Single-Cell RNA Sequencing Analysis Reveals Sequential Cell Fate Transition during Human Spermatogenesis

Abstract

Spermatogenesis generates mature male gametes and is critical for the proper transmission of genetic information between generations. However, the developmental landscapes of human spermatogenesis remain unknown. Here, we performed single-cell RNA sequencing (scRNA-seq) analysis for 2,854 testicular cells from donors with normal spermatogenesis and 174 testicular cells from one nonobstructive azoospermia (NOA) donor. A hierarchical model was established, which was characterized by the sequential and stepwise development of three spermatogonia subtypes, seven spermatocyte subtypes, and four spermatid subtypes. Further analysis identified several stage-specific marker genes of human germ cells, such as HMGA1, PIWIL4, TEX29, SCML1, and CCDC112. Moreover, we identified altered gene expression patterns in the testicular somatic cells of one NOA patient via scRNA-seq analysis, paving the way for further diagnosis of male infertility. Our work allows for the reconstruction of transcriptional programs inherent to sequential cell fate transition during human spermatogenesis and has implications for deciphering male-related reproductive disorders.

Listing candidate diagnostic markers and transcriptomic exploration of the molecular basis of a type of male infertility (Non-Obstructive Azoospermia), via next generation sequencing methods

“…Cell-type-specific enriched expression of genes is critical and this has been addressed well (20) and can help to explore cell-type specificity of key genes. Of the top 250 genes down-regulated with high StA score in the current study, 148 genes were common to the significant cell-typeenriched list from Wang et al, and 133 of these were enriched in spermatids.…”

“…That is why, even in cases such as NOA, where biopsy-based examinations are permitted, there have not been enough studies with a good number of samples. Recently single-cell RNA sequencing has been used to explore the transcriptome of specific cell types of testicular cells where tissue sample from a single NOA-donor has been used (20).…”

Meiosis related cancer-testis genes correlate with tumor aneuploidy and immune infiltration in 21 cancer types

“…CT genes, which are specifically expressed in testis and cancer tissues, were regarded as the molecular basis of the two processes. However, spermatogenesis is a tightly regulated developmental process and include multiple phases (10). In each phase of human spermatogenesis, various developmental germ cell subtypes can be identified.…”

Transcriptomic changes due to early, chronic alcohol exposure during cortical development implicate regionalization, cell-type specification, synaptogenesis and WNT signaling as primary determinants of fetal alcohol Spectrum Disorders

“…Recently developed single-cell sequencing techniques have begun to uncover the multitude of neuronal cell types making up the circuitry of the mammalian cortex (Wang 2018, Tang 2018).…”