A Taybi-Linder syndrome-related RTTN variant impedes neural rosette formation in human cortical organoids

PLoS Genet. 2024 Dec 16;20(12):e1011517. doi: 10.1371/journal.pgen.1011517. eCollection 2024 Dec.

Abstract

Taybi-Linder syndrome (TALS) is a rare autosomal recessive disorder characterized by severe microcephaly with abnormal gyral pattern, severe growth retardation and bone abnormalities. It is caused by pathogenic variants in the RNU4ATAC gene. Its transcript, the small nuclear RNA U4atac, is involved in the excision of ~850 minor introns. Here, we report a patient presenting with TALS features but no pathogenic variants were found in RNU4ATAC, instead the homozygous RTTN c.2953A>G variant was detected by whole-exome sequencing. After deciphering the impact of the variant on the RTTN protein function at centrosome in engineered RTTN-depleted RPE1 cells and patient fibroblasts, we analysed neural stem cells (NSC) derived from CRISPR/Cas9-edited induced pluripotent stem cells and revealed major cell cycle and mitotic abnormalities, leading to aneuploidy, cell cycle arrest and cell death. In cortical organoids, we discovered an additional function of RTTN in the self-organisation of NSC into neural rosettes, by observing delayed apico-basal polarization of NSC. Altogether, these defects contributed to a marked delay of rosette formation in RTTN-mutated organoids, thus impeding their overall growth and shedding light on mechanisms leading to microcephaly.

MeSH terms

  • CRISPR-Cas Systems
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Centrosome / metabolism
  • Cerebral Cortex / metabolism
  • Exome Sequencing
  • Humans
  • Induced Pluripotent Stem Cells / metabolism
  • Microcephaly* / genetics
  • Mutation
  • Neural Stem Cells* / metabolism
  • Organoids* / metabolism
  • RNA, Small Nuclear / genetics
  • RNA, Small Nuclear / metabolism

Substances

  • RTTN protein, human
  • Cell Cycle Proteins
  • RNA, Small Nuclear

Grants and funding

This work was supported by CNRS, Inserm and Université Claude Bernard Lyon 1 through recurrent funding; the Agence Nationale de la Recherche (no. ANR-18CE12-0007; no. ANR-22CE12-0007); the Fondation Jérôme Lejeune and the Fondation pour la recherche sur le Cerveau « Espoir en tête » (confocal microscope). J.G. was supported by the Ministère de l'Enseignement Supérieur et de la Recherche and by the Fondation pour la Recherche Médicale (grant number FDT202304016375), and S.Cu. by a Poste Accueil Inserm 2023. T-Y.C. was supported by a postdoctoral fellowship from Academia Sinica, Taiwan and T.K.T. by the National Science and Technology Council (NSTC 112-2326-B001-010) and Academia Sinica (AS-IA-109-L04), Taiwan. E.B. was supported by an EMBO long-term fellowship (ALTF-284-2019), and V.H. by the Swiss National Foundation (SNSF 310030_205087). S.T. was supported by Agence Nationale de la Recherche (no. ANR-17-CE16-0003-01). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.