Sperm DNA Methylation Alterations from Cannabis Extract Exposure Are Evident in Offspring

Overview

Journal Epigenetics Chromatin

Publisher Biomed Central

Specialties Biochemistry
Genetics

Date 2022 Sep 9

PMID 36085240

Authors

Rose Schrott

Jennifer L Modliszewski

Andrew B Hawkey

Carole Grenier

Zade Holloway

Janequia Evans

Erica Pippen

David L Corcoran

Edward D Levin

Susan K Murphy

Affiliations

Soon will be listed here.

Abstract

Background: Cannabis legalization is expanding and men are the predominant users. We have limited knowledge about how cannabis impacts sperm and whether the effects are heritable.

Results: Whole genome bisulfite sequencing (WGBS) data were generated for sperm of rats exposed to: (1) cannabis extract (CE) for 28 days, then 56 days of vehicle only (~ one spermatogenic cycle); (2) vehicle for 56 days, then 28 days of CE; or (3) vehicle only. Males were then mated with drug-naïve females to produce F1 offspring from which heart, brain, and sperm tissues underwent analyses. There were 3321 nominally significant differentially methylated CpGs in F0 sperm identified via WGBS with select methylation changes validated via bisulfite pyrosequencing. Significant methylation changes validated in F0 sperm of the exposed males at the gene 2-Phosphoxylose Phosphatase 1 (Pxylp1) were also detectable in their F1 sperm but not in controls. Changes validated in exposed F0 sperm at Metastasis Suppressor 1-Like Protein (Mtss1l) were also present in F1 hippocampal and nucleus accumbens (NAc) of the exposed group compared to controls. For Mtss1l, a significant sex-specific relationship between DNA methylation and gene expression was demonstrated in the F1 NAc. Phenotypically, rats born to CSE-exposed fathers exhibited significant cardiomegaly relative to those born to control fathers.

Conclusions: This is the first characterization of the effect of cannabis exposure on the entirety of the rat sperm methylome. We identified CE-associated methylation changes across the sperm methylome, some of which persisted despite a "washout" period. Select methylation changes validated via bisulfite pyrosequencing, and genes associated with methylation changes were involved in early developmental processes. Preconception CE exposure is associated with detectable changes in offspring DNA methylation that are functionally related to changes in gene expression and cardiomegaly. These results support that paternal preconception exposure to cannabis can influence offspring outcomes.

Citing Articles

Tackling somatic DNA contamination in sperm epigenetic studies.

Kumari A, Singh R Front Reprod Health. 2025; 7:1506117.

PMID: 39974749 PMC: 11835817. DOI: 10.3389/frph.2025.1506117.

Paternal impact on the developmental programming of sexual dimorphism.

Aljabali S, Pai S, Teperino R Front Cell Dev Biol. 2024; 12:1520783.

PMID: 39712575 PMC: 11659275. DOI: 10.3389/fcell.2024.1520783.

A new approach to study stochastic epigenetic mutations in sperm methylome of Vietnam war veterans directly exposed to Agent Orange.

Corsaro L, Sacco D, Corbetta C, Gentilini D, Faversani A, Ferrara F Environ Epigenet. 2024; 10(1):dvae020.

PMID: 39664489 PMC: 11631699. DOI: 10.1093/eep/dvae020.

Key insights into cannabis-cancer pathobiology and genotoxicity.

Reece A, Hulse G Addict Biol. 2024; 29(11):e70003.

PMID: 39538372 PMC: 11560801. DOI: 10.1111/adb.70003.

Influence of substance use on male reproductive health and offspring outcomes.

Lo J, Hedges J, Chou W, Tager K, Bachli I, Hagen O Nat Rev Urol. 2024; 21(9):534-564.

PMID: 38664544 DOI: 10.1038/s41585-024-00868-w.

References

Stelzer G, Rosen N, Plaschkes I, Zimmerman S, Twik M, Fishilevich S . The GeneCards Suite: From Gene Data Mining to Disease Genome Sequence Analyses. Curr Protoc Bioinformatics. 2016; 54:1.30.1-1.30.33. DOI: 10.1002/cpbi.5. View

Levin E, Hawkey A, Hall B, Cauley M, Slade S, Yazdani E . Paternal THC exposure in rats causes long-lasting neurobehavioral effects in the offspring. Neurotoxicol Teratol. 2019; 74:106806. DOI: 10.1016/j.ntt.2019.04.003. View

Holloway Z, Hawkey A, Torres A, Evans J, Pippen E, White H . Paternal cannabis extract exposure in rats: Preconception timing effects on neurodevelopmental behavior in offspring. Neurotoxicology. 2020; 81:180-188. DOI: 10.1016/j.neuro.2020.10.007. View

Dolzhenko E, Smith A . Using beta-binomial regression for high-precision differential methylation analysis in multifactor whole-genome bisulfite sequencing experiments. BMC Bioinformatics. 2014; 15:215. PMC: 4230021. DOI: 10.1186/1471-2105-15-215. View

Yagi S, Galea L . Sex differences in hippocampal cognition and neurogenesis. Neuropsychopharmacology. 2018; 44(1):200-213. PMC: 6235970. DOI: 10.1038/s41386-018-0208-4. View

Brunton P . Effects of maternal exposure to social stress during pregnancy: consequences for mother and offspring. Reproduction. 2013; 146(5):R175-89. DOI: 10.1530/REP-13-0258. View

Hodes G, Pfau M, Purushothaman I, Ahn H, Golden S, Christoffel D . Sex Differences in Nucleus Accumbens Transcriptome Profiles Associated with Susceptibility versus Resilience to Subchronic Variable Stress. J Neurosci. 2015; 35(50):16362-76. PMC: 4679819. DOI: 10.1523/JNEUROSCI.1392-15.2015. View

Schrott R, Acharya K, Itchon-Ramos N, Hawkey A, Pippen E, Mitchell J . Cannabis use is associated with potentially heritable widespread changes in autism candidate gene DNA methylation in sperm. Epigenetics. 2019; 15(1-2):161-173. PMC: 6961656. DOI: 10.1080/15592294.2019.1656158. View

Murphy S, Adigun A, Huang Z, Overcash F, Wang F, Jirtle R . Gender-specific methylation differences in relation to prenatal exposure to cigarette smoke. Gene. 2011; 494(1):36-43. PMC: 3627389. DOI: 10.1016/j.gene.2011.11.062. View

10.

Chan J, Morgan C, Leu N, Shetty A, Cisse Y, Nugent B . Reproductive tract extracellular vesicles are sufficient to transmit intergenerational stress and program neurodevelopment. Nat Commun. 2020; 11(1):1499. PMC: 7083921. DOI: 10.1038/s41467-020-15305-w. View

11.

Uysal F, Akkoyunlu G, Ozturk S . DNA methyltransferases exhibit dynamic expression during spermatogenesis. Reprod Biomed Online. 2016; 33(6):690-702. DOI: 10.1016/j.rbmo.2016.08.022. View

12.

de Winter J, Ottenheijm C . Sarcomere Dysfunction in Nemaline Myopathy. J Neuromuscul Dis. 2017; 4(2):99-113. PMC: 5467716. DOI: 10.3233/JND-160200. View

13.

Bara A, Ferland J, Rompala G, Szutorisz H, Hurd Y . Cannabis and synaptic reprogramming of the developing brain. Nat Rev Neurosci. 2021; 22(7):423-438. PMC: 8445589. DOI: 10.1038/s41583-021-00465-5. View

14.

Xi Y, Li W . BSMAP: whole genome bisulfite sequence MAPping program. BMC Bioinformatics. 2009; 10:232. PMC: 2724425. DOI: 10.1186/1471-2105-10-232. View

15.

Uhlen M, Fagerberg L, Hallstrom B, Lindskog C, Oksvold P, Mardinoglu A . Proteomics. Tissue-based map of the human proteome. Science. 2015; 347(6220):1260419. DOI: 10.1126/science.1260419. View

16.

Goldschmidt L, Richardson G, Willford J, Severtson S, Day N . School achievement in 14-year-old youths prenatally exposed to marijuana. Neurotoxicol Teratol. 2011; 34(1):161-7. PMC: 3260381. DOI: 10.1016/j.ntt.2011.08.009. View

17.

Lee K, Laviolette S, Hardy D . Exposure to Δ9-tetrahydrocannabinol during rat pregnancy leads to impaired cardiac dysfunction in postnatal life. Pediatr Res. 2021; 90(3):532-539. PMC: 8519775. DOI: 10.1038/s41390-021-01511-9. View

18.

Edwards J, Yarychkivska O, Boulard M, Bestor T . DNA methylation and DNA methyltransferases. Epigenetics Chromatin. 2017; 10:23. PMC: 5422929. DOI: 10.1186/s13072-017-0130-8. View

19.

Innocenzi E, De Domenico E, Ciccarone F, Zampieri M, Rossi G, Cicconi R . Paternal activation of CB cannabinoid receptor impairs placental and embryonic growth via an epigenetic mechanism. Sci Rep. 2019; 9(1):17034. PMC: 6863860. DOI: 10.1038/s41598-019-53579-3. View

20.

Bassil C, Huang Z, Murphy S . Bisulfite pyrosequencing. Methods Mol Biol. 2013; 1049:95-107. DOI: 10.1007/978-1-62703-547-7_9. View