ROLE OF HYPERTHERMIA IN BONE MARROW TRANSPLANTATION

热疗在骨髓移植中的作用

• 批准号: 3460142
• 负责人: NAHID F MIVECHI
• 金额: $ 9.9万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3460142
• 关键词: HeLa cells; acute leukemia; antineoplastics; bone marrow transplantation; clone cells; colony stimulating factor; cyclophosphamide; disease /disorder model; drug adverse effect; endonuclease; etoposide; gel electrophoresis; human therapy evaluation; human tissue; hyperthermia therapy; laboratory mouse; messenger RNA; neoplasm /cancer chemotherapy; neoplasm /cancer thermotherapy; polymerase chain reaction; southern blotting; stress proteins; tissue /cell culture; western blottings

Autologous bone marrow transplantation is used for many patients including those with acute myelogenous leukemias and acute lymphoblastic leukemias who lack appropriate donor marrow. Recurrence of malignancy, however, is a major problem for patients receiving autologous bone marrow. In this study we will explore and optimize the use of hyperthermia to purge leukemic cells from bone marrow specimens in vitro. Heat response of human leukemic cell lines, acute leukemias from patients and normal human bone marrow progenitors (CFU-GM, BFU-E, CFU-GEMM) will be determined using clonogenic assays. Single or fractionated doses of heat with or without chemotherapeutic agents now used in the clinic for in vitro purging of leukemias will be used. Fractionated doses of heat will be used to take advantage of the differential rate of decay of thermotolerance of heat sensitive leukemic cells vs normal bone marrow progenitors. The damage to the bone marrow stem cells and reconstitution of the purged human bone marrow will be measured using long-term bone marrow cultures. To ensure the in vivo reconstitution of the purged bone marrow and to measure the leukemia relapse, a murine leukemia model system will be used. Both AKR leukemia and murine bone marrow will be treated in vitro with single or fractionated hyperthermia with or without chemotherapeutic agents. Furthermore, following purging of leukemia:bone marrow mixtures, both in vivo reconstitution of the bone marrow and leukemia relapse will be measured. Finally, we will test the feasibility of using the expression of the inducible HSP-72 kDa as the predictor of heat response. To determine the expression of the HSP-72 mRNA, in vitro enzymatic amplification of the HSP- 72 mRNA (PCR) will be performed by the synthetic primers. The inducible HSP-72 will then be detected by southern blot analysis using synthetic probes. We will also use 1 & 2 dimensional PAGE to detect the expression of the HSP-72 kDa protein. The expression of the inducible HSP-72 mRNA and proteins will then be correlated with the heat response of human leukemia cells. If there is a positive correlation between the presence of the inducible HSP-72 kDa and thermal resistance, PCR could be used clinically to predict the thermal response of leukemic cells before in vitro purging with heat.

自体骨髓移植被用于许多患者，包括 急性髓系白血病和急性淋巴细胞性白血病 他们缺乏合适的捐赠者骨髓。然而，恶性肿瘤的复发是一种 接受自体骨髓移植的患者面临的主要问题。 在这项研究中，我们将探索和优化使用热疗通便 体外培养骨髓标本中的白血病细胞。人体的热反应 白血病细胞系、患者急性白血病和正常人骨骼 骨髓祖细胞(CFU-GM、BFU-E、CFU-GEMM)将用以下方法确定 克隆形成试验。单次或分次加热或不加热 目前临床上用于体外净化的化疗药物 白血病将被使用。分次剂量的热量将被用来吸收 热耐受差衰减率的优势 敏感的白血病细胞与正常的骨髓祖细胞。造成的损害 骨髓干细胞与净化人骨的重建 骨髓将使用长期骨髓培养进行测量。 以确保净化的骨髓在体内重建，并 测量白血病复发，将使用小鼠白血病模型系统。 AKR白血病和小鼠骨髓都将在体外用 单次或分次热疗加或不加化疗 探员们。此外，在净化白血病之后：骨髓混合物， 骨髓的体内重建和白血病的复发将是 量过了。 最后，我们将测试使用 诱导型HSP-72 kDa作为热反应的预测因子。要确定 HSP-72mRNA的表达，体外酶扩增HSP-72 用合成的引物进行72m RNA的聚合酶链式反应。诱导性 然后通过Southern杂交分析检测HSP-72，使用合成的 探测器。我们还将使用1维和2维页面来检测表情 HSP-72 kDa蛋白的表达。诱导型HSP-72基因的表达及 然后，蛋白质将与人类白血病的热反应相关 细胞。如果存在正相关性，则 诱导型HSP-72 kDa和耐热性，可用于临床 体外净化前白血病细胞热反应的预测 带着热气。