Understanding the Roles of the E6 and E7 Oncoproteins in MmuPV1 Induced Carcinogenesis

项目摘要/摘要 人类乳头瘤病毒(HPV)导致的癌症约占所有人类癌症的5%。当人乳头瘤病毒在转变时 在体外使用组织培养模型和在体内使用 表达HPV癌蛋白的转基因动物模型，这些功能在HPV诱导中的相关性 疾病还没有在真正的生理模型中得到验证。理解HPV癌蛋白功能的障碍 HPV不能感染小鼠细胞，这使得人们不能在实验室小鼠身上研究HPV疾病。这个 最近发现的小鼠乳头瘤病毒(MmuPV1)为我们提供了在临床前模型中进行研究的机会 自然感染和乳头瘤病毒通过其癌蛋白促进肿瘤疾病的能力， 特别是E6和E7。利用MmuPV1，我们建立了HPV诱导的小鼠肿瘤模型 小鼠的疾病使我们现在能够在自然环境中定义E6和E7癌蛋白的作用 乳头瘤病毒感染。MmuPV1感染皮肤和粘膜上皮，后者类似于肿瘤 由“高危”α-HPV引起的疾病。后一项发现很有趣，因为MmuPV1 E6和E7 癌蛋白在生物化学上模仿β-和γ-HPV。我们将研究MmuPV1 E6和E7的功能 癌蛋白在促进MmuPV1诱导的疾病中的作用，特别是E6‘S抑制Notch信号转导和 E7‘S对非典范Rb功能的影响。使用基因工程小鼠模型(GEMM)，我们将 使用细胞突变来严格测试这些功能在促进皮肤癌发生中的重要性 可以为E6和E7的这些特定功能提供补充。最后，我们将确定E6和E7是否 “高危”α-HPV16型癌蛋白可反式补体MmuPV1E6和E7癌蛋白 使用我们现有的HPV16转基因动物模型。这些研究将进一步加深我们对E6角色的理解 和E7在促进乳头瘤病毒诱导的疾病中发挥作用，并可用于开发新的治疗方法 以这些功能为目标。 这个博士后奖学金的主要目标是帮助我培养成为一名成功的 一家专注于研究的学术机构的教员。拟议的培训计划为我提供了一个 实现这一目标的一套独特方法，包括强大的指导计划、许多机会 发展我作为导师和教师的技能，以及一系列职业发展机会，包括一个 专注于帮助代表不足的少数族裔在学术界取得成功，成为教授。这 再加上我的赞助人保罗·兰伯特博士的良好培训记录，以及我出色的生产力记录 作为一名研究生，这些都是我应该成功实现职业目标的令人信服的理由。通过 完成研究目标和培训计划，我希望在乳头瘤病毒领域建立一个重要的利基 并培养建立一个成功的独立研究项目所必需的技能。 他是一个专注于研究的学术机构的成员。

Project Summary/Abstract Human papillomavirus (HPV) causes approximately 5% of all human cancers. While HPV's transforming and tumorigenic properties have been studied extensively in vitro using tissue culture models and in vivo using transgenic animal models that express the HPV oncoproteins, the relevance of these functions in HPV-induced disease have not been verified in a true physiological model. A barrier to understanding HPV oncoprotein function is that HPV cannot infect murine cells, which disallows one from studying HPV disease in laboratory mice. The recent discovery of murine papillomavirus (MmuPV1) provides us the opportunity to study in a preclinical model natural infections and the ability of papillomaviruses to promote neoplastic disease through their oncoproteins, specifically E6 and E7. Using MmuPV1, we have established a mouse model for HPV-induced neoplastic disease in mice allowing us to now define the roles of the E6 and E7 oncoproteins in the context of natural papillomavirus infection. MmuPV1 infects both cutaneous and mucosal epithelia, the later mimicking neoplastic disease caused by the “high-risk” α-HPVs. The latter finding is interesting because MmuPV1 E6 and E7 oncoproteins biochemically mimic the β- and γ-HPVs. We will examine the functions of the MmuPV1 E6 and E7 oncoprotein's in promoting MmuPV1-induced disease, specifically E6's ability to inhibit NOTCH signaling and E7's effects on non-canonical RB function. Using genetically engineered mouse models (GEMMs), we will rigorously test the importance of these functions in promoting skin carcinogenesis using cellular mutations that may complement for these specific functions of E6 and E7. Finally, we will determine if the E6 and E7 oncoproteins from the “high-risk” α-HPV, HPV16, can trans-complement for MmuPV1 E6 and E7 oncoproteins using our existing HPV16 transgenic animal models. These studies will further our understanding of the roles E6 and E7 play in promoting papillomavirus-induced disease and could be used to develop novel therapies that target these functions. The main goal of this postdoctoral fellowship is to help me develop the skill sets needed to become a successful faculty member at a research-focused academic institution. The proposed training plan provides me with a unique set of means by which to accomplish this goal including a strong mentoring plan, many opportunities to develop my skills as a mentor and teacher, and an array of career development opportunities including one focused specifically on helping under represented minorities be successful in academia as professors. This combined with the strong training record of my sponsor, Dr. Paul Lambert, and my strong record of productivity as a graduate student are compelling reasons why I should be successful in attaining my career goals. By accomplishing the research goals and training plan, I hope to establish an important niche in the papillomavirus field and to develop the skill sets necessary to establish a successful independent research program as a faculty member in a research-focused academic institution.