Alpha-crystallin mutations alter lens metabolites in mouse models of human cataracts.

Cheryl Frankfater,Fong-Fu Hsu,Stephanie L Bozeman,Usha P Andley,Ram Nagaraj

doi:10.1371/journal.pone.0238081

Cheryl Frankfater, Fong-Fu Hsu + Show 3 more

Open Access

https://doi.org/10.1371/journal.pone.0238081

Copy DOI

Abstract

Cataracts are a major cause of blindness worldwide and commonly occur in individuals over 70 years old. Cataracts can also appear earlier in life due to genetic mutations. The lens proteins, αA- and αB-crystallins, are chaperone proteins that have important roles maintaining protein solubility to prevent cataract formation. Mutations in the CRYAA and CRYAB crystallin genes are associated with autosomal dominant early onset human cataracts. Although studies about the proteomic and genomic changes that occur in cataracts have been reported, metabolomics studies are very limited. Here, we directly investigated cataract metabolism using gas-chromatography-mass spectrometry (GC-MS) to analyze the metabolites in adult Cryaa-R49C and Cryab-R120G knock-in mouse lenses. The most abundant metabolites were myo-inositol, L-(+)-lactic acid, cholesterol, phosphate, glycerol phosphate, palmitic and 9-octadecenoic acids, α-D-mannopyranose, and β-D-glucopyranose. Cryaa-R49C knock-in mouse lenses had a significant decrease in the number of sugars and minor sterols, which occurred in concert with an increase in lactic acid. Cholesterol composition was unchanged. In contrast, Cryab-R120G knock-in lenses exhibited increased total amino acid content including valine, alanine, serine, leucine, isoleucine, glycine, and aspartic acid. Minor sterols, including cholest-7-en-3-ol and glycerol phosphate were decreased. These studies indicate that lenses from Cryaa-R49C and Cryab-R120G knock-in mice, which are models for human cataracts, have unique amino acid and metabolite profiles.

Highlights

Cataracts are a major cause of blindness worldwide, and protein aggregation and insolubility are the key sources of lens opacification in human cataractogenesis [1]
The lens metabolites that were detectable by gas-chromatography-mass spectrometry (GC-mass spectrometry (MS)) included amino acids, organic acids, sugars and sugar alcohols, fatty acids, and sterols
Because we were most interested in dose-dependent effects of the mutated α-crystallin gene, we first analyzed lenses from the WT, Cryaa-R49C-het, and Cryaa-R49C-homo mice separate from the WT, Cryab-R120G-het, and Cryab-R120G-homo mice lenses

Summary

Introduction

Cataracts are a major cause of blindness worldwide, and protein aggregation and insolubility are the key sources of lens opacification in human cataractogenesis [1]. Congenital cataracts, which have been linked to crystallin gene mutations, appear early in life and account for approximately 30% of childhood blindness [2,3,4]. Metabolomics has been used in both clinical and animal studies of several diseases, including some ocular pathologies [5, 6]. Metabolomic studies focused on cataracts are very limited [7,8,9]. Proteomic and RNA-seq studies that investigated the biochemical mechanisms of congenital.

Methods

Results

Conclusion