由原发性免疫缺陷，骨髓移植和基本免疫生物学领域的专家组成。 核心B：细胞隔离和分析核心 导演：医学博士Linda Baum博士 修订的摘要部分 细胞隔离和分析核心（核心B）将由两个组成部分组成，即流式细胞仪和组织学，并将为两个项目提供技术和咨询支持。核心将帮助研究人员进行实验设计，以分离细胞分离和分析来自鼠和人类样品的流式细胞仪数据。核心B将由Linda Baum博士管理。核心将提供FACS分析和分类以及 组织切片的制备和特定组织样品的解释。 FACSARIA和BD LSRII流式细胞仪机将用于细胞分离和分析。组织学成分将通过低温恒温器和微型群制备整个坐骑和组织切片，并使用标准或免疫荧光技术进行染色，来支持项目1和2。 核心C：动物核心 主任：医学博士Donald B. Kohn 修订的摘要部分 动物核心实验室（ACL）的重点是为程序项目提供免疫缺陷的小鼠。嵌合小鼠人类杂种已成为评估造血和免疫学祖细胞种群重构和分化潜力的重要研究工具。 ACL将为程序中的项目提供以下服务：1）免疫缺陷小鼠的育种菌落的建立和维护3）提供幼崽和成人进行实验的提供3）帮助通过人体细胞移植免疫缺陷小鼠的帮助，4）对免疫缺陷小鼠的健康监测以及对免疫治疗的护理和5）对MOTICE培训的护理和5）进行了培训。该核心将在唐纳德·科恩（Donald Kohn）博士的指导下安置在加州大学洛杉矶分校的动物护理设施中。