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

• 批准号: 10180871
• 负责人: Sayo Erick McCowin
• 金额: $ 3.48万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10180871
• 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; Chromosome Positioning; Clinical; Collection; Colon; Color; Contracts; Cryptosporidiosis; Cytoskeleton; Data; Databases; Development; Diarrhea; Disease; Dominant-Negative Mutation; Epithelial; Epithelial Cells; Genetic; Genetic Polymorphism; Genome; Genotype; Genotype-Tissue Expression Project; Goals; Human; Human Genome; Image; Immunity; Immunocompetent; Immunocompromised Host; Immunoglobulin A; In Vitro; Incidence; Infant; Infection; Intestines; 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; Pharmacology; Phenotype; Phosphotransferases; Placebos; Population; Predisposition; Preparation; Protein Conformation; Quantitative Trait Loci; Regulation; Research Proposals; Risk; Role; Rotavirus; Severities; Severity of illness; Single Nucleotide Polymorphism; Small Intestines; Symptoms; Testing; Therapeutic; Time; Tissues; Transcript; Variant; Work; alpha Actin; base; 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/antagonist; insight; intestinal epithelium; link protein; mortality; nitazoxanide; parasite invasion; pathogen; protein function; risk variant; tool; transcriptome sequencing

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.

项目总结 隐孢子虫是一种腹泻病原体，对全球健康有重要影响。总体而言，腹泻病是 据估计，造成10%的5岁以下儿童死亡，以及最近的全球负担 疾病分析腹泻死亡率发现，隐孢子虫是仅次于轮状病毒的第二大致病原因 五岁以下儿童死亡(5)。硝唑尼特，目前唯一被批准用于治疗隐孢子虫的药物 感染，只对免疫功能正常的患者有中等疗效。此外，它只是 相当于免疫功能严重受损患者的安慰剂。隐孢子虫的临床表现 感染的严重程度是不同的；一些患者经历了严重的腹泻疾病，而一些感染仍然存在 亚临床的。尽管人们对广谱疾病的原因知之甚少，但有几项研究已经 证明了无症状和有症状感染的长期临床意义。我们表演了一场 对孟加拉国出生一年内的婴儿进行全基因组关联研究。我们评估了 人类基因组中650万个单核苷酸多态(SNPs)与 在三个独立的患者队列中出现症状的隐孢子虫感染。这一分析揭示了高度的 PRKCA基因多个SNP与隐孢子虫易感性的显著统计学关联 拉肚子。风险等位基因的每一个拷贝都会使感染隐孢子虫病的风险增加2.4倍 生命的一年。PRKCA中最重要的SNP是GTEx数据库中的eQTL，然而，这是一个明确的联系 PKC、α和隐孢子虫之间的关系尚未建立。蛋白激酶Cα激酶是一种主要的调节因子 在细胞内入侵过程中，隐孢子虫诱导宿主细胞肌动蛋白重塑。进一步调查 PrKCA基因变异与PKCα功能和隐孢子虫感染易感性的关系将如何 洞察一种重要的人类致病病原体的发病机制。我们假设 宿主PKCα是隐孢子虫通过肌动蛋白重塑侵袭肠上皮所必需的 细胞骨架。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 肠上皮侵袭的体内外研究(II)SNPs对PRKCA表达的影响 和下游功能，以及(Iii)确定PRKCA的表达是否调节儿童疾病的严重程度。至 要做到这一点，研究建议包括积极调查隐孢子虫在现场和在 长凳上。在这里，我们建议通过使用分子工具来连接两种科学发现的方法 检查PRKCA并监测基因分型人类儿童的临床结果。的首要目标是 这些独立但相互作用的目标将是东道主pkcα的翻译开发作为潜在的 目标是推进急需的隐孢子虫病治疗。