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Investigating the role of liquid-liquid phase separation in the interaction between Mycobacterium tuberculosis and macrophages

研究液-液相分离在结核分枝杆菌与巨噬细胞相互作用中的作用

基本信息

批准号：
10663318
负责人：
Samantha Lynn Bell
金额：
$ 46.61万
依托单位：
RUTGERS BIOMEDICAL AND HEALTH SCIENCES
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：
2021-08-10 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10663318
关键词：
Affect Antibacterial Response Antibiotic Resistance Automobile Driving Autophagocytosis Bacillus Biochemical Biochemistry Biological Biological Process Biophysical Process Biophysics Cell Nucleus Cell membrane Cells Cellular biology Communicable Diseases Complex Cytoplasm Data Defense Mechanisms Development Disease Disease Outcome Event Galactose Binding Lectin Health Human Immune Immune response Immune system In Vitro Individual Infection Infection Control Innate Immune Response Knowledge Link Liquid substance Macrophage Membrane Microbiology Mission Modification Molecular Molecular Biology Mycobacterium tuberculosis Natural Immunity Organelles Pathogenesis Patient-Focused Outcomes Patients Persons Phase Phenotype Phosphorylation Physical condensation Polyubiquitination Population Post-Translational Protein Processing Process Proteins Proteomics Reaction Recording of previous events Regulation Reporting Research Role Signal Transduction Stimulator of Interferon Genes Structure TBK1 gene Testing Therapeutic Training Tuberculosis Ubiquitination United States National Institutes of Health Work bacterial genetics combat design experimental study fighting fluorescence imaging genetic approach human pathogen in vivo innate immune pathways innovation interdisciplinary approach live cell imaging novel novel therapeutics optogenetics pathogen prevent response tool

项目摘要

PROJECT SUMMARY There is a fundamental gap in our understanding of the complex processes that govern the interactions between Mtb and macrophages. The overall objective of this application is to investigate how the novel biophysical phenomenon of phase separation impacts biological processes, specifically in the context of Mtb infection. A detailed knowledge of the molecules that recognize and respond to pathogens is required to reveal how cells fight infection; therefore, there is a critical need to understand how phase separation may influence or control innate immune responses. Mycobacterium tuberculosis (Mtb) is an incredibly successful and deadly human pathogen that infects one-quarter of the world's population. While interaction of Mtb bacilli and macrophages activates numerous innate immune pathways, we have a limited understanding of how these complex networks of host sensing molecules are regulated to work cooperatively. Furthermore, only a small subset of the many secreted effectors used by M. tuberculosis have well-characterized functions. Recent studies have illuminated the biological and cellular importance of liquid-liquid phase separation, a process by which proteins condense into discrete droplets to alter their localization and function in a cell. Several proteins involved in the host response to M. tuberculosis infection, like cGAS, TBK1, p62, and LC3, have been found to phase separate in vitro, but how in vivo phase separation impacts host responses to infection is unknown. Preliminary studies have found that these and other innate immune proteins form circular puncta in M. tuberculosis-infected cells that resemble phase separated droplets. The central hypothesis of this proposal is that upon infection, pathogen- sensing and post-translational modifications induce phase separation of host proteins and that Mtb modulates these condensation events with its own phase-separating PE/PPE proteins. Here, a combination of novel optogenetics tools, live cell fluorescent imaging, and host and bacterial genetics will be employed to probe the biological consequences of phase separation of host proteins (Aim 1) and Mtb proteins (Aim 2). In addition, directed and unbiased genetics approaches will be used to probe how post-translational modifications, and especially ubiquitination in particular, contributes to phase separation during Mtb infection (Aim 3). This approach is innovative in that it uses novel tools to specifically and precisely modulate phase separation in order to link this biophysical process with meaningful cellular phenotypes. The proposed research is significant because it will greatly expand our understanding of how macrophages destroy Mtb and advance efforts to combat Mtb infection via enhancing host responses.

项目总结