Engineered in vivo nuclear fusion: application to regenerative medicine

工程体内核聚变：在再生医学中的应用

• 批准号: 7833374
• 负责人: RICHARD C MULLIGAN
• 金额: $ 47.62万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7833374
• 关键词: Activities of Daily Living; Address; Adoptive Transfer; Affect; Allogenic; Animal Model; Animals; Applications Grants; Area; Biological Preservation; Cell Nucleus; Cell Transplantation; Cell Transplants; Cell fusion; Cell physiology; Cell surface; Cells; Clinical; Development; Disease; Disorder by Site; Engineering; Engraftment; Genetic; Genetic Materials; Goals; Health; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Homologous Transplantation; Human; Immunoglobulin-Secreting Cells; In Vitro; Inherited; Injection of therapeutic agent; Injury; Insulin; Lentivirus Vector; Liver; Maintenance; Mesenchymal; Mus; Muscle; Natural regeneration; Nuclear Fusion; Organ; Outcome; Pancreas; Phenotype; Population; Production; Proteins; Regenerative Medicine; Relative (related person); Research; Series; Source; Stem cells; T-Lymphocyte; Therapeutic; Tissues; Transplantation; Viral; Viral Envelope Gene; Wound Healing; base; cell type; cellular engineering; effective therapy; env Gene Products; gene therapy; genetic technology; in vivo; interest; islet; macrophage; mutant; new technology; novel; nuclear transfer; programs; public health relevance; receptor; regenerative; repaired; research study; response; therapeutic gene

DESCRIPTION (provided by applicant): Engineered in vivo nuclear fusion: application to regenerative medicine. This application addresses broad Challenge Area (11) Regenerative Medicine and specific Challenge Topic, 11-DK-104: Use of Hematopoietic Stem Cells (HSC) to regenerate or repair mesenchymal tissues. This application focuses on the development of a new cell-based strategy for regenerative medicine and gene therapy which depends upon the ability to genetically engineer hematopoietic stem cells or other cell populations to be capable of efficient fusion to cells within organs and tissues after their transplantation in vivo, using murine retroviral envelope or human viral-like fusogenic gene products. The approach seeks to build upon provocative recent studies of the ability of hematopoietic or other stem cells to transdifferentiate into unexpected differentiated cells which have suggested that spontaneous cell fusion involving circulating hematopoietic cells and cells within organs and tissues can occur, particularly in response to tissue injury, and may represent an important mechanism for the repair of organs and tissues. The hypothesis underlying this grant application is that genetic technologies can provide a means of dramatically increasing the efficiency of cell fusion of transplanted cells, and that such engineered cell-cell fusion in vivo can provide a broad and powerful platform for repairing tissues and for the delivery of useful therapeutics. A major goal of the two-year program is to rapidly assess the potential breadth of the new technology so that subsequent studies can immediately focus on the most promising therapeutic applications. In a series of in vitro experiments, effort will be made to further understand the experimental parameters governing engineered cell-cell fusion via fusogenic viral envelope gene products, with an emphasis on characterizing the viability and fate of fused cells, and determining how expression levels of either the fusogenic envelopes or their corresponding receptors affect the absolute efficiency of fusion and the average number of nuclei that are fused. In vivo studies will focus on a determination of the range of tissues and organs that are amenable to cell fusion after local or systemic delivery of different types of donor cells. In addition to determining the efficiency of nuclear transfer to muscle and other tissues and organs, the capacity of donor nuclei derived from different cell types to provide for the expression of therapeutic gene products, and the immunological consequences of the transplantation of allogeneic and xenogeneic cells, will be determined. A particularly important goal of the in vivo studies will be to understand how interactions between donor and recipient nuclei affect the extent of reprogramming and/or maintenance of the differentiated phenotype of the donor and recipient nuclei in different tissues. Of specific interest is whether in vivo nuclear fusion can be employed to reprogram cells in the pancreas or liver to the islet phenotype, and/or to enable the preservation of the specialized differentiated functions of specific cells, such as insulin or antibody secreting cells via introduction of nuclei from those specialized cells into muscle or other tissues. In efforts to systemically deliver nuclei to tissues via the transplantation of hematopoietic stem cells, an important objective is to directly determine whether the expression of fusogenic gene products can provide for increased levels of engraftment of donor derived nuclei and regenerative repair relative to untransduced cells. PUBLIC HEALTH RELEVANCE: The proposed research is highly relevant to human health, as the goal of the studies is to develop a novel cell transplantation approach to regenerative medicine. Such a therapy could have a major impact upon the treatment of many inherited and acquired diseases for which there are currently no effective treatments

描述（由申请人提供）：工程化体内核融合：应用于再生医学。本申请涉及广泛的挑战领域（11）再生医学和特定的挑战主题，11-DK-104：使用造血干细胞（HSC）再生或修复间充质组织。本申请集中于开发用于再生医学和基因治疗的新的基于细胞的策略，其依赖于使用鼠逆转录病毒包膜或人病毒样融合基因产物对造血干细胞或其它细胞群进行遗传工程改造以在其体内移植后能够有效融合到器官和组织内的细胞的能力。该方法试图建立在造血或其他干细胞转分化成意想不到的分化细胞的能力的最近的挑衅性研究的基础上，这些研究表明，涉及循环造血细胞和器官和组织内的细胞的自发细胞融合可以发生，特别是响应于组织损伤，并且可能代表器官和组织修复的重要机制。这项资助申请的假设是，遗传技术可以提供一种显著提高移植细胞的细胞融合效率的方法，并且这种体内工程化细胞-细胞融合可以为修复组织和递送有用的治疗剂提供广泛而强大的平台。这项为期两年的计划的一个主要目标是快速评估新技术的潜在广度，以便后续研究能够立即专注于最有前途的治疗应用。在一系列体外实验中，将努力进一步了解通过融合病毒包膜基因产物控制工程化细胞-细胞融合的实验参数，重点是表征融合细胞的活力和命运，并确定融合包膜或其相应受体的表达水平如何影响融合的绝对效率和融合的核的平均数量。体内研究将集中于确定在局部或全身递送不同类型的供体细胞后适合于细胞融合的组织和器官的范围。除了确定核转移到肌肉和其它组织和器官的效率之外，还将确定来源于不同细胞类型的供体核提供治疗性基因产物表达的能力，以及同种异体和异种细胞移植的免疫学后果。体内研究的一个特别重要的目标是了解供体和受体细胞核之间的相互作用如何影响不同组织中供体和受体细胞核的重编程程度和/或分化表型的维持。特别感兴趣的是体内核融合是否可以用于将胰腺或肝脏中的细胞重编程为胰岛表型，和/或通过将来自那些特化细胞的核引入肌肉或其他组织中来使得能够保留特定细胞（例如胰岛素或抗体分泌细胞）的特化分化功能。在通过造血干细胞移植将细胞核系统递送至组织的努力中，一个重要的目的是直接确定融合基因产物的表达是否可以提供相对于未转导的细胞的供体来源的细胞核的增加的植入水平和再生修复。 公共卫生关系：拟议的研究与人类健康高度相关，因为研究的目标是开发一种新的细胞移植方法用于再生医学。这种疗法可能会对许多目前没有有效治疗方法的遗传性和获得性疾病的治疗产生重大影响