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: 2771478
• 负责人: COLIN A SIEFF
• 金额: $ 29.16万
• 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/2771478
• 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（改编自申请人的摘要）长期目标 本研究的目的是确定造血干细胞的最佳来源， 用于移植、基因转移和预防移植的HSC 抗宿主病（GVHD）。虽然大多数移植手术 使用骨髓（BM）作为干细胞的来源， 越来越多的人对使用化疗和/或生长 因子动员的外周血祖细胞/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将是首选的供体，CB有许多 优点，包括创造冷冻保存脐带血的潜力 库和CB的免疫反应性可能降低 淋巴细胞，从而减少GVHD。在自体环境中 CB和动员的PB可能含有增加数量的循环HSC， 因此，构成了逆转录病毒介导的基因的更好靶点， 转移移植这两种选择的主要问题之一是， 干细胞的来源和基因转移效率的进展， 临床应用的最大障碍是缺乏最佳的人类干细胞测定系统。目的 1将干细胞含量和生长因子与基质细胞进行比较 来自BM的高度富集的原始静息G 0细胞的反应性， CB和PB，以检验CB和PB富含这些物质的假设 细胞这一目标将通过使用新的手段来实现， 这些细胞，并通过在体外和异种体内测试它们， 培养系统，以优化人类长期 再生干细胞目标2将检查干细胞的能力 从这些不同的来源进行基因改造， 逆转录病毒载体或假型化以表达生长因子的新载体 基因代替逆转录病毒包膜基因。GVHD仍然是一个来源 同种异体骨髓移植后的主要发病率和死亡率 移植（BMT）。我们可以假设PB和CB不同于 BM关于其Th 1或Th 2 T细胞亚群的功能， 可以解释在细胞移植中观察到的GVHD减少 目标3将研究替代人类 HSC来源的数量和功能的细胞是牵连 在GVHD。在小鼠GVHD模型中，初步数据显示， 通过培养供体T细胞产生干扰素-γ（IFN-γ） 在BMT之前的白细胞介素4（IL-4）预防这种炎症级联反应; G-CSF似乎具有与IL-4类似的作用。目标3将探讨 假设用G-CSF处理鼠供体细胞可以防止 GVHD的炎性细胞因子级联反应。通过这些手段，希望 选择干细胞用于移植和基因治疗的基本原理 在不同的临床背景下转移将被放置在一个更安全的 科学依据，通过了解和预防GVHD， 最终有可能更成功地扩展移植 穿越HLA屏障

项目成果

Granulocyte colony-stimulating factor-mobilized allogeneic stem cell transplantation maintains graft-versus-leukemia effects through a perforin-dependent pathway while preventing graft-versus-host disease.

粒细胞集落刺激因子动员的同种异体干细胞移植通过穿孔素依赖性途径维持移植物抗白血病效应，同时预防移植物抗宿主病。

• 发表时间: 1999
• 期刊: Blood
• 影响因子: 20.3
• 作者: Pan,L;Teshima,T;Hill,GR;Bungard,D;Brinson,YS;Reddy,VS;Cooke,KR;Ferrara,JL
• 通讯作者: Ferrara,JL