TRANSLATIONAL CORRELATES OF AN EPOTHILONE B ANALOG

BMS-247550的最佳剂量、临床毒性和抗肿瘤活性将在紫杉醇耐药晚期卵巢癌患者的剂量递增I期临床试验中确定。我们将评估BMS-24755对其分子靶点，微管，特别是β-微管蛋白的影响，并研究其在调控药物转运、细胞骨架完整性和细胞死亡的特定生物标志物的表达和功能中的作用。所有结果都将通过单变量和多变量分析与药代动力学、临床病理和患者反应相关联，这项初步研究的结果适用于大规模研究。肿瘤活组织检查将从治疗前和治疗后的患者身上采集，如果有的话，还将收集恶性积液。总RNA、DNA和蛋白质将从所有标本中分离出来，并保存在70摄氏度。BMS-247550剂量药代动力学与外周血单个核细胞微管束和抗癌激素形成的相关性将被确定。这项科学探索性研究的主要目的是：1.评估肿瘤反应与(A)I类β-微管蛋白亚型突变的发生和(B)β-微管蛋白亚型分布的关系。已经在对紫杉醇耐药的癌细胞系中分离到了β-微管蛋白的突变，并且在非小细胞肺癌患者中，β-微管蛋白的GTP结合区的突变与对紫杉醇单一治疗的不良反应/存活有关。2.探讨mdr1、mrp和cmoat基因表达与肿瘤对BMS-247550反应的预测作用。3.探讨bms-247550对bc1-2家族成员和凋亡抑制因子成员Survivin、P53等细胞凋亡调控蛋白表达的影响。线粒体膜电位的变化是细胞凋亡的早期指标，将通过流式细胞仪进行评估，并在治疗前后的肿瘤样本中进行比较，并与反应相关。4.确定Stathmin的表达和磷酸化状态之间的关系作为反应的函数。

The optimum dose, clinical toxicity and antitumor activity of BMS-247550 will be determined in a dose-escalation phase I clinical trial of patients with advanced Taxol-refractory ovarian cancer. We will evaluate the effects of BMS-24755 on its molecular target, the microtubule, specifically beta- tubulin and investigate its role in modulating the expression and function of defined biological markers that regulate drug transport, cytoskeletal integrity and cell death. All results will be correlated with pharmacokinetics, clinicopathology and patient response using univariate and multivariate analyses, and the findings from this pilot study applied to a large-scale study. Tumor biopsies will be harvested from pre- and post- treatment patients, together with malignant fluid, when available. Total RNA, DNA and protein will be isolated from all specimens and stored at 70 centigrade. Correlates of BMS-247550 dose pharmacokinetics with microtubule bundle and aster formation in peripheral blood mononuclear cells (PBMC's ) will be determined. The primary aims of this scientific exploratory study are: 1. To evaluate the relationship between tumor response and (a) the occurrence of mutation in the class I isotype of beta-tubulin and (b) beta- tubulin isotype distribution. Mutations in beta-tubulin have been isolated in Taxol-resistant cancer cell lines, and mutations in the GTP-binding regions of beta-tubulin have been associated with poor response/survival to Taxol monotherapy in NSCLC patients. 2. To investigate MDR1, MRP and cMOAT mRNA and protein expression as prognosticators of tumor response to BMS-247550. 3. To correlate the effects of BMS-247550 on the expression of proteins which regulate apoptosis, such as bcl-2 family members and IAP (inhitors of apoptosis) members, e.g. survivin and p53. Changes in mitochondrial membrane potential, an early indicator of apoptosis, will be evaluated by flow cytometry and compared in pre-and post-treatment tumor samples and correlated with response. 4. To determine the relationship between stathmin expression and phosphorylation status as a function of response.