Role of Host Protein Kinase C in Cryptosporidiosis

项目总结 影响：这项提案解决了对婴儿隐孢子虫病有效治疗的迫切需要。这个 治疗靶点是我们新近发现的宿主易感基因--人蛋白激酶C(PKCα) 治疗隐孢子虫病。这项工作的成功完成将广泛推进宿主靶向治疗领域 在疾病期间，通过机械验证，从正向基因筛查中受到打击。 意义：隐孢子虫病是中低收入家庭儿童腹泻的五大原因之一 国家。目前还没有疫苗，药物也极其有限，没有一种被批准用于婴儿。因此，在那里 迫切需要有效的治疗方法，特别是对儿童。这项提议开创了发展的先例 微生物的宿主目标，这是面对新出现的抗菌素耐药性的一个具有高度意义的领域。 创新：对来自低资源国家的婴儿进行全基因组扫描，以了解婴儿对传染病的易感性 在我们的研究团队开创这项技术之前，从来没有人这样做过。该提案融合了人类遗传学和 寄生细胞生物学将PKCα的发现发展为宿主定向治疗的靶点。我们之前的工作 确定了可以被阻断以防止内阿米巴寄生虫杀死细胞的新宿主因子。我们建议 将这一创新方法推广到隐孢子虫。这一战略从根本上不同于传统的 寄生虫靶向药物，基于PKCα是一种新的宿主易感因子的初步数据 隐孢子虫病。 方法：我们发现人类PKCα是隐孢子虫病易感性的关键成分。 使用全基因组扫描疾病相关基因的婴儿。我们假设pkcα是 隐孢子虫的细胞内感染，可作为治疗隐孢子虫病的靶点。我们将定义角色 一项多学科研究中PKCα的研究，该研究整合了婴儿自然感染期间PKCα的分析 (目标1)，有针对性地对PKCα基因座进行测序，以确定原因SNP和作用机制(目标2)， 体外和体内感染研究中宿主蛋白激酶Cα的分析，并结合FDA- 批准的PKCα拮抗剂(目标3)。成功完成此R01将产生一项新的调查 治疗隐孢子虫病的宿主靶向PKC药物的药物申请。 环境：调查人员带来了完成这项工作的强大和互补的技能。玛丽博士(圆周率) 是宿主和寄生虫细胞和分子生物学方面的专家。达格尔博士领导着一个享誉国际的项目 在人类对感染的遗传易感性方面，哈克博士在儿童实地研究方面带来了丰富的经验 在孟加拉国。重要的是，这个团队高度合作，他们过去的发现和合作证明了这一点 出版物。该团队汇集了三个国际公认的全球健康中心：加州大学 弗吉尼亚(玛丽博士)、ICDDR、B(哈克博士)和约翰·霍普金斯大学(杜格尔博士)。此次合作将 发生在一个令人兴奋和严谨的跨学科环境中。

PROJECT SUMMARY Impact: This proposal addresses the pressing need for effective treatments for cryptosporidiosis in infants. The therapeutic target is human protein kinase C (PKCα), which we recently discovered as a host susceptibility gene for cryptosporidiosis. Successful completion of this work will broadly advance the field of host-targeted therapy by taking a hit from a forward-genetic screen through mechanistic validation during disease. Significance: Cryptosporidiosis is one of the top 5 causes of diarrhea in children in low- and middle-income countries. There is no vaccine and the drugs are extremely limited with none approved for infants. Thus, there is an urgent need for effective therapies, particularly for children. This proposal sets a precedent for developing host targets for microorganisms, an area of high significance in the face of emerging antimicrobial resistance. Innovation: A genome-wide scan for infectious disease susceptibility in infants from low-resource countries had never been done until it was pioneered by our investigator team. The proposal integrates human genetics and parasite cell biology to develop the discovery of PKCα into a target for host directed therapy. Our previous work identified novel host factors that could be blocked to prevent cell-killing by Entamoeba parasites. We propose to expand this innovative approach to Cryptosporidium. This strategy is fundamentally different from traditional parasite-targeted drugs and is based on preliminary data that PKCα is a novel host susceptibility factor for cryptosporidiosis. Approach: We have discovered that human PKCα is a key component of susceptibility to cryptosporidiosis in infants using a genome wide scan for disease-associated loci. We hypothesize that PKCα is required for intracellular infection by Cryptosporidium and could be targeted to cure cryptosporidiosis. We will define the role of PKCα in a multi-disciplinary study that integrates an analysis of PKCα during natural infection in infants (Aim 1), targeted sequencing of the PKCα locus to identify the causal SNPs and mechanism of action (Aim 2), and an analysis of host PKCα during in vitro and in vivo infection studies in combination with testing of FDA- approved PKCα antagonists (Aim 3). Successful completion of this R01 will result in an Investigational New Drug Application for a host-targeted PKC drug for cryptosporidiosis. Environment: The Investigators bring strong and complementary skills to accomplish this work. Dr. Marie (PI) is an expert in host and parasite cell and molecular biology. Dr. Duggal leads an internationally regarded program in human genetic susceptibility to infection and Dr. Haque brings extensive experience in field studies in children in Bangladesh. Importantly this team is highly collaborative as evidenced by their past discoveries and co- publications. The team brings together three internationally recognized global health centers: The University of Virginia (Dr. Marie), the ICDDR,B (Dr. Haque), and John Hopkins University (Dr. Duggal). This collaboration will occur in an exciting and rigorous interdisciplinary environment.