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

研究 FOXO 转录因子在 GBM 和基底乳腺癌中的作用和调节

基本信息

批准号：
9883816
负责人：
Megan E Keniry
金额：
$ 10.9万
依托单位：
UNIVERSITY OF TEXAS RIO GRANDE VALLEY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9883816
关键词：
1-Phosphatidylinositol 3-Kinase Address Aging Apoptosis Binding Biological Breast Cancer cell line Bypass CRISPR/Cas technology Cancer Prognosis Cancer cell line Cell Cycle Progression Cell Nucleus Cells Clustered Regularly Interspaced Short Palindromic Repeats Confocal Microscopy Cytosine Data Development Diabetes Mellitus Environment Epigenetic Process FOXO1A gene FOXO3A gene Funding Gene Expression Gene Mutation Genes Genetic Models Genetic Models for Cancer Genetic Transcription Genomic approach Glioblastoma Goals Growth Hispanic-serving Institution Hispanics Histones Human Investigation Lead Lymphoma cell Malignant Neoplasms Maps Mediating Methylation Mission Modeling Modification Molecular Mus Mutate Mutation Nerve Degeneration Nuclear Output Pathway interactions Pharmacology Promoter Regions Proteins Proto-Oncogene Proteins c-akt Publishing Quantitative Reverse Transcriptase PCR RNA Interference RNA Polymerase II Regulation Reporting Research Research Personnel Role STAT3 gene Sampling Signal Transduction Students T-Lymphocyte Tail Testing Texas Therapeutic Intervention Treatment Efficacy Tumor Suppressor Proteins United States United States National Institutes of Health Universities Western Blotting Work cancer cell cancer therapy chromatin immunoprecipitation design effective therapy embryonic stem cell genetic manipulation genome editing human disease innovation large cell Diffuse non-Hodgkin&apos s lymphoma malignant breast neoplasm novel outcome forecast overexpression pluripotency programs promoter recruit stem cells transcription factor translational impact

项目摘要

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.

项目概要：该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 - 癌症中相关基因表达和信号转导。磷脂酰肌醇 3 激酶 (PI3K) 途径是在癌症中几乎普遍突变为激活状态以促进生长和生存。部分冗余，进化上保守的 FOXO -1、-3 和 -4 转录因子因阻碍细胞周期而闻名 PI3K 通路上的进展并诱导细胞凋亡。通常，PI3K 间接使 FOXO 因子失活。然而，我们在 PI3K 激活的癌细胞系（例如 U87MGs 和 BT549s，表明这些因素在这些环境中的新调节和作用。其他研究人员发现 FOXO 在激活 PI3K（胚胎干细胞和 DLBCL）的情况下在细胞核中发挥作用。为了解决为了了解 FOXO 因子在 PI3K 激活的癌症（例如 U87MG）中的作用，我们采用了一种创新方法（CRISPR、 RNAi 和过表达研究证实）。我们的初步证据表明 FOXO3 破坏降低 U87MG 中干细胞相关基因的表达，例如 OCT4 和 SOX2，而外源性 FOXO3 诱导这些基因。目标 1 将确定促进 FOXO 核定位的机制新颖的背景。目标 2 将定义 FOXO 因子利用的精确分子机制来诱导 PI3K 激活的癌症（如 U87MG 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来完成，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人类流行的疾病有广泛的影响，例如癌症、神经退行性疾病、糖尿病和衰老。拟议的研究和研究加强目标将在第二次会议上进行美国最大的拉美裔服务机构，德克萨斯大学里奥格兰德河谷分校 (UTRGV)，它并不是 NIH 支持的主要接受者。这些研究的资助和完成将会增加 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，从而服务于 SCORE 计划的使命。