Chondro/osteoblastic and cardiovascular gene modulation in human artery smooth muscle cells that calcify in the presence of phosphate and calcitriol or paricalcitol.
Vitamin D sterol administration, a traditional treatment for secondary hyperparathyroidism, may increase serum calcium and phosphorus, and has been associated with increased vascular calcification (VC). In vitro studies suggest that in the presence of uremic concentrations of phosphorus, vitamin D sterols regulate gene expression associated with trans-differentiation of smooth muscle cells (SMCs) to a chondro/osteoblastic cell type. This study examined effects of vitamin D sterols on gene expression profiles associated with phosphate-enhanced human coronary artery SMC (CASMC) calcification. Cultured CASMCs were exposed to phosphate-containing differentiation medium (DM) with and without calcitriol, paricalcitol, or the calcimimetic R-568 (10(-11)-10(-7) M) for 7 days. Calcification of CASMCs, determined using colorimetry following acid extraction, was dose dependently increased (1.6- to 1.9-fold) by vitamin D sterols + DM. In contrast, R-568 did not increase calcification. Microarray analysis demonstrated that, compared with DM, calcitriol (10(-8) M) + DM or paricalcitol (10(-8) M) + DM similarly and significantly (P < 0.05) regulated genes of various pathways including: metabolism, CYP24A1; mineralization, ENPP1; apoptosis, GIP3; osteo/chondrogenesis, OPG, TGFB2, Dkk1, BMP4, BMP6; cardiovascular, HGF, DSP1, TNC; cell cycle, MAPK13; and ion channels, SLC22A3 KCNK3. R-568 had no effect on CASMC gene expression. Thus, SMC calcification observed in response to vitamin D sterol + DM may be partially mediated through targeting mineralization, apoptotic, osteo/chondrocytic, and cardiovascular pathway genes, although some gene changes may protect against calcification. Further studies to determine precise roles of these genes in development of, or protection against VC and cardiovascular disease are required.
Shalhoub
V
V
Department of Metabolic Disorders, Amgen, Inc., Thousand Oaks, California, USA. shalhoub@amgen.com
Shatzen
E M
EM
Ward
S C
SC
Young
J-I
JI
Boedigheimer
M
M
Twehues
L
L
McNinch
J
J
Scully
S
S
Twomey
B
B
Baker
D
D
Kiaei
P
P
Damore
M A
MA
Pan
Z
Z
Haas
K
K
Martin
D
D
D021121:Branched DNA Signal Amplification Assay; D002113:Calcification, Physiologic / Q000187:drug effects / Q000235:genetics*; D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003331:Coronary Vessels / Q000166:cytology*; D003470:Culture Media / Q000494:pharmacology; D004872:Ergocalciferols / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010710:Phosphates / Q000494:pharmacology*; D018167:Receptors, Calcitriol / Q000235:genetics / Q000378:metabolism; D044169:Receptors, Calcium-Sensing / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D020218:Response Elements / Q000235:genetics; D014019:Tissue Donors; D014807:Vitamin D / Q000235:genetics
D003470:Culture Media; D004872:Ergocalciferols; D010710:Phosphates; D018167:Receptors, Calcitriol; D044169:Receptors, Calcium-Sensing; D014807:Vitamin D; C084656:paricalcitol; D002117:Calcitriol