Signaling Mechanism of TGF-1 in Prevention of Renal Inflammation: Role of Smad7 Wansheng Wang,* Xiao R. Huang,* Allen G. Li, Fang Liu,* Jin-Hua Li,* Luan D. Truong,* Xiao J. Wang, and Hui Y. Lan* *Departments of Medicine and Pathology, Baylor College of Medicine, Houston, Texas; Department of Dermatology, Oregon Health & Science University, Portland, Oregon; and Department of Pathology, The Methodist Hospital, Houston, Texas TGF-has been shown to play a critical role in anti-inflammation; however, the signaling mechanisms of TGF-in anti-inflammatory response remains largely unclear. This study reported that mice that overexpress latent TGF-1 on skin are protected against renal inflammation in a model of obstructive kidney disease and investigated the signaling mechanism of TGF-1 in inhibition of renal inflammation in vivo and in vitro. Seven days after urinary obstruction, wild-type mice developed severe renal inflammation, including massive T cell and macrophage infiltration and marked upregulation of IL-1, TNF-, and intercellular adhesion molecule-1 (all P < 0.001). Surprising, renal inflammation was prevented in transgenic mice. This was associated with an increase in latent TGF-1 in circulation (a 10-fold increase) and renal tissues (a 2.5-fold increase). Further studies showed that inhibition of renal inflammation in TGF-1 transgenic mice was also associated with a marked upregulation of renal Smad7 and IBand a suppression of NF-B activation in the diseased kidney (all P < 0.01). These in vivo findings suggested the importance of TGF-–NF-B cross-talk signaling pathway in regulating renal inflammation. This was tested in vitro in a doxycycline-regulated Smad7-expressing renal tubular cell line. Overexpression of Smad7 was able to upregulate IBdirectly in a time- and dose-dependent manner, thereby inhibiting NF-B activation and NF-B– driven inflammatory response. In conclusion, latent TGF-may have protective roles in renal inflammation. Smad7- mediated inhibition of NF-B activation via the induction of IkBmay be the central mechanism by which latent TGF- prevents renal inflammation. J Am Soc Nephrol 16: 1371–1383, 2005. doi: 10.1681/ASN.2004121070 T he TGF-family, including TGF-1, TGF-2, and TGF- 3, is a group of pleiotropic secreted cytokines with unique and potent anti-inflammatory and immuno- regulatory properties (1). They have a broad spectrum of bio- logic functions and act on a large variety of cell types. Several experimental and human studies reveal an important role for TGF-in renal fibrosis (2), which is crucial for the progression to ESRD. Mice with overexpression of active TGF-1 develop severe renal damage with progressive renal fibrosis (3). It is interesting that blockade of TGF-by anti–TGF-antibodies alleviates renal fibrosis in diabetic nephropathy in mice and puromycin aminonucleoside nephropathy in rats but worsens albuminuria and proteinuria in these disease models (4,5), sug- gesting a complex role of TGF-, beyond its profibrotic effects, in the progression of renal diseases. We speculated that these observations might be associated with protective roles of circu- lating latent TGF-in renal inflammation. Thus, blockade of circulating TGF-with a neutralizing TGF-antibody enhances proteinuria in these disease models. However, there is lack of evidence to support this hypothesis. The signaling mechanisms that are responsible for the potential anti-inflammatory effects of TGF-are yet largely unknown. Although the role of TGF- in fibrosis has been firmly established, little attention has been paid to the circulating levels of this cytokine in the disease conditions. TGF-functions in both autocrine and paracrine manners to regulate cell proliferation, apoptosis, differentia- tion, chemotaxis, extracellular matrix production, cell migra- tion, and immune response (6). Mice with targeted disruption of TGF-1 display no gross abnormalities at birth but succumb to wasting associated with multifocal inflammation at approx- imately day 20 after birth (7). Systemic administration of TGF- greatly alleviates the inflammatory response in streptococcus cell wall–induced erosive arthritis and prevents the relapse of autoimmune encephalomyelitis, insulitis, and systemic lupus erythematosus in the MRL/lpr/lpr murine model (8 –11), strongly suggesting the anti-inflammatory effect of circulating TGF-. Although it is known that TGF-plays an important role in inflammatory responses, the underlying signaling mechanisms by which TGF-exerts its anti-inflammatory function remain largely unclear. Because mice that overexpress the bioactive form of hepatic TGF-1 exhibit progressive renal injury associ- ated with a highly increased circulating active TGF-1 (12), our study sought to test the hypothesis that the increased circulat- Received December 10, 2004. Accepted February 16, 2005. Published online ahead of print. Publication date available at www.jasn.org. Address correspondence to: Dr. Hui Y. Lan, Department of Medicine-Nephrol- ogy, Baylor College of Medicine, One Baylor Plaza, Alkek N520, Houston, TX 77030. Phone: 713-798-1303; Fax: 713-798-5010; E-mail: hlan@bcm.tmc.edu Copyright © 2005 by the American Society of Nephrology ISSN: 1046-6673/1605-1371