Phenotype screens of Chlamydia Inclusions

衣原体包涵体表型筛选

基本信息

批准号：
9979523
负责人：
Heather Brown-Harding
金额：
$ 7.72万
依托单位：
WAKE FOREST UNIVERSITY
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：
2020-03-19 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9979523
关键词：
3-Dimensional Acrylates Aftercare Agonist Antibiotics Applications Grants Bacteria Cells Chlamydia Chlamydia Infections Chlamydia genome Chlamydia trachomatis Chloramphenicol Classification Clinical Computer software Confocal Microscopy Consumption Data Detection Development Developmental Gene Drug Costs Drug Screening Electron Microscopy Eye Infections Gel Genitourinary System Infection Grant Growth Health Hour Human Image Image Analysis Individual Iron Chelating Agents Lead Light Microscope Machine Learning Malaria Measurement Methods Microbiology Microscope Microscopy Morbidity - disease rate PF4 Gene Penicillins Pharmaceutical Preparations Pharmacotherapy Phenotype Polymers Preparation Research Resolution SIRT1 gene Sampling Sodium Techniques Time Work cell type chlamydia vaccine human pathogen inhibitor/antagonist innovation interest machine learning method novel novel therapeutics pathogen promoter screening small molecule small molecule inhibitor therapy development tool

3-Dimensional Acrylates Aftercare Agonist Antibiotics Applications Grants Bacteria Cells Chlamydia Chlamydia Infections Chlamydia genome Chlamydia trachomatis Chloramphenicol Classification Clinical Computer software Confocal Microscopy Consumption Data Detection Development Developmental Gene Drug Costs Drug Screening Electron Microscopy Eye Infections Gel Genitourinary System Infection Grant Growth Health Hour Human Image Image Analysis Individual Iron Chelating Agents Lead Light Microscope Machine Learning Malaria Measurement Methods Microbiology Microscope Microscopy Morbidity - disease rate PF4 Gene Penicillins Pharmaceutical Preparations Pharmacotherapy Phenotype Polymers Preparation Research Resolution SIRT1 gene Sampling Sodium Techniques Time Work cell type chlamydia vaccine human pathogen inhibitor/antagonist innovation interest machine learning method novel novel therapeutics pathogen promoter screening small molecule small molecule inhibitor therapy development tool

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项目摘要

Abstract Chlamydia trachomatis is a major health concern with over 200 million people with active urogenital or ocular infection each year worldwide. Chlamydia are obligate intracellular bacteria with a unique biphasic developmental cycle. A better understanding of that biphasic cycle can lead to inhibitors that are specific for chlamydial infection in order to avoid overuse of antibiotics. Individual Chlamydia are too small and tightly packed to be spatially separated with conventional light microscopes, and 3D SEM is too labor-intensive for inhibitor studies. We will use a new sample preparation method that physically expands the sample with polymers termed "Expansion Microscopy" or ExM. Expanded samples can then be imaged with a traditional confocal microscope, and high-content analysis performed automatically using machine learning methods such as pixel classification and novelty detection. Prepared samples can be imaged and analyzed in under an hour instead of the multiple days required for 3D SEM. This R03 grant will develop an innovative high-content screening platform, called Expansion Microscopy Aided Phenotyping (ExMAP), for the quantification of changes in Chlamydia development after treatment. ExMAP can be paired with Chlamydia transformed with promoters for EUO and IhtA (RB cell types) and the promoters for HctB and Tarp (EB cell types). The combination of expansion microscopy, machine learning, and chlamydial transformation will make ExMAP a powerful tool for research on both the developmental cycle and new therapy development.

抽象的沙眼衣原体是一个主要的健康问题，有超过2亿人有活跃每年全球泌尿生殖器或眼部感染。衣原体是义务细胞内细菌具有独特的双相发育周期。更好地了解该双相周期可以导致针对衣原体感染的抑制剂，以避免过度使用抗生素。单个衣原体太小，挤得紧密，无法在空间上与常规分离光显微镜和3D SEM对于抑制剂研究过于劳动密集型。我们将使用新的样品制备方法可以物理地用称为“膨胀的聚合物”样品扩展样品显微镜”或EXM。然后可以使用传统共焦点进行成像显微镜和使用机器学习方法自动执行的高含量分析例如像素分类和新颖性检测。可以对准备的样品进行成像和分析在一个小时的时间内，而不是3D SEM所需的数天。 R03赠款将发展一个创新的高含量筛选平台，称为扩展显微镜辅助表型（EXMAP），用于量化治疗后衣原体发育的变化。 exmap 可以与Chlamydia与EUO和IHTA的启动子（RB细胞类型）和启动子配对 HCTB和TARP的启动子（EB细胞类型）。扩展显微镜的组合，机器学习和衣原体转化将使Exmap成为研究的强大工具在发育周期和新疗法发展上。