Identification and initial characterization of 5000 expressed sequenced tags (ESTs) each from adult human normal and osteoarthritic cartilage cDNA libraries.
OBJECTIVE
To prepare, sequence and analyse adult human cartilage cDNA libraries to study the gene expression pattern between normal and osteoarthritic cartilage.


METHODS
Poly A(+)RNA from adult human normal and osteoarthritic articular cartilage was isolated and used to prepare cDNA libraries. Approximately 5000 ESTs from each library were sequenced and analysed using bioinformatic tools. The expression of select genes was confirmed by Northern blot and in situ hybridization analysis.


RESULTS
Multiple gene families including several classical cartilage matrix protein encoding genes were identified. Approximately 28-40% of the genes sequenced from these libraries were novel, while half of the genes encoded known proteins and 4-6% of the genes encoded novel homologs of known proteins. Several known genes, whose expression has not been reported previously in cartilage, were also identified. We have confirmed the cartilage expression of three known (CTGF, CTGF-L and clusterin) and two novel homologs of known genes (PCPE-2 and Gal-Nac transferase) by Northern blot and in situ hybridization analysis.


CONCLUSIONS
This is the first report of the preparation and sequencing of cDNA libraries from adult human normal and osteoarthritic articular cartilage. Further analysis of genes identified from these libraries may provide molecular targets for diagnosis and/or treatment of osteoarthritis (OA).
Kumar
S
S
Department of Musculoskeletal Diseases, GlaxoSmithKline Pharmaceuticals, 709 Swedeland Rd, King of Prussia, Pennsylvania 19406, USA. Sanjay_kumar-1@GSK.COM
Connor
J R
JR
Dodds
R A
RA
Halsey
W
W
Van Horn
M
M
Mao
J
J
Sathe
G
G
Mui
P
P
Agarwal
P
P
Badger
A M
AM
Lee
J C
JC
Gowen
M
M
Lark
M W
MW
D000328:Adult; D015152:Blotting, Northern / Q000379:methods; D002358:Cartilage, Articular / Q000502:physiology*; D016022:Case-Control Studies; D020224:Expressed Sequence Tags; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D008969:Molecular Sequence Data; D020370:Osteoarthritis, Knee / Q000235:genetics*