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.

项目摘要：该建议研究了FOXO转录因子在调节干细胞中的新鉴定的细胞作用癌症中相关的基因表达和信号转导。磷脂酰肌醇3激酶（PI3K）途径为几乎普遍变为癌症中激活状态，以促进生长和生存。部分多余，进化保守的FOXO -1，-3和-4转录因子是阻碍细胞周期的最著名的 PI3K途径上的进展并诱导凋亡。从规范上讲，PI3K间接使FOXO因素灭活。但是，我们在PI3K激活的癌细胞系（例如U87MG和）的细胞核中发现了FOXO因子 BT549在这些情况下提出了这些因素的新调节和作用。其他研究人员发现活化PI3K（胚胎干细胞和DLBCL）的情况下的核因子中的FOXO因子。解决 FOXO因素在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，用RNAi和过表达研究证实了）。我们的初步证据表明FOXO3中断降低了干细胞相关基因在U87MG（例如OCT4和SOX2）中的表达，而外源性 FOXO3诱导了这些基因。 AIM 1将确定在这些机制中促进FOXO核定位的机制新颖的环境。 AIM 2将定义FOXO因素用于诱导的精确分子机制 PI3K激活的癌症（如U87MG和BT549细胞）中与干细胞相关基因的表达。这些目标将通过使用QRT-PCR，Western印迹分析，染色质免疫沉淀分析来实现，基因组学方法和共聚焦显微镜。 FOXO因素对干细胞基因表达和信号转导对流行的人类疾病（例如癌症，神经退行性，，糖尿病和衰老。拟议的研究和研究增强目标将在第二个美国最大的西班牙裔服务机构，德克萨斯大学里奥格兰德大学（UTRGV），这不是NIH支持的主要接受者。这些研究的资金和完成将增加 UTRGV的研究能力，为27,000多名西班牙裔的学生提供服务，从而为分数计划的任务。