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: 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
• 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

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.

项目摘要 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 估计负责5岁以下儿童死亡率的10％，以及最近的全球负担 腹泻死亡率的疾病分析发现，隐孢子虫仅次于轮状病毒作为原因 五岁以下儿童死亡（5）。 Nitazoxanide，目前是唯一获得cryptosporidia的批准药物 感染仅适用于治疗免疫能力患者。此外，这只是 在严重免疫功能低下的患者中相当于安慰剂。隐孢子虫的临床表现 感染的严重程度是可变的。一些患者患有严重的腹泻病，而某些感染仍然存在 亚临床。尽管疾病广泛的原因知之甚少，但有几项研究已经 证明了渐近和症状感染的长期临床意义。我们执行了一个 全基因组协会研究（GWAS）在生命的第一年内对孟加拉国的婴儿。我们评估了 在人类基因组和 三个独立患者队列中的有症状隐孢子虫感染。该分析表明了高度 PRKCA基因中多个SNP的显着统计关联和对隐孢子虫的易感性 腹泻。风险等位基因的每个副本将收缩隐孢子虫病的风险增加2.4倍 生命年。 PRKCA中最重要的SNP是GTEX数据库中的EQTL 在PKCα和隐孢子虫之间尚未建立。 PKCα激酶是 肌动蛋白和细胞内侵袭期间，隐孢子虫会影响宿主细胞肌动蛋白的重塑。进一步调查 PRKCA基因变异与PKCα功能和对隐孢子虫感染的敏感性有关 洞悉引起疾病的重要人类病原体的发病机理。我们假设这一点 通过重塑肌动蛋白，宿主PKCα是肠道上皮侵入肠上皮所必需的 细胞骨架。基于此初步数据，我们旨在表征（i）PKCα在隐孢子虫中的作用 体外和体内肠上皮的入侵定义了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。到 做到这一点，研究建议包括对现场和AT的隐孢子虫的积极研究 长凳。在这里，我们提出了通过使用分子工具来桥接科学发现的两种方法 检查prkca并监测基因分型的人类儿童的临床结果。总体目标 这些独立但相互作用的目的将是宿主PKCα的翻译发展作为潜力 促进隐孢子虫病的迫切需要治疗的目标。