Investigating the Roles and Regulation of FOXO Transcription Factors in GBM and basal breast cancer

项目概要： 这项提案研究了新发现的FOXO转录因子在调节干细胞中的细胞作用， 肿瘤相关基因表达和信号转导。磷脂酰肌醇3激酶（PI 3 K）途径是 在癌症中几乎普遍突变为激活状态以驱动生长和存活。部分冗余， 进化上保守的FOXO-1、-3和-4转录因子以阻碍细胞周期而闻名 在PI 3 K通路上的进展和诱导凋亡。规范上，PI 3 K间接灭活FOXO因子。 然而，我们发现FOXO因子在PI 3 K激活的癌细胞系如U87 MG和U87 MG的细胞核中存在。 BT549，表明这些因素在这些环境中的新的调节和作用。其他研究人员发现 FOXO因子在细胞核中与活化的PI 3 K（胚胎干细胞和DLBCL）的背景。解决 FOXO因子在PI 3 K激活的癌症如U87 MG中的作用，我们利用了一种创新的方法（CRISPR， 通过RNAi和过表达研究证实）。我们的初步证据表明FOXO 3的破坏 降低了U87 MG中干细胞相关基因如OCT 4和SOX 2的表达，而外源性 FOXO 3诱导这些基因。目的1将确定促进FOXO核定位的机制， 小说背景目的2将定义FOXO因子用于诱导细胞凋亡的精确分子机制。 PI 3 K激活的癌症如U87 MG和BT549细胞中干细胞相关基因的表达。这些目标 将通过采用qRT-PCR，蛋白质印迹分析，染色质免疫沉淀分析， 基因组学方法和共聚焦显微镜。FOXO因子对干细胞基因表达的影响 信号转导对普遍存在的人类疾病如癌症，神经变性， 糖尿病和衰老建议的研究和研究加强目标将在第二次 美国最大的西班牙裔服务机构，德克萨斯大学格兰德河谷分校（UTRGV）， 它不是NIH支持的主要接受者。这些研究的资金和完成情况将增加 UTRGV的研究能力，为超过27，000名学生提供服务，其中89%是西班牙裔，从而为 SCORE计划的使命。

PROJECT SUMMARY: This proposal examines newly identified cellular roles for FOXO transcription factors in regulating stem cell- related gene expression and signal transduction in cancer. The phosphatidylinositol 3 kinase (PI3K) pathway is almost universally mutated to an activate state in cancer to drive growth and survival. Partially redundant, evolutionarily conserved FOXO -1, -3 and -4 transcription factors are best known for hindering cell cycle progression and inducing apoptosis on the PI3K pathway. Canonically, PI3K indirectly inactivates FOXO factors. However, we found FOXO factors in the nucleus of PI3K-activated cancer cell lines such as U87MGs and BT549s, suggesting novel regulation and roles for these factors in these settings. Other researchers have found FOXO factors in the nucleus in contexts with activated PI3K (embryonic stem cells and DLBCL). To address the role of FOXO factors in PI3K-activated cancers such as U87MG, we utilized an innovative approach (CRISPR, corroborated with RNAi and overexpression studies). Our preliminary evidence indicated that FOXO3 disruption reduced the expression of stem cell-related genes in U87MGs such as OCT4 and SOX2, whereas exogenous FOXO3 induced these genes. Aim 1 will identify mechanisms that promote FOXO nuclear localization in these novel contexts. Aim 2 will define precise molecular mechanisms that are utilized by FOXO factors to induce the expression of stem cell-related genes in PI3K-activated cancers such as U87MG and BT549 cells. These aims will be accomplished by employing qRT-PCR, western blot analyses, chromatin immuno-precipitation analyses, genomics approaches and confocal microscopy. The impact of FOXO factors on stem cell gene expression and signal transduction has broad ramifications to prevalent human diseases such as cancer, neurodegeneration, diabetes and aging. Proposed studies and research enhancement objectives will be conducted at the second largest Hispanic-serving Institution in the United States, the University of Texas Rio Grande Valley (UTRGV), which has not been a major recipient of NIH support. Funding and completion of these studies will increase research capabilities at UTRGV, which serves over 27,000 students who are 89% Hispanic thereby serving the mission of the SCORE Program.