Association of matrix metalloproteinase-3 with cardiogenic activity during Noggin-induced differentiation of mouse embryonic stem cells.
BACKGROUND
Despite the pluripotency of embryonic stem (ES) cells, their clinical applications have been hindered due to the lack of reliable differentiation methods. Recently, it was shown that Noggin could effectively induce cardiomyocyte differentiation by transient treatment of ES cells.


METHODS
To determine how Noggin may induce cardiac differentiation, we compared differentially expressed genes during Noggin-induced differentiation of ES cells using microarray analysis. We found Matrix metalloproteinase-3 (Mmp-3) expression was highly up-regulated by Noggin treatment. To understand the role of Mmp-3 in the cardiac differentiation of ES cells, we inhibited Mmp-3 activity by treating with a specific Mmp-3 inhibitor during Noggin-induced cardiac differentiation of ES cells. We also analyzed the expression levels of cardiac markers and the ratio of spontaneously beating embryoid bodies (EBs) in the presence of the Mmp-3 inhibitor.


RESULTS
We analyzed EB samples from zero, two, and four days with or without Noggin treatment, and found that the expression levels of 2 (0 day), 56 (2 days), and 805 (4 days) genes were altered with Noggin treatment. Up-regulation of Mmp-3 was closely associated with relative increases of cardiogenic, vasculogenic, and hematopoietic genes in EB treated with Noggin. By inhibiting Mmp-3 activity, we verified that Mmp-3 activation is partly responsible for both the expression of cardiac markers and the elevated ratio of spontaneously beating to non-beating EBs.


CONCLUSIONS
The concurrent expression of Mmp-3 with many cardiogenic genes and the specific inhibition of Mmp-3 revealed a critical role for Mmp-3 in Noggin-induced cardiac differentiation of ES cells.
Hong
Su
S
Macrogen Inc, World Meridian Venture Center, 60-24 Gasan-dong, Seoul 153-023, Republic of Korea.
Kang
Jae Ku
JK
Park
Jung Jun
JJ
Ryu
Eun Sook
ES
Choi
Sung Sik
SS
Lee
Sang Ho
SH
Lee
Jong Ho
JH
Seo
Jeong-Sun
JS
D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D006877:Hydroxamic Acids / Q000494:pharmacology; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis* / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D009842:Oligopeptides / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology; D000099176:Noggin Protein
D002352:Carrier Proteins; D006877:Hydroxamic Acids; D019278:Matrix Metalloproteinase 3; D061965:Matrix Metalloproteinase Inhibitors; D009842:Oligopeptides; D000099176:Noggin Protein; C095170:4-aminobenzoyl-glycyl-prolyl-leucyl-alanine hydroxamic acid