Molecular drivers of ageing and emerging longevity therapies

Abstract

Ageing is a gradual, multifactorial process that leads to the deterioration of physical and mental health, increasing the risk of disease and eventually death. Indicators of ageing manifest at the molecular level, including genomic instability, telomere attrition, epigenetic alterations, mitochondrial dysfunction, loss of proteostasis, and dysregulation of key signalling pathways such as the mechanistic target of rapamycin (mTOR) and insulin signalling. These molecular hallmarks of ageing are interconnected, amplifying one another over time. The resulting cellular stress triggers apoptosis or drives cells into a pathological state known as cellular senescence, in which they secrete inflammatory, pro-ageing factors. Consequently, there is a progressive decline in tissue function and regenerative capacity, accompanied by atrophy and stem cell exhaustion under a chronically inflamed microenvironment. Although functional decline with age is irreversible, research indicates it can be delayed. In this review, we discuss the hallmarks of ageing, conventional pharmacological interventions with demonstrated anti-ageing effects in cellular and animal models, and emerging therapeutic strategies being explored as ageing becomes increasingly recognized as a major risk factor for disease development.

Keywords

ageing / inflammation / senescence / hallmarks of ageing / therapeutic strategies

References

[1] Dattani S, Rodés—Guirao L, Ritchie H, Ortiz—Ospina E, Roser M. Life Expectancy [Internet]. [cited 2025 Aug 11]. Available from: https://ourworldindata.org/life—expectancy

[2] GBD 2021 Fertility and Forecasting Collaborators. Global fertility in 204 countries and territories, 1950—2021, with forecasts to 2100: a comprehensive demographic analysis for the Global Burden of Disease Study 2021. Lancet. 2024; 403: 2057-99.

[3] State of World Population 2023: 8 Billion Lives, Infinite Possibilities — The case for rights and choices [Internet]. reliefweb; c2026 [cited 2025 Jul 13]. Available from: https://reliefweb.int/report/world/state—world—population—2023—8—billion—lives—infinite—possibilities—case—rights—and—choices—enarru

[4] World Population Prospects 2022: Summary of Results [Internet]. [cited 2025 Jul 13]. Available from: https://www.un.org/development/desa/pd/content/World—Population—Prospects—2022

[5] Araujo de Carvalho I, Epping—Jordan J, Pot AM, Kelley E, Toro N, Thiyagarajan JA, et al. Organizing integrated health—care services to meet older people’s needs. Bull World Health Organ. 2017; 95: 756-63.

[6] López—Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 2013; 153: 1194-217.

[7] Prince MJ, Wu F, Guo Y, Gutierrez Robledo LM, O’Donnell M, Sullivan R, et al. The burden of disease in older people and implications for health policy and practice. Lancet. 2015; 385: 549-62.

[8] McGough M, Telesford I, Winger A, Cotter L, Cox C. How much is health spending expected to grow? [Internet]. PETERSON—KFF Health System Tracker; 2026 [cited 2025 Jul 20]. Available from: https://www.healthsystemtracker.org/chart—collection/how—much—is—health—spending—expected—to—grow/

[9] Global spending on health: Coping with the pandemic [Internet]. WHO; c2026 [cited 2025 Jul 25]. Available from: https://www.who.int/publications/i/item/9789240086746

[10] Malki ST, Johansson P, Andersson G, Andréasson F, Mourad G. Caregiver burden, psychological well—being, and support needs among Swedish informal caregivers. BMC Public Health. 2025; 25: 867.

[11] Kallestrup—Lamb M, Marin AOK, Menon S, Søgaard J. Aging populations and expenditures on health. J Econ Ageing. 2024; 29: 100518.

[12] Panier S, Wang S, Schumacher B. Genome Instability and DNA Repair in Somatic and Reproductive Aging. Annu Rev Pathol. 2024; 19: 261-90.

[13] Hoeijmakers JH. DNA damage, aging, and cancer. N Engl J Med. 2009; 361: 1475-85.

[14] Gorbunova V, Seluanov A, Mao Z, Hine C. Changes in DNA repair during aging. Nucleic Acids Res. 2007; 35: 7466-74.

[15] d’Adda di Fagagna F. Living on a break: cellular senescence as a DNA—damage response. Nat Rev Cancer. 2008; 8: 512-22.

[16] Lee J, Pellegrini MV. Biochemistry, Telomere And Telomerase. Treasure Island (FL): StatPearls Publishing; 2025.

[17] von Zglinicki T. Oxidative stress shortens telomeres. Trends Biochem Sci. 2002; 27: 339-44.

[18] d’Adda di Fagagna F, Reaper PM, Clay—Farrace L, Fiegler H, Carr P, Von Zglinicki T, et al. A DNA damage checkpoint response in telomere—initiated senescence. Nature. 2003; 426: 194—8.

[19] Campisi J, d’Adda di Fagagna F. Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol. 2007; 8: 729—40.

[20] Tsurumi A, Li WX. Global heterochromatin loss: a unifying theory of aging? Epigenetics. 2012; 7: 680—8.

[21] Booth LN, Brunet A. The Aging Epigenome. Mol Cell. 2016; 62: 728-44.

[22] Maslov AY, Vijg J. Genome instability, cancer and aging. Biochim Biophys Acta. 2009; 1790: 963-9.

[23] Hipp MS, Kasturi P, Hartl FU. The proteostasis network and its decline in ageing. Nat Rev Mol Cell Biol. 2019; 20: 421—35.

[24] Lim SM, Nahm M, Kim SH. Proteostasis and Ribostasis Impairment as Common Cell Death Mechanisms in Neurodegenerative Diseases. J Clin Neurol. 2023; 19: 101-14.

[25] Meller A, Shalgi R. The aging proteostasis decline: From nematode to human. Exp Cell Res. 2021; 399: 112474.

[26] Manczak M, Jung Y, Park BS, Partovi D, Reddy PH. Time—course of mitochondrial gene expressions in mice brains: implications for mitochondrial dysfunction, oxidative damage, and cytochrome c in aging. J Neurochem. 2005; 92: 494-504.

[27] Sun N, Youle RJ, Finkel T. The Mitochondrial Basis of Aging. Mol Cell. 2016; 61: 654-66.

[28] Payne BA, Chinnery PF. Mitochondrial dysfunction in aging: Much progress but many unresolved questions. Biochim Biophys Acta. 2015; 1847: 1347—53.

[29] Trifunovic A, Wredenberg A, Falkenberg M, Spelbrink JN, Rovio AT, Bruder CE, et al. Premature ageing in mice expressing defective mitochondrial DNA polymerase. Nature. 2004; 429: 417—23.

[30] Youle RJ, van der Bliek AM. Mitochondrial fission, fusion, and stress. Science. 2012; 337: 1062—5.

[31] Fiorese CJ, Schulz AM, Lin YF, Rosin N, Pellegrino MW, Haynes CM. The Transcription Factor ATF5 Mediates a Mammalian Mitochondrial UPR. Curr Biol. 2016; 26: 2037—43.

[32] Panwar V, Singh A, Bhatt M, Tonk RK, Azizov S, Raza AS, et al. Multifaceted role of mTOR (mammalian target of rapamycin) signaling pathway in human health and disease. Signal Transduct Target Ther. 2023; 8: 375.

[33] Saxton RA, Sabatini DM. mTOR Signaling in Growth, Metabolism, and Disease. Cell. 2017; 168: 960—76.

[34] Xu S, Cai Y, Wei Y. mTOR Signaling from Cellular Senescence to Organismal Aging. Aging Dis. 2014; 5: 263-73.

[35] Liu X, Guo B, Li Q, Nie J. mTOR in metabolic homeostasis and disease. Exp Cell Res. 2024; 441: 114173.

[36] Antikainen H, Driscoll M, Haspel G, Dobrowolski R. TOR—mediated regulation of metabolism in aging. Aging Cell. 2017; 16: 1219—33.

[37] Kolb H, Kempf K, Martin S. Insulin and aging — a disappointing relationship. Front Endocrinol (Lausanne). 2023; 14: 1261298.

[38] Guo X, Asthana P, Gurung S, Zhang S, Wong SKK, Fallah S, et al. Regulation of age—associated insulin resistance by MT1—MMP—mediated cleavage of insulin receptor. Nat Commun. 2022; 13: 3749.

[39] Lu M, Li C. Nutrient sensing in pancreatic islets: lessons from congenital hyperinsulinism and monogenic diabetes. Ann N Y Acad Sci. 2018; 1411: 65-82.

[40] Lee CG, Boyko EJ, Strotmeyer ES, Lewis CE, Cawthon PM, Hoffman AR, et al. ; Osteoporotic Fractures in Men Study Research Group. Association between insulin resistance and lean mass loss and fat mass gain in older men without diabetes mellitus. J Am Geriatr Soc. 2011; 59: 1217—24.

[41] Tessari P. Role of insulin in age—related changes in macronutrient metabolism. Eur J Clin Nutr. 2000; 54: S126-30.

[42] Johnson SC. Nutrient Sensing, Signaling and Ageing: The Role of IGF—1 and mTOR in Ageing and Age—Related Disease. Subcell Biochem. 2018; 90: 49-97.

[43] Salminen A, Kaarniranta K, Kauppinen A. Insulin/IGF—1 signaling promotes immunosuppression via the STAT3 pathway: impact on the aging process and age—related diseases. Inflamm Res. 2021; 70: 1043-61.

[44] Li WJ, Wang CW, Tao L, Yan YH, Zhang MJ, Liu ZX, et al. Insulin signaling regulates longevity through protein phosphorylation in Caenorhabditis elegans. Nat Commun. 2021; 12: 4568.

[45] Webb AE, Brunet A. FOXO transcription factors: key regulators of cellular quality control. Trends Biochem Sci. 2014; 39: 159-69.

[46] Zheng L, He S, Wang H, Li J, Liu Y, Liu S. Targeting Cellular Senescence in Aging and Age—Related Diseases: Challenges, Considerations, and the Emerging Role of Senolytic and Senomorphic Therapies. Aging Dis. 2024; 15: 2554-94.

[47] Kroemer G, Maier AB, Cuervo AM, Gladyshev VN, Ferrucci L, Gorbunova V, et al. From geroscience to precision geromedicine: Understanding and managing aging. Cell. 2025; 188: 2043-62.

[48] Pereira B, Correia FP, Alves IA, Costa M, Gameiro M, Martins AP, et al. Epigenetic reprogramming as a key to reverse ageing and increase longevity. Ageing Res Rev. 2024; 95: 102204.

[49] Papadopoli D, Boulay K, Kazak L, Pollak M, Mallette FA, Topisirovic I, et al. mTOR as a central regulator of lifespan and aging. F1000Res. 2019; 8: 998.

[50] Miwa S, Kashyap S, Chini E, von Zglinicki T. Mitochondrial dysfunction in cell senescence and aging. J Clin Invest. 2022; 132: e158447.

[51] Tchkonia T, Zhu Y, van Deursen J, Campisi J, Kirkland JL. Cellular senescence and the senescent secretory phenotype: therapeutic opportunities. J Clin Invest. 2013; 123: 966-72.

[52] Coppé JP, Desprez PY, Krtolica A, Campisi J. The senescence—associated secretory phenotype: the dark side of tumor suppression. Annu Rev Pathol. 2010; 5: 99-118.

[53] Olascoaga S, Konigsberg M, Espinal—Enríquez J, Tovar H, Matadamas—Martínez F, Pérez—Villanueva J, et al. Transcriptomic signatures and network—based methods uncover new senescent cell anti—apoptotic pathways and senolytics. FEBS J. 2025; 292: 1950-71.

[54] Chen S, Gan D, Lin S, Zhong Y, Chen M, Zou X, et al. Metformin in aging and aging—related diseases: clinical applications and relevant mechanisms. Theranostics. 2022; 12: 2722—40.

[55] Oh J, Lee YD, Wagers AJ. Stem cell aging: mechanisms, regulators and therapeutic opportunities. Nat Med. 2014; 20: 870—80.

[56] Tenchov R, Sasso JM, Wang X, Zhou QA. Aging Hallmarks and Progression and Age—Related Diseases: A Landscape View of Research Advancement. ACS Chem Neurosci. 2024; 15: 1-30.

[57] Campbell JM, Bellman SM, Stephenson MD, Lisy K. Metformin reduces all—cause mortality and diseases of ageing independent of its effect on diabetes control: A systematic review and meta—analysis. Ageing Res Rev. 2017; 40: 31-44.

[58] Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk—Melody J, et al. Role of AMP—activated protein kinase in mechanism of metformin action. J Clin Invest. 2001; 108: 1167-74.

[59] Long YC, Zierath JR. AMP—activated protein kinase signaling in metabolic regulation. J Clin Invest. 2006; 116: 1776-83.

[60] Foretz M, Even PC, Viollet B. AMPK Activation Reduces Hepatic Lipid Content by Increasing Fat Oxidation In Vivo. Int J Mol Sci. 2018; 19: 2826.

[61] Srivastava RA, Pinkosky SL, Filippov S, Hanselman JC, Cramer CT, Newton RS. AMP—activated protein kinase: an emerging drug target to regulate imbalances in lipid and carbohydrate metabolism to treat cardio—metabolic diseases. J Lipid Res. 2012; 53: 2490-514.

[62] Zilov AV, Abdelaziz SI, AlShammary A, Al Zahrani A, Amir A, Assaad Khalil SH, et al. Mechanisms of action of metformin with special reference to cardiovascular protection. Diabetes Metab Res Rev. 2019; 35: e3173.

[63] Mohammed I, Hollenberg MD, Ding H, Triggle CR. A Critical Review of the Evidence That Metformin Is a Putative Anti—Aging Drug That Enhances Healthspan and Extends Lifespan. Front Endocrinol (Lausanne). 2021; 12: 718942.

[64] Kulkarni AS, Gubbi S, Barzilai N. Benefits of Metformin in Attenuating the Hallmarks of Aging. Cell Metab. 2020; 32: 15-30.

[65] Liu XD, Li YG, Wang GY, Bi YG, Zhao Y, Yan ML, et al. Metformin protects high glucose—cultured cardiomyocytes from oxidative stress by promoting NDUFA13 expression and mitochondrial biogenesis via the AMPK signaling pathway. Mol Med Rep. 2020; 22: 5262-70.

[66] Zhong T, Men Y, Lu L, Geng T, Zhou J, Mitsuhashi A, et al. Metformin alters DNA methylation genome—wide via the H19/SAHH axis. Oncogene. 2017; 36: 2345-54.

[67] Barzilai N, Crandall JP, Kritchevsky SB, Espeland MA. Metformin as a Tool to Target Aging. Cell Metab. 2016; 23: 1060—5.

[68] Kukidome D, Nishikawa T, Sonoda K, Imoto K, Fujisawa K, Yano M, et al. Activation of AMP—activated protein kinase reduces hyperglycemia—induced mitochondrial reactive oxygen species production and promotes mitochondrial biogenesis in human umbilical vein endothelial cells. Diabetes. 2006; 55: 120-7.

[69] Rehman A, Satyam SM, El—Tanani M, Prabhakar S, Kumari R, Shetty P, et al. Metformin Beyond Diabetes: A Precision Gerotherapeutic and Immunometabolic Adjuvant for Aging and Cancer. Cancers (Basel). 2025; 17: 2466.

[70] Zhu Y, Tchkonia T, Pirtskhalava T, Gower AC, Ding H, Giorgadze N, et al. The Achilles’ heel of senescent cells: from transcriptome to senolytic drugs. Aging Cell. 2015; 14: 644—58.

[71] Dai H, Chen R, Gui C, Tao T, Ge Y, Zhao X, et al. Eliminating senescent chondrogenic progenitor cells enhances chondrogenesis under intermittent hydrostatic pressure for the treatment of OA. Stem Cell Res Ther. 2020; 11: 199.

[72] Fan T, Du Y, Zhang M, Zhu AR, Zhang J. Senolytics Cocktail Dasatinib and Quercetin Alleviate Human Umbilical Vein Endothelial Cell Senescence via the TRAF6—MAPK—NF—κB Axis in a YTHDF2—Dependent Manner. Gerontology. 2022; 68: 920—34.

[73] Yakubo S, Abe H, Li Y, Kudo M, Kimura A, Wakabayashi T, et al. Dasatinib and Quercetin as Senolytic Drugs Improve Fat Deposition and Exhibit Antifibrotic Effects in the Medaka Metabolic Dysfunction—Associated Steatotic Liver Disease Model. Diseases. 2024; 12: 317.

[74] Rodríguez—Agustín A, Casanova V, Grau—Expósito J, Sánchez—Palomino S, Alcamí J, Climent N. Immunomodulatory Activity of the Tyrosine Kinase Inhibitor Dasatinib to Elicit NK Cytotoxicity against Cancer, HIV Infection and Aging. Pharmaceutics. 2023; 15: 917.

[75] Islam MT, Tuday E, Allen S, Kim J, Trott DW, Holland WL, et al. Senolytic drugs, dasatinib and quercetin, attenuate adipose tissue inflammation, and ameliorate metabolic function in old age. Aging Cell. 2023; 22: e13767.

[76] Wang Y, Che L, Chen X, He Z, Song D, Yuan Y, et al. Repurpose dasatinib and quercetin: Targeting senescent cells ameliorates postmenopausal osteoporosis and rejuvenates bone regeneration. Bioact Mater. 2023; 25: 13-28.

[77] Nieto M, Konigsberg M, Silva—Palacios A. Quercetin and dasatinib, two powerful senolytics in age—related cardiovascular disease. Biogerontology. 2024; 25: 71-82.

[78] Xu M, Pirtskhalava T, Farr JN, Weigand BM, Palmer AK, Weivoda MM, et al. Senolytics improve physical function and increase lifespan in old age. Nat Med. 2018; 24: 1246—56.

[79] Georgiou N, Kakava MG, Routsi EA, Petsas E, Stavridis N, Freris C, et al. Quercetin: A Potential Polydynamic Drug. Molecules. 2023; 28: 8141.

[80] Targeting Cellular Senescence With Senolytics to Improve Skeletal Health in Older Humans [Internet]. [cited 2025 Sep 11]. Available from: https://www.clinicaltrials.gov/study/NCT04313634?term=NCT04313634&rank=1

[81] Millar CL, Iloputaife I, Baldyga K, Norling AM, Boulougoura A, Vichos T, et al. A pilot study of senolytics to improve cognition and mobility in older adults at risk for Alzheimer’s disease. EBioMedicine. 2025; 113: 105612.

[82] Espinoza SE, Khosla S, Baur JA, de Cabo R, Musi N. Drugs Targeting Mechanisms of Aging to Delay Age—Related Disease and Promote Healthspan: Proceedings of a National Institute on Aging Workshop. J Gerontol A Biol Sci Med Sci. 2023; 78: 53-60.

[83] Justice JN, Nambiar AM, Tchkonia T, LeBrasseur NK, Pascual R, Hashmi SK, et al. Senolytics in idiopathic pulmonary fibrosis: Results from a first—in—human, open—label, pilot study. EBioMedicine. 2019; 40: 554-63.

[84] Alejandra WP, Miriam Irene JP, Fabio Antonio GS, Patricia RR, Elizabeth TA, Aleman—Aguilar JP, et al. Production of monoclonal antibodies for therapeutic purposes: A review. Int Immunopharmacol. 2023; 120: 110376.

[85] Squarzoni S, Schena E, Sabatelli P, Mattioli E, Capanni C, Cenni V, et al. Interleukin—6 neutralization ameliorates symptoms in prematurely aged mice. Aging Cell. 2021; 20: e13285.

[86] Giuliani A, Prattichizzo F, Micolucci L, Ceriello A, Procopio AD, Rippo MR. Mitochondrial (Dys)Function in Inflammaging: Do MitomiRs Influence the Energetic, Oxidative, and Inflammatory Status of Senescent Cells? Mediators Inflamm. 2017; 2017: 2309034.

[87] Rašková M, Lacina L, Kejík Z, Venhauerová A, Skaličková M, Kolář M, et al. The Role of IL—6 in Cancer Cell Invasiveness and Metastasis—Overview and Therapeutic Opportunities. Cells. 2022; 11: 3698.

[88] Soler MF, Abaurrea A, Azcoaga P, Araujo AM, Caffarel MM. New perspectives in cancer immunotherapy: targeting IL—6 cytokine family. J Immunother Cancer. 2023; 11: e007530.

[89] Tyrrell DJ, Goldstein DR. Ageing and atherosclerosis: vascular intrinsic and extrinsic factors and potential role of IL—6. Nat Rev Cardiol. 2021; 18: 58-68.

[90] Coppé JP, Patil CK, Rodier F, Sun Y, Muñoz DP, Goldstein J, et al. Senescence—associated secretory phenotypes reveal cell—nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol. 2008; 6: e301.

[91] Wang Y, Zhu S, Wei W, Tu Y, Chen C, Song J, et al. Interleukin—6 knockout reverses macrophage differentiation imbalance and alleviates cardiac dysfunction in aging mice. Aging (Albany NY). 2020; 12: 20184-97.

[92] Tanaka T, Narazaki M, Ogata A, Kishimoto T. A new era for the treatment of inflammatory autoimmune diseases by interleukin—6 blockade strategy. Semin Immunol. 2014; 26: 88-96.

[93] Nowak KL, Kakkar R, Devalaraja M, Lo L, Park W, Gobburu J, et al. A Phase 1 Randomized Dose—Escalation Study of a Human Monoclonal Antibody to IL—6 in CKD. Kidney360. 2021; 2: 224-35.

[94] Specker C, Aringer M, Burmester GR, Killy B, Hofmann MW, Kellner H, et al. The safety and effectiveness of tocilizumab in elderly patients with rheumatoid arthritis and in patients with comorbidities associated with age. Clin Exp Rheumatol. 2022; 40: 1657—65.

[95] Devauchelle—Pensec V, Carvajal—Alegria G, Dernis E, Richez C, Truchetet ME, Wendling D, et al. Effect of Tocilizumab on Disease Activity in Patients With Active Polymyalgia Rheumatica Receiving Glucocorticoid Therapy: A Randomized Clinical Trial. JAMA. 2022; 328: 1053-62.

[96] Law S, Chaudhuri S. Mesenchymal stem cell and regenerative medicine: regeneration versus immunomodulatory challenges. Am J Stem Cells. 2013; 2: 22-38.

[97] Yang YK. Aging of mesenchymal stem cells: Implication in regenerative medicine. Regen Ther. 2018; 9: 120—2.

[98] Ermolaeva M, Neri F, Ori A, Rudolph KL. Cellular and epigenetic drivers of stem cell ageing. Nat Rev Mol Cell Biol. 2018; 19: 594-610.

[99] Steinert AF, Rackwitz L, Gilbert F, Nöth U, Tuan RS. Concise review: the clinical application of mesenchymal stem cells for musculoskeletal regeneration: current status and perspectives. Stem Cells Transl Med. 2012; 1: 237-47.

[100] FDA Approves First Mesenchymal Stromal Cell Therapy to Treat Steroid—refractory Acute Graft—versus—host Disease [Internet]. Cision US Inc.; c2026 [cited 2025 Sep 20]. Available from: https://www.prnewswire.com/news—releases/fda—approves—first—mesenchymal—stromal—cell—therapy—to—treat—steroid—refractory—acute—graft—versus—host—disease—302335542.html

[101] Kitada M, Dezawa M. Parkinson’s disease and mesenchymal stem cells: potential for cell—based therapy. Parkinsons Dis. 2012; 2012: 873706.

[102] Ward MR, Abadeh A, Connelly KA. Concise Review: Rational Use of Mesenchymal Stem Cells in the Treatment of Ischemic Heart Disease. Stem Cells Transl Med. 2018; 7: 543—50.

[103] Hall B, Cho A, Limaye A, Cho K, Khillan J, Kulkarni AB. Genome Editing in Mice Using CRISPR/Cas9 Technology. Curr Protoc Cell Biol. 2018; 81: e57.

[104] Pelletier S, Gingras S, Green DR. Mouse genome engineering via CRISPR—Cas9 for study of immune function. Immunity. 2015; 42: 18-27.

[105] Qin W, Dion SL, Kutny PM, Zhang Y, Cheng AW, Jillette NL, et al. Efficient CRISPR/Cas9—Mediated Genome Editing in Mice by Zygote Electroporation of Nuclease. Genetics. 2015; 200: 423—30.

[106] Shin HY, Wang C, Lee HK, Yoo KH, Zeng X, Kuhns T, et al. CRISPR/Cas9 targeting events cause complex deletions and insertions at 17 sites in the mouse genome. Nat Commun. 2017; 8: 15464.

[107] Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, et al. Multiplex genome engineering using CRISPR/Cas systems. Science. 2013; 339: 819—23.

[108] Quadros RM, Miura H, Harms DW, Akatsuka H, Sato T, Aida T, et al. Easi—CRISPR: a robust method for one—step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins. Genome Biol. 2017; 18: 92.

[109] Davis DJ, McNew JF, Maresca—Fichter H, Chen K, Telugu BP, Bryda EC. Efficient DNA knock—in using AAV—mediated delivery with 2—cell embryo CRISPR—Cas9 electroporation. Front Genome Ed. 2023; 5: 1256451.

[110] Wang W, Zheng Y, Sun S, Li W, Song M, Ji Q, et al. A genome—wide CRISPR—based screen identifies KAT7 as a driver of cellular senescence. Sci Transl Med. 2021; 13: eabd2655.

[111] Huang S, Lau CH, Tin C, Lam RHW. Extended replicative lifespan of primary resting T cells by CRISPR/dCas9—based epigenetic modifiers and transcriptional activators. Cell Mol Life Sci. 2024; 81: 407.

[112] Mostoslavsky R, Chua KF, Lombard DB, Pang WW, Fischer MR, Gellon L, et al. Genomic instability and aging—like phenotype in the absence of mammalian SIRT6. Cell. 2006; 124: 315—29.

[113] Roichman A, Elhanati S, Aon MA, Abramovich I, Di Francesco A, Shahar Y, et al. Restoration of energy homeostasis by SIRT6 extends healthy lifespan. Nat Commun. 2021; 12: 3208.

[114] You W, Zheng W, Weiss S, Chua KF, Steegborn C. Structural basis for the activation and inhibition of Sirtuin 6 by quercetin and its derivatives. Sci Rep. 2019; 9: 19176.

[115] Michishita E, McCord RA, Berber E, Kioi M, Padilla—Nash H, Damian M, et al. SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature. 2008; 452: 492—6.

[116] Mao Z, Hine C, Tian X, Van Meter M, Au M, Vaidya A, et al. SIRT6 promotes DNA repair under stress by activating PARP1. Science. 2011; 332: 1443—6.

[117] Toiber D, Erdel F, Bouazoune K, Silberman DM, Zhong L, Mulligan P, et al. SIRT6 recruits SNF2H to DNA break sites, preventing genomic instability through chromatin remodeling. Mol Cell. 2013; 51: 454-68.

[118] Wang X, Li X, Zhou J, Lei Z, Yang X. Fisetin suppresses chondrocyte senescence and attenuates osteoarthritis progression by targeting sirtuin 6. Chem Biol Interact. 2024; 390: 110890.

[119] Zhong L, D’Urso A, Toiber D, Sebastian C, Henry RE, Vadysirisack DD, et al. The histone deacetylase Sirt6 regulates glucose homeostasis via Hif1α. Cell. 2010; 140: 280-93.

[120] Tao R, Xiong X, DePinho RA, Deng CX, Dong XC. FoxO3 transcription factor and Sirt6 deacetylase regulate low density lipoprotein (LDL)—cholesterol homeostasis via control of the proprotein convertase subtilisin/kexin type 9 (Pcsk9) gene expression. J Biol Chem. 2013; 288: 29252—9.

[121] Wang X, Chen X, Zhang X, Su C, Yang M, He W, et al. A small—molecule inhibitor of PCSK9 transcription ameliorates atherosclerosis through the modulation of FoxO1/3 and HNF1α. EBioMedicine. 2020; 52: 102650.

[122] Ramteke P, Watson B, Toci M, Tran VA, Johnston S, Tsingas M, et al. Sirt6 deficiency promotes senescence and age—associated intervertebral disc degeneration in mice. Bone Res. 2025; 13: 50.

[123] Liu X, Ren S, Li Z, Hao D, Zhao X, Zhang Z, et al. Sirt6 mediates antioxidative functions by increasing Nrf2 abundance. Exp Cell Res. 2023; 422: 113409.

[124] Guo Z, Li P, Ge J, Li H. SIRT6 in Aging, Metabolism, Inflammation and Cardiovascular Diseases. Aging Dis. 2022; 13: 1787-822.

[125] McHugh D, Sun B, Gutierrez—Muñoz C, Hernández—González F, Mellone M, Guiho R, et al. COPI vesicle formation and N—myristoylation are targetable vulnerabilities of senescent cells. Nat Cell Biol. 2023; 25: 1804-20.

[126] Aman Y. Targeting COPIng liabilities of senescent cells. Nat Aging. 2024; 4: 5.

[127] Ko A, Han SY, Song J. Dynamics of ARF regulation that control senescence and cancer. BMB Rep. 2016; 49: 598-606.

[128] Lueg GA, Zhang J, Faronato M, Gorelik A, Kallemeijn WW, Falciani F, et al. MYC deregulation sensitizes cancer cells to N—myristoyltransferase inhibition. Cell Rep. 2025; 44: 116180.

[129] Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double—stranded RNA in Caenorhabditis elegans. Nature. 1998; 391: 806—11.

[130] Bernstein E, Caudy AA, Hammond SM, Hannon GJ. Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature. 2001; 409: 363-6.

[131] Hammond SM, Bernstein E, Beach D, Hannon GJ. An RNA—directed nuclease mediates post—transcriptional gene silencing in Drosophila cells. Nature. 2000; 404: 293-6.

[132] Buj R, Leon KE, Anguelov MA, Aird KM. Suppression of p16 alleviates the senescence—associated secretory phenotype. Aging (Albany NY). 2021; 13: 3290-312.

[133] Takaya K, Asou T, Kishi K. Downregulation of senescence—associated secretory phenotype by knockdown of secreted frizzled—related protein 4 contributes to the prevention of skin aging. Aging (Albany NY). 2022; 14: 8167—78.

[134] Bleau AM, Jimenez AI, Gramis V, Ruz V, Segerros RA, Vargas B. Clinical trial for SYL1801, a new siRNA—Based Treatment for Neovascular AMD. Invest Ophthalmol Vis Sci. 2025; 66: 697.

[135] Hong SW, Park JH, inventors; Olix Pharmaceuticals Inc, assignee. RNAi AGENT TARGETING MYD88 AND USE THEREOF. United States patent US20240093188A1. 2024 Mar 21.

[136] Grogan M, Sheikh FH, Sperry BW, Sweetser MT, Capocelli K, Roblin S, et al. Realizing the therapeutic potential of rapid knockdown of transthyretin via RNA interference in transthyretin amyloidosis. Mol Ther Nucleic Acids. 2025; 36: 102590.

[137] Lathe R, Sapronova A, Kotelevtsev Y. Atherosclerosis and Alzheimer—diseases with a common cause? Inflammation, oxysterols, vasculature. BMC Geriatr. 2014; 14: 36.

[138] Adams D, Wixner J, Polydefkis M, Berk JL, Conceição IM, Dispenzieri A, et al. ; patisiran Global OLE study group. Five—Year Results With Patisiran for Hereditary Transthyretin Amyloidosis With Polyneuropathy: A Randomized Clinical Trial With Open—Label Extension. JAMA Neurol. 2025; 82: 228-36.

[139] Wang Y, Zhou J, Yang Q, Li X, Qiu Y, Zhang Y, et al. Therapeutic siRNA targeting PLIN2 ameliorates steatosis, inflammation, and fibrosis in steatotic liver disease models. J Lipid Res. 2024; 65: 100635.

[140] Justice JN, Ferrucci L, Newman AB, Aroda VR, Bahnson JL, Divers J, et al. A framework for selection of blood—based biomarkers for geroscience—guided clinical trials: report from the TAME Biomarkers Workgroup. Geroscience. 2018; 40: 419-36.

[141] Horvath S, Raj K. DNA methylation—based biomarkers and the epigenetic clock theory of ageing. Nat Rev Genet. 2018; 19: 371-84.

[142] Belsky DW, Caspi A, Arseneault L, Baccarelli A, Corcoran DL, Gao X, et al. Quantification of the pace of biological aging in humans through a blood test, the DunedinPoAm DNA methylation algorithm. Elife. 2020; 9: e54870.

[143] Wagner KH, Cameron—Smith D, Wessner B, Franzke B. Biomarkers of Aging: From Function to Molecular Biology. Nutrients. 2016; 8: 338.

[144] Ho AD, Wagner W, Mahlknecht U. Stem cells and ageing. The potential of stem cells to overcome age—related deteriorations of the body in regenerative medicine. EMBO Rep. 2005; 6: S35-8.

[145] Hodrosky N, Cacho G, Ahmed F, Mudireddy R, Wen Y, Nembhard K, et al. The Pharmaceutical Price Regulation Crisis: Implications on Antidepressant Access for Low—Income Americans. arXiv:2506.14849 [Preprint]. 2025 [cited 2025 Sep 25]. Available from: https://doi.org/10.48550/arXiv.2506.14849

[146] Gallini J, Baucom Z, Tripodis Y. Cognitive factor—based selection increases power in Alzheimer’s dementia randomized clinical trials. arXiv:2503.16044 [Preprint]. 2025 [cited 2025 Oct 1]. Available from: https://doi.org/10.48550/arXiv.2503.16044

[147] Cabañero—Garcia E, Martinez—Lacoba R, Pardo—Garcia I, Amo—Saus E. Barriers to health, social and long—term care access among older adults: a systematic review of reviews. Int J Equity Health. 2025; 24: 72.

[148] How Might Longevity Therapies Affect Social and Economic Inequality? [Internet]. Sustainability Directory; c2024 [cited 2025 Oct 1]. Available from: https://lifestyle.sustainability—directory.com/question/how—might—longevity—therapies—affect—social—and—economic—inequality/