Sperm DNA methylation alterations from cannabis extract exposure are evident in offspring

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.

Epigenomic and Other Evidence for Cannabis-Induced Aging Contextualized in a Synthetic Epidemiologic Overview of Cannabinoid-Related Teratogenesis and Cannabinoid-Related Carcinogenesis

“…Cannabis exposure was recently shown to be causally related to all four indices of mental dysfunction (depressive symptoms, any mental illness, severe mental illness and suicidal thinking) tracked by the annual nationwide massive National Survey of Drug Use and Health in a space time and causal inferential analysis [ 106 ]. Cannabis exposure has also been linked with the development of autism-like and ADHD-like syndromes in children [ 117 , 174 ] in spacetime and causal inferential studies [ 175 ] and in epigenomic studies [ 26 , 159 , 176 , 177 ]. An extensive literature and many meta-analyses strongly connect cannabis use and the development of schizophrenia by many mechanisms [ 17 , 178 , 179 , 180 , 181 , 182 , 183 , 184 , 185 , 186 , 187 , 188 , 189 , 190 , 191 , 192 , 193 , 194 , 195 , 196 ].…”

Clinical Epigenomic Explanation of the Epidemiology of Cannabinoid Genotoxicity Manifesting as Transgenerational Teratogenesis, Cancerogenesis and Aging Acceleration

“…Similar results in many different studies are clearly mutually supportive and strengthen the overall quality of the body of evidence. Similarly, there is a striking concordance between the many epigenomic studies of gestational cannabis exposure in relation to global DNA hypomethylation and the disruption of DNA methylation levels at key promoter and enhancer sites, which control the regulation of many critical genes [ 42 , 101 , 102 , 103 , 105 , 106 , 107 , 108 , 233 , 234 ].…”

The Biological Characteristics and Differential Expression Patterns of TSSK1B Gene in Yak and Its Infertile Hybrid Offspring

“…The purified DNA samples were treated with the EpiArt ® DNA Methylation Bisulfite Kit (EM101, Vazyme, Nanjing, China) by following the instructions of the manufacturer. The bisulfite treatment converted the unmethylated cytosine bases to thymine bases (C-T), whereas the methylated cytosine bases remained unchanged [24].…”