Involvement of circulating factors in the transmission of paternal experiences through the germline

• DOI: 10.15252/embj.2020104579
• Publication Date: 2020-10-09T11:41:11Z
• Authors: Gretchen van Stee...; Katharina Gapp; Ali Jawaid; Pierre‐Luc Germain; Francesca Manuella; Deepak K Tanwar; Nicola Zamboni; Niharika Gaur; Anastasiia Efimova; Kristina M Thumfart; Eric A Miska; Isabelle M Mansuy
• Keywords: Epigenomics; Male; 0301 basic medicine; paternal experiences; blood serum; paternal experiences; PPAR; sperm; transmission; 610 Medicine & health; Genetics and Molecular Biology; PPAR; sperm; Epigenesis, Genetic; Fathers; Mice; 03 medical and health sciences; 1300 General Biochemistry, Genetics and Molecular Biology; 2400 General Immunology and Microbiology; 1312 Molecular Biology; Animals; Humans; Molecular Biology; 0303 health sciences; 10242 Brain Research Institute; General Immunology and Microbiology; General Neuroscience; Reproduction; transmission; 2800 General Neuroscience; Articles; Spermatozoa; 3. Good health; Mice, Inbred C57BL; Blood; Germ Cells; Phenotype; blood serum; General Biochemistry; Paternal Exposure; 570 Life sciences; biology; Wounds and Injuries; Transcriptome

Article9 October 2020Open Access Transparent process Involvement of circulating factors in the transmission paternal experiences through germline Gretchen van Steenwyk orcid.org/0000-0001-8225-3818 Laboratory Neuroepigenetics, Brain Research Institute, Medical Faculty University Zurich, Switzerland Institute for Neuroscience, Department Health Sciences and Technology, ETH Zurich Neuroscience Center, Search more papers by this author Katharina Gapp Molecular Behavioral Gurdon Cambridge, UK Wellcome Trust Sanger Hinxton, Genetics, Ali Jawaid Translational Neuropsychiatric Disorders, BRAINCITY Nencki-EMBL Center Excellence Neural Plasticity Warsaw, Poland Pierre-Luc Germain orcid.org/0000-0003-3418-4218 Statistical Bioinformatics Group, Swiss Bioinformatics, Zürich, Francesca Manuella Deepak K Tanwar orcid.org/0000-0001-8036-1989 Nicola Zamboni Systems Biology, Niharika Gaur Anastasiia Efimova Kristina M Thumfart orcid.org/0000-0002-7065-4347 Eric A Miska orcid.org/0000-0002-4450-576X Isabelle Mansuy Corresponding Author [email protected] orcid.org/0000-0001-7785-5371 Information Steenwyk1,2,3,‡, Gapp2,3,4,5,6,7,‡, Jawaid1,2,3,8, Germain1,2,9, Manuella1,2,3, Tanwar1,2,3,9, Zamboni10, Gaur1,2,3, Efimova1,2,3, Thumfart1,2,3, Miska5,6,7 *,1,2,3 1Laboratory 2Institute 3Zurich 4Laboratory 5Gurdon 6Wellcome 7Department 8Laboratory 9Statistical 10Institute ‡These authors contributed equally to work *Corresponding author. Tel: [phone]; E-mail: The EMBO Journal (2020)39:e104579https://doi.org/10.15252/embj.2020104579 See also: AJ Hannan (November 2020) PDFDownload PDF article text main figures. Peer ReviewDownload a summary editorial decision including letters, reviewer comments responses feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Environmental can change phenotypes exposed individuals offspring involve germline, likely via biological signals periphery that communicate with germ cells. Here, using mouse model exposure traumatic stress, we identify involving peroxisome proliferator-activated receptor (PPAR) pathways effects germline. We show alters metabolic functions pathways, particularly lipid-derived metabolites, fathers their offspring. collected data human cohort childhood trauma observed similar alterations circulation, suggesting conserved effects. Chronic injection serum from trauma-exposed males into controls recapitulates lipid-activated nuclear receptors PPARs as potential mediators father Pharmacological PPAR activation vivo reproduces dysfunctions grand-offspring injected affects sperm transcriptome sons. In germ-like cells vitro, both agonist induce activation. Together, these results highlight role communication vectors between Synopsis Exposure mice stress early life leads phenotypic changes are transmitted progeny mechanisms remain poorly characterized. vitro findings humans implicate serum-induced proliferating-activated signaling environmentally-induced traits Lipid metabolism altered plasma adult male postnatal is also saliva orphan children. Serum activates spermatogonial vitro. trauma-induced RNA fathers. Injection control symptoms Introduction events have long-lasting consequences individuals, some cases, they impact Transmission environmentally induced features diseases has been overlooked decades. But today, evidence diet, or endocrine disruptors across generations accumulated experimental animals (Bohacek Mansuy, 2015; Nilsson et al, 2018; Panzeri Pospisilik, 2018). These known depend on epigenetic constitute an important aetiological component many diseases. When parent not depending maternal care social factors, thought They therefore represent form heredity. how affect which body reach known. may vary type exposure, its time window, chronicity, etc. common ability Circulating tissues body. postulate carry contribute progeny. Blood metabolites particular strong candidates being such carriers because potent signalling molecules, e.g. hormones, lipids, organic acids antioxidants. Further, dynamically regulated physiological states mammals. Several previously implicated regulation genome different (Donohoe Bultman, 2012; Kaelin 2013; Sharma Rando, 2017). Results examined contribution used established based unpredictable separation combined (MSUS) (Fig 1A B). Mice MSUS behavioural deficits several (Franklin 2010; 2014b, 2016b; conducted unbiased metabolomic analyses time-of-flight mass spectrometry (TOF-MS). showed polyunsaturated fatty acid (PUFA) metabolism, particular, involved α-linolenic/linoleic (ALA/LA), arachidonic (AA) significantly upregulated 1C). PUFAs, eicosapentaenoic (EPA) dihomo-gamma-linoleic (DGLA), hydroxyeicosatetraenoic (HETEs), were most within enrichments 1D). addition, bile biosynthesis well steroidogenesis steroidogenic ligand aldosterone downregulated 1C, full table Appendix Figs S1 S2). Altered consistent previous observation steroid mineralocorticoid (MR) downregulated. Its pharmacological blockade mimics (Gapp 2014b). Remarkably, except AA when Fig S1). focused since bodyweight affected but female (Appendix S3). Figure 1. Effects Paradigms humans. mice, consists (MSUS). humans, involves loss (PLMS) (age 6–12). Scheme illustrating showing blood collection breeding generate For (symbolized yellow blitz), newborn pups separated mother unpredictably 3 h/day day (PND) 1–14. During separation, dam stressors unpredictably9. was prepared 3-month-old males. Differential pathway enrichment compared (each group n = 5), (PLMS, 20; control, 14) 25; PLMS Asterisk hashtag FDR after multiple testing corrections Benjamini–Hochberg (BH) test. Columns indicate significance positive (+) negative (−) enrichment. Individual ALA/LA PLMS. Numbers fold according heat scale (right). Data information: #FDR < 0.1, *FDR 0.05, **FDR 0.01, ***FDR 0.001, ****FDR 0.0001. (/) symbolizes non-significance. FDR, false discovery rate. ALA/LA, alpha-linolenic acid/linoleic acid. AA, HETE, Download figure PowerPoint children assessed relevance conducting (6- 12-year-old girls boys) SOS Children's Village Lahore, Pakistan. lost (paternal PLMS) during preceding year 1A). conditions closely resemble model. This highly relevant our study comparable at age, key correlative data. All live same orphanage, so differences lifestyle minimal. Control schoolmates living parents any trauma. groups matched index gender S4A–C). attended school, equal access playground facilities physical exercise. Pakistani population advantageous consanguinity high Pakistan (Bittles 1991) S4D), making genetically homogeneous than other populations. Body index, diet ethnicity account 5% variance healthy 6 European populations (Lau 2018), samples indeed following ones over avoid interference clotting factors. had modest controls, EPA, DGLA HETEs strongly 1C D) similarly MSUS. saliva, S5), indicating fluids. Among enrichments, individual comparably S6). activated While consider all be potentially transmission, acids, especially PUFA, metabolites. PUFAs modulate inflammation cognitive functions. Fatty bind various receptors, ligands (PPARs). widely expressed regulate gene expression chromatin structure, act forming transcription factor complexes retinoid X (RXR). interact modifying enzymes (Yu Reddy, 2007; Romagnolo 2014). Further MSUS, farsenoid (FXR) liver (LXR). FXR LXR belong family RXR, them (Chawla 2001). Since ligands, next if linked activity. target genes tissues. white adipose tissue, PPARγ abundant regulates adipocyte differentiation (Lee Ge, measured binding assay increased tissue S7A). liver, PPARα activity (Rakhshandehroo 2010), targets differentially expressed, S7B). passed offspring, PPAR. PPARγ, isotype gametes (Aquila 2006), 2A). then asked influence stem cell-like (GC-1 spg), diploid early-stage cells, assess Spermatogonial chosen primary present developing testes (first 2 weeks birth). GC-1 spg culture medium enriched 10% 2B). Prior transfected plasmid expressing luciferase under response element (PPRE). Luciferase luminescence higher 2C), serum. 2. Analyses mRNA mice. Control, 15; 13, two-tailed Student's t-test, P 0.029, t 2.30, df 26. Schematic treatment On 1, PPRE normalization plasmids. mixed concentration Cells harvested 24 h later (Day 3), measured. Relative correspond applied cell wells. 17; 22, 0.042, 2.1, 37. reported mean ± SEM. Paternal induces Although studies environmental phenotype (Lillycrop 2008; Carone Zeybel Martínez 2014; Baptissart none tested causal involvement. mimicking chronic intraperitoneal (i.p.) dual PPARα/γ tesaglitazar (10 μg/kg) 3A). chose activate better mimic Following 46-day delay last allow spermatogenesis cycle eliminate transient drug, bred females adult, naïve produce grand-offspring. Both tesaglitazar-injected reduced weight vehicle-injected 3B; S8A), despite initial increase PND8 S9). effect due difference fathers' injections S10). glucose tolerance test (GTT) 3C; S8B), insulin sensitivity 2014a). However, restraint altered, unlike 2014a; S11A). does generations. 3. Tesaglitazar either (grey syringe) vehicle twice per week 4 weeks. Males paired Adult (Tesa-inj, 16) (Vehicle-inj, 23). Two-tailed 0.019, 2.44, Glucose level Tesa-inj (n Vehicle-inj 21) Repeated-measures ANOVA, 0.0083, F (1, 33) 7.877, 0.0001, (4, 132) 347.7, interaction 0.0776, 2.155 Conc.; concentration. opposite Because knockout models cannot viable (Barak 1999; Kubota 1999) lifelong alteration physiology (Bensinger Tontonoz, 2008), antagonist (T0070907), suitable existing drug could identify. T0070907 saline males, obtain did GTT S12A B), inhibition adulthood sufficient effectively compensated for. MSUS-induced shift payload reflected causally 2014a, 2020), Deep sequencing revealed differential dysregulation transposable elements (TEs) S13A). observations tesaglitazar-treated (Ferguson 2018) 2020). targeted analysis, 630 TE > 4,000 mRNA/lincRNA transcripts annotated there significant correlation TEs (MSUS versus controls), long terminal repeat (LTRs) 4A; S13B). mRNAs/lincRNAs