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

• 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
• 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' s lymphoma; malignant breast neoplasm; novel; outcome forecast; overexpression; pluripotency; programs; promoter; recruit; stem cells; transcription factor; translational impact

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.

项目总结： 这项建议研究了新发现的FOXO转录因子在调节干细胞中的细胞角色。 肿瘤相关基因表达与信号转导。磷脂酰肌醇3激酶(PI3K)途径是 在癌症中，几乎所有人都突变到激活状态，以推动生长和生存。部分冗余， 进化上保守的FOXO-1、-3和-4转录因子最为人所知的是阻碍细胞周期 PI3K通路的进展和诱导细胞凋亡。典型地，PI3K间接地灭活FOXO因子。 然而，我们在PI3K激活的癌细胞系如U87Mgs和U87Mgs的细胞核中发现了FOXO因子 BT549，提示了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 细胞核中的FOXO因子与激活的PI3K(胚胎干细胞和DLBCL)相关。要解决这个问题 FOXO因子在PI3K激活的癌症中的作用，例如U87 MG，我们利用了一种创新的方法(CRISPR， 与RNAi和过度表达研究相印证)。我们的初步证据表明FOXO_3的破坏 降低OCT4和SOX2等干细胞相关基因在U87MGs中的表达，而外源性 FOXO_3诱导了这些基因。目标1将确定促进FOXO核局部化的机制 新奇的背景。目标2将定义FOXO因子用来诱导 干细胞相关基因在PI3K激活的肿瘤如U87 MG和BT549细胞中的表达这些目标 将通过使用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 信号转导对常见的人类疾病有广泛的影响，如癌症、神经变性、 糖尿病和衰老。建议的研究和研究提升目标将在第二次会议上进行 美国最大的拉美裔服务机构，德克萨斯大学格兰德河谷大学(UTRGV)， 它并不是NIH支持的主要接受者。这些研究的资金和完成情况将会增加 UTRGV的研究能力，为超过27,000名拉美裔学生提供服务，从而为 SCORE计划的使命。