Preventing liver fibrosis in alcoholic hepatitis by enhancing liver regenerative capacity via transient telomere extension using lipid nanoparticle-encapsulated TERT mRNA

使用脂质纳米颗粒封装的 TERT mRNA 进行短暂端粒延伸，增强肝脏再生能力，从而预防酒精性肝炎中的肝纤维化

• 批准号: 10082259
• 负责人: John Ramunas
• 金额: $ 27.24万
• 依托单位: REJUVENATION TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-10 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10082259
• 关键词: Acute; Albumins; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcoholic beverage heavy drinker; Alzheimer' s Disease; Animals; Apoptosis; Bile Acids; Bilirubin; Biological Assay; Blood; Blood Chemical Analysis; CDKN2A gene; Cardiovascular Diseases; Cell Aging; Cell Death; Cell division; Cells; Cerebrovascular Disorders; Chromosomes; Chronic; Cirrhosis; Clinical; DNA; DNA Damage; DNA Sequence; Data; Development; Dose; Economic Burden; Encapsulated; Exhibits; Fibrosis; Frequencies; Future; Heavy Drinking; Hepatic Stellate Cell; Hepatocyte; Histologic; Histology; Hospitals; Hour; Human; Hybrids; Immunologic Deficiency Syndromes; Impairment; In Situ Nick-End Labeling; Individual; Infection; Inflammatory; Intravenous; Knock-out; Knockout Mice; Lead; Length; Liver; Liver Cirrhosis; Liver Failure; Liver Fibrosis; Liver diseases; Measures; Messenger RNA; Methods; Modeling; Mus; Mutation; Myocardial Infarction; Nucleosides; Osteoporosis; Pancytopenia; Partial Hepatectomy; Patient-Focused Outcomes; Patients; Phase; Phenotype; Preparation; Proliferating; Proteins; Pulmonary Fibrosis; RNA-Directed DNA Polymerase; Regimen; Rejuvenation; Role; Safety; Stains; Steroids; Symptoms; Technology; Telomerase; Telomere Shortening; Testing; Therapeutic; Time; Toxicology; Translating; Visit; Wild Type Mouse; anti-cancer; beta-Galactosidase; cancer type; chronic liver injury; combat; cytokine; feeding; health care economics; improved; lipid nanoparticle; liver cell proliferation; mortality; mouse model; novel; novel therapeutics; preclinical safety; prednisolone; preservation; prevent; regenerative; response; senescence; standard care; stellate cell; telomere

Abstract Rejuvenation Technologies Inc. (RTI) aims to prevent liver fibrosis in alcoholic hepatitis (AH) by enhancing liver regenerative capacity. AH is an acute form of alcoholic liver disease with mortality of up to 50% within 1 month of presentation. Most AH patients exhibit advanced fibrosis/cirrhosis, which contributes to acute-on-chronic liver failure. RTI will ameliorate/prevent this fibrosis by implementing a method for the therapeutic extension of telomeres, the DNA sequences that protect chromosome ends. Telomeres naturally shorten over time and with cell division, eventually exposing the DNA end and triggering a DNA damage response that induces cell senescence and death. Accelerated telomere attrition has been identified as a plausible driver of fibrosis in AH and other fibrotic liver diseases. Thus, to combat AH-related fibrosis, RTI will use lipid nanoparticles (LNPs) to encapsulate nucleoside-modified mRNA (modRNA) encoding the telomerase reverse transcriptase (TERT) protein to transiently extend telomeres in proliferating hepatocytes. Animal studies have demonstrated the potential of this approach, as telomere extension reduces hepatocyte loss and fibrosis in a mouse model of liver cirrhosis. Moreover, RTI's preliminary results have shown that a single intravenous dose of TERT LNPs in mice extends liver telomeres by an average of 230 bp, reversing the equivalent of 5 years of telomere shortening in humans. Notably, TERT LNPs only increase telomerase activity for about 24 hours, after which the extended telomeres resume shortening at their normal rate, leaving the important anti-cancer telomere shortening mechanism intact. In this Phase I project, RTI will: 1) confirm that TERT knockout (TERT KO) and the resulting shortened telomeres exacerbate AH symptoms in the hybrid Tsukamoto-French (HTF) mouse model, the model that most closely reproduces the histologic and clinical features of AH, and 2) assess whether TERT LNPs ameliorate liver fibrosis and AH in TERT KO and wild-type mice in the HTF model. Completion of this project will demonstrate the key role of shortened telomeres in AH-related fibrosis, as well as the efficacy of TERT LNPs at extending telomeres and preventing fibrosis in a mouse model. This will pave the way for future toxicology testing to establish the preclinical safety of TERT LNPs in preparation of an IND application. Ultimately, successful development of TERT LNPs will lead to improved treatment and survival of patients with AH.

摘要 Rejuvenation Technologies Inc. (RTI)目的是通过增强肝脏功能来预防酒精性肝炎（AH）的肝纤维化。 再生能力AH是一种急性酒精性肝病，1个月内死亡率高达50%。 的介绍。大多数AH患者表现出晚期纤维化/肝硬化，这有助于慢性肝急性发作 失败RTI将通过实施一种用于治疗扩展的方法来改善/预防这种纤维化。 端粒，保护染色体末端的DNA序列。端粒随着时间的推移自然缩短， 细胞分裂，最终暴露DNA末端并引发DNA损伤反应， 衰老和死亡加速端粒磨损已被确定为AH纤维化的合理驱动因素 和其他肝纤维化疾病。因此，为了对抗AH相关的纤维化，RTI将使用脂质纳米颗粒（LNP）， 编码端粒酶逆转录酶（TERT）的包裹核苷修饰mRNA（modRNA） 在增殖的肝细胞中瞬时延长端粒的蛋白质。动物研究表明， 这种方法的潜力，因为端粒延伸减少了小鼠肝脏模型中的肝细胞损失和纤维化。 肝硬化此外，RTI的初步结果表明，在小鼠中单次静脉注射TERTLNP， 延长肝脏端粒平均230 bp，逆转相当于5年的端粒缩短， 人类值得注意的是，TERTLNP仅在约24小时内增加端粒酶活性，之后延长的端粒酶活性持续时间。 端粒以正常速率恢复缩短，留下重要的抗癌端粒缩短。 机制完好。在这个I期项目中，RTI将：1）确认TERT敲除（TERTKO）和由此产生的 缩短的端粒加剧了Tsukamoto-French（HTF）杂交小鼠模型中的AH症状，该模型 最接近地再现AH的组织学和临床特征，和2）评估TERTLNP是否 改善HTF模型中的TERT KO和野生型小鼠中的肝纤维化和AH。该项目的完成将 证明了端粒缩短在AH相关纤维化中的关键作用，以及TERTLNP在AH相关纤维化中的疗效。 在小鼠模型中延长端粒和预防纤维化。这将为未来的毒理学测试铺平道路 确定TERT LNP在IND申请准备中的临床前安全性。最终，成功 TERTLNP的开发将导致AH患者的治疗和存活的改善。