ROLE OF HYPERTHERMIA IN BONE MARROW TRANSPLANTATION

热疗在骨髓移植中的作用

• 批准号: 3460139
• 负责人: NAHID F MIVECHI
• 金额: $ 9.21万
• 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/3460139
• 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; natural gene amplification; neoplasm /cancer chemotherapy; neoplasm /cancer thermotherapy; nucleic acid hybridization; polymerase chain reaction; 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.

自体骨髓移植用于许多患者，包括 急性骨髓性白血病和急性淋巴细胞白血病患者 缺乏合适的骨髓捐赠者 然而，恶性肿瘤的复发是一种 这是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高温净化 白血病细胞从骨髓标本在体外。 人体热反应 白血病细胞系、来自患者的急性白血病和正常人骨 将使用以下方法测定骨髓祖细胞（CFU-GM、BFU-E、CFU-GEMM）： 克隆形成测定。 单次或分次剂量的热量，有或没有 目前临床上用于体外清除 将使用白血病。 将使用分次剂量的热量 耐热性不同衰减速率的优点 敏感的白血病细胞与正常骨髓祖细胞。 的损坏 骨髓干细胞和净化的人骨的重建 将使用长期骨髓培养物测量骨髓。 为了确保净化骨髓的体内重建， 为了测量白血病复发，将使用鼠白血病模型系统。 AKR白血病和鼠骨髓都将在体外用 单次或分次热疗，伴或不伴化疗 剂. 此外，净化白血病后：骨髓混合物， 骨髓的体内重建和白血病复发都将被 测定了 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。 确定 HSP-72 mRNA的表达，HSP-72 mRNA的体外酶促扩增， 72 mRNA（PCR）将通过合成引物进行。 诱导型 HSP-72然后将通过使用合成的DNA印迹分析来检测。 probes. 我们还将使用1&2维PAGE来检测表达 HSP-72 kDa蛋白质。 诱导型HSP-72 mRNA和蛋白质的表达 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， 诱导型HSP-72 kDa和耐热性，PCR可用于临床 预测白血病细胞体外净化前的热反应 加热。