Evaluation of SPARC as a candidate gene of juvenile-onset primary open-angle glaucoma by mutation and copy number analyses.
OBJECTIVE
To investigate the involvement of SPARC (secreted protein acidic and rich in cysteine) mutations and copy number variation in juvenile-onset primary open-angle glaucoma (JPOAG).


METHODS
This study involved the 27 family members from the GLC1M (glaucoma 1, open angle, M)-linked Philippine pedigree with JPOAG, 46 unrelated Chinese patients with JPOAG and 95 controls. Mutation screening of the SPARC sequence, covering the promoter, 5'-untranslated region (UTR), entire coding regions, exon-intron boundaries, and part of the 3'-UTR, was performed using polymerase chain reaction and direct DNA sequencing. Copy number of the gene was analyzed by three TaqMan copy number assays.


RESULTS
No putative SPARC mutation was detected in the Philippine family. In the Chinese participants, 11 sequence variants were detected. Two were novel: IVS2+8G>T and IVS2+32C>T. For the 9 known SNPs, one was synonymous (rs2304052, p.Glu22Glu) and the others were located in noncoding regions. No individual SNP was associated with JPOAG. Five of the most common SNPs, i.e., rs2116780, rs1978707, rs7719521, rs729853, and rs1053411, were contained in a LD (linkage disequilibrium) block. Haplotype-based analysis showed that no haplotype was associated with the disorder. Copy number analysis revealed that all study subjects had two copies of the gene, suggesting no correlation between the copy number of SPARC and JPOAG.


CONCLUSIONS
We have excluded SPARC as the causal gene at the GLC1M locus in the Philippine pedigree and, for the first time, revealed that the coding sequences, splice sites and copy number of SPARC do not contribute to JPOAG. Further investigations are warranted to unravel the involvement of SPARC in the pathogenesis of other forms of glaucoma.
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong, China.
Li Jia
LJ
Chen
Pancy O S
PO
Tam
Clement C Y
CC
Tham
Xiao Ying
XY
Liang
Sylvia W Y
SW
Chiang
Oscar
O
Canlas
Robert
R
Ritch
Douglas J
DJ
Rhee
Chi Pui
CP
Pang
D000293:Adolescent; D000328:Adult; D017668:Age of Onset; D000368:Aged; D000369:Aged, 80 and over; D044466:Asian People / Q000235:genetics; D001483:Base Sequence; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D002681:China; D004252:DNA Mutational Analysis; D003710:Demography; D005260:Female; D018628:Gene Dosage / Q000235:genetics*; D005902:Glaucoma, Open-Angle / Q000453:epidemiology* / Q000235:genetics*; D006801:Humans; D015810:Linkage Disequilibrium / Q000235:genetics; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D015676:Osteonectin / Q000235:genetics*; D010375:Pedigree; D010679:Philippines; D055815:Young Adult
D015676:Osteonectin