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Tobacco-induced microRNAs profile alterations in human spermatozoa: a preliminary study for further knowledge of toxical spermatogenesis impairment

ABSTRACT: Tobacco smoking generates deleterious effects on human semen quality but mechanisms by which cigarettes smoking can impact spermatogenesis are poorly understood. Recent works have shown that spermatozoa RNAs can be used to understand mechanisms involved in tobacco induced spermatogenesis impairment. We performed a prospective study of 8 smoker and 8 non-smoker patients in an university hospital. All patients were selected according to an occupational exposure standardized questionnaire and the sperm parameters. We performed gene expression and miRNA microarrays using RNA extracts from spermatozoa of 8 smokers and 8 non-smokers. Quantification of selected miRNA was performed using quantitative RT-PCR. We show that 16 genes were differentially expressed between smokers and non-smokers, of which 5 were upregulated and 11 were down regulated in smokers. 23 microRNAs were differentially expressed, of which 16 were upregulated and 7 were down regulated in smokers. Quantitative RT-PCR confirmed the down regulation in smokers for 3 microRNAs. Moreover in smokers, one of the upregulated genes is a putative target for one down regulated microRNA. This is a preliminary and innovating study on spermatozoa RNA extracts in the field of infertility. This preliminary study shows that large scale approaches are non invasive diagnostic tools that may help elucidate the mechanisms that mediate the response to tobacco smoke exposure on human spermatogenesis. This could be used to design biomarkers of human spermatogenetic damage, and help elucidate the fine regulatory mechanisms that mediate responses to environmental agent exposure during human spermatogenesis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE44134 | GEO | 2015/01/01

SECONDARY ACCESSION(S): PRJNA188744

REPOSITORIES: GEO

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Project description:Maternal smoking has a severe negative effect on all stages of pregnancy that in consequence impairs fetal growth and development. Tobacco smoke-related defects are well established at the clinical level; however, little is known about molecular mechanisms underlying these pathological conditions. We thus employed a genomic approach to determine transcriptome alterations induced by maternal smoking in pregnancy. We assayed gene expression profiles in peripheral blood (M) leukocytes and placentas (PL) of pregnant smokers and those without significant exposure, and in cord blood (D) leukocytes of their babies. Comparative analyses defined significant deregulation of 193 genes in M cells, 329 genes in placentas, and 49 genes in D cells of smokers. These genes were mainly involved in xenobiotic metabolism, oxidative stress, inflammation, immunity, hematopoiesis, trophoblast differentiation, and vascularization. Functional annotation of the deregulated genes outlined processes and pathways affected by tobacco smoke. In smoker newborns, we identified several deregulated pathways associated with autoimmune diseases. The study demonstrates a limited ability of placenta to modulate toxic effects of maternal tobacco use at the gene expression level. Blood and placental samples were obtained from 91 women who gave birth to a baby in the Ceske Budejovice Hospital from November 2008 to March 2009. The study was approved by the Local Institutional Review Board. All participants provided written informed consent and completed an extensive questionnaire on pregnancy history (medication, diseases), delivery course, newborn characteristics and life style (smoking, diet, alcohol drinking, area of residency). Only subjects who underwent vaginal delivery were included in the study. Based on smoking history, the women were divided into two groups, “smokers” (N=20) and “non-smokers” (N=52), while passive smokers (N=19) were excluded from the study. Smoking status was further confirmed by detection of excessive plasma levels of cotinine (the major nicotine metabolite) in maternal blood.

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Tobacco-induced microRNAs profile alterations in human spermatozoa: a preliminary study for further knowledge of toxical spermatogenesis impairment

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