Polymorphisms link Protein Kinase C-α to Cryptosporidia susceptibility in 1st year of life

多态性将蛋白激酶 C-α 与生命第一年的隐孢子虫易感性联系起来

基本信息

批准号：
10649620
负责人：
Sayo Erick McCowin
金额：
$ 5.27万
依托单位：
UNIVERSITY OF VIRGINIA
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：
2020-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10649620
关键词：
5 year old Actins Agonist Alleles Bangladesh Biological Biological Assay Biopsy CRISPR/Cas technology Cause of Death Cell physiology Cells Child Child Development Child Health Child Mortality Childhood Chromosomes Clinical Collection Colon Color Contracts Cryptosporidiosis Cytoskeleton Data Databases Development Diarrhea Disease Dominant-Negative Mutation Epithelial Cells Epithelium Genetic Genetic Polymorphism Genome Genotype Goals Human Human Genome Image Immunity Immunocompetent Immunocompromised Host Immunoglobulin A In Vitro Incidence Infant Infection Intestines Invaded Investigation Life Link Linkage Disequilibrium Malnutrition Manuscripts Measures Mediating Messenger RNA Molecular Monitor Mus Neurocognitive Outcome PRKCA gene Parasites Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Phenotype Phosphotransferases Placebos Population Positioning Attribute Predisposition Preparation Protein Conformation Proteins Quantitative Trait Loci Regulation Research Proposals Risk Role Rotavirus Severities Severity of illness Single Nucleotide Polymorphism Small Intestines Symptoms Testing Therapeutic Tissues Transcript Variant Work burden of illness cohort cytokine diarrheal disease effective therapy enteric infection experience genome wide association study genome-wide global health human pathogen in vivo inhibitor insight intestinal epithelium link protein mortality nitazoxanide parasite invasion pathogen pharmacologic protein function risk variant tool transcriptome sequencing

项目摘要

PROJECT SUMMARY Cryptosporidia is a diarrheal pathogen with an important impact on global health. Overall, diarrheal disease is estimated to be responsible for 10% of under 5-year old child mortality, and the recent Global Burden of Diseases analysis of diarrheal mortality found that Cryptosporidia was second only to rotavirus as a cause of death in children under five years old (5). Nitazoxanide, currently the only approved drug for Cryptosporidia infection, is only moderately effective for treatment of immunocompetent patients. Furthermore, it is only equivalent to placebo in severely immunocompromised patients. Clinical presentation of Cryptosporidia infection is variable in severity; some patients experience severe diarrheal illness while some infections remain subclinical. Although the cause for the wide spectrum of disease is poorly understood, several studies have demonstrated long-term clinical implications of both asymptomatic and symptomatic infection. We performed a genome-wide association study (GWAS) on infants in Bangladesh within the first year of life. We evaluated the association between 6.5 million single nucleotide polymorphisms (SNPs) across the human genome and symptomatic Cryptosporidia infection in three independent patient cohorts. This analysis revealed a highly significant statistical association of multiple SNPs in the PRKCA gene and susceptibility to Cryptosporidia diarrhea. Each copy of the risk allele increased the risk of contracting cryptosporidiosis by 2.4 times in the first year of life. The most significant SNP in PRKCA is an eQTL in the GTex database, however, a definitive link between PKCα and Cryptosporidia has not yet been established. The PKCα kinase is a primary regulator of actin and during intracellular invasion, Cryptosporidia induces host cell actin remodeling. Further investigation into how the PRKCA gene variation relates to PKCα function and susceptibility to Cryptosporidia infection will provide insight into the pathogenesis of an important human pathogen to cause disease. We hypothesize that host PKCα is required for Cryptosporidia invasion of intestinal epithelium through remodeling of actin cytoskeleton. Based on this preliminary data, we aim to characterize (i) the role of PKCα in Cryptosporidia invasion of the intestinal epithelium in vitro and in vivo (ii) define the impact of the SNPs on PRKCA expression and downstream function, and (iii) identify if PRKCA expression mediates severity of disease in children. To accomplish this, the research proposal includes active investigation of Cryptosporidia both in the field and at the bench. Here, we propose bridging two approaches for scientific discovery by using molecular tools to examine PRKCA and monitoring clinical outcomes in genotyped human children. The overarching goal of these independent yet interacting aims will be the translational development of host PKCα as a potential target to advance critically needed treatments for cryptosporidiosis.

项目概要隐孢子虫是一种腹泻病原体，对全球健康具有重要影响。据估计，儿童死亡率占 5 岁以下儿童死亡率的 10%，而最近的全球负担腹泻死亡率的疾病分析发现，隐孢子虫是仅次于轮状病毒的第二大腹泻病死因。五岁以下儿童死亡 (5) 硝唑尼特，目前唯一批准治疗隐孢子虫的药物。感染，对于免疫功能正常的患者的治疗仅具有中等效果。相当于严重免疫功能低下患者的安慰剂。感染的严重程度各不相同；有些患者会出现严重的腹泻症状，而有些患者仍然存在感染。尽管对广泛疾病的病因知之甚少，但一些研究已经表明。我们对无症状和有症状感染的长期临床影响进行了研究。我们对孟加拉国婴儿出生后一年内进行的全基因组关联研究（GWAS）进行了评估。人类基因组中 650 万个单核苷酸多态性 (SNP) 之间的关联三个独立患者队列中的症状性隐孢子虫感染显示出高度的隐孢子虫感染。 PRKCA 基因中多个 SNP 与隐孢子虫易感性的显着统计学关联每个风险等位基因拷贝都会使感染隐孢子虫病的风险增加 2.4 倍。然而，PRKCA 中最重要的 SNP 是 GTex 数据库中的 eQTL。 PKCα 和隐孢子虫之间的关系尚未确定。 PKCα 激酶是隐孢子虫的主要调节因子。肌动蛋白和细胞内侵袭过程中，隐孢子虫诱导宿主细胞肌动蛋白重塑。研究 PRKCA 基因变异如何与 PKCα 功能和对隐孢子虫感染的易感性相关提供对引起疾病的重要人类病原体的发病机制的深入了解。隐孢子虫通过肌动蛋白重塑入侵肠上皮需要宿主 PKCα 基于这些初步数据，我们的目标是描述 (i) PKCα 在隐孢子虫中的作用。体外和体内肠上皮的侵袭 (ii) 确定 SNP 对 PRKCA 表达的影响和下游功能，(iii) 确定 PRKCA 表达是否介导儿童疾病的严重程度。为了实现这一目标，该研究计划包括在现场和现场对隐孢子虫进行积极调查在这里，我们建议通过使用分子工具来桥接两种科学发现方法。检查 PRKCA 并监测基因分型人类儿童的临床结果。这些独立但相互作用的目标将是将宿主 PKCα 转化为潜在的目标是推进急需的隐孢子虫病治疗方法。