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

项目总结/摘要 人乳头瘤病毒（HPV）导致约5%的人类癌症。当HPV在转化， 已经使用组织培养模型在体外广泛研究了肿瘤发生特性， 表达HPV癌蛋白的转基因动物模型，这些功能在HPV诱导的 疾病尚未在真实的生理模型中得到验证。理解HPV癌蛋白功能的障碍 HPV不能感染鼠细胞，这使得人们不能在实验室小鼠中研究HPV疾病。的 最近发现的小鼠乳头瘤病毒（MmuPV 1）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MmuPV 1，我们建立了HPV诱导的肿瘤小鼠模型， 小鼠中的疾病，使我们现在能够定义E6和E7癌蛋白在自然环境中的作用。 乳头瘤病毒感染MmuPV 1感染皮肤和粘膜上皮，后者模仿肿瘤 由“高危”α-HPV引起的疾病。后一个发现是有趣的，因为MmuPV 1 E6和E7 癌蛋白在生物化学上模拟β-和γ-HPV。我们将研究MmuPV 1 E6和E7的功能， 癌蛋白在促进MmuPV 1诱导的疾病中的作用，特别是E6抑制NOTCH信号传导的能力， E7对非典型RB功能的影响。使用基因工程小鼠模型（GEMM），我们将 使用细胞突变严格测试这些功能在促进皮肤癌发生中的重要性， 可以补充E6和E7的这些特定功能。最后，我们将确定E6和E7是否 来自“高危”α-HPV的癌蛋白HPV 16可以反式互补MmuPV 1 E6和E7癌蛋白 使用我们现有的HPV 16转基因动物模型。这些研究将进一步加深我们对E6的作用的理解， 和E7在促进乳头瘤病毒诱导的疾病中发挥作用，并可用于开发新的疗法， 针对这些功能。 这个博士后奖学金的主要目标是帮助我发展成为一个成功的 在一个以研究为重点的学术机构工作。建议的培训计划为我提供了一个 实现这一目标的一套独特的方法，包括一个强大的指导计划， 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 特别侧重于帮助代表性不足的少数群体在学术界成为成功的教授。这 再加上我的赞助人保罗·兰伯特博士的良好训练记录， 作为一名研究生，我有令人信服的理由来说明为什么我应该成功地实现我的职业目标。通过 完成研究目标和培训计划，我希望建立一个重要的利基在乳头瘤病毒 领域，并发展必要的技能，以建立一个成功的独立研究计划，作为一个教师 一个以研究为中心的学术机构的成员。

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.