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Identification of Differentially Expressed Genes in Human Testis Biopsies with Defective Spermatogenesis

ABSTRACT: The study was designed to identify differences in the expressed transcriptome of the two pathological human testis sample groups compared to normal spermatogenesis samples, with the objective of discovering genes related to the pathological condition. Transcriptional differences in human testis biopsies collected from men with normal spermatogenesis (NSP, n=3; intact germ cells, clinical score - 10), spermatid arrest (SDA, n=2; contains somatic cells and germ cells up to round spermatids, clinical score – 4-5) and Sertoli cell-only (SCO, n=2; complete absence of germ cells, clinical score - 0) phenotypes were assessed by RNA sequencing. A total of over 49 million reads were generated in NSP samples, over 24 million reads in SDA samples, and over 13 million reads in SCO samples. Differentially expressed genes (DEG) were identified based on a mean minimum count => 5, FDR <= 0.05 and log2FC fold change of >0.585 or <-0.585. Genes differentially expressed in pathological groups SDA and SCO compared to NSP, and genes differentially expressed in between the two pathological groups SDA and SCO were filtered. The study discovered a number of significantly differentially expressed genes (DEG) between groups as follows: NSP vs SDA n=1,873; SDA vs SCO n= 4,017; and NSP vs SCO n=10,253. A number of genes (n=10) were selected for further analysis by qRT-PCR and the detection of the protein (n=2) localization in testis by immunohistochemistry.

ORGANISM(S): Homo sapiens

PROVIDER: GSE224929 | GEO | 2024/12/05

REPOSITORIES: GEO

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Project description:Testicular germ cell tumours (TGCTs) of young adult men, seminoma (SEM) and nonseminoma (NSEM), develop from a precursor cell, carcinoma in situ (CIS), which resembles foetal gonocytes and retains embryonic pluripotency. We used microarrays to analyse microRNA (miRNA) expression in 12 human testis samples with CIS cells and compared it to the miRNA expression profiles of normal adult testis, testis with Sertoli-cell-only (SCO) that lack germ cells, testis tumours (SEM and embryonal carcinoma, EC; an undifferentiated component of NSEM) and foetal male and female gonads. Principal component analysis revealed distinct miRNA expression profiles characteristic for each of the different tissue types. We identified several miRNAs that were unique to testis with CIS cells, foetal gonads, and testis tumours. These included miRNAs from the hsa-miR-371~373 and -302~367 clusters that have previously been reported in germ cell tumours and three miRNAs (hsa-miR-96, -141 and -200c) that were also expressed in human epididymis. We found several miRNAs that were upregulated in testis tumours: hsa-miR-9, -105 and the hsa-miR-182~183~96 cluster were highly expressed in SEM, while the hsa-miR-515~526 cluster was high in EC. We conclude that miRNA expression profile changes during testis development and that the miRNA profile of adult testis with CIS cells shares characteristic similarities with the expression in foetal gonocytes. We used Agilent microarrays to determine the expression of microRNAs in samples from testis. We first used Agilent-016436 Human miRNA Microarray 1.0 G4472A (GPL9081; miRNA ID version) on 4 samples: normal testis, testis with 30% carcinoma in situ (CIS) tubules, testis with 50% CIS tubules and testis with 99-100% CIS tubules. This allowed us to determine the correlation between miRNA expressiona and the content of CIS cells in the samples. This is the file with data from those four arrays. Next, we used Agilent-019118 Human miRNA Microarray 2.0 G4470B (GPL8936; Probe Name version) on 24 samples that included normal testis, testis with the precursor for testis cancer carcinoma in situ (CIS), Sertoli-cell-only testis (contains the somatic testis cells but no germ cells), seminom and non-seminom tumours, and foetal gonads. This constitute the main part of the experiment and the publication, but the data was submitted to GEO in an independent file.

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Identification of Differentially Expressed Genes in Human Testis Biopsies with Defective Spermatogenesis

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