The role of phosphorylation of isocitrate dehydrogenase in breast cancer

异柠檬酸脱氢酶磷酸化在乳腺癌中的作用

基本信息

批准号：
10574569
负责人：
Chenguang Fan
金额：
$ 21.36万
依托单位：
UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10574569
关键词：
Address Arkansas Bacteria Bioenergetics Breast Cancer Cell Breast Cancer Patient Breast Epithelial Cells Cancerous Cell Proliferation Cells Citrate (si)-Synthase Citric Acid Cycle Data Data Analyses Data Science Data Science Core Detection Development Diagnosis Disease Enzymes Generations Genetic Code Glutamates Goals Heterogeneity Homeostasis Homologous Gene Human Human Cell Line Image Individual Investigation Isocitrate Dehydrogenase Kinetics Knowledge Link Malate Dehydrogenase Malignant Neoplasms Measures Mentors Metabolic Metabolism Methods Molecular Mutation Oncogenic Oxidation-Reduction Pathway Analysis Pathway interactions Phosphoamino Acids Phosphorylation Phosphorylation Site Phosphotransferases Play Production Protein Isoforms Protein Kinase Proteins Recombinants Reporting Research Role Signal Pathway Site Somatic Mutation Spectrum Analysis Structural Models Structure Substrate Specificity System Techniques Testing Tissues Variant Woman Women&apos s Health Work breast cancer diagnosis cancer cell cancer therapy cofactor data integration experimental study innovation malignant breast neoplasm metabolic imaging novel marker phosphoproteomics repaired therapeutic target

项目摘要

Project Summary – Project Leader Chenguang Fan Breast cancer is one of the top threats to women’s health. About 1 in 8 US women (~12%) will develop invasive breast cancer over the course of her lifetime. Previous studies have shown the important role of a key tricarboxylic acid cycle (TCA) enzyme isocitrate dehydrogenase (IDH) in breast cancer. A variety of IDH alternations including somatic mutations, aberrant expression, and altered activities have been observed in breast cancer patients. Recently, phosphoproteomic studies of breast cancer cells have identified several unique phosphorylation sites in IDH isoforms, and those sites are specific for different types of breast cancer. Such heterogeneity poses challenges for effective diagnosis and treatment, so there is a critical need to discover specific mechanisms of IDH phosphorylation for breast cancer. The overall goal of this project is to identify the role of IDH phosphorylation in breast cancer. As IDH homologues in bacteria are known to be regulated by phosphorylation, the hypothesis is that phosphorylation of IDH in human cells alters IDH functions and cellular metabolism, thus facilitating cancerous transformations. To test this hypothesis, three specific aims are proposed: Aim 1 is to identify the effects of phosphorylation on IDH enzyme functions at the molecular level. Aim 2 is to identify the influences of IDH phosphorylation on cellular metabolism and redox status at the metabolic level. Aim 3 is to identify mechanisms or pathways in which IDH phosphorylation plays roles in breast cancer at the cellular level. The oncogenic transformation test on normal human mammary epithelial cells will be performed to identify whether IDH phosphorylation is an initiator for breast cancer or a cellular adaption to facilitate cancer formation. This proposal is innovative because: 1) the functions of IDH phosphorylation in human remain largely unknown, and this proposal will be the first to systematically study the phosphorylation of all the three IDH isoforms in human cells; 2) To address the problem that the classic glutamate-substitution method for phosphorylation studies is not always effective, the genetic code expansion technique will be applied in this proposal to co-translationally incorporate phosphoamino acid (pAA) at a controlled site in target proteins in order to produce site-specifically phosphorylated IDH variants which have been identified in breast cancer. This proposal will be the first to introduce pAA-incorporation systems into human cell lines to study phosphorylation in living cells. The proposed research is significant because it will provide key evidence for the influences and roles of IDH phosphorylation in breast cancer. Given that protein phosphorylation is usually involved in signaling pathways through different kinase cascades, the role of IDH phosphorylation in breast cancer could be different from other IDH alternations. Ultimately, the proposed studies on IDH phosphorylation are expected to identify novel biomarker and targets for breast cancer diagnosis and treatment.

项目总结 – 项目负责人范晨光乳腺癌是对女性健康的最大威胁之一，大约八分之一的美国女性（约 12%）会患上侵袭性乳腺癌。先前的研究表明乳腺癌在她一生中的重要作用。三羧酸循环 (TCA) 酶异柠檬酸脱氢酶 (IDH) 乳腺癌中的多种 IDH。已观察到包括体细胞突变、异常表达和活性在内的改变最近，乳腺癌细胞的磷酸化蛋白质组学研究发现了几种独特的乳腺癌细胞。 IDH 亚型中的磷酸化位点，这些位点对于不同类型的乳腺癌具有特异性。异质性给有效诊断和治疗带来了挑战，因此迫切需要发现 IDH 磷酸化在乳腺癌中的具体机制该项目的总体目标是确定 IDH 磷酸化的具体机制。 IDH 磷酸化在乳腺癌中的作用众所周知，细菌中的 IDH 同系物受到调节。磷酸化，假设是人体细胞中 IDH 的磷酸化会改变 IDH 功能和细胞为了检验这一假设，提出了三个具体目标：目标 1 是在分子水平上确定磷酸化对 IDH 酶功能的影响。确定 IDH 磷酸化对细胞代谢和代谢水平氧化还原状态的影响。目标 3 是确定 IDH 磷酸化在乳腺癌中发挥作用的机制或途径将在正常人乳腺上皮细胞上进行致癌转化测试。确定 IDH 磷酸化是否是乳腺癌的引发剂或促进癌症的细胞适应该提议具有创新性，因为：1）IDH磷酸化在人体中的功能在很大程度上仍然存在。未知，该提案将是第一个系统研究所有三个 IDH 磷酸化的提案 2) 解决经典谷氨酸替代法无法解决的问题磷酸化研究并不总是有效，遗传密码扩展技术将应用于此建议在目标蛋白的受控位点共翻译掺入磷酸氨基酸 (PAA)，以便产生已在乳腺癌中发现的位点特异性磷酸化 IDH 变体。该提案将是第一个将 PAA 掺入系统引入人类细胞系以研究磷酸化的提案拟议的研究意义重大，因为它将提供影响和影响的关键证据。 IDH 磷酸化在乳腺癌中的作用鉴于蛋白质磷酸化通常涉及信号传导。通过不同激酶级联途径，IDH磷酸化在乳腺癌中的作用可能不同最终，拟议的 IDH 磷酸化研究有望确定。乳腺癌诊断和治疗的新型生物标志物和靶点。