THE DNA DAMAGE RESPONSE, IMMUNITY AND AGING 10 - 13 October 2022 Singapore EMBO WORKSHOP 2022 Prof Vinay Tergaonkar Institute of Molecular and Cell Biology (IMCB), A*STAR, Singapore Prof Eric Gilson Institute for Research on Cancer and Aging, France Organisers DNA damage and inflammation are two major contributing factors of aging that are intimately connected. Indeed, recent ground-breaking studies revealed how DDR can mobilize the immune system by inducing the expression of pro-inflammatory factors as well as ligands for immune receptors. The activation of immune response is induced by different DDR components including DNA damage sensors, transducer kinases, and effectors. For instance, the release of DNA fragments in the cytosol activates the cGAS-STING pathway, the expression of interferon genes and the inflammatory response. Conversely, chronic inflammation has recently emerged as an important mutagenic and cancer promoting process by generating reactive oxygen and nitrogen reactive species. These discoveries have profound implications not only in aging mechanisms but also in cancer therapy and in strategies to prevent age-related diseases. Thus, linking DNA damage response and immunity in the aging process is a novel concept with rapid intellectual advances that deserve intensive interaction between researchers of different fields. To ensure in-depth coverage of all topics relating to DNA damage, immunity and aging, the following thematic scientific sessions are planned: · Session 1: DNA damage and aging · Session 2: Inflammation, immunity and aging · Session 3: Generation and signaling of cytosolic DNA · Session 4: Cancer immunity This EMBO Workshop on this topic represents the ideal environment to fuel intellectual exchange and build-up of long-term, extensive synergies amongst immunologists and researchers on DNA damage response and aging. About The Workshop Table Of Contents Program Schedule Session 1: Immunity & Aging I Session 2: Immunity & Aging II Session 3: DNA Damage & Aging I Session 4: DNA Damage & Aging II Session 5: DNA Damage & Aging III P 7 - P 10 P 13 - P 16 P 19 - P 24 P 27 - P 30 P 33 - P 38 P 41 - P 44 Session 6: Cancer I Session 7: Cancer II Session 8: Cancer III Abstract Submissons Short Oral Presentations Abstract Submissons Poster Presentations Notes Acknowledgements P 47 - P 52 P 55 - P 56 P 59 - P 60 P 63 - P 72 P 75 - P 110 P 111 - P 120 P 121 - 128 PROGRAM SCHEDULE P 7 Program Schedule (Day 1 - 2) Day 1 - 10 October 2022, Monday 8.30am 9.00am 9.10am SESSION 1: Immunity & Aging I 9.20am 10.10am 10.40am 11.10am 11.40am 12.10pm SESSION 2: Immunity & Aging II 1.25pm 1.55pm 2.25pm 2.55pm 3.25pm 3.55pm 4.05pm 4.15pm 4.45pm 5.55pm Registration / Arrival of guests (Coffee | Mixer) Welcome Remarks: Vinay Tergaonkar Opening Address: Barry Halliwell Chairman, BMRC Advisory Council, A*STAR, Singapore Senior Advisor, Office of the Senior Deputy President and Provost, NUS, Singapore Distinguished Professor, Yong Loo Lin School of Medicine, NUS, Singapore Chaired by: Ashok Venkitaraman Cancer Science Institute, Singapore Fabrizio d’Adda di Fagagna T he FITC Institute of Molecular Oncology, Milan, Italy Keynote: Telomere Biology in Aging and Disease Jan Rehwinkel University of Oxford, UK Viral Targeting Of The cGAS Pathway COFFEE BREAK Nicolas Manel Curie Institute, Paris, France Nuclear Envelope Disruption Triggers Hallmarks Of Aging In Lung Alveolar Macrophages Vinay Tergaonkar Institute of Molecular and Cell Biology, Singapore Molecular Determinants Of Telomere Mediated Inflammation LUNCH Chaired by: Vincent Géli Marseille Cancer Research Centre, Marseille, France Florent Ginhoux Institut Gustave Roussy, Villejuif, France Understanding Macrophage Heterogeneity Bing Su Shanghai Institute of Immunology, Shanghai, China Sin1 Mediated mTOR Signal In Lymphocyte Growth And Metabolism Violeta Serra Vall d’Hebron Institute of Oncology, Barcelona, Spain Advances Using Functional HRD To Identify PARP Inhibitor Sensitive Tumours Nadine Laguette Institut Genetique Humaine, Montpellier, France STING Is A Central Rheostat of Metabolic And Inflammatory Homeostasis COFFEE BREAK Microglial Dysfunction in Ataxia-Telangiectasia: Role Of The Cystolic DNA Sensing cGAS-STING Pathway Emily Talbot Determining The Molecular Mechanism Of Cellular Aging In The Premature Aging Syndrome Hutchinson-Gilford Progeria Mattheus Foo Toshio Suda Cancer Science Institute of Singapore, Singapore Self-Renewal And Expansion Of Hematopoietic Stem Cells Under The Stress DNA Damage, Lipids Mediators and Aging Aditi Uday, Gurkar Eric Gilson Institute for Research on Cancer and Aging, Nice, France The Role Of The Shelterin TRF2 Subunit In Cellular Aging: From Dividing To Post- Mitotic Cells EMBO WORKSHOP 2022 P 8 THE DNA DAMAGE RESPONSE, IMMUNITY AND AGING Day 2 - 11 October 2022, Tuesday 9.15am SESSION 3: DNA Damage & Aging I 10.00am 10.50am 11.00am 11.30am 12.00pm 12.10pm 12.20pm 12.50pm SESSION 4: DNA Damage & Aging II 2.35pm 3.05pm 3.35pm 4.05pm 4.55pm 5.25pm 6.00pm POSTERS (Coffee served) Chaired by: John Sedivy Brown University, Providence, USA Marco Foiani IFOM, Milan, Italy Keynote: An Integrated Mechano-Response Mediated By ATR And ATM cGAS Accelerates Mitotic Cell Death Through The NLRP3 Inflammasome Enaam Alghamdi COFFEE BREAK Ming Lei Institute for Precision Medicine, Jiaotong University, Shanghai, China A Novel Primate-Specific X-Linked Amplicon Maintains Spermatogonia Self-Renewal And Genome New Anti-Aging Targets In Progeria Identified Through Synthetic Rescue Genome Wide Arrayed CRISPR Screening Delphine Larrieu Ganglioside Based Immune Checkpoint Controls Senescent Cell Fate: A Paradigm Shift In Cancer & Aging? Julien Cherfils-Vicini Brian Kennedy National University of Singapore, Singapore Pillars And Hallmarks Of Aging From The Perspective Of Longevity Interventions LUNCH / POSTER SESSION 1 [01 - 16] Chaired by: Eric Gilson Institute for research on Cancer and Aging, Nice, France Jing Ye Personalized Aging Medicine Center, Ruijin Hospital, Jiaotong University, Shanghai, China The Evolutionary Trajectory Of Telomeric Protective Proteins In Zebrafish Sophie Postel-Vinay Institut Gustave Roussy, Villejuif, France Harnessing Genetic Vulnerabilities In Immuno-oncology: Novel Therapeutic Opportunities COFFEE BREAK James Chen Howard Hughes Medical Institute, University of Texas Southwestern Medical Center Dallas, USA Keynote: Igniting An Immune Response To DNA With cGAS Hyeseong Cho Ajou University of Medicine, Suwon, South Korea DNA Damage Response And Mitochondrial Dynamics Eiji Hara Osaka University, Osaka, Japan Cellular Senescence And Cancer: Relevance To Microorganisms End of Day 2 Depart for Dinner MIXER / DISCUSSION (Meeting rooms #1-06, #2-08) End of Day 1 Depart for dinner 5.25pm 6:00pm PROGRAM SCHEDULE P 9 Program Schedule (Day 3 - 4) Day 3 - 12 October 2022, Wednesday 9.15am SESSION 5: DNA Damage & Aging III 10.00am 10.50am 11.00am 11.30am 12.00pm 12.30pm SESSION 6: Cancer I 2.00pm 2.30pm 3.00pm 3.30pm 4.10pm 4.40pm 4.50pm 5.00pm 6:00pm POSTERS (Coffee served) Chaired by: Hyunsook Lee Seoul National University, Seoul, South Korea Judith Campisi Buck Institute, Novato, USA Keynote: TBD Nuclear Translocation Of Protein Arginine Methyltransferase 5 (PRMT5) Regulates The Immune Response To Replication Stress Anand D Jeyasekharan COFFEE BREAK Joao Passos Mayo Clinic, Rochester, USA Targeting Senescence For Healthier Aging: All Roads Lead To Mitochondria George Garinis IMBB-FORTH, Heraklion, Greece DNA Damage And Innate Immune Responses In Health And Disease LUNCH / POSTER SESSION 2 [17 - 32] Chaired by: Fabrizio d’Adda di Fagagna The FIRC Institute of Molecular Oncology & IGM-CNR (National Research Council), Milan, Italy Vincent Géli Marseille Cancer Research Centre, Marseille, France Telomerase Preserves Lung Function In Old Mice By Promoting Endothelial Cell Renewal And Preventing Cellular Senescence Hyunsook Lee Seoul National University, Seoul, South Korea Dynamic Interaction Of BRCA2 With The Telomeric G-Quadruplexes Karen Crasta National University of Singapore, Singapore Small Extracellular Vesicles From Therapy-Induced Senescent Breast Cancer Cells Elicit Paracrine Anti-Tumour Effects COFFEE BREAK Sunny Songyang Zhou Sun Yat-sen University Guangzhou Key Laboratory of Healthy Aging Research, Guangzhou, China Identification Of Key Regulators Of DNA Repair In Human Cells By Inducible CRISPR/ Cas9 Screens ATR Inhibition As A Therapeutic Strategy To Boost Antitumour Immune Reponses In Triple Negative Breast Cancer Giulia Bastianello ZNF524 Directly Interacts With Telomeric DNA And Supports Telomere Integrity Dennis Kappei Andrea Ablasser EPFL, Lausanne, Switzerland Keynote: The cGAS-STING Pathway In Aging-Associated Neurodegeneration End of Day 3 Free and Easy Dinner EMBO WORKSHOP 2022 P 10 THE DNA DAMAGE RESPONSE, IMMUNITY AND AGING Day 4 - 13 October 2022, Thursday 9.15am SESSION 7: Cancer II 10.00am 10.50am 11.00am 11.30am 12.00pm 12.10pm 12.20pm SESSION 8: Cancer III 1.50pm 2.00pm 2.30pm 3.20pm 3.30pm POSTERS (Coffee served) Chaired by: Jan Karlseder Salk Institute, La Jolla, USA John Sedivy Brown University, Providence, USA Keynote: Involvement of Retrotransposons in Aging and Age-Related Diseases The Relationship Between Spliceosome Mutations And Immune Activation In Cancer Katrina Lappin COFFEE BREAK Ashok Venkitaraman Cancer Science Institute, Singapore Regulation Of DNA Recombination By The BRCA2 Cancer Suppressor And Its Modulation By Drug-Like Small Molecules Cisplatin-Induced DNA Damage Triggers MRE11a Proteasomal Degradation Mediated By UBQLN4 In Solid Tumours Dave Hoon Potassium Efflux By Nigericin Triggers ZAKa-Driven Ribotoxic Stress And NLRP1 Inflammasome Activation Pritisha Rozario LUNCH / POSTER SESSION 3 [33 - 46] Chaired by: Violeta Serra Vall d’Hebron Institute of Oncology, Barcelona, Spain Cdkn1a-Driven Expression Of Telomerase Improves Insulin-Resistance By Preventing Obesity Associated Senescence Phenotype In Adipose Tissue Laura Braud Chris Lord The Institute of Cancer Research, London, UK Using Forward And Reverse Translation Approaches To Understand DDR Inhibitor Response Jan Karlseder Salk Institute, La Jolla, USA Closing Keynote: Telomere To Mitochondria Signaling Prevents Cancer Initiation Thank You Note Hong Wanjin Institute of Molecular and Cell Biology, A*STAR, Singapore Closing Remarks Eric Gilson End of Workshop Session 1 Immunity & Aging I Chaired by: Ashok Venkitaraman SESSION 1: IMMUNITY & AGING I P 13 SPEAKERS Dr Fabrizio is a molecular and cell biologist with twenty years of experience as a group leader. He has devoted his professional life to understanding the critical biological mechanisms underlying aging and cancer. Dr Fabrizio works at IFOM (Milan) and IGM-CNR (Pavia) in Italy. Together with his group, they study the connection between DNA damage and disease. Dr Fabrizio is generally recognized for his discoveries in telomere biology and cellular senescence. He has been awarded the European Research Council (ERC) advanced grant twice and is a member of the European Molecular Biology Organization (EMBO). Recently, his group has recently identified what they believe is the main cause of several diseases of aging: loss of integrity of the telomeres, the protective caps at the ends of our chromosomes. Following that discovery, they have developed a universal inhibitor of cellular aging based on RNA therapeutics and have validated it in several in vivo models of human diseases. Telomere Biology In Aging And Disease During aging, senescent cells accumulate in tissues and organs, which become unable to proliferate and function properly. Telomeric shortening and damage trigger most, if not all, the downstream pathways leading to senescence and disease. The DNA damage response (DDR) pathways activated by short or damaged telomeres are necessary for cellular senescence enforcement. We have shown that damage-induced long non-coding RNAs (dilncRNAs) are generated at sites of DNA damage, including telomeres. Antisense oligonucleotides (ASOs) against such RNAs allow site-specific DDR inhibition (Francia et al. Nature 2012, Michelini et al. Nature Cell Biology 2017, Pessina et al. 2019). Telomeric ASOs inhibit DDR activation at dysfunctional telomeres in cultured cells and in vivo in mice (Rossiello et al . Nature Communications 2017). In a mouse model of Hutchinson-Gilford Progeria Syndrome (HGPS), selective DDR inhibition at telomeres improve tissue homeostasis, reduce inflammation and extend lifespan (Aguado et al. Nature Communications 2019). Lung fibrosis and altered hematopoiesis are human conditions associated with short telomeres and telomerase mutations. Telomerase knockout mice recapitulate these pathologies. In this model, tASO improves respiratory and hematopoietic systems. In addition, we recently observed that telomeric shortening or dysfunction cause an increase in the levels of ACE2, the SARS-CoV-2 receptor, in humans and in mice and that selective tDDR inhibition prevents it in vivo PRINCIPAL INVESTIGATOR (PI) IFOM & IGM-CNR Dr. Fabrizio d’Adda di Fagagna EMBO WORKSHOP 2022 P 14 THE DNA DAMAGE RESPONSE, IMMUNITY AND AGING Jan is interested in the molecular mechanisms underlying host-pathogen interactions, especially how cells detect virus infection. His work lies at the intersection of immunology, virology and molecular biology. After his undergrad at the University of Heidelberg, Jan joined the European Molecular Biology Laboratory (EMBL) as a PhD student. Under supervision of Elisa Izaurralde, Jan studied post-transcriptional control of messenger RNA, and obtained a PhD in 2007. This background in RNA biology led him to develop an interest in nucleic acids in innate immunity. As a postdoctoral fellow, he joined the group of Caetano Reis e Sousa, then at the Cancer Research UK London Research Institute (London, UK). Jan investigated how RNA viruses are recognised by innate immune sensors. In 2012, Jan moved to the University of Oxford, UK. His laboratory is part of the MRC Human Immunology Unit and the MRC Weatherall Institute of Molecular Medicine. Jan’s research dissects nucleic acid sensing by innate receptors in the context of virus infection, autoinflammatory disease and cancer. Viral Targeting Of The cGAS Pathway Nucleic acid sensing is a fundamental process in the immune system. Viruses introduce DNA or RNA genomes into cells, which often serve as molecular signatures of infection, detected by nucleic acid sensors. cGAS is a sensor for double-stranded DNA which detects many viral pathogens and ‘self’ DNA, thereby driving sterile inflammation. cGAS produces cGAMP, activating STING, which signals for innate immune activation. Viruses antagonize the cGAS pathway to dampen innate immunity and facilitate their replication. We found that the cGAS pathway was required for type I interferon (T1-IFN) induction during Varicella-Zoster virus (VZV) infection. Viral gene overexpression screening identified the VZV tegument protein ORF9 as a cGAS antagonist. Ectopically and virally expressed ORF9 bound to cGAS and reduced the T1-IFN response to transfected DNA. ORF9 and cGAS also interacted directly in a cell-free system and phase- separated together with DNA. Taken together, we uncovered the importance of the cGAS pathway for VZV recognition and identified a viral cGAS antagonist. cGAMP not only activates STING but also transfers to other cells. We report that infection with multiple viruses depleted cGAMP channels from cells. This included HSV-1 that targeted the VRAC subunits LRRC8A/C, SLC46A2, and P2X7R for degradation. The HSV-1 protein UL56 was required and sufficient for these effects. We propose this limits innate immunity by reducing cell-to-cell communication via cGAMP. GROUP LEADER, MRC HUMAN IMMUNOLOGY UNIT MRC Weatherall Institute of Molecular Medicine, University of Oxford, UK Prof. Jan Rehwinkel SESSION 1: IMMUNITY & AGING I P 15 SPEAKERS Nicolas Manel received his PhD from University of Montpellier, France in 2005. He then received training as a postdoctoral fellow in the lab of Dan Littman at NYU. In 2010, he became an INSERM principal investigator at Institut Curie in Paris, France. The Manel lab focuses on understanding how cells discriminate self from non-self or altered self. The lab pioneered the study of the cGAS-STING pathway in the context of HIV infection. The lab currently studies the regulation of innate sensors in response to viruses and self, their evolution during aging, and their use in immuno-oncology. Nuclear Envelope Disruption Triggers Hallmarks Of Aging In Lung Alveolar Macrophages Aging is characterized by gradual immune dysfunction and increased risk for many diseases, including respiratory infections. Genomic instability is thought to play a central role in the aging process but the mechanisms that damage nuclear DNA in aging are insufficiently defined. Cells that migrate or reside within confined environments experience forces applied to their nucleus, leading to transient nuclear envelope (NE) ruptures. NE ruptures are associated with DNA damage, and Lamin A/C is required to limit these events. Here, we show that Lamin A/C protects lung alveolar macrophages from NE rupture and hallmarks of aging. Lamin A/C ablation in immune cells results in a selective depletion of lung alveolar macrophages (AM) and a heightened susceptibility to influenza infection. Lamin A/C-deficient AM that persist display constitutive nuclear envelope rupture marks, DNA damage and p53-dependent senescence. In wild-type mice, we found that AM migrates within constricted spaces in vivo , at heights that induce NE rupture and DNA damage. AM from aged wild-type mice and from Lamin A/C-deficient mice share an upregulated lysosomal signature with CD63 expression, and we find that CD63 is required to clear damaged DNA in macrophages. We propose that induction of genomic instability by NE disruption represents a mechanism of aging in alveolar macrophages. RESEARCH DIRECTOR Institut Curie Prof. Nicolas Manel EMBO WORKSHOP 2022 P 16 THE DNA DAMAGE RESPONSE, IMMUNITY AND AGING Vinay Tergaonkar obtained his Ph.D. (2001) from NCBS Bangalore, through an international cancer society (UICC) fellowship for collaborative research at Tufts University, Boston, USA. He has been a fellow (2001-2004) and a special fellow (2004-present) of the Leukemia and Lymphoma Society of America and conducted his postdoctoral studies at the Salk Institute for Biological Studies, La Jolla, California. He currently serves as Research Director at Institute for Molecular and Cell Biology (IMCB), Singapore, and a Professor at School of Medicine at National University of Singapore. He serves on Editorial Boards of 1) Science Advances (AAAS), 2) Molecular and Cellular Biology (American Society for Molecular Biology), 3) Biochemical Journal (Portland Press). Work from his lab has received international recognition including the British council development award (2014), the Premiers’ fellowship from Government of South Australia (2015) and University of Macau Distinguished Professorship (2019). Molecular Determinants Of Telomere Mediated Inflammation Over 90% of human cancers attain immortality by transcriptional reactivation of an enzyme called telomerase reverse transcriptase (TERT). How and why TERT is reactivated in the vast majority of human cancers remain as fundamental unsolved problems in biology. I will present unpublished and unexpected set of results which suggest why TERT is reactivated in human cancers. RESEARCH DIRECTOR Institute of Molecular and Cell Biology, A*STAR Prof. Vinay Tergaonkar Session 2 Immunity & Aging II Chaired by: Vincent Géli SESSION 2: IMMUNITY & AGING II P 19 SPEAKERS Florent Ginhoux graduated in Biochemistry from the University Pierre et Marie CURIE (UPMC), Paris VI, obtained a Masters degree in Immunology from the Pasteur Institute in 2000 and his PhD in 2004 from UPMC, Paris VI. As a postdoctoral fellow, he joined the Laboratory of Miriam Merad in the Mount Sinai School of Medicine (MSSM), New York, where he studied the ontogeny and the homeostasis of cutaneous dendritic cell populations, with a strong focus on Langerhans cells and Microglia. In 2008, he became an Assistant Professor in the Department of Gene and Cell Medicine, MSSM and member of the Immunology Institute of MSSM. He joined the Singapore Immunology Network (SIgN), A*STAR in May 2009 as a Junior Principal Investigator and became Senior Principal Investigator in 2014. He joined the EMBO Young Investigator (YIP) program in 2013 and is a Web of Science Highly Cited Researcher since 2016. He is also an Adjunct Visiting Associate Professor in the Shanghai Immunology Institute, Jiao Tong University, in Shanghai, China since 2015 and Adjunct Associate Professor in the Translational Immunology Institute, SingHealth and Duke NUS, Singapore since 2016. He is now a Laboratory Director in Gustave Roussy focusing on pediatric cancers and the role of myeloid cells in tumor progression and became an EMBO member in 2022. Understanding Macrophage Heterogeneity Resident tissue macrophages (RTMs) have a broad spectrum of immune- and non-immune-related tissue-supporting activities. The roots of this heterogeneity and versatility are only beginning to be understood. Here, we propose a conceptual framework for considering the RTM heterogeneity that organizes the factors shaping RTM identity within four cardinal points: (1) ontogeny and the view that adult RTM populations comprise a defined mixture of cells that arise from either embryonic precursors or adult monocytes; (2) local factors unique to the niche of residence, evolving during development and aging; (3) inflammation status; and (4) the cumulative effect of time spent in a specific tissue that contributes to the resilient adaptation of macrophages to their dynamic environment. Prof. Florent Ginhoux LABORATORY DIRECTOR Gustave Roussy Senior PI & SIgN A*STAR EMBO WORKSHOP 2022 P 20 THE DNA DAMAGE RESPONSE, IMMUNITY AND AGING Dr. Bing Su is the Director of Shanghai Institute of Immunology and Chair of Department of Immunology and Microbiology at Shanghai JiaoTong University School of Medicine, Co-Founding Director of Shanghai JiaoTong University School of Medicine-Yale University Institute for Immune Metabolism (SYIIM) and an Adjunct Professor at Department of Immunobiology at Yale University School of Medicine. Dr. Su also holds a KC Wong Chair Professorship at Shanghai JiaoTong University. Dr. Su’s research focuses on the intracellular signal transduction pathways controlled by the mammalian target of rapamycin (mTOR) and the mitogen activated protein kinases (MAPKs). He studies the roles of these intracellular signaling cascades in immune regulation and vascular function. Dr. Su has published over 110 peer-reviewed papers, many of which were in high-impact journals such as Nature, Cell, Nature Genetics, Nature Immunology, Immunity, Molecular Cell, EMBO J, etc. Sin1 Mediated mTOR Signal In Lymphocyte Growth And Metabolism The mammalian target of rapamycin (mTOR) is an evolutionarily conserved protein kinase with a central role in cell growth and metabolism. Multiple mTOR-containing protein complexes (mTORCs) exist to mediate mTOR function via controlling many cellular targets including the members of the protein kinase (PK)A/PKG/PKC (AGC) family. Sin1 is an essential component of mTORC2 with crucial roles in regulating immune cell growth and metabolism. The function of Sin1-mTORC2 has been explored in immune cell growth and metabolism but its roles in tumor growth and anti-tumor immunity is still largely unclear. We will present data on Sin1-mTORC2’s roles in these processes. DIRECTOR; CHAIR Shanghai Institute of Immunology; Shanghai JiaoTong University School of Medicine Prof. Bing Su