A Precision Targeted Therapeutics for Nonalcoholic Fatty Liver Disease

非酒精性脂肪肝的精准靶向治疗

• 批准号: 10614050
• 负责人: Wanqing Liu
• 金额: $ 42万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614050
• 关键词: Acetylgalactosamine; Alcoholic Liver Diseases; Alleles; Automobile Driving; Binding; Biological Assay; Classification; Clinical; DNA Sequence Alteration; Data; Development; Disease; Dominant-Negative Mutation; Down-Regulation; Drug Delivery Systems; FDA approved; Fatty Liver; Fibrosis; Gene Deletion; Genes; Genetic; Genetic Medicine; Genetic Transcription; Goals; Health; Hepatic; Histology; Human; Hydrolase; Hypertriglyceridemia; In Vitro; Inflammation; Investigational Drugs; Lead; Legal patent; Lipids; Lipolysis; Liver; Liver diseases; Mediating; Medicine; Messenger RNA; Metabolic; Modeling; Modification; Mutation; Names; Oligonucleotides; Patients; Pharmaceutical Preparations; Pharmacodynamics; Pharmacotherapy; Phenotype; Phospholipase; Precision therapeutics; Prevention; Property; Protein Isoforms; Proteins; RNA Interference; RNA delivery; Role; Safety; Series; Small Interfering RNA; Specificity; Testing; Therapeutic; Therapeutic Agents; Time; Toxic effect; Toxicokinetics; Translations; Triglycerides; Variant; chronic liver disease; comorbidity; comparative efficacy; design; disease phenotype; disorder subtype; drug development; effective therapy; efficacy evaluation; first-in-human; genetic risk factor; genome wide association study; hepatic lipase; human disease; humanized mouse; improved; in vivo; knock-down; mouse model; mutant; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; novel strategies; overexpression; pharmacologic; pre-clinical; precision genetics; preclinical evaluation; prototype; screening; social; success; targeted treatment; therapeutic candidate; therapeutic target; tool; treatment strategy

Abstract The ultimate goal of this project is to explore the feasibility and mechanism of an allele-specific, RNAi-based genetic medicine approach to treating nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). NAFLD/NASH is the most common chronic liver disease worldwide. However, no proven drug for NAFLD/NASH is available. There is an urgent need to identify novel targets and develop promising drugs for both NASH prevention and treatment. Recent GWAS has identified that a genetic mutation (148I>M) in the patatin-like phospholipase domain containing 3 (PNPLA3) gene is the single largest genetic risk factor contributing to the progression of NAFLD to NASH. Our and others’ studies demonstrated that the PNPLA3148M- associated NASH should be classified as a unique disease. Mechanistically, our team recently demonstrated that accumulated PNPLA3148M protein isoform on lipid droplets competes with PNPLA2 (also known as ATGL) for α/β-hydrolase-domain-containing 5 (ABHD5, or CGI-58), an important activator for PNPLA2, blocking the ABHD5-mediated, PNPLA2-driving lipolysis and leading to triglycerides accumulation in lipid droplets. We thus hypothesize that transcriptional downregulation of the PNPLA3148M isoform is a cause-targeting strategy for the treatment of NAFLD/NASH among patients carrying the PNPLA3148M allele. To this end, we have developed and patented an allele-specific small interfering RNA (siRNA) that possesses excellent potency and specificity for downregulating the PNPLA3148M mutant isoform but not its wild-type isoform (148I). We have further discovered two more modified oligonucleotides with excellent pharmacological properties. To facilitate the preclinical evaluation of these agents, we have developed a humanized PNPLA3148I/M mouse model, which recapitulates the typical NASH phenotype and related comorbidities. Delivering our siRNA into this mouse model with N- Acetylgalactosamine (GalNAc)-conjugation, a well-established, clinically validated, and liver-specific RNAi drug delivery strategy effectively knocked down hepatic hPNPLA3148M mRNA and ameliorated the liver histology. With this success, we propose to advance our drug development by evaluating the efficacy, toxicity, and mechanism of action of our therapeutics for both NASH prevention and treatment, with the final goal to generate the first precision genetic medicine for PNPLA3148M-specific NASH. We aim in this study 1) to evaluate and compare the efficacy of our 3 hPNPLA3148M-targeting, GalNAc-conjugated oligonucleotide therapeutics; 2) to evaluate and compare the safety and toxicity of our therapeutic candidates; and 3) to explore the mechanism of action of our therapeutics. We expect that via this study, we will eventually develop the first genetic medicine for NAFLD/NASH prevention/treatment, which could also serve as a new prototype for drug development for other common human diseases. The collected data in this study will generate the most promising therapeutic lead for late-stage pre- clinical and the first-in-human studies.

摘要 本项目的最终目的是探索等位基因特异性的、基于RNAi的基因工程技术的可行性和作用机制。 治疗非酒精性脂肪性肝病（NAFLD）和非酒精性脂肪性肝炎的遗传医学方法 （NASH）。NAFLD/NASH是全球最常见的慢性肝病。然而，没有经过验证的药物 NAFLD/NASH可用。迫切需要确定新的靶点并开发有前途的药物， NASH的预防和治疗。最近的GWAS已经确定，在细胞中的一个基因突变（148 I>M）， patatin-like phospholipase domain containing 3（PNPLA 3）基因是最大的遗传危险因子 促进NAFLD向NASH的进展。我们和其他人的研究表明，PNPLA 3148 M- 因此，NASH应该被归类为一种独特的疾病。从机制上讲，我们的团队最近证明了 脂滴上积累的PNPLA 3148 M蛋白同种型与PNPLA 2（也称为ATGL）竞争， 对于含有α/β-水解酶结构域的5（ABHD 5或CGI-58），其是PNPLA 2的重要激活剂，阻断了 ABHD 5介导的、PNPLA 2驱动的脂解并导致甘油三酯在脂滴中积累。我们因此 假设PNPLA 3148 M同种型转录下调是一种病因靶向策略， 在携带PNPLA 3148 M等位基因的患者中治疗NAFLD/NASH。为此，我们制定了 获得专利的等位基因特异性小干扰RNA（siRNA），具有优异的效力和特异性， 下调PNPLA 3148 M突变体同种型，但不下调其野生型同种型（148 I）。我们进一步发现 另外两种具有优异药理学性质的修饰寡核苷酸。为了促进临床前 为了评价这些药物，我们开发了人源化PNPLA 3148 I/M小鼠模型，其概括了 典型NASH表型和相关合并症。将我们的siRNA输送到这个小鼠模型中， 乙酰半乳糖胺（GalNAc）结合，一种成熟的，临床验证的肝脏特异性RNAi药物 递送策略有效地敲低肝hPNPLA 3148 M mRNA并改善肝组织学。与 这一成功，我们建议通过评估疗效，毒性和机制来推进我们的药物开发 我们的治疗药物对NASH预防和治疗的作用，最终目标是产生第一个 PNPLA 3148 M特异性NASH的精准基因药物。我们在这项研究中的目的是1）评估和比较 我们的3 hPNPLA 3148 M靶向的GalNAc缀合的寡核苷酸治疗剂的功效; 2）评估和 比较我们的治疗候选药物的安全性和毒性; 3）探索我们的作用机制， 治疗学我们希望通过这项研究，我们最终将开发出第一种用于NAFLD/NASH的基因药物 预防/治疗，这也可以作为一个新的原型药物开发，为其他共同的人类 疾病本研究中收集的数据将为晚期前 临床和首次人体研究。

项目成果

Phosphatidylethanolamines Are Associated with Nonalcoholic Fatty Liver Disease (NAFLD) in Obese Adults and Induce Liver Cell Metabolic Perturbations and Hepatic Stellate Cell Activation.

• DOI: 10.3390/ijms24021034
• 发表时间: 2023-01-05
• 期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
• 影响因子: 5.6
• 作者: Shama, Samaa;Jang, Hyejeong;Wang, Xiaokun;Zhang, Yang;Shahin, Nancy Nabil;Motawi, Tarek Kamal;Kim, Seongho;Gawrieh, Samer;Liu, Wanqing
• 通讯作者: Liu, Wanqing

Mendelian Randomization Analysis Reveals No Causal Relationship Between Nonalcoholic Fatty Liver Disease and Severe COVID-19.

• DOI: 10.1016/j.cgh.2022.01.045
• 发表时间: 2022-07
• 期刊: Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association
• 作者: Li J;Tian A;Zhu H;Chen L;Wen J;Liu W;Chen P
• 通讯作者: Chen P