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Rtp801, a suppressor of mTOR signaling, is an essential mediator of cigarette smoke–induced pulmonary injury and emphysema

Nature Medicine volume 16, pages 767–773 (2010)Cite this article

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Abstract

Rtp801 (also known as Redd1, and encoded by Ddit4), a stress-related protein triggered by adverse environmental conditions, inhibits mammalian target of rapamycin (mTOR) by stabilizing the TSC1-TSC2 inhibitory complex and enhances oxidative stress–dependent cell death. We postulated that Rtp801 acts as a potential amplifying switch in the development of cigarette smoke–induced lung injury, leading to emphysema. Rtp801 mRNA and protein were overexpressed in human emphysematous lungs and in lungs of mice exposed to cigarette smoke. The regulation of Rtp801 expression by cigarette smoke may rely on oxidative stress–dependent activation of the CCAAT response element in its promoter. We also found that Rtp801 was necessary and sufficient for nuclear factor-κB (NF-κB) activation in cultured cells and, when forcefully expressed in mouse lungs, it promoted NF-κB activation, alveolar inflammation, oxidative stress and apoptosis of alveolar septal cells. In contrast, Rtp801 knockout mice were markedly protected against acute cigarette smoke–induced lung injury, partly via increased mTOR signaling, and, when exposed chronically to cigarette smoke, against emphysema. Our data support the notion that Rtp801 may represent a major molecular sensor and mediator of cigarette smoke–induced lung injury.

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Figure 1: Enhanced expression of RTP801 in human emphysematous lungs.
Figure 2: Cigarette smoke–induced upregulation of Rtp801 expression occurs in lung septal but not resident or infiltrating inflammatory cells and relies on oxidative stress–dependent activation of the CCAAT promoter region.
Figure 3: Rtp801-dependent NF-κB activation by cigarette smoke.
Figure 4: Forced in vivo overexpression of human RTP801 in mouse lungs enhances oxidative stress, inflammation and alveolar cell apoptosis.
Figure 5: Rtp801-knockout mice are protected against cigarette smoke–induced pulmonary inflammation, apoptosis and emphysema.
Figure 6: mTOR regulates lung homeostasis and inflammatory responses due to cigarette smoke in wild-type (WT) and Rtp801-knockout mice.

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Acknowledgements

We thank T. Stevens (University of South Alabama) for providing the primary rat lung microvascular endothelial cells, J. Whitsett (University of Cincinnati) for allowing us to use the MLE-15 cells, T. Flotte and P. Cruz (Universities of Florida and Massachusetts, respectively) for preparing the AAV5 vectors, Y. Chen (University of Indiana) for the DDIT4 promoter constructs, I. Rahman (University of Rochester) for sharing cigarette smoke–sensitive and cigarette smoke–resistant mouse lung specimens, D. Brenner (University of California, San Diego) for the adenovirus IκB superrepressor, L. Ellison (Dana-Farber Cancer Center) for providing the DDIT4-RPAA construct and S. Reynolds (National Jewish Health) for the mouse-specific clara cell antigen 10 antibody. This work was supported by the grants R01 US National Heart, Lung, and Blood Institute (NHLBI) RO1 66554, Alpha 1 Foundation, Flight Attendant Medical Research Institute and Quark Pharmaceuticals (to R.M.T.); R01 NHLBI 077328 (to I.P.), COPD SCCOR P01–NHLBI 085609, Project 1 (to R.M.T. and I.P.) and Project 4 (to S.B.), RO1 NHLBI RO1 081205 to (S.B.) and NHLBI R01 HL082541–01 to (S.D.S.).

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Authors and Affiliations

  1. Division of Cardiopulmonary Pathology, Department of Pathology, Johns Hopkins University, Baltimore, Maryland, USA

    Toshinori Yoshida, Anil K Bhunia, Lijie Zhen, Ugonma Chukwueke, Tianzhi Mao, Amy Richter, Emile Brown, Irina Petrache & Rubin M Tuder

  2. Quark Pharmaceutical Inc, Fremont, California, USA

    Igor Mett, Hagit Ashush, Natalie Notkin, Anna Gelfand & Elena Feinstein

  3. Division of Pulmonary Sciences and Critical Care Medicine, Department of Medicine, Program in Translational Lung Research, University of Colorado, Denver School of Medicine, Denver, Colorado, USA

    Joel Bowman, Mario Perez, Li Zhang, Aneta Gandjeva & Rubin M Tuder

  4. Department of Environmental Health Sciences, Bloomberg School of Public Health, Baltimore, Maryland, USA

    Rajesh K Thimmulappa, Tirumalai Rangasamy, Thomas Sussan & Shyam Biswal

  5. National Jewish Health, Denver, Colorado, USA

    Gregory Cosgrove

  6. Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA

    Majd Mouded & Steven D Shapiro

  7. Division of Pulmonary, Department of Medicine, Allergy, Critical Care and Occupational Medicine, School of Medicine, Indiana University, Indianapolis, Indiana, USA

    Irina Petrache

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Contributions

T.Y., E.F. and R.M.T. designed the experiments, analyzed the data and composed the manuscript; T.Y., I.M., A.K.B., J.B., M.P., L. Zhang, A. Gandjeva, L. Zhen, U.C., T.M., A.R., E.B., H.A., N.N., A. Gelfand and I.P. performed in vitro and in vivo experiments; R.K.T., T.R., T.S. and S.B. provided mouse lung samples for Rtp801 expression studies; G.C. provided normal human lung samples for Rtp801 expression studies; M.M. and S.D.S. performed the chronic exposure to cigarette smoke.

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Correspondence to Elena Feinstein or Rubin M Tuder.

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Yoshida, T., Mett, I., Bhunia, A. et al. Rtp801, a suppressor of mTOR signaling, is an essential mediator of cigarette smoke–induced pulmonary injury and emphysema. Nat Med 16, 767–773 (2010). https://doi.org/10.1038/nm.2157

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