Transcriptome analysis of the human corneal endothelium.
OBJECTIVE
To comprehensively characterize human corneal endothelial cell (HCEnC) gene expression and age-dependent differential gene expression and to identify expressed genes mapped to chromosomal loci associated with the corneal endothelial dystrophies posterior polymorphous corneal dystrophy (PPCD)1, Fuchs endothelial corneal dystrophy (FECD)4, and X-linked endothelial dystrophy (XECD).


METHODS
Total RNA was isolated from ex vivo corneal endothelium obtained from six pediatric and five adult donor corneas. Complementary DNA was hybridized to the Affymetrix GeneChip 1.1ST array. Data analysis was performed using Partek Genomics Suite software, and differentially expressed genes were validated by digital molecular barcoding technology.


RESULTS
Transcripts corresponding to 12,596 genes were identified in HCEnC. Nine genes displayed the most significant differential expression between pediatric and adult HCEnC: CAPN6, HIST1H3A, HIST1H4E, and HSPA2 were expressed at higher levels in pediatric HCEnC, while ITGBL1, NALCN, PREX2, TAC1, and TMOD1 were expressed at higher levels in adult HCEnC. Analysis of the PPCD1, FECD4 and XECD loci demonstrated transcription of 53/95 protein-coding genes in the PPCD1 locus, 27/40 in the FECD4 locus, and 35/68 in the XECD locus.


CONCLUSIONS
An analysis of the HCEnC transcriptome reveals the expression of almost 13,000 genes, with less than 1% mapped to chromosomal loci associated with PPCD1, FECD4, and XECD. At least nine genes demonstrated significant differential expression between pediatric and adult HCEnC, defining specific functional properties distinct to each age group. These data will serve as a resource for vision scientists investigating HCEnC gene expression and can be used to focus the search for the genetic basis of the corneal endothelial dystrophies for which the genetic basis remains unknown.
The Jules Stein Eye Institute, David Geffen School of Medicine at the University of California-Los Angeles, Los Angeles, California, United States.
Ricardo F
RF
Frausto
The Jules Stein Eye Institute, David Geffen School of Medicine at the University of California-Los Angeles, Los Angeles, California, United States.
Cynthia
C
Wang
The Jules Stein Eye Institute, David Geffen School of Medicine at the University of California-Los Angeles, Los Angeles, California, United States.
Anthony J
AJ
Aldave
D000328:Adult; D003317:Corneal Dystrophies, Hereditary / Q000235:genetics* / Q000378:metabolism; D004728:Endothelium, Corneal / Q000378:metabolism*; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D005642:Fuchs' Endothelial Dystrophy / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D059467:Transcriptome
D005136:Eye Proteins; D012333:RNA, Messenger
HCEnC; corneal endothelial dystrophy; corneal endothelium; gene expression; transcriptome