TERT mRNA lipid nanoparticles to extend telomeres to treat pulmonary fibrosis

摘要 短端粒，染色体的DNA末端，驱动在人类中确定的多种关键致病机制。 特发性肺纤维化（IPF）患者。Rejuvenation Technologies正在开发第一个安全有效的 干预以延长肺中的短端粒，从而延长IPF患者的生存期。IPF的特征是 肺组织的进行性瘢痕形成，导致血液中缺氧，并最终导致呼吸道疾病。 失败IPF影响多达200，000名美国人，每年有多达50，000例新发病例。IPF患者的中位 从诊断之日起生存期少于5年，即使采用标准护理治疗。增加 然而，有证据表明端粒缩短在IPF的病因学中具有致病作用。功能丧失 在2-5%的IPF患者和高达15%的家族性PF患者中发现端粒酶突变。小鼠 在IPF小鼠模型中，缩短的端粒表现出对纤维化的易感性增加。此外，端粒 使用TERTDNA在小鼠中的延伸（由于基因组整合的风险，其对人类是不安全的）减少了 纤维化和改善肺功能。在IPF患者中确定的几种关键致病机制也是 严重短端粒的后果，包括细胞衰老，TGFβ升高和其他炎症 介质、慢性炎症、肌成纤维细胞活化、祖细胞损失和增殖性降低 剩余祖细胞的能力。这些发现为开发一种安全的方法提供了强有力的理论基础， 延长端粒以治疗IPF。RTI建议使用脂质纳米颗粒（LNP）包封TERTmRNA（TERT LNP）来延长肺中的端粒以治疗IPF。RTI专有的LNP肺输送载体的成功率>90% 肺上皮细胞的端粒，以及小鼠体内单次静脉注射TERTmRNA， 平均230 bp，逆转了人类端粒缩短数年的效果。重要的是为了安全， TERTmRNA仅在一天左右增加端粒酶活性，之后延长的端粒恢复 端粒以正常速率缩短，使重要的抗癌端粒缩短机制完好无损。RTI 证明i. v. -注射的TERTmRNA LNP使存活率增加210%，使纤维化减少68%， 在IPF的人源化端粒长度（TERT KO）小鼠博来霉素模型中，肺功能改善58%。到 在IND批准之前，该快速通道项目将完成以下具体目标。第一阶段：1） i. v. -100的药代动力学（PK）和剂量确定注射的TERTmRNA LNP。2)药效学（PD）， 生物标志物和与FDA批准的IPF药物的比较研究。3)IPF患者细胞中的药理学。第二阶段： 4)在第二个小鼠模型中测定疗效（二氧化硅）。5)用于生产和放大TERT的CMC活动 mRNA LNP产生。6)进行IND启用毒理学和药理学研究。如果成功，这些 研究将提供一种新方法的概念证明，以保护肺功能，减少纤维化， IPF的生存率。TERT LNP的商业化将为IPF患者和临床医生提供急需的治疗药物， 选择改善结果和生存。

Abstract Short telomeres, the DNA tips of chromosomes, drive multiple key pathogenic mechanisms identified in idiopathic pulmonary fibrosis (IPF) patients. Rejuvenation Technologies is developing the first safe and effective intervention to extend short telomeres in lung and thereby extend IPF patient survival. IPF is characterized by progressive scarring of lung tissue, leading to a lack of oxygen in the blood, and ultimately resulting in respiratory failure. IPF affects up to 200,000 Americans, with up to 50,000 new cases each year. IPF patients have a median survival of less than 5 years from the time of diagnosis, even with standard of care treatment. Increasing evidence, however, points to a causative role of shortened telomeres in the etiology of IPF. Loss-of-function mutations in telomerase are found in 2–5% of IPF patients and up to 15% of familial PF patients. Mice with shortened telomeres exhibit increased susceptibility to fibrosis in a mouse model of IPF. Moreover, telomere extension in mice using TERT DNA (which is not safe for humans due to the risk of genomic integration) reduces fibrosis and improves lung function. Several key pathogenic mechanisms identified in IPF patients are also consequences of critically short telomeres, including cellular senescence, elevated TGFβ and other inflammatory mediators, chronic inflammation, myofibroblast activation, loss of progenitor cells, and reduced proliferative capacity of remaining progenitor cells. These findings provide a strong rationale for developing a safe method to extend telomeres to treat IPF. RTI proposes to use lipid nanoparticles (LNPs) encapsulating TERT mRNA (TERT LNPs) to extend telomeres in the lung to treat IPF. RTI’s proprietary LNP lung delivery vehicle transfects >90% of lung epithelial cells, and a single intravenous dose of TERT mRNA in mice extends telomeres in vivo by an average of 230 bp, reversing the equivalent of years of telomere shortening in humans. Importantly for safety, TERT mRNA only increases telomerase activity for about a day, after which the extended telomeres resume shortening at their normal rate, leaving the important anti-cancer telomere shortening mechanism intact. RTI demonstrated that i.v.-injected TERT mRNA LNPs increase survival by 210%, reduce fibrosis by 68%, and improve lung function by 58% in the humanized telomere length (TERT KO) mouse bleomycin model of IPF. To advance to IND approval, this Fast Track project will complete the following Specific Aims. Phase I: 1) Pharmacokinetics (PK) and dose determination of i.v.-injected TERT mRNA LNPs. 2) Pharmacodynamics (PD), biomarker, and comparative studies to FDA approved IPF drugs. 3) Pharmacology in IPF patient cells. Phase II: 4) Determine efficacy in second mouse model (silica). 5) CMC activities for manufacturing and scale-up of TERT mRNA LNP production. 6) Perform IND-enabling toxicology and pharmacology studies. If successful, these studies will provide proof of concept of a novel approach to preserve lung function, reduce fibrosis, and extend survival in IPF. Commercialization of TERT LNPs will give IPF patients and clinicians a much-needed therapeutic option to improve outcomes and survival.