Single-cell analysis of developing and azoospermia human testicles reveals central role of Sertoli cells

Abstract

Clinical efficacy of treatments against non-obstructive azoospermia (NOA), which affects 1% of men, are currently limited by the incomplete understanding of NOA pathogenesis and normal spermatogenic microenvironment. Here, we profile >80,000 human testicular single-cell transcriptomes from 10 healthy donors spanning the range from infant to adult and 7 NOA patients. We show that Sertoli cells, which form the scaffold in the testicular microenvironment, are severely damaged in NOA patients and identify the roadmap of Sertoli cell maturation. Notably, Sertoli cells of patients with congenital causes (Klinefelter syndrome and Y chromosome microdeletions) are mature, but exhibit abnormal immune responses, while the cells in idiopathic NOA (iNOA) are physiologically immature. Furthermore, we find that inhibition of Wnt signaling promotes the maturation of Sertoli cells from iNOA patients, allowing these cells to regain their ability to support germ cell survival. We provide a novel perspective on the development of diagnostic methods and therapeutic targets for NOA.

Genome medicine in male infertility: From karyotyping to single‐cell analysis

“… 29 Tan et al listed most of the single‐cell transcriptome analyses that have been conducted on mouse, human, and macaque testes 30 . By modifying their report, single‐cell transcriptome analysis performed using human testicular samples is summarized in Table 1, 31–38 which included eight publications and our current project. The majority of them have focused on germ cells, and the major method to isolate and harvest a single cell type, which is the most important and difficult part of single‐cell analysis, was enzymatic digestion combined with flow cytometric sorting or physical filtering.…”

Infertility network and hub genes for nonobstructive azoospermia utilizing integrative analysis

“…Currently, except for traditional microarray analysis, several studies have used weighted gene co-expression network analysis (WGCNA) analysis [ 14 ], whole-exome sequencing [ 15 ], and single-cell transcriptome sequencing (scRNA-seq) [ 16 , 17 ] to screen for novel infertility causative genes in NOA. The resulting data provide a basis for further studies on the pathogenesis of NOA, but these existing studies do not integrate multiple sequencing data, which makes the results obtained by a single analytical approach less convincing.…”

“…Since its development, single-cell transcriptome sequencing has been widely used in cancer and bio-developmental fields with excellent results. Although two studies performed single-cell transcriptome sequencing on testicular tissue from NOA patients [ 16 , 17 ], no study has yet applied these open resources to an integrated analysis of NOA; this omission lays the foundation for our study.…”

A Novel Organoid Model of In Vitro Spermatogenesis Using Human Induced Pluripotent Stem Cells

“…[1] Pinpointing the causes of infertility therefore requires an understanding of the cellular niche where sperm are generated. Advances in single cell sequencing of testicular tissue have begun to shed light on the transcriptomic signatures of the cell types within the niche, [2][3][4][5][6][7][8] however the development of in vitro tools to study their interactions and functionalities remains elusive. [9] Historically, animal models have been the standard in modeling human diseases, however there are specifically human biological processes such as infertility whose complexity and interindividual variability cannot be accurately modeled using animals.…”