Novel Approach to Cervical Cancer Prevention

宫颈癌是世界范围内妇女最常见的癌症和死亡原因之一。在美国，每年有15，000名妇女患上宫颈癌，大约4，500人死于这种疾病。人乳头瘤病毒（HPV）感染是宫颈癌的主要危险因素。宫颈癌在HPV感染和浸润性癌之间有很长的临床前阶段，这使得预防成为可能。我的长期目标是了解HPV诱导肿瘤的机制，并制定消除宫颈癌的预防策略。HPV编码蛋白E6，其对于病毒复制和HPV诱导的细胞转化是必需的。E6是一种多功能蛋白质，通过与细胞蛋白质相互作用来发挥其功能。我们最近已经确定了一个α螺旋E6结合结构域，是保守的E6结合蛋白。我们的突变分析已经确定了对E6结合重要的一致基序内的氨基酸。这些氨基酸和E6结合结构域的三维结构定义了第一代药效团-结合E6所需原子的空间分布。我们假设，模拟药效团的小分子将结合E6并阻断E6与其细胞结合蛋白的相互作用，从而抑制E6导致HPV感染细胞转化的生物活性。我们已经使用这种基于结构的药物设计来选择可以抑制E6与其细胞伴侣相互作用的小分子。我们的初步体外研究已经确定了多种潜在的E6抑制剂。在本提案中，我们将描述进一步表征这些化合物的实验。我们的具体目标是1）潜在E6抑制剂的体外特异性试验; 2）在组织培养试验中检查潜在E6抑制剂破坏E6生物活性的能力。这些研究可能导致开发治疗HPV感染和预防宫颈癌的药物。

Cervical cancer is one of the most frequent causes of cancer and death in women worldwide. Each year in the United States, 15,000 women develop cervical cancer and approximately 4,500 die from the disease. Cervical infection with human papillomaviruses (HPV) is the primary risk factor for cervical cancer. Cervical cancer has a long preclinical phase between HPV infection and invasive carcinoma that makes its prevention possible. My long-term goal is to understand the mechanisms by which HPV induces tumors and to develop preventive strategies for elimination of cervical cancer. HPV encodes a protein E6 that is essential for viral replication and HPV-induced cellular transformation. E6 is a multi-functional protein that performs its functions through interaction with cellular proteins. We have recently identified an alpha helical E6 binding domain that is conserved among E6-binding proteins. Our mutational analysis has identified amino acids within the consensus motif that are important for E6 binding. These amino acids and the three- dimensional structure of the E6-binding domain defined a first generation pharmacophore-the spatial distribution of the atoms needed to bind E6. We hypothesize that small molecules mimicking the pharmacophore will bind E6 and block the interaction of E6 with its cellular binding proteins and thus inhibit E6 biological activities that lead to the transformation of HPV-infected cells. We have used this structure-based drug design to select small molecules that could inhibit the interaction of E6 with its cellular partners. Our preliminary in vitro studies have identified multiple potential E6 inhibitors. In this proposal, we will describe experiments to further characterize these compounds. Our specific aims are 1) In vitro specificity test for the potential E6 inhibitors; 2) To examine the ability of potential E6 inhibitors to disrupt biological activities of E6 in tissue culture assays. These studies may lead to the development of drugs to treat HPV infection and to prevent cervical cancer.