Global Experts Meeting on Infectious Diseases
Chengdu Jinxin Research Institute for Reproductive Medicine, China
Title: Transmission of hepatitis B virus genes via patients’ spermatozoa and regulation of their expression by host microRNAs in sperm-derived embryos
Biography: Tianhua Huang
Statement of the Problem: Hepatitis B virus (HBV) genes are able to integrate into sperm genome and can transcribe viral mRNA in spermatozoa. Sperm nucleus contains diverse RNA populations. We explored whether HBV genes could be transmitted vertically via patients’ spermatozoa and aimed to screen and identify host microRNAs regulating HBV gene expression in spermatozoa and sperm-derived embryos. Methodology & findings: Using fluorescence in situ hybridization, positive signals for HBV DNA were detected in both nuclei of 2-cell embryos derived from patients’ spermatozoa but not in control embryos. There was a significant difference in the percentage of embryos with positive signals between the test and control groups. Using reverse transcription polymerase chain reaction (RT–PCR), positive bands for HBV S and X genes were observed only in the test group but not in controls. Using immunofluorescence assays, clear HBV surface antigen signals were detected in the cytoplasm of embryos derived from patients’ spermatozoa but not in those derived from donor spermatozoa. Using microarrays, 336 miRs were found to be differentially expressed. After validation using real-time quantitative RT–PCR (RT-qPCR), four miRs were selected as targets. Using RT–qPCR and enzyme-linked immunosorbent assays, when patients’ spermatozoa were treated with mimics or inhibitors specific for hsa-miR-19a-3p and hsa-miR-29c-3p, the S gene transcription in spermatozoa and translation in sperm-derived embryos was downregulated or upregulated. There were significant differences in transcriptional and translational levels of the S gene between the test and control groups.