SBIR-STTR Award

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.

Public Health Relevance Statement:
Narrative Alcoholic hepatitis (AH) is a severe and acute form of alcoholic liver disease that develops in approximately one- third of chronic and heavy drinkers, is associated with high mortality, and represents a considerable healthcare and economic burden in the U.S. The standard treatment for AH, prednisolone, has not changed in over 40 years, only benefits ~50% of patients, and is associated with a significant increase in the rate of serious infections. Rejuvenation Technologies, Inc.'s novel platform for therapeutic telomere extension has the potential to translate into a new and improved treatment for AH, improving patient outcomes and survival, as well as serving as a basis for a number of other therapeutic applications in which shortened telomeres have been implicated, including bone marrow failure, pulmonary fibrosis, immunodeficiency, osteoporosis, cardiovascular disease, Alzheimer's disease, and multiple types of cancer.

Project Terms:
Acute; Albumins; Alcoholic beverage heavy drinker; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alzheimer's Disease; Animals; anti-cancer; Apoptosis; beta-Galactosidase; Bile Acids; Bilirubin; Biological Assay; Blood; Blood Chemical Analysis; cancer type; Cardiovascular Diseases; CDKN2A gene; Cell Aging; Cell Death; Cell division; Cells; Cerebrovascular Disorders; Chromosomes; Chronic; chronic liver injury; Cirrhosis; Clinical; combat; cytokine; Data; Development; DNA; DNA Damage; DNA Sequence; Dose; Economic Burden; Encapsulated; Exhibits; feeding; Fibrosis; Frequencies; Future; health care economics; Heavy Drinking; Hepatic Stellate Cell; Hepatocyte; Histologic; Histology; Hospitals; Hour; Human; Hybrids; Immunologic Deficiency Syndromes; Impairment; improved; In Situ Nick-End Labeling; Individual; Infection; Inflammatory; Intravenous; Knock-out; Knockout Mice; Lead; Length; lipid nanoparticle; Liver; liver cell proliferation; Liver Cirrhosis; Liver diseases; Liver Failure; Liver Fibrosis; Measures; Messenger RNA; Methods; Modeling; mortality; mouse model; Mus; Mutation; Myocardial Infarction; novel; novel therapeutics; Nucleosides; Osteoporosis; Pancytopenia; Partial Hepatectomy; Patient-Focused Outcomes; Patients; Phase; Phenotype; preclinical safety; prednisolone; Preparation; preservation; prevent; Proliferating; Proteins; Pulmonary Fibrosis; regenerative; Regimen; Rejuvenation; response; RNA-Directed DNA Polymerase; Role; Safety; senescence; Stains; standard care; stellate cell; Steroids; Symptoms; Technology; Telomerase; telomere; Telomere Shortening; Testing; Therapeutic; Time; Toxicology; Translating; Visit; Wild Type Mouse

Rejuvenation Technologies Inc. (RTI) is taking the next translational step following their recent highly successfulproof-of-concept rescue of multiple mouse models that mimic key pathological features of alcoholic hepatitis(AH). AH is an acute form of alcoholic liver disease in which the clinical response to loss of the liver's regenerativecapacity from long-term damage includes liver failure. AH occurs in ~1/3 of heavy and chronic drinkers, and itsseverity increases with the amount and duration of alcohol consumption. AH patients overall have poorprognoses and high short-term mortality, with a 1-year survival rate of 25-50%, while those with severe AH mayshow features of acute-on-chronic liver failure and exhibit a 40-50% mortality rate at 1 month from presentation.There were over 300,000 hospital admissions for AH in 2017, and AH hospital admissions increased by over50% during the COVID pandemic due to increased alcohol consumption that is expected to impact AHepidemiology for years to come. The only treatments are steroids, which are ineffective at reducing patientmortality and can increase risk of infection. There is strong evidence that short telomeres play a causal role inAH. Telomeres are the protective DNA tips of chromosomes essential for cell and liver health. Chronic liver injurycaused by excessive alcohol consumption induces continuous hepatocyte turnover, which causes rapid telomereshortening. When telomeres become too short, hepatocytes senesce and secrete senescence-associatedsecretory phenotype factors that activate hepatic stellate cells, causing them to become fibrogenic. Transienttelomerase to extend telomeres in the liver represents a promising strategy to modify AH progression. RTI hasinvented a breakthrough treatment comprising: 1) the telomere-extending biologic telomerase (TERT) mRNAand 2) a lipid nanoparticle (LNP) vehicle that delivers mRNA to the liver with world-leading efficiency, transfecting>99% of hepatocytes at very low doses (0.05 mg/kg), even in cirrhotic livers. During the Phase I SBIR project,RTI demonstrated that intravenously (i.v.) injected TERT mRNA LNPs reduced high-grade inflammation by 60%,senescence by 30%, and liver fibrosis by 38% and increased median survival by 42% in humanized telomerelength (TERT KO) mice with thioacetamide (TAA)-induced liver disease. TERT mRNA LNPs reduced liverfibrosis by 74%, infiltrating T-cells by 33%, and DNA damage in hepatocytes by 73% in a preliminary study of anacute-on-chronic model of AH. For Phase II, RTI will advance this product by undertaking three specific aims: 1)investigating drug pharmacology in an acute-on-chronic liver disease model, mimicking the proposed AH clinicaltreatment scenario, 2) optimizing drug product and scaling up manufacturing, and 3) piloting large animaltoxicology, pharmacokinetics, and biodistribution studies. This will provide critical efficacy and toxicology data tosupport a pre-IND application, paving the way for IND-enabling studies and RTI's first-in-human clinical trials ofTERT mRNA LNPs in AH patients. Should these prove successful, RTI will expand to other liver indications, asshortened telomeres are a pathological hallmark of cirrhotic liver disease of any etiology.

Public Health Relevance Statement:
Narrative Alcoholic hepatitis (AH) is an acute form of alcoholic liver disease (ALD) with high short-term mortality, with over 450,000 hospital admissions per year and a 40-50% mortality rate (for severe AH) at 1 month from presentation. Chronic liver injury from excessive alcohol consumption leads to telomere shortening in the liver, and there is strong evidence that telomere extension may improve AH patient survival. This project will advance Rejuvenation Technologies' breakthrough treatment for AH, comprised of telomerase mRNA that extends telomeres transiently, the same mechanism used by stem cells, and a lipid nanoparticle vehicle that delivers the telomerase mRNA to the liver with high efficiency and low toxicity.

Project Terms: