Accumulation of senescent cells may drive age-associated alterations and pathologies. Senolytics are promising therapeutics that can preferentially eliminate senescent cells. Here, we performed a high-throughput automatized screening (HTS) of the commercial LOPAC®Pfizer library on aphidicolin-induced senescent human fibroblasts, to identify novel senolytics. We discovered the nociceptin receptor FQ opioid receptor (NOP) selective ligand 1-[1-(1-methylcyclooctyl)-4-piperidinyl]-2-[(3R)-3-piperidinyl]-1H-benzimidazole (MCOPPB, a compound previously studied as potential anxiolytic) as the best scoring hit. The ability of MCOPPB to eliminate senescent cells in in vitro models was further tested in mice and in C. elegans. MCOPPB reduced the senescence cell burden in peripheral tissues but not in the central nervous system. Mice and worms exposed to MCOPPB also exhibited locomotion and lipid storage changes. Mechanistically, MCOPPB treatment activated transcriptional networks involved in the immune responses to external stressors, implicating Toll-like receptors (TLRs). Our study uncovers MCOPPB as a NOP ligand that, apart from anxiolytic effects, also shows tissue-specific senolytic effects.

Department of Anatomy and Cell Biology Research Institute of the Medical University of Varna Varna Bulgaria

Department of Biology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Biomedical and Biotechnological Sciences University of Catania Catania Italy

Department of Health Promotion Mother and Child Care Internal Medicine and Medical Specialties Pathologic Anatomy Unit University of Palermo Palermo Italy

Department of Molecular and Computational Biology Arts and Sciences Dornsife College of Letters University of Southern California Los Angeles CA USA

Department of Translational Stem Cell Biology Research Institute of the Medical University of Varna Varna Bulgaria

Division of Genome Biology Department of Medical Biochemistry and Biophysics Science for Life Laboratory Karolinska Institute Stockholm Sweden

Genome Integrity Unit Danish Cancer Society Research Center Copenhagen Denmark

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

International Clinical Research Center St Anne's University Hospital Brno Czech Republic

Laboratory of Bioinformatics Fondazione IRCCS Casa Sollievo Della Sofferenza San Giovanni Rotondo Italy

Leonard Davis School of Gerontology University of Southern California Los Angeles CA USA

Norris Comprehensive Cancer Center Keck School of Medicine University of Southern California Los Angeles CA USA

Psychogenics Inc Tarrytown NY USA

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Campisi J. Aging, cellular senescence, and cancer. Annu Rev Physiol. 2013 doi: 10.1146/annurev-physiol-030212-183653. PubMed DOI PMC

Hodes RJ, et al. Disease drivers of aging. Ann N Y Acad Sci. 2016 doi: 10.1111/nyas.13299. PubMed DOI PMC

Inouye SK, et al. Geriatric syndromes: clinical, research, and policy implications of a core geriatric concept. J Am Geriatr Soc. 2007 doi: 10.1111/j.1532-5415.2007.01156.x. PubMed DOI PMC

McHugh D, Gil J. Senescence and aging: causes, consequences, and therapeutic avenues. J Cell Biol. 2018 doi: 10.1083/jcb.201708092. PubMed DOI PMC

Childs BG, et al. Senescent cells: an emerging target for diseases of ageing. Nat Rev Drug Discov. 2017 doi: 10.1038/nrd.2017.116. PubMed DOI PMC

Schafer MJ, et al. The senescence-associated secretome as an indicator of age and medical risk. JCI Insight. 2020 doi: 10.1172/jci.insight.133668. PubMed DOI PMC

Gorgoulis V, et al. Cellular senescence: defining a path forward. Cell. 2019 doi: 10.1016/j.cell.2019.10.005. PubMed DOI

Bartek J, Bartkova J, Lukas J. DNA damage signalling guards against activated oncogenes and tumour progression. Oncogene. 2007 doi: 10.1038/sj.onc.1210881. PubMed DOI

Pignolo RJ, et al. Reducing senescent cell burden in aging and disease. Trends Mol Med. 2020 doi: 10.1016/j.molmed.2020.03.005. PubMed DOI PMC

Kirkland JL, et al. The clinical potential of senolytic drugs. J Am Geriatr Soc. 2017 doi: 10.1111/jgs.14969. PubMed DOI PMC

Kirkland JL, Tchkonia T. Cellular senescence: a translational perspective. EBioMedicine. 2017 doi: 10.1016/j.ebiom.2017.04.013. PubMed DOI PMC

Calimport SRG, et al. To help aging populations, classify organismal senescence. Science. 2019 doi: 10.1126/science.aay7319. PubMed DOI PMC

Ogrodnik M, et al. Cellular senescence drives age-dependent hepatic steatosis. Nat Commun. 2017 doi: 10.1038/ncomms15691. PubMed DOI PMC

Zhu Y, et al. The Achilles’ heel of senescent cells: from transcriptome to senolytic drugs. Aging Cell. 2015 doi: 10.1111/acel.12344. PubMed DOI PMC

Volochnyuk DM, et al. Evolution of commercially available compounds for HTS. Drug Discov Today. 2019 doi: 10.1016/j.drudis.2018.10.016. PubMed DOI

Fuhrmann-Stroissnigg H, et al. Identification of HSP90 inhibitors as a novel class of senolytics. Nat Commun. 2017 doi: 10.1038/s41467-017-00314-z. PubMed DOI PMC

Wakita M, et al. A BET family protein degrader provokes senolysis by targeting NHEJ and autophagy in senescent cells. Nat Commun. 2020 doi: 10.1038/s41467-020-15719-6. PubMed DOI PMC

Cho HJ, et al. Identification of SYK inhibitor, R406 as a novel senolytic agent. Aging (Albany NY). 2020. 10.18632/aging.103135. PubMed PMC

Ozsvari B, et al. Azithromycin and Roxithromycin define a new family of “senolytic” drugs that target senescent human fibroblasts. Aging (Albany NY). 2018. 10.18632/aging.101633. PubMed PMC

Bucknall RA, et al. Antiviral effects of aphidicolin, a new antibiotic produced by Cephalosporium aphidicola. Antimicrob Agents Chemother. 1973 doi: 10.1128/aac.4.3.294. PubMed DOI PMC

Huberman JA. New views of the biochemistry of eucaryotic DNA replication revealed by aphidicolin, an unusual inhibitor of DNA polymerase alpha. Cell. 1981 doi: 10.1016/0092-8674(81)90426-8. PubMed DOI

Glover TW, et al. DNA polymerase alpha inhibition by aphidicolin induces gaps and breaks at common fragile sites in human chromosomes. Hum Genet. 1984 doi: 10.1007/BF00272988. PubMed DOI

Mazouzi A, et al. A Comprehensive Analysis of the Dynamic Response to Aphidicolin-Mediated Replication Stress Uncovers Targets for ATM and ATMIN. Cell Rep. 2016 doi: 10.1016/j.celrep.2016.03.077. PubMed DOI

Levenson VV, et al. A combination of genetic suppressor elements produces resistance to drugs inhibiting DNA replication. Somat Cell Mol Genet. 1999 doi: 10.1023/b:scam.0000007136.49230.b3. PubMed DOI

Marusyk A, et al. p53 mediates senescence-like arrest induced by chronic replicational stress. Mol Cell Biol. 2007 doi: 10.1128/MCB.01316-06. PubMed DOI PMC

Novakova Z, et al. Cytokine expression and signaling in drug-induced cellular senescence. Oncogene. 2010 doi: 10.1038/onc.2009.318. PubMed DOI

Kovacovicova K, et al. Senolytic cocktail dasatinib+quercetin (D+Q) does not enhance the efficacy of senescence-inducing chemotherapy in liver cancer. Front Oncol. 2018 doi: 10.3389/fonc.2018.00459. PubMed DOI PMC

Terzian AL, et al. The dopamine and cannabinoid interaction in the modulation of emotions and cognition: assessing the role of cannabinoid CB1 receptor in neurons expressing dopamine D1 receptors. Front Behav Neurosci. 2011 doi: 10.3389/fnbeh.2011.00049. PubMed DOI PMC

Stark T, et al. Peripubertal cannabidiol treatment rescues behavioral and neurochemical abnormalities in the MAM model of schizophrenia. Neuropharmacology. 2019 doi: 10.1016/j.neuropharm.2018.11.035. PubMed DOI

Pamplona FA, et al. Prolonged fear incubation leads to generalized avoidance behavior in mice. J Psychiatr Res. 2011 doi: 10.1016/j.jpsychires.2010.06.015. PubMed DOI

Hazane F, et al. Behavioral perturbations after prenatal neurogenesis disturbance in female rat. Neurotox Res. 2009 doi: 10.1007/s12640-009-9035-z. PubMed DOI

Murgatroyd C, et al. Dynamic DNA methylation programs persistent adverse effects of early-life stress. Nat Neurosci. 2009 doi: 10.1038/nn.2436. PubMed DOI

Direnberger S, et al. Biocompatibility of a genetically encoded calcium indicator in a transgenic mouse model. Nat Commun. 2012 doi: 10.1038/ncomms2035. PubMed DOI

Micale V, Kucerova J, Sulcova A. Leading compounds for the validation of animal models of psychopathology. Cell Tissue Res. 2013 doi: 10.1007/s00441-013-1692-9. PubMed DOI

Takeda D, et al. Effect of preoperative chemotherapy on postoperative liver regeneration following hepatic resection as estimated by liver volume. World J Surg Oncol. 2013 doi: 10.1186/1477-7819-11-65. PubMed DOI PMC

Raffaele M, et al., Senescence-like phenotype in post-mitotic cells of mice entering middle age. Aging (Albany NY), 2020. 10.18632/aging.103637. PubMed PMC

Anders S, Pyl PT, Huber W. HTSeq–a Python framework to work with high-throughput sequencing data. Bioinformatics. 2015 doi: 10.1093/bioinformatics/btu638. PubMed DOI PMC

Escorcia W, et al. Quantification of lipid abundance and evaluation of lipid distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining. J Vis Exp. 2018 doi: 10.3791/57352. PubMed DOI PMC

Vesela E, et al. Common chemical inductors of replication stress: focus on cell-based studies. Biomolecules. 2017 doi: 10.3390/biom7010019. PubMed DOI PMC

Rossiello F, et al. Irreparable telomeric DNA damage and persistent DDR signalling as a shared causative mechanism of cellular senescence and ageing. Curr Opin Genet Dev. 2014 doi: 10.1016/j.gde.2014.06.009. PubMed DOI PMC

Hirao A, et al. Pharmacological characterization of the newly synthesized nociceptin/orphanin FQ-receptor agonist 1-[1-(1-methylcyclooctyl)-4-piperidinyl]-2-[(3R)-3-piperidinyl]-1H-benzimidazole as an anxiolytic agent. J Pharmacol Sci. 2008 doi: 10.1254/jphs.fp0071742. PubMed DOI

Holanda VAD, et al. Modulation of the NOP receptor signaling affects resilience to acute stress. J Psychopharmacol. 2019 doi: 10.1177/0269881119864942. PubMed DOI

Kovacovicova K, Vinciguerra M. Isolation of senescent cells by iodixanol (OptiPrep) density gradient-based separation. Cell Prolif. 2019 doi: 10.1111/cpr.12674. PubMed DOI PMC

Tchkonia T, et al. Fat tissue, aging, and cellular senescence. Aging Cell. 2010 doi: 10.1111/j.1474-9726.2010.00608.x. PubMed DOI PMC

Palmer AK, et al. Targeting senescent cells alleviates obesity-induced metabolic dysfunction. Aging Cell. 2019 doi: 10.1111/acel.12950. PubMed DOI PMC

Sheedfar F, et al. Liver diseases and aging: friends or foes? Aging Cell. 2013 doi: 10.1111/acel.12128. PubMed DOI

Miura K, Ishioka M, Iijima K. The roles of the gut microbiota and toll-like receptors in obesity and nonalcoholic fatty liver disease. J Obes Metab Syndr. 2017 doi: 10.7570/jomes.2017.26.2.86. PubMed DOI PMC

Behmoaras J, Gil J. Similarities and interplay between senescent cells and macrophages. J Cell Biol. 2021 doi: 10.1083/jcb.202010162. PubMed DOI PMC

Ye X, et al. A pharmacological network for lifespan extension in Caenorhabditis elegans. Aging Cell. 2014 doi: 10.1111/acel.12163. PubMed DOI PMC

Nhan JD, Curran SP. Metabolic assessment of lipid abundance and distribution. Methods Mol Biol. 2020 doi: 10.1007/978-1-0716-0592-9_9. PubMed DOI

Laranjeiro R, et al. Single swim sessions in C. elegans induce key features of mammalian exercise. BMC Biol. 2017. 10.1186/s12915-017-0368-4. PubMed PMC

Apfeld J, et al. The AMP-activated protein kinase AAK-2 links energy levels and insulin-like signals to lifespan in C. elegans. Genes Dev. 2004. 10.1101/gad.1255404. PubMed PMC

Lee S, Dong HH. FoxO integration of insulin signaling with glucose and lipid metabolism. J Endocrinol. 2017 doi: 10.1530/JOE-17-0002. PubMed DOI PMC

Jia K, Chen D, Riddle DL. The TOR pathway interacts with the insulin signaling pathway to regulate C. elegans larval development, metabolism and life span. Development. 2004. 10.1242/dev.01255. PubMed

Lin K, et al. Regulation of the Caenorhabditis elegans longevity protein DAF-16 by insulin/IGF-1 and germline signaling. Nat Genet. 2001 doi: 10.1038/88850. PubMed DOI

Lee SS, et al. DAF-16 target genes that control C. elegans life-span and metabolism. Science. 2003. 10.1126/science.1083614. PubMed

Kaletsky R, et al. The C. elegans adult neuronal IIS/FOXO transcriptome reveals adult phenotype regulators. Nature. 2016. 10.1038/nature16483. PubMed PMC

Nhan JD, et al. Redirection of SKN-1 abates the negative metabolic outcomes of a perceived pathogen infection. Proc Natl Acad Sci U S A. 2019 doi: 10.1073/pnas.1909666116. PubMed DOI PMC

Xu M, et al. Senolytics improve physical function and increase lifespan in old age. Nat Med. 2018 doi: 10.1038/s41591-018-0092-9. PubMed DOI PMC

Baker DJ, et al. Clearance of p16Ink4a-positive senescent cells delays ageing-associated disorders. Nature. 2011 doi: 10.1038/nature10600. PubMed DOI PMC

Blagosklonny MV. Paradoxes of senolytics. Aging (Albany NY). 2018. 10.18632/aging.101750. PubMed PMC

Hayashi S, et al. Novel non-peptide nociceptin/orphanin FQ receptor agonist, 1-[1-(1-Methylcyclooctyl)-4-piperidinyl]-2-[(3R)-3-piperidinyl]-1H-benzimidazole: design, synthesis, and structure-activity relationship of oral receptor occupancy in the brain for orally potent antianxiety drug. J Med Chem. 2009 doi: 10.1021/jm7012979. PubMed DOI

Zhu Y, et al. Identification of a novel senolytic agent, navitoclax, targeting the Bcl-2 family of anti-apoptotic factors. Aging Cell. 2016 doi: 10.1111/acel.12445. PubMed DOI PMC

Wang Y, et al. Discovery of piperlongumine as a potential novel lead for the development of senolytic agents. Aging (Albany NY). 2016. 10.18632/aging.101100. PubMed PMC

Chang J, et al. Clearance of senescent cells by ABT263 rejuvenates aged hematopoietic stem cells in mice. Nat Med. 2016 doi: 10.1038/nm.4010. PubMed DOI PMC

Jenck F, et al. Orphanin FQ acts as an anxiolytic to attenuate behavioral responses to stress. Proc Natl Acad Sci U S A. 1997 doi: 10.1073/pnas.94.26.14854. PubMed DOI PMC

Jenck F, et al. A synthetic agonist at the orphanin FQ/nociceptin receptor ORL1: anxiolytic profile in the rat. Proc Natl Acad Sci U S A. 2000 doi: 10.1073/pnas.090514397. PubMed DOI PMC

Perna G, et al. Are anxiety disorders associated with accelerated aging? A focus on neuroprogression Neural Plast. 2016 doi: 10.1155/2016/8457612. PubMed DOI PMC

Zhou QG, et al. Hippocampal telomerase is involved in the modulation of depressive behaviors. J Neurosci. 2011 doi: 10.1523/JNEUROSCI.0805-11.2011. PubMed DOI PMC

Ogrodnik M, et al. Obesity-induced cellular senescence drives anxiety and impairs neurogenesis. Cell Metab. 2019 doi: 10.1016/j.cmet.2018.12.008. PubMed DOI PMC

Zaveri NT, Meyer ME. NOP-targeted nonpeptide ligands. Handb Exp Pharmacol. 2019 doi: 10.1007/164_2019_213. PubMed DOI

Toll L, et al. Nociceptin/orphanin FQ receptor structure, signaling, ligands, functions, and interactions with opioid systems. Pharmacol Rev. 2016 doi: 10.1124/pr.114.009209. PubMed DOI PMC

Burton DGA, Stolzing A. Cellular senescence: immunosurveillance and future immunotherapy. Ageing Res Rev. 2018 doi: 10.1016/j.arr.2018.02.001. PubMed DOI

Li Z, Weinman SA. Regulation of hepatic inflammation via macrophage cell death. Semin Liver Dis. 2018 doi: 10.1055/s-0038-1670674. PubMed DOI

Ferreira-Gonzalez S, et al. Cellular senescence in liver disease and regeneration. Semin Liver Dis. 2021 doi: 10.1055/s-0040-1722262. PubMed DOI

Mangan DF, Wahl SM. Differential regulation of human monocyte programmed cell death (apoptosis) by chemotactic factors and pro-inflammatory cytokines. J Immunol. 1991. PubMed

Perera LP, Waldmann TA. Activation of human monocytes induces differential resistance to apoptosis with rapid down regulation of caspase-8/FLICE. Proc Natl Acad Sci U S A. 1998 doi: 10.1073/pnas.95.24.14308. PubMed DOI PMC

Bellingan GJ, et al. In vivo fate of the inflammatory macrophage during the resolution of inflammation: inflammatory macrophages do not die locally, but emigrate to the draining lymph nodes. J Immunol. 1996. PubMed

Harmsen AG, et al. The role of macrophages in particle translocation from lungs to lymph nodes. Science. 1985 doi: 10.1126/science.4071052. PubMed DOI

MacPhee PJ, Schmidt EE, Groom AC. Evidence for Kupffer cell migration along liver sinusoids, from high-resolution in vivo microscopy. Am J Physiol. 1992 doi: 10.1152/ajpgi.1992.263.1.G17. PubMed DOI

Franceschi C, et al. Inflamm-aging. An evolutionary perspective on immunosenescence. Ann N Y Acad Sci. 2000. 10.1111/j.1749-6632.2000.tb06651.x. PubMed

Prattichizzo F, et al. Senescence associated macrophages and “macroph-aging”: are they pieces of the same puzzle? Aging (Albany NY). 2016. 10.18632/aging.101133. PubMed PMC

Cheong MC, et al. An opioid-like system regulating feeding behavior in C. elegans. Elife. 2015. 10.7554/eLife.06683. PubMed PMC

Wang D, et al. Genetic behavioral screen identifies an orphan anti-opioid system. Science. 2019 doi: 10.1126/science.aau2078. PubMed DOI PMC

Micioni Di Bonaventura MV, et al. N/OFQ-NOP system in food intake. Handb Exp Pharmacol. 2019. 10.1007/164_2019_212. PubMed

Matsushita H, et al. Chronic intracerebroventricular infusion of nociceptin/orphanin FQ produces body weight gain by affecting both feeding and energy metabolism in mice. Endocrinology. 2009 doi: 10.1210/en.2008-1515. PubMed DOI

Webster CM, et al. Genome-wide RNAi screen for fat regulatory genes in C. elegans identifies a proteostasis-AMPK axis critical for starvation survival. Cell Rep. 2017. 10.1016/j.celrep.2017.06.068. PubMed PMC

Azazmeh N, et al. Chronic expression of p16(INK4a) in the epidermis induces Wnt-mediated hyperplasia and promotes tumor initiation. Nat Commun. 2020 doi: 10.1038/s41467-020-16475-3. PubMed DOI PMC

Yosef R, et al. Directed elimination of senescent cells by inhibition of BCL-W and BCL-XL. Nat Commun. 2016 doi: 10.1038/ncomms11190. PubMed DOI PMC

Lim S, et al. Local delivery of senolytic drug inhibits intervertebral disc degeneration and restores intervertebral disc structure. Adv Healthc Mater. 2021 doi: 10.1002/adhm.202101483. PubMed DOI

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