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Mechanism of Lupus La:miR-122 ribonucleoprotein sorting into breast cancer-derived exosomes

狼疮 La:miR-122 核糖核蛋白分选至乳腺癌来源的外泌体的机制

基本信息

批准号：
10751552
负责人：
Jordan Matthew Ngo
金额：
$ 4.42万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-18 至 2025-07-17
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751552
关键词：
Antigens Binding Biochemical Biogenesis Biological Assay Breast Cancer Cell Breast Cancer cell line Breast cancer metastasis Cancer Biology Categories Cell membrane Cell secretion Cells Classification Complex Cytoplasm Data Disease Progression Dissection Electrophoretic Mobility Shift Assay Endosomes Eukaryotic Cell Event Extracellular Space Fluorescence Microscopy Foundations Fractionation Future Genetic Goals Imaging Techniques In Vitro Integral Membrane Protein Jordan Label Lipids Lupus MDA MB 231 Malignant Neoplasms Mediating Membrane Metastatic breast cancer MicroRNAs Molecular Multivesicular Body National Research Service Awards Neoplasm Metastasis Nucleic Acids Pathway interactions Pharmaceutical Preparations Play Positioning Attribute Process Proteins Proteomics RNA-Protein Interaction Reaction Recombinants Research Research Personnel Research Project Grants Resolution Ribonucleoproteins Role Small RNA Sorting Tertiary Protein Structure Therapeutic Training Transfer RNA Vesicle candidate identification cellular imaging delivery vehicle density diagnostic tool efficacy evaluation engineered exosomes exosome extracellular extracellular vesicles glucose metabolism in vivo insight interest late endosome malignant breast neoplasm microvesicles neoplastic cell novel nucleic acid delivery overexpression receptor reconstitution stem cells therapeutic target tumor progression

项目摘要

PROJECT SUMMARY / ABSTRACT Extracellular vesicles (EVs) comprise a heterogeneous pool of membrane-enclosed compartments secreted to the extracellular milieu of cells. Eukaryotic cells release a variety of EVs subpopulations that can be classified broadly into two categories on the basis of their membrane of origin. Microvesicles are EVs that form by direct outwards budding from the plasma membrane. Exosomes are EVs that originate from the endocytic pathway. Upon fusion of a multivesicular body (MVB) with the plasma membrane, the intraluminal vesicles (ILVs) are exported to the extracellular space as exosomes. Exosomes have elicited broad interest as their intraluminal contents (e.g. miRNAs, yRNAs, and tRNAs) are distinct from their progenitor cells. An accumulating repertoire of evidence has suggested that EV-mediated intercellular propagation of miRNAs plays important roles in various aspects of cancer biology. In particular, several studies have suggested that the intercellular propagation of miR-122 through breast cancer-derived EVs promotes breast cancer metastasis by reprogramming glucose metabolism in the pre-metastatic niche in vivo. Consistent with this line of experimental evidence, our lab demonstrated previously, both in cells and a cell-free reaction, that the Lupus La antigen (La) mediates the selective sorting and enrichment of miR-122 into exosomes derived from a metastatic breast cancer cell line. However, the molecular mechanism(s) by which the La:miR-122 ribonucleoprotein (RNP) complex itself is selectively incorporated into breast cancer-derived exosomes remains unknown. In this proposed research project, Jordan Ngo seeks to elucidate the molecular mechanism(s) by which the La:miR-122 RNP is selectively sorted into ILVs. In Specific Aim #1, Jordan will assess the efficacy by which a panel of candidate endosomal receptor proteins (identified by unbiased proximity labeling proteomics) are able to invoke the capture of cytoplasmic La into ILVs prior to exosome secretion. In Specific Aim #2, Jordan will identify the domains of La required for (i) its unconventional secretion within exosomes and (ii) its interaction with miR-122. In Specific Aim #3, Jordan will establish a cell-free reaction that recapitulates the selective sorting of La into ILVs, allowing further biochemical dissection of this high-fidelity sorting mechanism. Completion of the proposed research will provide novel insights into the molecular mechanisms by which specific cytoplasmic constituents are efficiently and selectively packaged into exosomes, and by extension, identify putative therapeutic targets for metastatic breast cancer and inform efforts to engineer exosomes into efficacious delivery vehicles for therapeutic compounds and nucleic acids. At the conclusion of Jordan’s NRSA-sponsored training, Jordan will have a rigorous intellectual foundation, scientific independence, and a uniquely broad technical toolkit that will make him an outstanding candidate for an independent investigator position in the future.

项目摘要/摘要