Host-directed Therapeutics for Amebiasis

阿米巴病的宿主导向治疗

基本信息

批准号：
9247887
负责人：
Chelsea S. Marie
金额：
$ 19.75万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9247887
关键词：
Acanthamoeba Acute Amebiasis Amebic colitis Amoeba genus Apical Balamuthia mandrillaris Biological Assay CRISPR/Cas technology Cell Death Cell physiology Cells Cessation of life Child Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Colon Data Development Disease Drug Targeting Entamoeba histolytica Enteral Environment Epithelial Epithelium Event Gene Deletion Genes Genetic Screening Goals Human In Vitro Infection Inflammation Integration Host Factors Intestines Invaded Ion Transport Ions Knock-out Laboratories Light Measurement Measures Mediating Mediator of activation protein Modeling Molecular Morbidity - disease rate Naegleria fowleri Nature Parasites Parasitic Diseases Pathogenicity Pathway interactions Pharmaceutical Preparations Pharmacology Physiological Postdoctoral Fellow Potassium Potassium Channel Predisposition Prevention RNA Interference Research Resistance Resources Role Stem cells Surface Susceptibility Gene Techniques Technology Testing Therapeutic Tissues Toxic effect Universities Virginia Virulence base career development cell killing chemokine collaborative environment cytokine cytotoxicity drug testing efficacy testing experimental study genetic variant genome editing genome wide association study ginsenoside M1 graduate student inhibitor/antagonist innovation insight killings knock-down meetings microbial monolayer mortality new therapeutic target novel novel therapeutics prevent programs public health relevance response success targeted treatment therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Hypothesis: Activation of human K+ channels during amebic trogocytosis is the causal mediator of cell death. Preliminary data: K+ channels were identified in a novel forward genetic screen for host factors required for trogocytic killing by Entamoeba histolytica. Key preliminary experimental data also support the importance of K+ channels including: (1) E. histolytica activated K+ channels prior to killing target cells; and (2) inhibition of K+ channels protected cells from amebic killing. Approach: We will extend this discovery by analyzing specific colonic K+ channels as potential drug targets in the human enteroid model of amebiasis. We will test CRISPR-Cas9 deletions of high confidence candidate K+ channels genes in human enteroids. Gene deletion studies will be paired with pharmacologic inhibitor testing in enteroids with the goal of providing proof-of-principle for K+ channel inhibitrs as novel therapeutics. Successful completion of these studies will determine if human K+ channels are novel host-directed, druggable targets for amebiasis. Significance: E. histolytica is a diarrheal disease with significant global morbidity and mortality, especially in children in the developing world. There is a single class of drugs for invasive disease and toxicity and the emergence of resistance are clinical concerns. The highly pathogenic free-living amebae (Naegleria fowleri, Acanthamoeba sp., and Balamuthia mandrillaris) also kill human cells via trogocytosis, therefore our studies aiming to prevent trogocytic death in enteric amebiasis, may also offer therapy for the free-living amebae. Innovative aspects of the proposal are: the forward genetic screen that identified human K+ channels as a potential host therapeutic target; the focus on amebic trogocytic killing, a unique cellular process shared by parasitic E. histolytica and the free-living amebae; and by the use of cutting edge technology (a human colonic enteroid model and validated CRISPR gene deletions). The environment for the work is Dr. William A. Petri's laboratory at The University of Virginia. Dr. Petri has been a leader in the fied of amebiasis research for 25 years and his laboratory is an intellectually exciting and rigorous environment for the study of enteric infections. There are 7 postdoctoral fellows and 5 graduate students, with independent yet complementary projects. The laboratory contains all the necessary resources and expertise to accomplish high-impact research and great emphasis is placed on collaboration and sharing of ideas and techniques, as well as independence and career development for the fellows and graduate students. Weekly laboratory meetings and daily informal discussions (facilitated by the presence of Dr. Petri's office within the laboratory serve to promote the ongoing research program.

描述（由申请人提供）：假设：阿米巴盘胞作用期间人类 K+ 通道的激活是细胞死亡的因果介质。初步数据：K+ 通道是在溶组织内阿米巴杀灭盘胞所需的宿主因子中鉴定的。初步实验数据也支持K+通道的重要性，包括：（1）溶组织内阿米巴在杀死靶细胞之前激活K+通道；和（2）方法：我们将通过分析作为人类肠样阿米巴病模型中潜在药物靶点的特定结肠 K+ 通道来扩展这一发现。基因缺失研究将与肠类药物抑制剂测试相结合，目的是为 K+ 通道抑制剂作为新型疗法提供原理证明，这些研究的成功完成将决定是否成功。人类 K+ 通道是阿米巴病的新型宿主导向药物靶点意义：溶组织阿米巴是一种在全球范围内发病率和死亡率很高的腹泻病，特别是在发展中国家的儿童中。针对侵袭性疾病和毒性的药物只有一类。高致病性自由生活阿米巴（福氏耐格里阿米巴、棘阿米巴和山魈）也会导致人类死亡。因此，我们的研究旨在预防肠道阿米巴病中的细胞死亡，也可能为自由生活的阿米巴提供治疗方法。重点关注阿米巴胞体杀灭，这是寄生性溶组织内阿米巴和自由生活的阿米巴共有的一种独特的细胞过程，并通过使用尖端技术（a人类结肠肠样模型和经过验证的 CRISPR 基因删除）。该工作环境是弗吉尼亚大学 William A. Petri 博士的实验室。Petri 博士在阿米巴病研究领域已担任 25 年的领导者，他的实验室是该实验室拥有 7 名博士后研究员和 5 名研究生，拥有独立但互补的项目，拥有完成高影响力研究所需的所有必要资源和专业知识。实验室非常重视合作和分享想法和技术，以及研究员和研究生的独立性和职业发展。每周的实验室会议和每日的非正式讨论（由实验室内的 Petri 博士办公室提供便利）有助于促进。正在进行的研究计划。