NF-KB MEDIATED DRUG RESISTANCE IN OVARIAN CANCER

本提案的实验假设是NF-kB DNA结合的激活是卵巢癌获得性CDDP耐药的共同特征。在本提案中，我们在体外和体内研究NF-kB激活及其药理学抑制，将实验室研究（目标1和3）与临床研究（目标2和4）与实验室相关性联系起来。研究NF-kappaB活化对CDDP紫杉醇、美法兰、阿霉素、拓扑替康等药物的细胞毒性抗性的影响。我们将描述与获得性CDDP耐药/敏感性相关的相关生物学事件以及与NF-kappaB激活的关联。通过转染实验，我们将检测转录激活因子NF-kappaB及其对化疗药物敏感性和耐药性的影响（阳性和阴性）。这些观察结果将通过对ansamycin抗生素（MSKCC正在开发的一类新型药物）和蛋白酶体抑制剂PS-341的研究扩展到新药开发中。提出了抑制剂对药物反应影响的机制研究，这些影响将与研究药物治疗后患者来源组织的研究联系起来。具体目标如下：特异性目的1：研究NF-kB在体外获得性化疗耐药中的作用，包括耐药谱、激活过程和NF-kB活性增加的细胞后果。特异性目的2：探讨卵巢癌患者体内IkB / IkB激酶与CDDP耐药组织的关系。特异性目的3：探讨两种可能的NF-kB活化临床抑制剂（Herbimycin A和PS-341）对体外化疗敏感性的影响及其机制。特异性目标4：开展两种新药PS-341和17烯丙基氨基格达那霉素的临床试验，并研究卵巢癌女性体内NF-kB活性增加、IkB水平降低与CDDP耐药之间的关系。这一提议提供了一个独特的机会，将NF-kB介导的获得性耐药的实验室研究与两种新药的初步临床研究结合起来，这两种新药可能很好地克服了晚期癌症患者的这种耐药机制。

The experimental hypothesis of this proposal is that the activation of NF-kB DNA binding is a common feature of acquired CDDP resistance in ovarian cancer. In this proposal, we examine NF-kB activation and its pharmacologic inhibition, both in vitro and in vivo, linking laboratory studies (aims 1 and 3) to clinical studies (aims 2 and 4) with laboratory correlates. The effect of NF-kappaB activation on resistance to the cytotoxicity of CDDP paclitaxel, melphalan, doxorubicin, topotecan and other agents will be examined. We will delineate the related biologic events associated with acquired CDDP resistance / sensitivity and the association to NF-kappaB activation. Through transfection experiments, we will examine the transcriptional activator, NF-kappaB and its effects (positive and negative) on chemotherapy drug sensitivity and resistance. These observations will be extended into new drug development through investigations of the ansamycin antibiotics, a novel class of agents under development at the MSKCC and the proteosome inhibitor PS-341. Mechanistic studies relating the effect of the inhibitors to drug response are proposed and these effects will be linked to studies of patient derived tissues after investigational drug treatment. The detailed objectives are: Specific Aim 1: To examine the role of NF-kB in acquired chemotherapy resistance in vitro, including the spectrum of resistance, the activation process and the cellular consequences of increased NF-kB activity. Specific Aim 2: To explore the in vivo association between the IkB / IkB kinase and CDDP resistance tissues from patients with ovarian cancer. Specific Aim 3: To explore the effect and mechanism of two potential clinical inhibitors of NF-kB activation (Herbimycin A and PS-341) on chemotherapy sensitivity in vitro. Specific Aim 4: To perform clinical trials of two new agents PS-341 and 17 allylaminogeldanamycin and examine the association between increased NF-kB activity, decreased IkB levels and CDDP resistance in vivo for women with ovarian cancer. This proposal represents a unique opportunity to integrate laboratory studies of NF-kB mediated acquired drug resistance with the initial clinical studies of two new agents which may very well overcome this mechanism of resistance in patients with advanced cancer.