Role of Host Protein Kinase C in Cryptosporidiosis

项目摘要 影响：该提案解决了对婴儿隐孢子虫病有效治疗的迫切需求。的 治疗靶点是人类蛋白激酶C（PKCα），我们最近发现它是一个宿主易感基因 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 通过在疾病期间通过机械验证从正向遗传筛选中获得命中。 意义：隐孢子虫病是低收入和中等收入家庭儿童腹泻的5大原因之一。 国家没有疫苗，药物也非常有限，没有一种被批准用于婴儿。因此 迫切需要有效的治疗方法，特别是对儿童。这一提议为发展 微生物的宿主靶标，这是面对新出现的抗菌素耐药性的一个重要领域。 创新：对低资源国家婴儿的传染病易感性进行全基因组扫描 直到我们的调查团队率先开展这项工作。该提案将人类遗传学和 寄生虫细胞生物学，以发展PKCα的发现成为宿主定向治疗的靶点。我们以前的工作 确定了新的宿主因子，这些因子可以被阻断以防止内阿米巴寄生虫杀死细胞。我们建议 将这种创新方法扩展到隐孢子虫。这一战略与传统的 该研究基于初步数据，即PKCα是一种新的宿主易感因子， 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， 婴儿使用全基因组扫描疾病相关基因座。我们假设PKCα是 隐孢子虫细胞内感染，并可靶向治疗隐孢子虫病。我们将定义角色 在一项多学科研究中，整合了婴儿自然感染期间PKCα的分析， (Aim 1），PKCα基因座的靶向测序以鉴定致病SNP和作用机制（目的2）， 以及在体外和体内感染研究中结合FDA-1000检测对宿主PKCα的分析。 批准的PKCα拮抗剂（Aim 3）。成功完成此R 01将导致研究性新 一种针对隐孢子虫病的宿主靶向PKC药物的药物申请。 环境：调查人员具有强大的互补技能来完成这项工作。玛丽博士（PI） 是宿主和寄生虫细胞和分子生物学专家。Duggal博士领导着一个国际知名的项目 在人类遗传易感性感染和哈克博士带来了广泛的经验，在实地研究的儿童 在孟加拉国。重要的是，这个团队是高度合作的，这一点可以从他们过去的发现和合作中得到证明。 出版物。该团队汇集了三个国际公认的全球健康中心： 弗吉尼亚（玛丽博士），ICDDR，B（哈克博士）和约翰霍普金斯大学（杜加尔博士）。这项合作将 发生在一个令人兴奋的和严格的跨学科的环境。

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.