Loss of the crumbs cell polarity complex disrupts epigenetic transcriptional control and cell cycle progression in the developing retina

• Published in: The Journal of pathology Vol. 259; no. 4; pp. 441 - 454
• Main Authors: Owen, Nicholas, Toms, Maria, Tian, Yuan, Toualbi, Lyes, Richardson, Rose, Young, Rodrigo, Tracey‐White, Dhani, Dhami, Pawan, Beck, Stephan, Moosajee, Mariya
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-04-2023; Wiley Subscription Services, Inc
• Subjects: Animals; Bone morphogenetic proteins; Cell Cycle; Cell Polarity - genetics; Danio rerio; DNA methylation; Epigenesis, Genetic; Epigenetics; epigenome; Eye Proteins - genetics; Eye Proteins - metabolism; Gene regulation; Genotypes; Humans; iPSC; Leber congenital amaurosis; Membrane Proteins - metabolism; Morphogenesis; Nerve Tissue Proteins - metabolism; Organoids; Original; Phenotypes; Polarity; polarity complex; Proteins; Retina; Retina - metabolism; Retinitis; Retinitis pigmentosa; RNA‐seq; Signal transduction; Smad protein; transcriptome; Transcriptomics; Transforming growth factor-b; zebrafish; Zebrafish - genetics; Zebrafish Proteins - genetics; Zebrafish Proteins - metabolism; polarity complex; epigenome; RNA-seq; retina; transcriptome; iPSC; DNA methylation; zebrafish
• ISSN: 0022-3417; 1096-9896
• DOI: 10.1002/path.6056

The crumbs cell polarity complex plays a crucial role in apical–basal epithelial polarity, cellular adhesion, and morphogenesis. Homozygous variants in human CRB1 result in autosomal recessive Leber congenital amaurosis (LCA) and retinitis pigmentosa (RP), with no established genotype–phenotype correlation. The associated protein complexes have key functions in developmental pathways; however, the underlying disease mechanism remains unclear. Using the oko meduzym289/m289 (crb2a−/−) zebrafish, we performed integrative transcriptomic (RNA‐seq data) and methylomic [reduced representation bisulphite sequencing (RRBS)] analysis of whole retina to identify dysregulated genes and pathways. Delayed retinal cell specification was identified in both the crb2a−/− zebrafish and CRB1 patient‐derived retinal organoids, highlighting the dysfunction of cell cycle modulation and epigenetic transcriptional control. Differential DNA methylation analysis revealed novel hypermethylated pathways involving biological adhesion, Hippo, and transforming growth factor β (TGFβ) signalling. By integrating gene expression with DNA methylation using functional epigenetic modules (FEM), we identified six key modules involving cell cycle control and disturbance of TGFβ, bone morphogenetic protein (BMP), Hippo, and SMAD protein signal transduction pathways, revealing significant interactome hotspots relevant to crb2a function and confirming the epigenetic control of gene regulation in early retinal development, which points to a novel mechanism underlying CRB1‐retinopathies. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.