The role of signal transduction cascades in mediating papillomavirus infection.

描述(由申请人提供)：感染高危致癌人乳头瘤病毒(HPV)16和18已被确定为宫颈癌和肛门癌的主要危险因素。为了实现这一点，致癌的HPV首先与基底上皮细胞的细胞表面受体结合。结合可能导致信号级联反应的诱导，该信号级联反应可能介导依赖动力蛋白的胞膜蛋白/小窝介导的病毒内吞作用，从而可能导致感染。随后使用宿主细胞机制复制病毒可能会导致宫颈癌和肛门癌。我的长期目标是破译哪些信号转导分子参与了结合后介导致癌HPV感染的过程，并确定这些分子或它们的效应器是否可以作为预防致癌HPV感染的广泛靶向策略。我的目标是1)确定HPV与硫酸肝素的初级结合和与α6β4整合素复合体受体的络合是否诱导了介导HPV感染所需的信号分子。2.)确定介导HPV感染动力蛋白依赖的内吞作用所需的信号分子。为了接近这项研究，我计划使用重新表达的野生型(WT)β4角质形成细胞和β4缺失角质形成细胞来确定感染是否需要AIM 1中的病毒受体相互作用。对于目标2，1计划使用免疫沉淀和免疫荧光技术显示磷酸化信号分子的蛋白表达。人乳头瘤病毒通过与细胞结合，进入细胞，利用细胞的机制繁殖，从而感染宫颈和肛门内的细胞，从而导致癌症。我们的研究目标是调查病毒结合和利用人类细胞繁殖的过程。最终，我们的目标是破译终止HPV感染从而预防癌症的方法。

DESCRIPTION (provided by applicant): Infection with high risk oncogenic Human Papillomavirus (HPV) 16 and 18 has been identified as the major risk factor for cervical cancer and anal cancer. In order for this to occur, oncogenic HPV first binds to cell surface receptors of basal epithelial cells. Binding may lead to induction of signaling cascades that could mediate the dynamin dependent clathrin/caveolae- mediated endocytosis of the virus which may lead to infection. Subsequent replication of the virus using the host cell machinery may lead to cervical and anal cancer. My long-term objective is to decipher which signal transduction molecules are involved in mediating oncogenic HPV infections upon binding, and to determine if the molecules or their effectors can be used as a broad target strategy in preventing oncogenic HPV infection. My aims would be to 1.)To determine whether primary binding of HPV to heparan sulfate and complexing to the alpha6 beta4 integrin complex receptor induces the signaling molecules needed to mediate HPV infection. 2.) To identify the signaling molecules needed to mediate the dynamin-dependent endocytosis of HPV infection. To approach this study, I plan to use re-expressed Wild-Type (WT) beta 4 keratinocytes and beta 4 null keratinocytes to determine if the virus receptor interaction in aim 1 is needed for infection. For Aim 2,1 plan to show the protein expression of phosphorylated signaling molecules using immunoprecipitation and immunofluorescence techniques. The human papilloma virus infects the cells lining the cervix and anus by binding to the cells, entering them, using their machinery to multiply consequently causing cancer. The goal of our research is to investigate the process by which the virus binds and utilizes the human cells to multiply. Ultimately, our goal is to decipher means of terminating HPV infection thus preventing cancer.