Chemoresistance and Stem Cell Selection

化疗耐药性和干细胞选择

• 批准号: 7049120
• 负责人: DAVID A WILLIAMS
• 金额: $ 33.29万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7049120
• 关键词: DNA repair; antineoplastics; biotechnology; blood disorder chemotherapy; bone marrow transplantation; carmustine; combination therapy; congenital aplastic anemia; cytogenetics; cytoprotection; disease /disorder model; drug adverse effect; dyserythropoietic anemia; gene delivery system; gene mutation; gene therapy; hematopoietic stem cells; laboratory mouse; leukemia; methylguanine DNA methyltransferase; nonhuman therapy evaluation; stem cell transplantation; therapy adverse effect; therapy design /development; transfection /expression vector

DESCRIPTION (provided by applicant): The success of the gene therapy trial in treatment of severe combined immunodeficiency (SCID) provides evidence of the power and utility of in vivo selection of transduced hematopoietic cell populations. Over the past decade, we and others have demonstrated the effectiveness of a combined gene transfer and pharmacologic approach for in vivo selection of hematopoietic stem cells utilizing chemotherapy-resistance genes. In particular, transfer of mutants of O6 methylguanine DNA methyltransferase (MGMT) in combination with inhibitors of this protein, such as 6-benzylguanine (6-BG), and treatment with alkylating agents have been demonstrated to be highly effective for in vivo selection of hematopoietic cells, specifically hematopoietic stem cells. Gene transfer of MGMT has wide application, not only in stem cell selection, but also in generating chemo-resistant hematopoiesis that may allow dose intensification in cancer therapies. While this approach is effective, little is known neither about the potential damage to the functional stem cell compartment nor the long term effects of residual DNA damage present in long-lived cells after such selection. In addition, although the risk of insertional mutagenesis resulting from integrating retroviruses has received wide-spread recent attention and is an important issue facing application of gene transfer to human diseases, we believe that these risks are related not simply to the integration event, but are likely also dependent upon the specific transgene expressed and other factors involving the manipulated cells, such as proliferative stress and proliferative history. In this interactive, multi-investigator grant application, we propose to examine the effectiveness of in vivo stem cell selection, the effects of such selection on long-term stem cell function, the quantity and quality of residual DNA damage that is present after in vivo selection and finally the pathophysiological relevance of the combination of these factors in addition to vector integration on the incidence of leukemia transformation of hematopoietic cells in vivo. The proposed work includes collaborative efforts of a number of accomplished and productive investigators with expertise in hematopoiesis, virus vector design, detection and characterization of DNA damage and vector integration identification/bioinformatics. The research plan directly models and evaluates the effects of in vivo selection and DNA damage on a critical, sensitive, and therapeutically relevant biological compartment (hematopoietic stem cells). The results from these studies will provide powerful insight into the application of virus vectors to genetic diseases and cancer as well as detailed consequences and benefits of protecting hematopoietic stem cells from DNA damage.

描述（由申请人提供）：基因治疗试验在治疗严重联合免疫缺陷（SCID）中的成功为体内选择转导的造血细胞群的能力和实用性提供了证据。在过去的十年中，我们和其他人已经证明了联合基因转移和药理学方法在体内选择造血干细胞利用化疗耐药基因的有效性。特别地，已经证明，O 6甲基鸟嘌呤DNA甲基转移酶（MGMT）的突变体与该蛋白质的抑制剂如6-苄基鸟嘌呤（6-BG）组合的转移以及用烷化剂处理对于造血细胞，特别是造血干细胞的体内选择是高度有效的。MGMT的基因转移具有广泛的应用，不仅在干细胞选择中，而且在产生可允许癌症治疗中的剂量强化的化学抗性造血中。虽然这种方法是有效的，但对功能性干细胞区室的潜在损伤以及在这种选择后存在于长寿细胞中的残留DNA损伤的长期影响知之甚少。此外，尽管整合逆转录病毒引起的插入突变的风险最近受到广泛关注，并且是基因转移应用于人类疾病所面临的重要问题，但我们认为这些风险不仅与整合事件有关，而且可能还取决于表达的特定转基因和涉及操纵细胞的其他因素，如增殖性应激和增殖史。在这个互动的，多研究者资助申请中，我们建议检查体内干细胞选择的有效性，这种选择对长期干细胞功能的影响，在体内选择后存在的残留DNA损伤的数量和质量，以及最后这些因素的组合加上载体整合对造血干细胞白血病转化发生率的病理生理学相关性。体内细胞。拟议的工作包括一些有成就和富有成效的研究人员在造血，病毒载体设计，DNA损伤和载体整合鉴定/生物信息学的检测和表征的专业知识的合作努力。该研究计划直接模拟和评估体内选择和DNA损伤对关键，敏感和治疗相关的生物区室（造血干细胞）的影响。这些研究的结果将为病毒载体在遗传疾病和癌症中的应用提供强有力的见解，以及保护造血干细胞免受DNA损伤的详细后果和益处。