LIPOPHILIC PROBES FOR TUMOR CELL SURFACES

对于许多白血病或播散性实体瘤患者 在标准治疗后复发， 化疗/放疗后进行骨髓移植， 只有现实的机会长期无病生存。 使用患者自身（自体）骨髓移植 在那些不适合接受化疗的患者中， 异基因骨髓移植，因为他们的年龄或因为 他们缺乏组织相容的骨髓捐赠者 但全面 自体骨髓移植的潜力无法实现 除非我们能找到方法去除自体移植物中的 肿瘤细胞 这一建议是围绕着这样一个假设提出的， 光敏化由亲脂性汞染料介导， 因为MC 540可用于选择性地杀死体内残留的肿瘤细胞， 自体骨髓移植 虽然MC 540介导 光敏化降低了实验性 白血病细胞大于100，000倍， 多能干细胞区室的损害，它可能不是 对一些自发性肿瘤同样有效， 足以净化含有高浓度 肿瘤细胞 因此，我们想确定是否使用MC 540 与另一种清除剂（例如抗体，药物， 热疗）可以增强其抑制作用， 增加干细胞毒性。 第二组实验旨在 在确定细胞和分子机制方面， 调节MC 540与细胞的结合并介导 通过MC 540和光灭活细胞。 我们还将完成一个 启动MC 540介导的II期临床试验 用于净化自体骨髓移植物的光敏化。 在本试验中，我们将研究染料的影响- 对造血干细胞介导的光敏化， 移植物中的辅助细胞及其对植入和 免疫功能的恢复。 提案工作将 对MC 540及其 与核DNA和质膜脂质的光产物， 蛋白质，脂质过氧化和蛋白质交联在 MC 540介导的细胞毒性，以及 汞染料。 （方法：克隆分析正常和 肿瘤细胞，骨髓移植，染料介导 光敏化，热疗，流式细胞术，计算机 膜脂的模拟、操作和分析， 蛋白质，紫外/维斯和荧光光谱，体细胞 遗传学）。

For many patients with leukemia or disseminated solid tumors who have relapsed after standard therapy, very intensive chemo/radiotherapy followed by a marrow transplant offers the only realistic chance of long-term disease-free survival. Transplantation of the patient's own (autologous) marrow is used with increasing frequency in patients who are not candidates for allogeneic marrow transplants because of their age or because they lack histocompatible marrow donors. However, the full potential of autologous marrow transplantation cannot be realized unless we find ways to rid autologous grafts of occult clonogenic tumor cells. This proposal is molded around the hypothesis that photosensitization mediated by lipophilic merocyanine dyes such as MC 540 can be used to selectively kill residual tumor cells in autologous bone marrow grafts. Although MC 540-mediated photosensitization reduces the concentration of experimental leukemia cells greater than 100,000-fold without excessive damage to the pluripotent stem cell compartment, it may not be equally effective against some spontaneous tumors or not be sufficient for the purging of marrows with very high levels of neoplastic cells. We therefore want to determine if using MC 540 in combination with another purging agent (e.g. antibody, drug, hyperthermia) can augment its antineoplastic effect without increasing stem cells toxicity. A second set of experiments aims at identifying the cellular and molecular mechanisms that regulate the binding of MC 540 to cells and mediate the inactivation of cells by MC 540 and light. We will also complete a phase I and initiate a phase II clinical trial of MC 540-mediated photosensitization for the purging of autologous marrow grafts. In the context of this trial we will study the effects of dye- mediated photosensitization on hematopoietic stem cells and accessory cells in the graft, and its effects on engraftment and the recovery of immune function. The proposal work will generate new insights into the interactions of MC 540 and its photoproducts with nuclear DNA and plasma membrane lipids and proteins, the role of lipid peroxidation and protein crosslinking in MC 540-mediated cytotoxicity, and the clinical usefulness of merocyanine dyes. (Methods: Clonal assays of normal and neoplastic cells, marrow transplantation, dye-mediated photosensitization, hyperthermia, flow cytometry, computer simulation, manipulation and analysis of membrane lipids and proteins, UV/VIS- and fluorescence spectroscopy, somatic cell genetics.)