Mechanistic insights into lysosomal nutrient efflux in cancer

癌症中溶酶体营养物流出的机制见解

基本信息

批准号：
10341120
负责人：
Kacper Rogala
金额：
$ 11.51万
依托单位：
WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
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：
2021-02-04 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10341120
关键词：
Affect Amino Acids Area Arginine Autophagocytosis Binding Biochemical Biological Assay Biology Cell physiology Chemicals Choking Coupled Cryoelectron Microscopy Cytosol Cytotoxic Chemotherapy DNA Development Eating Environment Essential Amino Acids Extracellular Protein Face Food Future Gatekeeping Goals Growth Human Impairment In Vitro Institutes Integral Membrane Protein Laboratories Lead Learning Leucine Libraries Lipids Lysosomes Malignant Neoplasms Malignant neoplasm of pancreas Mammalian Cell Mediating Medicine Membrane Proteins Membrane Transport Proteins Mentors Metabolic Molecular Molecular Conformation Molecular Machines Motivation Mutation Normal Cell Nutrient Oncogenic Outcome Pathway interactions Pharmaceutical Chemistry Pharmaceutical Preparations Phase Phosphotransferases Positioning Attribute Process Proteins Protocols documentation Recording of previous events Research Research Personnel Role Scientist Signal Transduction Starvation Structure Testing Training Transport Process Validation Work X-Ray Crystallography analog base cancer cell cancer type career chemotherapy diagnostic tool drug development drug discovery effective therapy expectation experimental study extracellular fighting improved innovation insight lysosomal proteins mTOR protein metabolomics mutant nanobodies nanodisk novel novel therapeutics pancreatic cancer cells prevent programs proteoliposomes rational design reconstitution screening small molecule small molecule inhibitor small molecule libraries structural biology success therapeutic target tumor tumor metabolism tumor microenvironment

Affect Amino Acids Area Arginine Autophagocytosis Binding Biochemical Biological Assay Biology Cell physiology Chemicals Choking Coupled Cryoelectron Microscopy Cytosol Cytotoxic Chemotherapy DNA Development Eating Environment Essential Amino Acids Extracellular Protein Face Food Future Gatekeeping Goals Growth Human Impairment In Vitro Institutes Integral Membrane Protein Laboratories Lead Learning Leucine Libraries Lipids Lysosomes Malignant Neoplasms Malignant neoplasm of pancreas Mammalian Cell Mediating Medicine Membrane Proteins Membrane Transport Proteins Mentors Metabolic Molecular Molecular Conformation Molecular Machines Motivation Mutation Normal Cell Nutrient Oncogenic Outcome Pathway interactions Pharmaceutical Chemistry Pharmaceutical Preparations Phase Phosphotransferases Positioning Attribute Process Proteins Protocols documentation Recording of previous events Research Research Personnel Role Scientist Signal Transduction Starvation Structure Testing Training Transport Process Validation Work X-Ray Crystallography analog base cancer cell cancer type career chemotherapy diagnostic tool drug development drug discovery effective therapy expectation experimental study extracellular fighting improved innovation insight lysosomal proteins mTOR protein metabolomics mutant nanobodies nanodisk novel novel therapeutics pancreatic cancer cells prevent programs proteoliposomes rational design reconstitution screening small molecule small molecule inhibitor small molecule libraries structural biology success therapeutic target tumor tumor metabolism tumor microenvironment

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项目摘要

ABSTRACT Many RAS-transformed aggressive cancer cells are able to escape cytotoxic chemotherapy and survive in near-starvation conditions. One adaptation making them hard to kill is their ability to scavenge extracellular proteins and recycle the cellular components using autophagy, both of which are then digested in lysosomes to recover free amino acids. The process of scavenging and internalization is known as macropinocytosis, and cancer cells aquire mutations to upregulate it when faced with nutrient-poor conditions. To fight such cancers, researchers are currently targeting the macropinocytosis machinery; however, because this process is not very well studied, and likely involves hundreds of proteins with redundant functions, such therapy might prove challenging to exectute without involving multiple drugs. We propose a better way of targeting nutrient-scavenging cancers by focusing on a downstream process of releasing digested nutrients from lysosomes to cytosol. The Sabatini Lab showed that the release of digested amino acids from lysosomes is orchestrated by the mTORC1 pathway, and specifically by SLC38A9. This lysosomal membrane protein senses the rising levels of digested amino acids in lysosomes by directly binding arginine. Our lab found that this sensing is coupled to activation of the transporter function, and results in the efflux of essential non-polar amino-acids, such as leucine, from lysosomes to cytosol. Importantly, RAS-transformed pancreatic cancer cells that feed on extracellular protein were unable to efficiently form tumors in the absence of SLC38A9. These results present a novel therapeutic idea of targeting a metabolic vulnerability in cancers transformed by oncogenic RAS signaling. In this five-year project we will elucidate the molecular mechanism of releasing digested amino acids from lysosomes to cytosol via SLC38A9, and therefore provide a rational approach to drug discovery. In parallel to that, we will screen for small molecules that specifically bind to SLC38A9, and develop them into chemical probes that modulate its transport activity. Impaired efflux function of SLC38A9 will lead to entrapment of macropinocytosis-derived amino-acids within the lysosomes, and our expectation is that this treatment will impair the growth of RAS-mutant and other tumors addicted to protein scavenging, while sparing normal cells that lack this requirement. Over the first two years of the mentored phase, I will be based at the Whitehead Institute, where I will learn cell signaling and metabolomics approaches from the experts in the field. I will also venture into a completely new research area to me, chemical biology, working with experts at the Broad Institute. After the completion of my K99 training, my aspiration is to lead a laboratory that combines cell signaling, structural biology, and chemical biology to study membrane transporters and their role in cancer metabolism. In parallel to understanding basic biology, I want my lab to develop specific small-molecule modulators that adjust transport activities of those proteins, facilitating further research in the field, and in long term – new medicines.

抽象的许多RAS转化的侵袭性癌细胞能够逃脱细胞毒性化疗，并在近饥饿的条件。一种使其难以杀死的改编是他们清除细胞外的能力蛋白质并使用自噬回收细胞成分，然后在溶酶体中消化它们恢复游离氨基酸。清除和内在化的过程称为大型细胞增多症，和癌细胞面对营养贫困的条件时，会在上调其上调。与这样的癌症作斗争，研究人员目前正在针对大型细胞增多症机制。但是，因为这个过程不是很井研究，可能涉及数百种具有多余功能的蛋白质，这种疗法可能证明挑战执行而无需多种药物。我们提出了一种更好的定位方式通过关注从中释放消化营养素的下游过程，从而溶酶体至细胞质。 Sabatini实验室表明，从溶酶体中释放消化的氨基酸是由MTORC1途径策划，尤其是SLC38A9。该溶酶体膜蛋白通过直接结合精氨酸，感觉到溶酶体中消化氨基酸的水平上升。我们的实验室发现这种灵敏度与转运蛋白功能的激活相连，并导致必需的非极性从溶酶体到细胞质的氨基酸，例如亮氨酸。重要的是，RAS转化的胰腺癌在没有SLC38A9的情况下，以细胞外蛋白为食的细胞无法有效形成肿瘤。这些结果提出了针对癌症中代谢脆弱性的新型热思想。致癌RAS信号传导。在这个五年的项目中，我们将阐明释放的分子机制通过SLC38A9从溶酶体到细胞质的消化氨基酸，因此提供了一种合理的方法药物发现。与此并联，我们将筛选专门结合SLC38A9的小分子，并且将它们发展为调节其运输活性的化学问题。 SLC38A9的外排功能受损将导致诱捕大胞胞病质衍生的氨基酸溶酶体，我们的期望是这种治疗将损害RAS突变剂的生长和其他肿瘤的生长，而添加到蛋白质清除中，而保留缺乏这种要求的正常细胞。在修补阶段的头两年中，我将依靠在怀特黑德研究所（Whitehead Institute），我将学习细胞信号传导和代谢组学的方法领域。我还将冒险进入一个全新的研究领域，化学生物学，与专家合作在布罗德学院。在完成K99培训后，我的愿望是领导一个实验室结合细胞信号传导，结构生物学和化学生物学来研究膜转运蛋白及其作用在癌症代谢中。与理解基本生物学并行，我希望我的实验室开发特定的调节这些蛋白质的运输活性的小分子调节剂，支持进一步研究长期和长期 - 新药物。