Unknown,Transcriptomics,Genomics,Proteomics

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Comparison of global gene expression profiles of microdissected human foetal Leydig cells with their normal and hyperplastic adult equivalents

ABSTRACT: Leydig cells (LCs) are the primary androgen producing cells in males throughout development and appear in chronologically distinct populations; fetal Leydig cells (FLCs), neonatal LCs and adult Leydig cells (ALCs). ALCs are responsible for progression through puberty and for maintenance of reproductive functions in adulthood. In patients with reproductive problems, such as infertility or testicular cancer, and especially in men with high gonadotrophin levels, LC function is often impaired, and LCs may cluster abnormally into hyperplastic micronodules. The purpose of this study was to investigate whether ALCs within hyperplastic micronodules display characteristics of FLCs. Transcriptome profiling was performed on LCs microdissected from 12 frozen human tissue samples, including morphologically normal foetal (gestational week 10-11) and adult testis samples, and adult specimens with LC micronodules or LC micronodules adjacent to chorionic gonadotrophin (hCG)-producing testicular germ cell tumours. Fifteen additional foetal and adult testis tissue specimens were used for validation of the expression data at the protein level. The study revealed the the gene expression profiles of FLCs and all ALC subgroups were clearly different, but there were no significant differences in expressed genes between the normally clustered and hyperplastic ALCs. Close similarity between transcriptome profiles of the normally clustered and hyperplastic ALCs suggest that the changes in LC function and morphology observed in patients with reproductive disorders possibly reflect subtle changes in the expression of many genes rather than regulatory changes of single genes or pathways. The transcriptome of FLCs revealed several genes not known previously to be expressed in this cell type during early development, including SULT2A1, WISP2, HPGD, and IGF2BP1, and their expression changes were validated at the protein level.

ORGANISM(S): Homo sapiens

SUBMITTER: Kirstine Belling

PROVIDER: E-MTAB-5453 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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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Comparison of global gene expression profiles of microdissected human foetal Leydig cells with their normal and hyperplastic adult equivalents

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