Differential gene expression in pulmonary artery endothelial cells exposed to sickle cell plasma.
Clinical variability in sickle cell disease (SCD) suggests a role for extra-erythrocytic factors in the pathogenesis of vasoocclusion. We hypothesized that endothelial cell (EC) dysfunction, one possible modifier of disease variability, results from induction of phenotypic changes by circulating factors. Accordingly, we analyzed gene expression in cultured human pulmonary artery ECs (HPAEC) exposed to plasma from 1) sickle acute chest syndrome (ACS) patients, 2) SCD patients at steady state, 3) normal volunteers, and 4) serum-free media, using whole genome microarrays (U133A-B GeneChip, Affymetrix). Data were analyzed by Bayesian analysis of differential gene expression (BADGE). Differential expression was defined by the probability of >1.5 fold change in signal intensity greater than 0.999 and a predicted score of 70-100, measured by cross-validation. Compared with normal plasma, plasma from SCD patients (steady state) resulted in differential expression of 50 genes in HPAEC. Of these genes, molecules involved in cholesterol biosynthesis and lipid transport, the cellular stress response, and extracellular matrix proteins were most prominent. Another 58 genes were differentially expressed in HPAEC exposed to plasma from ACS patients. The pattern of altered gene expression suggests that plasma from SCD patients induces an EC phenotype which is anti-apoptotic and favors cholesterol biosynthesis. An altered EC phenotype elicited by SCD plasma may contribute to the pathogenesis of sickle vasoocclusion.
Klings
Elizabeth S
ES
The Pulmonary Center, Boston University School of Public Health, Boston, Massachusetts 02118, USA. eklings@lung.bumc.bu.edu
Safaya
Surinder
S
Adewoye
Adeboye H
AH
Odhiambo
Adam
A
Frampton
Garrett
G
Lenburg
Marc
M
Gerry
Norman
N
Sebastiani
Paola
P
Steinberg
Martin H
MH
Farber
Harrison W
HW
D000755:Anemia, Sickle Cell / Q000097:blood / Q000235:genetics*; D001483:Base Sequence; D001499:Bayes Theorem; D002478:Cells, Cultured; D017931:DNA Primers; D018076:DNA, Complementary; D004730:Endothelium, Vascular / Q000502:physiology* / Q000503:physiopathology*; D005786:Gene Expression Regulation; D006451:Hemoglobin, Sickle / Q000235:genetics; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011651:Pulmonary Artery; D012313:RNA / Q000235:genetics; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D018076:DNA, Complementary; D006451:Hemoglobin, Sickle; D012313:RNA