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The c.119-123dup5bp mutation in human gamma-C-crystallin destabilizes the protein and activates the unfolded protein response to cause highly variable cataracts

Hejtmancik, James1

Research facility: National Eye Institute
Published Aug 23, 2024 on Dryad. https://doi.org/10.5061/dryad.rn8pk0pmm

Data files

Aug 23, 2024 version files 33.29 GB

Abstract

Ordered cellular architecture and high concentrations of stable crystallins are required for the lens to maintain transparency. Here we investigate the molecular mechanism of cataractogenesis of the CRYGC c.119-123dupGCGGC (p.Cys42AlafsX63) (CRYGC5bpd) mutation. Lenses were extracted from wild-type and transgenic mice carrying the CRYGC5bpdup minigene and RNA was isolated and converted into cDNA. Expression of genes in the unfolded protein response (UPR) pathways was estimated by qRT-PCR and RNA seq and pathway analysis was carried out using the Qiagen IPA website. P3W Transgenic mice exhibited phenotypic diversity with a dimorphic population of severe and clear lenses. PCA of RNA seq data showed separate clustering of wild-type, clear CRYGC5bpd, and severe CRYGC5bpd lenses. Transgenic mice showed differential upregulation in Master regulator Grp78 (Hspa5) and downstream targets in the PERK-dependent UPR pathway including Atf4 and Chop (Ddit3), but not GADD34. Thus, high levels of CRYGC5bpd transgene expression in severely affected lenses induce UPRer and UPRmt stress responses primarily through the PERK-dependent and Atf4/Atf5/Ddit3 pathways respectively, inducing autophagy and apoptosis and thence congenital nuclear cataracts. This effect is correlated to CRYGC5bpd transgene expression, offering insight into cataract pathogenic pathways and variation in cataract severity in humans.