TGF-β/Smad3 activates mammalian target of rapamycin complex-1 to promote collagen production by increasing HIF-1α expression

Benaya Rozen-Zvi, Tomoko Hayashida, Susan C. Hubchak, Christian Hanna, Leonidas C. Platanias, H. William Schnaper

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)F485-F494
JournalAmerican Journal of Physiology - Renal Physiology
Volume305
Issue number4
DOIs
StatePublished - May 8 2013

Keywords

  • Collagen
  • Fibrosis
  • HIF
  • Rapamycin
  • Smad3
  • TGF
  • mTOR

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