In vitro cellular reprogramming to model gonad development and its disorders

Nitzan Gonen (1, 2) , Caroline Eozenou (3) , Richard Mitter (2) , Maëva Elzaiat (3) , Isabelle Stévant (1) , Rona Aviram (1) , Andreia Sofia Bernardo (2, 4) , Almira Chervova (5) , Somboon Wankanit (3) , Emmanuel Frachon (6) , Pierre-Henri Commère (7) , Sylvie Brailly-Tabard (8) , Léo Valon (9) , Laura Barrio Cano (7) , Romain Levayer (9) , Inas Mazen (10) , Samy Gobaa (6) , James Smith (2) , Kenneth Mcelreavey (3) , Robin Lovell-Badge (2) , Anu Bashamboo (3)

1 Bar-Ilan University [Israël]
2 The Francis Crick Institute [London]
3 Génétique du Développement humain - Human developmental genetics
4 Imperial College London
5 Département de Biologie du Développement et Cellules souches - Department of Developmental and Stem Cell Biology
6 Biomatériaux et Microfluidiques (plateforme) - Biomaterials and Microfluidics (platform)
7 UTechS CB - Cytometrie et Biomarqueurs – Cytometry and Biomarkers
8 Hôpital Bicêtre [AP-HP, Le Kremlin-Bicêtre]
9 Mort cellulaire et homéostasie des épithéliums / Cell death and epithelial homeostasis
10 National Research Center [Caire, Egypte]

During embryonic development, mutually antagonistic signaling cascades determine gonadal fate toward a testicular or ovarian identity. Errors in this process result in disorders of sex development (DSDs), characterized by discordance between chromosomal, gonadal, and anatomical sex. The absence of an appropriate, accessible in vitro system is a major obstacle in understanding mechanisms of sex-determination/DSDs. Here, we describe protocols for differentiation of mouse and human pluripotent cells toward gonadal progenitors. Transcriptomic analysis reveals that the in vitro–derived murine gonadal cells are equivalent to embryonic day 11.5 in vivo progenitors. Using similar conditions, Sertoli-like cells derived from 46,XY human induced pluripotent stem cells (hiPSCs) exhibit sustained expression of testis-specific genes, secrete anti-Müllerian hormone, migrate, and form tubular structures. Cells derived from 46,XY DSD female hiPSCs, carrying an NR5A1 variant, show aberrant gene expression and absence of tubule formation. CRISPR-Cas9–mediated variant correction rescued the phenotype. This is a robust tool to understand mechanisms of sex determination and model DSDs.

Domaines: Sciences du Vivant [q-bio]

Nitzan Gonen, Caroline Eozenou, Richard Mitter, Maëva Elzaiat, Isabelle Stévant, et al.. In vitro cellular reprogramming to model gonad development and its disorders. Science Advances , 2023, 9 (1), pp.eabn9793. ⟨10.1126/sciadv.abn9793⟩. ⟨hal-04150365⟩