Molecular signaling mechanisms controlling Cryptosporidium proliferation and development

控制隐孢子虫增殖和发育的分子信号机制

基本信息

批准号：
10548206
负责人：
Sumiti Vinayak Alam
金额：
$ 38.45万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
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：
2021-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10548206
关键词：
Biochemical Genetics Biological Biological Assay Biological Models Biological Process Biology Calcium Candidate Disease Gene Cessation of life Child Clustered Regularly Interspaced Short Palindromic Repeats Complex Cryptosporidiosis Cryptosporidium Cryptosporidium parvum Dependence Development Diarrhea Disease Disease Outbreaks Drug Targeting Epithelial Cells Epitopes FDA approved Fertilization Future Gametogenesis Genetic Genome Germ Cells Goals Homologous Gene Human Immunocompromised Host In Vitro Individual Infant Infection Intestines Knowledge Laboratories Life Cycle Stages Light Link Mediating Meiosis Methods Modeling Molecular Molecular Genetics Mus Nature Oocysts Parasite Control Parasites Pharmaceutical Preparations Phenotype Phosphorylation Phosphotransferases Plants Plasmodium Play Proliferating Protein-Serine-Threonine Kinases Recreation Resource-limited setting Role Rotavirus Sexual Development Signal Pathway Signal Transduction Signaling Protein System Technology Testing Toddler Toxoplasma Transgenic Organisms Vaccines Water asexual calcium-dependent protein kinase candidate identification chemical genetics diarrheal disease drug development effective therapy enteric infection genetic approach genetic manipulation genome editing immunological status in vivo insight intestinal epithelium kinase inhibitor male mouse model mutant nitazoxanide novel novel therapeutics novel vaccines parasite genome pathogen prevent programs protein degradation reverse genetics stem superresolution microscopy transmission process vaccine development waterborne outbreak

项目摘要

Project Abstract Cryptosporidium is a leading cause of diarrheal disease (cryptosporidiosis) and death among young children living in resource-poor settings. In the US, Cryptosporidium is the major cause of waterborne outbreaks linked to recreational water use. Currently, there is no fully effective drug and no vaccine to treat or prevent cryptosporidiosis. The only available US FDA approved drug, nitazoxanide has no proven efficacy in young children with weak immune status and immunocompromised individuals. Therefore, there is an urgent need to develop new drugs and vaccine to reduce the burden of cryptosporidiosis. Progress in anti-cryptosporidial drug and vaccine development has been hampered due to our limited understanding of parasite biology. The underlying reasons for this slow progress have been the unavailability of a robust method to continuously propagate Cryptosporidium, and the absence of molecular genetics to manipulate the parasite genome. We have overcome these hurdles by developing a powerful technology to manipulate the Cryptosporidium genome and propagate these genetically modified parasites in an immunocompromised mouse model system. The key advantage of this genetic system is that the entire life cycle of Cryptosporidium (both asexual and sexual stages) is completed in the mouse intestine, allowing us to unravel parasite biology (Vinayak et al 2015, Nature 523:477). We lack an understanding of the molecular signaling mechanisms that control development of parasite stages for successful completion of the complex life cycle. Signaling pathway components such as the plant-like calcium-dependent protein kinases (CDPKs) have emerged as attractive drug targets in Cryptosporidium and related parasites, due to the absence of their homologues in human host. Taking advantage of our genetic system, we have demonstrated the efficacy of selective bumped kinase inhibitors against calcium-dependent protein kinase-1 (CDPK1), thus indicating a critical role of this signaling kinase in C. parvum. Utilizing the conditional protein degradation system recently developed in our laboratory, we have demonstrated the essential role of CDPK1 in asexual proliferation and parasite survival. Moreover, we have compelling preliminary evidence that sheds light on the role of two signaling kinases in sexual developmental stages. The goal of this project is to elucidate the mechanistic role of these signaling proteins in regulating development of asexual and sexual stages in C. parvum required for parasite proliferation and transmission. Elucidation of these mechanisms will provide novel insights into the fundamental biology of Cryptosporidium, and open new avenues for development of effective therapies.

项目摘要隐孢子虫是腹泻病（隐孢子虫病）和幼儿死亡的主要原因生活在资源贫乏的环境中。在美国，隐孢子虫是连接水上爆发的主要原因用于休闲用水。目前，没有完全有效的药物，也没有疫苗可以治疗或预防隐孢子虫病。 Nitazoxanide是唯一可用的美国FDA批准的药物，在Young中没有可靠的功效免疫状态弱和免疫功能低下的个体的儿童。因此，迫切需要开发新药和疫苗，以减轻隐孢子虫病的负担。抗孢子虫药的进展由于我们对寄生虫生物学的了解有限，因此疫苗开发受到阻碍。这这种缓慢进步的根本原因一直是一种坚固的方法的不可用传播隐孢子虫，以及没有分子遗传学来操纵寄生虫基因组。我们有通过开发强大的技术来操纵隐孢子虫基因组和在免疫功能低下的小鼠模型系统中传播这些基因修饰的寄生虫。钥匙这种遗传系统的优势是隐孢子虫的整个生命周期（无论是无性阶段还是性阶段）在小鼠肠道中完成，使我们能够揭开寄生虫的生物学（Vinayak等，2015年，自然523：477）。我们对控制寄生虫阶段发展的分子信号传导机制缺乏了解为了成功完成复杂的生命周期。信号通路途径组件，例如植物样钙依赖性蛋白激酶（CDPK）已成为隐孢子虫中的有吸引力的药物靶标相关的寄生虫，由于其在人类宿主中缺乏同源物。利用我们的遗传系统，我们已经证明了选择性碰撞激酶抑制剂针对钙依赖性的功效蛋白激酶-1（CDPK1），因此表明该信号激酶在C. parvum中的关键作用。利用有条件的蛋白质降解系统最近在我们的实验室开发，我们证明了必不可少的 CDPK1在无性增殖和寄生虫存活中的作用。而且，我们有令人信服的初步证据这阐明了两个信号激酶在性发育阶段的作用。这个项目的目标是阐明这些信号蛋白在调节无性和性的发展中的机械作用寄生虫增殖和传播所需的C. parvum阶段。阐明这些机制将提供有关隐孢子虫基本生物学的新颖见解，并开放新的发展途径有效的疗法。