TRANSPLANT BIOLOGY, GENE TRANSFER, AND STEM CELL SOURCES

移植生物学、基因转移和干细胞来源

• 批准号: 2519548
• 负责人: COLIN A SIEFF
• 金额: $ 28.3万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2519548
• 关键词: bone marrow transplantation; disease /disorder model; gene therapy; graft versus host disease; hematopoietic stem cells; interferon gamma; interleukin 4; laboratory mouse; tissue /cell culture; transplantation immunology; umbilical cord; virus envelope; xenotransplantation

DESCRIPTION (Adapted from the applicant's abstract) The long-term goal of this study is to determine the optimal source of hematopoietic stem cells (HSC) for transplantation, gene transfer, and prevention of graft versus host disease (GVHD). While the majority of transplants are carried out using bone marrow (BM) as the source of stem cells, there is increasing interest in the use of both chemotherapy and/or growth factor-mobilized peripheral blood progenitor/stem cells (PB) and umbilical cord blood (CB) as alternative stem cell sources. In the allogeneic setting PB would be preferable for the donor, and CB has many advantages, including the potential to create cryopreserved cord blood banks and the possible reduced immunological reactivity of CB lymphocytes with consequent decrease in GVHD. In the autologous setting CB and mobilized PB may contain increased numbers of cycling HSC and, therefore, constitute better targets for retrovirally mediated gene transfer. One of the major problems in transplanting both alternative sources of stem cells and advances in gene transfer efficiency to clinical use is the lack of optimal human stem cell assay systems. Aim 1 will compare the stem cell content and growth factor and stromal cell responsiveness of highly enriched primitive resting G0 cells from BM, CB, and PB, to test the hypothesis that CB and PB are enriched for these cells. This aim will be achieved by using novel means to enrich for these cells and by testing them in in vitro and xenogeneic in vivo culture systems, in an effort to optimize an assay for human long-term repopulating stem cells. Aim 2 will examine the capacity of stem cells from these different sources to be genetically modified by conventional retroviral vectors or novel vectors pseudotyped to express growth factor genes in place of the retroviral envelope gene. GVHD remains a source of major morbidity and mortality after allogeneic bone marrow transplantation (BMT). One can hypothesize that PB and CB differ from BM with respect to the function of their Th1 or Th2 T cell subsets that could account for the reduced GVHD observed in transplants with cells obtained from these sources, and Aim 3 will examine alternative human HSC sources for the number and function of the cells that are implicated in GVHD. In a murine GVHD model, preliminary data show that reduction of interferon-gamma (IFN- gamma) production by culturing donor T cells in interleukin 4 (IL-4) before BMT prevents this inflammatory cascade; G-CSF appears to have a similar effect to IL-4. Aim 3 will explore the hypothesis that treatment of murine donor cells with G-CSF prevents the inflammatory cytokine cascade of GVHD. By these means it is hoped that the rationale for selecting stem cells for transplantation and for gene transfer in different clinical contexts will be placed on a more secure scientific basis, and that by understanding and preventing GVHD it will eventually be possible to extend transplantation more successfully across HLA barriers.

描述（根据申请人的摘要改编）长期目标 这项研究是确定造血茎的最佳来源 细胞（HSC）用于移植，基因转移和预防移植 与宿主疾病（GVHD）。虽然大多数移植是 用骨髓（BM）作为干细胞来源进行 对化疗和/或生长的使用越来越兴趣 因子润滑的外周血祖细胞/干细胞（PB）和 脐带血（CB）作为替代干细胞来源。在 同种异体设置PB比捐赠者更可取，并且CB有很多 优点，包括产生冷冻脐带血的潜力 银行和CB的免疫反应性可能降低 淋巴细胞随之而来的GVHD降低。在自动环境中 CB和动员PB可能包含增加循环HSC的数量，并且 因此，构成了逆转录病毒介导的基因的更好靶标 转移。移植两种替代方案的主要问题之一 干细胞的来源以及基因转移效率提高到 临床用途是缺乏最佳的人类干细胞测定系统。目的 1将比较干细胞含量和生长因子和基质细胞 来自BM的高度富集原始静息G0细胞的响应性， CB和PB，以测试CB和PB富含这些的假设 细胞。通过使用新颖的手段丰富来实现此目标 这些细胞并通过在体内进行体外和异构化测试 培养系统，以优化人类长期测定法 重新流动干细胞。 AIM 2将检查干细胞的能力 从这些不同的来源通过常规修饰 逆转录病毒载体或新型载体，以表达生长系数 基因代替逆转录病毒包膜基因。 GVHD仍然是来源 同种异体骨髓后的主要发病率和死亡率 移植（BMT）。可以假设PB和CB不同 BM关于其Th1或Th2 T细胞子集的功能 可以考虑在细胞移植中观察到的GVHD降低 从这些来源获得，AIM 3将检查其他人类 HSC来源的涉及单元格的数量和功能 在GVHD中。在鼠GVHD模型中，初步数据表明还原 通过培养供体T细胞生产干扰素 - 伽马（IFN-GAMMA）的生产 在BMT之前，在白介素4（IL-4）中可以防止这种炎症性级联反应； G-CSF似乎具有与IL-4相似的作用。 AIM 3将探索 假设用G-CSF处理鼠供体细胞可防止 GVHD的炎症细胞因子级联反应。通过这些意味着希望 选择用于移植和基因的干细胞的理由 在不同的临床环境中转移将放在更安全的 科学基础，通过理解和预防GVHD将会 最终有可能更成功地扩展移植 跨越HLA障碍。