TY - JOUR
T1 - TGF-β/Smad3 activates mammalian target of rapamycin complex-1 to promote collagen production by increasing HIF-1α expression
AU - Rozen-Zvi, Benaya
AU - Hayashida, Tomoko
AU - Hubchak, Susan C.
AU - Hanna, Christian
AU - Platanias, Leonidas C.
AU - Schnaper, H. William
PY - 2013/5/8
Y1 - 2013/5/8
N2 - Transforming growth factor (TGF)- β is a major mediator of kidney fibrosis. In the past decade it was recognized that, besides canonical Smad signaling, many other signaling pathways participate in the process of TGF- β - induced fibrogenesis. One such pathway involves mammalian target of rapamycin complex (mTORC)1. We recently reported that the hypoxia-inducible factor (HIF)-1 is essential for TGF- β -induced collagen expression regardless of ambient oxygen tension. A modulator of HIF expression other than oxygen tension is mTORC1. We therefore sought to evaluate a possible role for mTORC1 activity in TGF- β -induced fibrogenesis. mTORC1 activity was increased in human mesangial cells treated with TGF- β in a TGF- β receptordependent manner. Short hairpin (sh)RNA to Smad3 decreased, while overexpression of Smad3 increased, the mTORC1 activity, suggesting that TGF-β stimulation of mTORC1 also requires Smad3. Pretreatment with rapamycin or shRNA for a regulatory molecule of mTORC1, Raptor, reduced TGF- β -induced COL1A2-luc activity and collagen I protein expression. mTORC1 inhibition also prevented the TGF-β-stimulated increase in both hypoxia-responsive element (HRE) activity and HIF-1 protein expression, while activation of mTORC1 by active Rheb increased basal but not TGF--induced HRE activity. shRNA to Smad3 reduced HRE activity, while overexpression of Smad3 increased HIF-1 protein expression and activity in an mTORC1-dependent manner. Lastly, overexpression of HIF-1 bypassed the inhibitory effect of mTORC1 blockade on collagen expression. These results suggest that Smad3/mTORC1 interaction to promote HIF-1α expression is a key step in normoxic kidney fibrogenesis.
AB - Transforming growth factor (TGF)- β is a major mediator of kidney fibrosis. In the past decade it was recognized that, besides canonical Smad signaling, many other signaling pathways participate in the process of TGF- β - induced fibrogenesis. One such pathway involves mammalian target of rapamycin complex (mTORC)1. We recently reported that the hypoxia-inducible factor (HIF)-1 is essential for TGF- β -induced collagen expression regardless of ambient oxygen tension. A modulator of HIF expression other than oxygen tension is mTORC1. We therefore sought to evaluate a possible role for mTORC1 activity in TGF- β -induced fibrogenesis. mTORC1 activity was increased in human mesangial cells treated with TGF- β in a TGF- β receptordependent manner. Short hairpin (sh)RNA to Smad3 decreased, while overexpression of Smad3 increased, the mTORC1 activity, suggesting that TGF-β stimulation of mTORC1 also requires Smad3. Pretreatment with rapamycin or shRNA for a regulatory molecule of mTORC1, Raptor, reduced TGF- β -induced COL1A2-luc activity and collagen I protein expression. mTORC1 inhibition also prevented the TGF-β-stimulated increase in both hypoxia-responsive element (HRE) activity and HIF-1 protein expression, while activation of mTORC1 by active Rheb increased basal but not TGF--induced HRE activity. shRNA to Smad3 reduced HRE activity, while overexpression of Smad3 increased HIF-1 protein expression and activity in an mTORC1-dependent manner. Lastly, overexpression of HIF-1 bypassed the inhibitory effect of mTORC1 blockade on collagen expression. These results suggest that Smad3/mTORC1 interaction to promote HIF-1α expression is a key step in normoxic kidney fibrogenesis.
KW - Collagen
KW - Fibrosis
KW - HIF
KW - Rapamycin
KW - Smad3
KW - TGF
KW - mTOR
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U2 - 10.1152/ajprenal.00215.2013
DO - 10.1152/ajprenal.00215.2013
M3 - Article
C2 - 23761672
AN - SCOPUS:84881619024
SN - 1931-857X
VL - 305
SP - F485-F494
JO - American Journal of Physiology - Renal Physiology
JF - American Journal of Physiology - Renal Physiology
IS - 4
ER -