Investigating the sex-specific roles of PKLR and CHCHD6 in insulin resistance and NAFLD

研究 PKLR 和 CHCHD6 在胰岛素抵抗和 NAFLD 中的性别特异性作用

• 批准号: 10323755
• 负责人: Karthickeyan Chella Krishnan
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-12 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10323755
• 关键词: Address; Atherosclerosis; Automobile Driving; Award; Bioenergetics; Candidate Disease Gene; Cardiovascular Diseases; Cells; Cirrhosis; Complex; Development; Development Plans; Disease; Disease Resistance; Etiology; Fatty Liver; Fatty acid glycerol esters; Female; Fibrosis; Frequencies; Genes; Genetic; Goals; Hepatic; Hepatocyte; Heterogeneity; Hybrids; Individual; Insulin; Insulin Resistance; Investigation; Knowledge; Lead; Link; Liver; Liver Failure; Liver Fibrosis; Liver Mitochondria; Malignant neoplasm of liver; Measures; Mediating; Mentors; Metabolic; Metabolic Pathway; Metabolic syndrome; Mitochondria; Modeling; Molecular; Mus; Obesity; Obesity Epidemic; Outcome; Pathway interactions; Phase; Phenotype; Physiology; Population; Population Study; Prevalence; Protein Isoforms; Public Health; Publishing; Pyruvate Kinase; Regulation; Research; Research Personnel; Role; Sex Differences; System; Talents; Testing; Therapeutic; Training; Triglycerides; United States; Validation; base; blood glucose regulation; cardiovascular disorder risk; cardiovascular risk factor; career; career development; chronic liver disease; cohort; disease phenotype; experience; experimental analysis; experimental study; fasting glucose; follow-up; genetic analysis; genotypic sex; glucose tolerance; improved; insulin sensitivity; insulin tolerance; intrahepatic; knock-down; lipid metabolism; liver metabolism; liver transplantation; male; men; metabolic phenotype; mitochondrial dysfunction; mitochondrial metabolism; mouse model; multiple omics; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; overexpression; personalized therapeutic; prevent; sex; single cell sequencing; therapy design; trait; transcriptome; transcriptomics

PROJECT SUMMARY/ABSTRACT Non-alcoholic fatty liver disease (NAFLD) is increasingly being recognized as the hepatic manifestation of metabolic syndrome that can lead to cardiovascular diseases. Currently, very little is known about the genetic and molecular factors underlying NAFLD. Besides, no known therapeutic strategies are available to prevent or treat the condition. To address this problem, we applied an integrative multi-omics approach using an extensively phenotyped mouse cohort, Hybrid Mouse Diversity Panel (HMDP) and identified many known and several novel candidate genes regulating NAFLD. We further validated two novel candidates, pyruvate kinase, liver isoform (PKLR) and coiled-coil-helix-coiled-coil-helix domain containing 6 (CHCHD6). When these two genes were knockdown in male mouse models of steatosis, both NAFLD and insulin sensitivity improved as measured by intrahepatic triglyceride accumulation levels, glucose- and insulin- tolerance tests, and fasting glucose and insulin levels. Follow-up bioenergetic analyses revealed these two genes regulated mitochondrial metabolism in developing steatosis and insulin resistance. When we further expanded this to include the female HMDP strains, we found that these two genes have no effect. Specifically, correlation analyses revealed PKLR and CHCHD6 do not develop NAFLD and insulin resistance in female strains, respectively. To further our understanding on the gene-by-sex interactions mediated by these two genes in regulating mitochondrial metabolism and in developing NAFLD and insulin resistance, I have proposed three interrelated aims to examine 1) mechanisms underlying PKLR- and CHCHD6- mediated lipid metabolism, 2) metabolic consequences of liver-specific overexpression or knockdown in female mouse models of steatosis, 3) gene networks and cellular heterogeneity altered by liver-specific overexpression or knockdown in mouse models of advanced NASH and fibrosis. Furthermore, I have developed a detailed five-year career development plan that includes two-year mentored phase (K99 phase) followed by three-year independent phase (R00 phase) for my transition into an independent academic career. I have included a highly talented and distinguished panel of advisors including Drs. Aldons J. Lusis, Orian Shirihai, Neil Kaplowitz, Andrea Hevener, Peter Tontonoz, and Steven Bensinger, to assist in my transition to independence. I have significant experience in studying how host genetics and sex differences dictate the outcomes of complex disease traits using systems genetics analyses. I now plan to expand my training in liver and mitochondrial physiology, lipid metabolism, metabolic phenotyping and single cell sequencing. Overall, the goal is to advance my training to become an independent investigator and perform research bridging systems genetics, mitochondria and liver metabolism.

项目概要/摘要 非酒精性脂肪性肝病（NAFLD）越来越被认为是以下疾病的肝脏表现： 可导致心血管疾病的代谢综合征。目前，人们对遗传基因知之甚少 以及 NAFLD 的分子因素。此外，尚无已知的治疗策略可用于预防或 治疗病情。为了解决这个问题，我们应用了综合多组学方法，广泛使用 对小鼠群体进行表型分析，混合小鼠多样性面板 (HMDP) 并鉴定了许多已知的和一些新颖的 调节 NAFLD 的候选基因。我们进一步验证了两种新的候选物，丙酮酸激酶，肝脏亚型 (PKLR) 和含有 6 的卷曲螺旋-螺旋-卷曲螺旋-螺旋结构域 (CHCHD6)。当这两个基因 在雄性小鼠脂肪变性模型中进行基因敲除后，NAFLD 和胰岛素敏感性均得到改善（测量结果为） 肝内甘油三酯蓄积水平、葡萄糖和胰岛素耐量试验以及空腹血糖和 胰岛素水平。后续的生物能分析表明这两个基因调节线粒体代谢 发生脂肪变性和胰岛素抵抗。当我们进一步扩大范围以包括雌性 HMDP 品系时， 我们发现这两个基因没有作用。具体来说，相关性分析揭示了 PKLR 和 CHCHD6 雌性菌株中分别不会出现 NAFLD 和胰岛素抵抗。为了进一步加深我们对 这两个基因介导的基因与性别相互作用在调节线粒体代谢和 针对 NAFLD 和胰岛素抵抗的发展，我提出了三个相互关联的目标来研究 1) 机制 潜在的 PKLR 和 CHCHD6 介导的脂质代谢，2) 肝脏特异性的代谢后果 雌性脂肪变性小鼠模型中的过度表达或敲低，3）基因网络和细胞异质性 在晚期 NASH 和纤维化小鼠模型中，肝脏特异性过度表达或敲低会改变。 此外，我还制定了详细的五年职业发展计划，其中包括两年 指导阶段（K99 阶段），然后是三年独立阶段（R00 阶段），以帮助我过渡到 独立的学术生涯。我组建了一个才华横溢、杰出的顾问小组，其中包括 博士。 Aldons J. Lusis、Orian Shirihai、Neil Kaplowitz、Andrea Hevener、Peter Tontonoz 和 Steven Bensinger， 帮助我过渡到独立。我在研究宿主遗传学和性别如何影响方面拥有丰富的经验 使用系统遗传学分析，差异决定了复杂疾病特征的结果。我现在计划 扩大我在肝脏和线粒体生理学、脂质代谢、代谢表型和单细胞方面的训练 测序。总体而言，目标是推进我的培训，成为一名独立调查员并执行 研究桥接系统遗传学、线粒体和肝脏代谢。