CReATe Fertility Centre, Toronto, ON, Canada.
Racine IVF Unit, Lis Maternity Hospital, Tel Aviv Sourasky Medical Center, Affiliated to the Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Mol Hum Reprod. 2022 Jun 30;28(7). doi: 10.1093/molehr/gaac022.
Cannabis is increasingly consumed by women of childbearing age, and the reproductive and epigenetic effects are unknown. The purpose of this study was to evaluate the potential epigenetic implications of cannabis use on the female ovarian follicle. Whole-genome methylation was assessed in granulosa cells from 14 matched case-control patients. Exposure status was determined by liquid chromatography-mass spectrometry (LC-MS/MS) measurements of five cannabis-derived phytocannabinoids in follicular fluid. DNA methylation was measured using the Illumina TruSeq Methyl Capture EPIC kit. Differential methylation, pathway analysis and correlation analysis were performed. We identified 3679 differentially methylated sites, with two-thirds affecting coding genes. A hotspot region on chromosome 9 was associated with two genomic features, a zinc-finger protein (ZFP37) and a long non-coding RNA (FAM225B). There were 2214 differentially methylated genomic features, 19 of which have been previously implicated in cannabis-related epigenetic modifications in other organ systems. Pathway analysis revealed enrichment in G protein-coupled receptor signaling, cellular transport, immune response and proliferation. Applying strict criteria, we identified 71 differentially methylated regions, none of which were previously annotated in this context. Finally, correlation analysis revealed 16 unique genomic features affected by cannabis use in a concentration-dependent manner. Of these, the histone methyltransferases SMYD3 and ZFP37 were hypomethylated, possibly implicating histone modifications as well. Herein, we provide the first DNA methylation profile of human granulosa cells exposed to cannabis. With cannabis increasingly legalized worldwide, further investigation into the heritability and functional consequences of these effects is critical for clinical consultation and for legalization guidelines.
越来越多的育龄妇女开始使用大麻，但其对生殖和表观遗传的影响尚不清楚。本研究旨在评估大麻使用对女性卵巢卵泡的潜在表观遗传影响。通过对 14 对匹配的病例对照患者的卵泡液进行液相色谱-质谱（LC-MS/MS）测量，评估了五种大麻衍生植物大麻素的暴露状况。使用 Illumina TruSeq Methyl Capture EPIC 试剂盒测量 DNA 甲基化。进行了差异甲基化、通路分析和相关性分析。我们鉴定出 3679 个差异甲基化位点，其中三分之二影响编码基因。9 号染色体上的一个热点区域与两个基因组特征相关，一个锌指蛋白（ZFP37）和一个长非编码 RNA（FAM225B）。存在 2214 个差异甲基化的基因组特征，其中 19 个先前与其他器官系统中与大麻相关的表观遗传修饰有关。通路分析显示，G 蛋白偶联受体信号、细胞转运、免疫反应和增殖途径富集。应用严格的标准，我们鉴定出 71 个差异甲基化区域，这些区域在这方面以前没有被注释过。最后，相关性分析显示，有 16 个独特的基因组特征受大麻使用的影响，呈浓度依赖性。其中，组蛋白甲基转移酶 SMYD3 和 ZFP37 呈低甲基化状态，这可能暗示着组蛋白修饰也参与其中。本研究首次提供了暴露于大麻的人类颗粒细胞的 DNA 甲基化图谱。随着大麻在全球范围内越来越合法化，进一步研究这些影响的遗传性和功能后果对于临床咨询和合法化指南至关重要。
