Roles and regulation of p53

描述(由申请人提供)：这是我们的计划项目的第二次续签，该计划已经存在超过10年，并且一直具有很高的生产力和互动性。该计划的目标将通过跨学科和协作的方法来实现，以了解突变的p53、p53家族成员的p63和p73以及新的肿瘤抑制基因在癌症中的作用。我们的方法包括细胞生物学、蛋白质组学、显微镜、生物信息学、功能基因组学、小鼠建模和人类和小鼠病理学。随着这个项目的进展，我们的研究变得更具翻译性，与人类疾病相关，现在专注于乳腺癌、膀胱癌和淋巴瘤的肿瘤发生。Carol Prives将使用3D培养方案来研究突变型P53和P53同源物(p63和p73)在乳腺细胞形态和致癌转化中的作用。她还将描述在这些情况下ANp63蛋白质周转的机制。阿诺德·莱文将继续观察带有突变型p53的乳腺癌细胞具有干细胞基因表达特征，并将测试通过新的生物信息学方法确定的用于治疗癌细胞的突变型p53等位基因特异性药物。莱文还将研究p63或p73中影响雌性卵子DNA修复系统的SNPs的作用，以及影响后代癌症的拷贝数变化。Scott Lowe将继续研究淋巴瘤的遗传和分子基础。他将使用EP-Myc B细胞模型进行shRNA筛选，以确定新的肿瘤抑制基因。他还将研究与P53共同缺失的基因在17p染色体上的淋巴瘤中的作用，检验这样的基因可能也具有肿瘤抑制活性的假设。此外，Lowe还将研究通过下调MDM2来重新激活潜在的p53的影响，以及进行shRNA筛选，以确定其抑制促进依赖突变的p53的肿瘤死亡的基因。Cordon-Cardo将重点研究p63在小鼠和人类正常和癌变膀胱组织中的表达和作用。他将使用特定的shRNAs来表征p63亚型在小鼠尿路上皮发育过程中的影响，并确定p63亚型在膀胱癌中的表达。Cordon-Cardo还将致力于鉴定膀胱干细胞及其耐药机制。该计划高度依赖于支持管理、组织病理学、shRNAs、小鼠建模和生物信息学的核心功能，这是拟议的实验所需的。这些项目甚至比以前更加相互依赖和互动，因此，如果没有这个项目的支持，许多拟议的研究就不能有效地完成。

DESCRIPTION (provided by applicant): This is the second renewal of our Program Project that has been in existence for over 10 years and which has been highly productive and interactive. The goals of this program will be realized through an interdisciplinary and collaborative approach to understand the roles of mutant p53, p53 family member's p63 and p73 and new tumor suppressor genes in cancer. Our approaches include cell biology, proteomics, microscopy, bioinformatics, functional genomics, mouse modeling and human and mouse pathology. As this program has progressed, our research has become more translational and relevant to human disease, now focusing on breast, bladder and lymphoma tumor genesis. Carol Prives will employ the 3D culture protocol to examine the roles of mutant p53 and p53 homologues (P63 and p73) in mammary cell morphology and oncogenic transformation. She will also characterize the mechanisms of ANp63 protein turnover in these contexts. Arnold Levine will pursue the observation that breast cancer cells with mutant p53 have a stem cell gene expression signature, and will test mutant p53 allele-specific drugs that were identified by novel bioinformatic approaches for treatment of cancer cells. Levine will also study the roles of SNPs in p63 or p73 that influence DNA repair systems in female eggs and copy number variation that affect cancer in offspring Scott Lowe will continue to study the genetic and molecular basis of lymphoma. He will perform shRNA screens using the Ep-Myc B cell model to identify new tumor suppressor genes. He will also examine the role of genes co-deleted with p53 in lymphoma on chromosome 17p testing the hypothesis that such genes may also have tumor suppressive activity. Additionally Lowe will study the impact of reactivation of latent p53 by down-regulation of Mdm2, as well as perform an shRNA screen to identify genes whose inhibition facilitates death of mutant-p53 dependent tumors. Cordon-Cardo will focus on p63 expression and roles in normal and cancerous bladder tissue in mice and humans. He will use specific shRNAs to characterize the impact of p63 isoforms during urothelial development in mice, and determine the expression of p63 isoforms in bladder carcinoma. Cordon-Cardo will also pursue identification of bladder stem cells and the mechanisms by which they are chemoresistant. This program is highly dependent on the functioning of cores that support the administration, histopathology, shRNAs, mouse modeling and bioinfomatics that is required for the proposed experiments. The projects are even more interdependent and interactive than before and as a result much of the proposed research cannot be done effectively without the support of this program.