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Defining actin-based mechanisms driving basic cell functions and pathogenic behaviors in Naegleria

定义基于肌动蛋白的机制，驱动耐格里虫的基本细胞功能和致病行为

基本信息

批准号：
10213600
负责人：
Katrina Velle
金额：
$ 7.04万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-25 至 2023-08-24
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10213600
关键词：
Actins Actomyosin Address Amoeba genus Animals Automobile Driving Behavior Biology Brain Caliber Category B pathogen Cell Survival Cell division Cell membrane Cell physiology Cells Cellular biology Complement Complex Core Facility Cytokinesis Cytometry Cytoskeleton Data Defect Disease Drug Targeting Eating Encephalitis Ensure Environment Equilibrium Eukaryota F-Actin Fatality rate Flow Cytometry Foundations Future Gene Expression Profiling Gene Targeting Goals Health Human Image Impairment Individual Infection Measures Mentorship Microscopy Microtubules Mission Mitotic Molecular Myosin ATPase Myosin Type II Naegleria Naegleria fowleri National Institute of Allergy and Infectious Disease Organelles Organism Osmolar Concentration Osmotic Pressure Pathogenesis Pathogenicity Phenotype Play Population Postdoctoral Fellow Process Protein Family Proteins Pump Research Research Personnel Research Training Resolution Resource Development Resources Role Signal Transduction Solid Testing Training Transcript Translations United States National Institutes of Health Universities Vacuole Virulence Water Work base career career development cell motility cellular imaging chemokine driving force migration single cell analysis small molecule inhibitor success therapeutic development

项目摘要

Project Summary The “brain-eating amoeba” Naegleria fowleri is an NIAID Priority Category B Pathogen that carries a 95% fatality rate, yet the mechanisms underlying its basic biology and pathogenic behaviors remain largely unstudied. Because understanding the cell biology of Naegleria is critical to the development of therapeutics, my long-term goal is to define the cellular and molecular basis of Naegleria pathogenesis. Unlike human cells, from which Naegleria diverged 1-2 billion years ago, these amoebae do not possess cytoplasmic microtubules. This suggests that the actin cytoskeleton, assembled by the Arp2/3 complex and formin family proteins, is the primary driving force for many cellular processes essential to cell survival and pathogenesis. Therefore, my overall objective in this application is to determine how actin cytoskeletal rearrangements promote cell motility, which is important for establishing infection, contractile vacuole pumping, which is required for surviving osmotic pressure, and cell division, which is critical for robust colonization in the brain. To achieve this objective, I propose the following specific aims: (Aim 1) determine the Arp2/3 complex activators driving cell motility, (Aim 2) define the formin-based mechanisms governing contractile vacuole dynamics, and (Aim 3) identify the actin nucleators and molecular mechanisms responsible for cytokinesis. I will address these aims using orthogonal cellular perturbations (small molecule inhibitors to impair the Arp2/3 complex and formin family proteins, as well as individual gene targeting using morpholinos) and environmental perturbations by changing external osmotic pressure. I will measure phenotypes of single cells in detail with microscopy, and complement this by collecting quantitative data on cell populations with flow cytometry and gene expression assays. Because the cell biology underlying pathogenesis and basic functions in Naegleria is severely understudied, defining how actin orchestrates motility, division, and contractile vacuole dynamics is critical for uncovering drug targets to treat these deadly infections. Completing this project will not only advance the mission of the NIH, but it will also prepare me for a career as an independent investigator at a research-intensive university studying the cell biology of Naegleria. The career development resources available to postdocs and the excellent core facility trainings at UMass create an ideal institutional environment for my training. Further, the mentorship of Dr. Fritz- Laylin, an expert on Naegleria biology, and Dr. Wadsworth, who specializes in cell division, will ensure the success of this project as well as my transition to a career as an independent researcher.

项目总结