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.

项目摘要 结直肠癌（CRC）的特征在于具有需要增殖和增殖的分级组织。 去分化干细胞以维持肿瘤生长和进展。结直肠癌的细胞可塑性 癌症是在肿瘤微环境的选择性压力下发生的过程所必需的 和化疗药物。结肠肿瘤微环境的特征在于由于结肠癌的发生而导致的极度缺氧。 缺氧腔缺氧促进代谢重新布线，癌细胞利用这种过程来支持 生物合成、细胞存活和细胞命运的动态改变。细胞代谢的一个关键特征是 细胞器的相互作用和协调，但这些如何有助于CRC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）是最常见的 丰富的细胞器间相互作用。我生成了一组ERMCS报告基因CRC细胞系，并通过 无偏见的高内容成像和CRISPR屏幕，我已经确定了ER所需的基本机制， CRC中的线粒体相互作用。此外，我显示了肿瘤缺氧在调节ERMCS中的关键作用。 缺氧抑制线粒体复合物III和IV，降低ERMCS。用线粒体处理细胞 电子载体辅酶（CoQ）可恢复缺氧后ERMCS的抑制。我假设肿瘤 缺氧通过改变线粒体呼吸和辅酶Q氧化还原来调节内质网-线粒体接触（ERMCS）， 代谢适应和生存。在目标1（F99阶段）中，我将着重于确定 缺氧依赖性ERMCS抑制和用我们的新型ERMCS报告小鼠扩展到体内模型中 模型在K00阶段，我将应用研究生院在癌症代谢方面获得的知识， 细胞器相互作用到一个独立的博士后项目。大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 博士后研究员是为了了解细胞器相互作用的动态变化和要求， CRC中细胞命运改变期间的代谢区室化。我计划用遗传学的鼠类和 CRC患者类器官模型，体积电子显微镜，体内细胞器代谢组学，以及 功能性CRISPR筛选来回答这些问题。最后，除了拟议的研究， 计划包括对职业发展、指导、网络和科学重要的活动。 沟通，以准备我成功过渡到博士后奖学金和我的职业生涯作为一个 研究癌症代谢的独立研究员