Inter-organellar communication in metabolic reprogramming of colorectal cancer

结直肠癌代谢重编程中的细胞器间通讯

• 批准号: 10743454
• 负责人: Brandon Chen
• 金额: $ 4.23万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10743454
• 关键词: Anabolism; Automobile Driving; Bioenergetics; CRISPR screen; Cancer Etiology; Cell Differentiation process; Cell Survival; Cells; Cessation of life; Coenzymes; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Colorectal Neoplasms; Common Neoplasm; Communication; Data; Development; Electron Microscopy; Electron Transport; Electron Transport Complex III; Electrons; Endoplasmic Reticulum; Enzymes; Epithelium; Fellowship; Foundations; Genetic; Genetic Screening; Genetically Engineered Mouse; Goals; Homeostasis; Hypoxia; Image; In Vitro; Intestines; Ions; Knowledge; Lipids; Maintenance; Malignant Neoplasms; Mentorship; Metabolic; Metabolic Pathway; Metabolism; Metaplasia; Methods; Mitochondria; Modeling; Molecular; Monitor; Mus; Organelles; Organoids; Oxidation-Reduction; Oxidative Phosphorylation; Pathway interactions; Patients; Phase; Postdoctoral Fellow; Pre-Clinical Model; Principal Investigator; Process; Proliferating; Pyrimidine; Reaction; Regulation; Reporter; Repression; Research; Research Personnel; Research Proposals; Respiration; Role; Sampling; Scanning Electron Microscopy; Science; Shapes; Site; Sterols; Stimulus; System; Therapeutic; Training; Ubiquinone; Work; cancer cell; career; career development; colon cancer cell line; colon cancer patients; colorectal cancer progression; complex IV; genome-wide; graduate school; improved outcome; in vivo; in vivo Model; inducible Cre; innovation; knowledge base; lipid biosynthesis; lipidomics; metabolic fitness; metabolomics; mouse model; neoplastic cell; new therapeutic target; novel; novel therapeutics; oxidation; pharmacologic; pressure; response; stem cell expansion; stem cells; stressor; treatment response; tumor; tumor growth; tumor hypoxia; tumor metabolism; tumor microenvironment; tumor progression

PROJECT SUMMARY Colorectal cancers (CRC) are characterized as having a hierarchical organization requiring proliferating and de-differentiated stem cells to maintain tumor growth and progression. Cellular plasticity underlying colorectal cancer is essential for a process which occurs following selective pressures of the tumor microenvironment and chemotherapeutics. The colonic tumor microenvironment is characterized by extreme hypoxia due to the anoxic lumen. Hypoxia promotes metabolic rewiring, and such processes are utilized by cancer cells to support biosynthesis, cell survival and dynamic alteration in cell fates. A critical feature of cellular metabolism is organellar interaction and coordination, yet how these contribute to CRC plasticity, survival, progression and treatment response are unclear. Endoplasmic reticulum-mitochondria contact sites (ERMCS) are the most abundant inter-organellar interaction. I generated a panel of ERMCS reporter CRC cell lines, and through unbiased high content imaging and CRISPR screens, I have identified essential mechanisms required for ER- mitochondrial interactions in CRC. Moreover, I show a key role of tumor hypoxia in modulating ERMCS. Hypoxia inhibited mitochondrial complex III and IV to decrease ERMCS. Treating cells with the mitochondrial electron carrier, coenzyme (CoQ) rescued ERMCS suppression following hypoxia. I hypothesize that tumor hypoxia regulates ER-mitochondrial contacts (ERMCS) by altering mitochondrial respiration and CoQ redox for metabolic adaptation and survival. In aim 1 (F99 phase), I will focus on identifying the molecular mechanism of hypoxia dependent ERMCS inhibition and expand into in vivo models with our novel ERMCS reporter mouse model. During the K00 phase, I will apply knowledge gained during graduate school in cancer metabolism and organellar interaction to an independent postdoctoral project. The plasticity of colorectal tumor epithelium depends on integration of organellar functions to sustain metabolic demands. Therefore, my goal as a postdoctoral fellow is to understand the dynamic changes and requirement for organellar interactions and metabolic compartmentalization during cell fates alterations in CRC. I plan to use genetic murine and primary patient organoid models of CRC, volumetric electron microscopy, in vivo organellar metabolomics, and functional CRISPR screens to answer these questions. Lastly, in addition to the proposed studies, this training plan includes activities important for career development, mentorship, networking, and scientific communication to prepare me for successful transition to a postdoctoral fellowship and my career as an independent investigator studying cancer metabolism.

项目概要 结直肠癌 (CRC) 的特点是具有需要增殖和增殖的分层组织。 去分化干细胞维持肿瘤生长和进展。结直肠细胞的可塑性 癌症对于肿瘤微环境的选择性压力后发生的过程至关重要 和化疗。结肠肿瘤微环境的特点是极度缺氧，这是由于 缺氧管腔。缺氧促进代谢重新布线，癌细胞利用这些过程来支持 生物合成、细胞存活和细胞命运的动态改变。细胞代谢的一个重要特征是 细胞器的相互作用和协调，但它们如何促进结直肠癌的可塑性、生存、进展和 治疗反应尚不清楚。内质网-线粒体接触位点（ERMCS）是最多的 丰富的细胞器间相互作用。我生成了一组 EMCS 报告基因 CRC 细胞系，并通过 通过公正的高内涵成像和 CRISPR 筛选，我已经确定了 ER- 所需的基本机制 结直肠癌中线粒体的相互作用。此外，我还展示了肿瘤缺氧在调节 EMCS 中的关键作用。 缺氧抑制线粒体复合物 III 和 IV，从而降低 EMCS。用线粒体处理细胞 电子载体、辅酶 (CoQ) 挽救了缺氧后的 EMCS 抑制。我推测肿瘤 缺氧通过改变线粒体呼吸和 CoQ 氧化还原来调节 ER 线粒体接触 (ERMCS) 代谢适应和生存。在目标 1（F99 阶段）中，我将重点关注确定 缺氧依赖性 EMCS 抑制并利用我们的新型 EMCS 报告小鼠扩展到体内模型 模型。在K00阶段，我将应用在研究生院获得的癌症代谢知识和 独立博士后项目的细胞器相互作用。结直肠肿瘤上皮的可塑性 依赖于细胞器功能的整合来维持代谢需求。因此，我的目标是 博士后研究员的目的是了解细胞器相互作用的动态变化和要求， CRC 细胞命运改变期间的代谢区室化。我计划使用基因小鼠和原代 CRC 患者类器官模型、体积电子显微镜、体内细胞器代谢组学和 功能性 CRISPR 筛选可以回答这些问题。最后，除了拟议的研究之外，本次培训 计划包括对职业发展、指导、网络和科学重要的活动 沟通，让我为成功过渡到博士后奖学金和我的职业生涯做好准备 研究癌症代谢的独立研究者。