PGC1α drives NAD biosynthesis linking oxidative metabolism to renal protection (original) (raw)

References

  1. Thadhani, R., Pascual, M. & Bonventre, J. V. Acute renal failure. N. Engl. J. Med. 334, 1448–1460 (1996)
    CAS PubMed Google Scholar
  2. Lewington, A. J., Cerda, J. & Mehta, R. L. Raising awareness of acute kidney injury: a global perspective of a silent killer. Kidney Int. 84, 457–467 (2013)
    PubMed PubMed Central Google Scholar
  3. Puigserver, P. et al. A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis. Cell 92, 829–839 (1998)
    CAS PubMed Google Scholar
  4. Ruas, J. L. et al. A PGC-1α isoform induced by resistance training regulates skeletal muscle hypertrophy. Cell 151, 1319–1331 (2012)
    CAS PubMed PubMed Central Google Scholar
  5. Hanson, J. et al. Nicotinic acid- and monomethyl fumarate-induced flushing involves GPR109A expressed by keratinocytes and COX-2-dependent prostanoid formation in mice. J. Clin. Invest. 120, 2910–2919 (2010)
    CAS PubMed PubMed Central Google Scholar
  6. Pagliarini, D. J. et al. A mitochondrial protein compendium elucidates complex I disease biology. Cell 134, 112–123 (2008)
    CAS PubMed PubMed Central Google Scholar
  7. Tran, M. et al. PGC-1α promotes recovery after acute kidney injury during systemic inflammation in mice. J. Clin. Invest. 121, 4003–4014 (2011)
    CAS PubMed PubMed Central Google Scholar
  8. Arany, Z. et al. HIF-independent regulation of VEGF and angiogenesis by the transcriptional coactivator PGC-1α. Nature 451, 1008–1012 (2008)
    ADS CAS PubMed Google Scholar
  9. Agudelo, L. Z. et al. Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates resilience to stress-induced depression. Cell 159, 33–45 (2014)
    CAS PubMed Google Scholar
  10. Arany, Z. et al. Transverse aortic constriction leads to accelerated heart failure in micelacking PPAR-γ coactivator 1α. Proc. Natl Acad. Sci. USA 103, 10086–10091 (2006)
    ADS CAS PubMed Google Scholar
  11. St-Pierre, J. et al. Suppression of reactive oxygen species and neurodegeneration by the PGC-1 transcriptional coactivators. Cell 127, 397–408 (2006)
    CAS PubMed Google Scholar
  12. Weidemann, M. J. & Krebs, H. A. The fuel of respiration of rat kidney cortex. Biochem. J. 112, 149–166 (1969)
    CAS PubMed PubMed Central Google Scholar
  13. Collins, P. B. & Chaykin, S. The management of nicotinamide and nicotinic acid in the mouse. J. Biol. Chem. 247, 778–783 (1972)
    CAS PubMed Google Scholar
  14. Traykova-Brauch, M. et al. An efficient and versatile system for acute and chronic modulation of renal tubular function in transgenic mice. Nature Med. 14, 979–984 (2008)
    CAS PubMed Google Scholar
  15. Revollo, J. R., Grimm, A. A. & Imai, S. The NAD biosynthesis pathway mediated by nicotinamide phosphoribosyltransferase regulates Sir2 activity in mammalian cells. J. Biol. Chem. 279, 50754–50763 (2004)
    CAS PubMed Google Scholar
  16. Bai, P. et al. PARP-1 inhibition increases mitochondrial metabolism through SIRT1 activation. Cell Metab. 13, 461–468 (2011)
    CAS PubMed PubMed Central Google Scholar
  17. Gomes, A. P. et al. Declining NAD+ induces a pseudohypoxic state disrupting nuclear-mitochondrial communication during aging. Cell 155, 1624–1638 (2013)
    CAS PubMed PubMed Central Google Scholar
  18. Garcia Soriano, F. et al. Diabetic endothelial dysfunction: the role of poly(ADP-ribose) polymerase activation. Nature Med. 7, 108–113 (2001)
    CAS PubMed Google Scholar
  19. Jesinkey, S. R. et al. Formoterol restores mitochondrial and renal function after ischemia-reperfusion injury. J. Am. Soc. Nephrol. 25, 1157–1162 (2014)
    CAS PubMed PubMed Central Google Scholar
  20. Boström, P. et al. A PGC1-α-dependent myokine that drives brown-fat-like development of white fat and thermogenesis. Nature 481, 463–468 (2012)
    ADS PubMed PubMed Central Google Scholar
  21. Breyer, M. D., Jacobson, H. R. & Breyer, R. M. Functional and molecular aspects of renal prostaglandin receptors. J. Am. Soc. Nephrol. 7, 8–17 (1996)
    CAS PubMed Google Scholar
  22. Papanicolaou, N., Callard, P., Bariety, J. & Milliez, P. The effect of indomethacin and prostaglandin (PGE2) on renal failure due to glycerol in saline-loaded rats. Clin. Sci. Mol. Med. 49, 507–510 (1975)
    CAS PubMed Google Scholar
  23. Mauk, R. H., Patak, R. V., Fadem, S. Z., Lifschitz, M. D. & Stein, J. H. Effect of prostaglandin E administration in a nephrotoxic and a vasoconstrictor model of acute renal failure. Kidney Int. 12, 122–130 (1977)
    CAS PubMed Google Scholar
  24. Sketch, M. H., Jr et al. Prevention of contrast media-induced renal dysfunction with prostaglandin E1: a randomized, double-blind, placebo-controlled study. Am. J. Ther. 8, 155–162 (2001)
    PubMed Google Scholar
  25. Feldkamp, T. et al. Preservation of complex I function during hypoxia-reoxygenation-induced mitochondrial injury in proximal tubules. Am. J. Physiol. Renal Physiol. 286, F749–F759 (2004)
    CAS PubMed Google Scholar
  26. Morigi, M. et al. Sirtuin 3-dependent mitochondrial dynamic improvements protect against acute kidney injury. J. Clin. Invest. 125, 715–726 (2015)
    PubMed PubMed Central Google Scholar
  27. Ebrahimkhani, M. R. et al. Aag-initiated base excision repair promotes ischemia reperfusion injury in liver, brain, and kidney. Proc. Natl Acad. Sci. USA 111, E4878–E4886 (2014)
    CAS PubMed Google Scholar
  28. Rajakumar, S. V. et al. Deficiency or inhibition of CD73 protects in mild kidney ischemia-reperfusion injury. Transplantation 90, 1260–1264 (2010)
    CAS PubMed PubMed Central Google Scholar
  29. Wang, G. et al. P7C3 neuroprotective chemicals function by activating therate-limiting enzyme in NAD salvage. Cell 158, 1324–1334 (2014)
    CAS PubMed PubMed Central Google Scholar
  30. Kraus, D. et al. Nicotinamide _N_-methyltransferase knockdown protects against diet-induced obesity. Nature 508, 258–262 (2014)
    ADS CAS PubMed PubMed Central Google Scholar
  31. Tran, M. et al. PGC-1α promotes recovery after acute kidney injury during systemic inflammation in mice. J. Clin. Invest. 121, 4003–4014 (2011)
    CAS PubMed PubMed Central Google Scholar
  32. Traykova-Brauch, M. et al. An efficient and versatile system for acute and chronic modulation of renal tubular function in transgenic mice. Nature Med. 14, 979–984 (2008)
    CAS PubMed Google Scholar
  33. Rask-Andersen, M., Almen, M. S. & Schioth, H. B. Trends in the exploitation of novel drug targets. Nature Rev. Drug Discov. 10, 579–590 (2011)
    CAS Google Scholar
  34. Singh, V. et al. Mycobacterium tuberculosis-driven targeted recalibration of macrophage lipid homeostasis promotes the foamy phenotype. Cell Host Microbe 12, 669–681 (2012)
    CAS PubMed Google Scholar
  35. Feingold, K. R., Moser, A., Shigenaga, J. K. & Grunfeld, C. Inflammation stimulates niacin receptor (GPR109A/HCA2) expression in adipose tissue and macrophages. J. Lipid Res. 55, 2501–2508 (2014)
    CAS PubMed PubMed Central Google Scholar
  36. Zsengellér, Z. K. et al. Cisplatin nephrotoxicity involves mitochondrial injury with impaired tubular mitochondrial enzyme activity. J. Histochem. Cytochem. 60, 521–529 (2012)
    PubMed PubMed Central Google Scholar
  37. Rhee, E. P. et al. A genome-wide association study of the human metabolome in a community-based cohort. Cell Metab. 18, 130–143 (2013)
    CAS PubMed PubMed Central Google Scholar
  38. Kang, C. & Ji, L. L. Muscle immobilization and remobilization downregulates PGC-1α signaling and the mitochondrial biogenesis pathway. J. Appl. Physiol. (1985) 115, 1618–1625 (2013)
    CAS Google Scholar
  39. Ruas, J. L. et al. A PGC-1α isoform induced by resistance training regulates skeletal muscle hypertrophy. Cell 151, 1319–1331 (2012)
    CAS PubMed PubMed Central Google Scholar
  40. Nakahata, Y., Sahar, S., Astarita, G., Kaluzova, M. & Sassone-Corsi, P. Circadian control of the NAD+ salvage pathway by CLOCK-SIRT1. Science 324, 654–657 (2009)
    ADS CAS PubMed PubMed Central Google Scholar
  41. Agudelo, L. Z. et al. Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates resilience to stress-induced depression. Cell 159, 33–45 (2014)
    CAS PubMed Google Scholar
  42. Liu, L. et al. Nutrient sensing by the mitochondrial transcription machinery dictates oxidative phosphorylation. J. Clin. Invest. 124, 768–784 (2014)
    PubMed PubMed Central Google Scholar
  43. Hasmann, M. & Schemainda, I. FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis. Cancer Res. 63, 7436–7442 (2003)
    CAS PubMed Google Scholar
  44. Qi, Z. et al. Serial determination of glomerular filtration rate in conscious mice using FITC-inulin clearance. Am. J. Physiol. Renal Physiol. 286, F590–F596 (2004)
    CAS PubMed Google Scholar
  45. Antonica, F. et al. Generation of functional thyroid from embryonic stem cells. Nature 491, 66–71 (2012)
    ADS CAS PubMed PubMed Central Google Scholar

Download references

Acknowledgements

The authors thank B. Spiegelman (Harvard Medical School, Dana Farber Cancer Institute) for support from this project’s inception; M. Zeidel (Harvard Medical School, BIDMC) for advice; Z. Arany (University of Pennsylvania) for the VE-cadherin-tTA × TRE-PGC1α mice; A. Agarwal (University of Alabama, NIH P30-DK079337) for LC–MS measurements of serum creatinine; A. Kurmann and A. Hollenberg (Harvard Medical School, BIDMC) for thyroxine measurements; and P. Pacher (NIH) for cisplatin-treated kidneys for microscopy. This work was supported by R01-DK095072 and philanthropic funds to S.M.P.; K08-DK090142 and a grant from Satellite Healthcare to E.P.R.; and K08-DK101560 to E.V.K.

Author information

Authors and Affiliations

  1. Division of Nephrology and Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, 02215, Massachusetts, USA
    Mei T. Tran, Zsuzsanna K. Zsengeller, Eliyahu V. Khankin, S. Ananth Karumanchi & Samir M. Parikh
  2. Center for Vascular Biology Research, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, 02215, Massachusetts, USA
    Mei T. Tran, Zsuzsanna K. Zsengeller, Eliyahu V. Khankin, Manoj K. Bhasin, S. Ananth Karumanchi & Samir M. Parikh
  3. Division of Clinical Chemistry, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, 02215, Massachusetts, USA
    Zsuzsanna K. Zsengeller & Anders H. Berg
  4. Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, 02215, Massachusetts, USA
    Anders H. Berg & Isaac E. Stillman
  5. Bioinformatics and Systems Biology Core, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, 02215, Massachusetts, USA
    Manoj K. Bhasin
  6. Nephrology and Endocrine Divisions, Massachusetts General Hospital and Harvard Medical School, Boston, 02114, Massachusetts, USA
    Wondong Kim & Eugene P. Rhee
  7. Broad Institute of MIT and Harvard, Cambridge, 02139, Massachusetts, USA
    Clary B. Clish & Eugene P. Rhee
  8. Howard Hughes Medical Institute, Chevy Chase, 20815, Maryland, USA
    S. Ananth Karumanchi

Authors

  1. Mei T. Tran
  2. Zsuzsanna K. Zsengeller
  3. Anders H. Berg
  4. Eliyahu V. Khankin
  5. Manoj K. Bhasin
  6. Wondong Kim
  7. Clary B. Clish
  8. Isaac E. Stillman
  9. S. Ananth Karumanchi
  10. Eugene P. Rhee
  11. Samir M. Parikh

Contributions

M.T.T. designed experiments, performed the breeding, genotyping, renal injury models, cellular studies, analysed data, and wrote the manuscript. Z.K.Z. and I.E.S. performed and analysed histopathology, enzyme histochemistry, electron microscopy, and the human biopsy immunohistochemistry studies. A.H.B. created LC–MS assays, measured metabolites for cellular experiments, and analysed metabolic results. E.V.K. and S.A.K. performed micro-ultrasounds and analysed flow results. M.K.B. analysed raw RNA sequencing data. W.K., C.B.C. and E.P.R. performed metabolomics, follow-up metabolite measurements, and in vivo experiments with cisplatin and NAM. S.M.P. designed the experiments, analysed results, and wrote the manuscript with input from all authors.

Corresponding author

Correspondence toSamir M. Parikh.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Regulation of PGC1α and other features of post-ischaemic kidneys.

a, Serum creatinine 24 h after sham or IRI (n = 5 versus 14 mice); ***P < 0.001. b, Absence of class-wide changes in intrarenal phospholipids 24 h after IRI versus sham operation (n = 6 per group; NS, not significant). Each bar represents one lipid species. P value calculated using two-way ANOVA. c, Renal PGC1α expression 24 h after sham or IRI (n = 5 animals per group); **P < 0.01. d, Correlation of LC–MS method for serum creatinine and serum cystatin C (measured by ELISA). e, Glomerular filtration rate in controls or 24 h after IRI was determined by two-phase exponential decay curves of fluorescently labelled inulin as described in Methods (n = 5 per group); *P < 0.05. f, Correlation of LC–MS method for serum creatinine with clearance of FITC–inulin. Curve fit according to formula sCr = κ/GFR where κ is a constant. Error bars, s.e.m.

Extended Data Figure 2 Exacerbation of fat accumulation and tubular injury in post-ischaemic _Pgc1α_−/− kidneys.

a–d, Low- (top) and high- (bottom) power photomicrographs 24 h after IRI in wild-type (WT; a, c) versus _Pgc1α_−/− (KO; b, d) mice. Scale bars, 200 and 100 μm (top and bottom, respectively). e, f, Blinded scoring of tubular injury in cortex and outer stripe of outer medulla (OSOM) on 4-point injury scale as described in Methods (n = 8 wild-type versus 12 knockout mice); *P < 0.05. g, Di-/tri-acylglycerols (DAGs, TAGs) in renal homogenates of knockout mice at baseline and 24 h after injury (n = 6 per group). Each bar represents one lipid species. P value calculated using two-way ANOVA. Error bars, s.e.m.

Extended Data Figure 3 NAM reduction from IRI and PGC1α deficiency.

a, Heat maps (red, higher; green, lower) of Bonferroni-corrected significantly different metabolites in sham versus IRI kidneys and wild-type (WT) versus knockout (KO) kidneys. Metabolites listed in purple are shared between settings. b, Total ion chromatogram of polar, positive-ion mode method for representative wild-type IRI sample, with NAM peak at retention time of 3.88 min. Inset shows representative NAM peaks for kidney extracts from wild-type control (Ctrl) and wild-type IRI (IRI) mice. c–e, Relative renal NAM abundance in kidneys of knockout mice versus wild-type littermates; wild-type littermates at baseline and 24 h after IRI; and knockout mice at baseline and 24 h after IRI (n = 6 per group). f, Relative renal NAM concentrations in kidneys of mice following vehicle (Veh) versus NAM treatment (400 mg kg−1 intraperitoneal for 4 days) with and without IRI 24 h before tissue collection (n = 6 per group). P values calculated with two-way ANOVA. g, h, Oil-Red-O stain (pink) for fat accumulation 24 h after IRI with or without NAM pre-treatment (400 mg kg−1 intraperitoneal for 4 days); scale bar, 20 μm. Error bars, s.e.m.; *P < 0.05, **P < 0.01, ***P < 0.001.

Extended Data Figure 4 Increased mitochondrial abundance and post-ischaemic protection in renal tubular epithelial transgenic mice (iNephPGC1α).

a, Schematic for generating iNephPGC1α mice. b, Relative renal PGC1α expression in control versus iNephPGC1α mice with and without 4 weeks of doxycycline in drinking water (n = 5 per group; **P < 0.01 versus all other groups). c, Ratio of kidney weight to total body weight (note body weights statistically indistinguishable as well, n = 4 per group). d, Example gross images with 1 cm scale of control versus iNephPGC1α kidney. e, Renal mitochondrial DNA (mtDNA) copy number as described in Methods. f, Relative renal gene expression of PGC1α targets (Ndufs1, Cycs, Atp5o), partnering transcription factors (Nrf1), and the mitochondrial transcription factor, Tfam. Results analysed by two-way ANOVA with P value for genotype as noted. n = 8 per group. *P < 0.05 versus control after Bonferroni correction. g, Western blot analysis of kidney lysates for transcription factor a, mitochondrial (TFAM)38 and loading control. h, i, Transmission electron microscopy of mitochondria sectioned perpendicular and parallel to long axis demonstrating normal morphology in iNephPGC1α mice (representative of n = 4 per group); scale bar, 500 nm. j, k, Blinded scoring of tubular injury in cortex and outer stripe of outer medulla (n = 8 control; 12 iNephPGC1α). Error bars s.e.m.; *P < 0.05, **P < 0.01; NS, not significant.

Extended Data Figure 5 Renal protection in systemic inflammation conferred by renal tubular epithelial, but not endothelial, PGC1α.

a, Serum creatinine 24 h after bacterial endotoxin injection (LPS O111:B4), n = 9 per group. b, Serum creatinine 24 h after bacterial endotoxin (LPS O111:B4) in endothelial-specific (VEC, VE-cadherin) PGC1α transgenic mice (VEC-tTA × TRE-PGC1α), n = 5 per group. Error bars, s.e.m., *P < 0.05; NS, not significant.

Extended Data Figure 6 PGC1α-dependent de novo NAD biosynthesis and NAD-dependent accumulation of β-OHB and PGE2.

a, Gene expression for de novo NAD biosynthetic pathway in control renal tubular cells versus 48 h after PGC1α knockdown (n = 3 per condition). The gene expression set corresponds to the eight transcripts whose abundance was measured in kidney homogenates in Fig. 3. P = 0.0001 by two-way ANOVA with Bonferroni-corrected comparisons as indicated. b, Correlation of renal NAM versus renal NAD in mice treated with vehicle or different doses of NAM (one intraperitoneal dose of 100–400 mg kg−1). Arbitrary units on x and y axes. c, Renal β-OHB concentrations in kidneys of mice following vehicle (Veh) versus NAM treatment (400 mg kg−1 intraperitoneal for 4 days) with and without IRI 24 h before tissue collection (n = 5 per group). P value calculated with two-way ANOVA. Dashed line indicates normal circulating concentration of β-OHB. d, Dosing for siRNA against HCAR2 in renal tubular cells. e, Dose–inhibition curve in renal tubular cells for PGE2 release following 24 h of mepenzolate bromide at the indicated concentrations (n = 3 replicates per concentration)33,34,35. f, g, Intracellular NAM and secreted β-OHB for renal tubular cells following treatment with NAM (1 μM for 24 h) with or without pre-treatment with the NAMPT inhibitor FK866 (10 nM, n = 6 per condition). h, PGE2 in conditioned media of renal tubular cells after control versus PGC1α knockdown and with and without exogenous β-OHB application (+, 5 mM, n = 6 per condition, P values versus control group). Error bars, s.e.m.; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Extended Data Figure 7 Effects of PGC1α on renal metabolites and features of cisplatin nephrotoxicity.

a–c, Relative renal NAD, β-OHB, and PGE2 concentrations in wild-type (WT) littermates versus _Pgc1α_−/− (KO) mice (n = 6 per group). d, Serum creatinine in genetic control mice for iNephPGC1α 24 h after IRI with vehicle versus mepenzolate (MPN, 10 mg kg−1 intraperitoneal) treatment (n = 5 per group). e, Serum creatinine in genetic control mice for iNephPGC1α 24 h after IRI with vehicle versus indomethacin (INDO, 10 mg kg−1 intraperitoneal) treatment (n = 6 per group). f, Transmission electron microscopy with cytochrome c oxidase enzyme histochemistry of proximal tubular cell 24 h following cisplatin exposure (25 mg kg−1 intraperitoneal) demonstrating mitochondrial injury. Scale bar, 500 nm. g, Relative renal NAM concentrations following cisplatin as in f. Error bars, s.e.m.; *P < 0.05, **P < 0.01, ***P < 0.001; NS, not significant.

Extended Data Figure 8 Renal immunostaining for PGC1α declines in human chronic kidney disease.

a–d, Low- (a, b) and high- (c, d) power photomicrographs of PGC1α immunoreactivity (brown) in wild-type (WT, left) littermates and _Pgc1α_−/− (KO, right) kidneys. Scale bars, 100 and 50 μm. e, f, Representative results of peptide competition attenuating PGC1α immunoreactivity against human kidney (n = 4) as described in Methods. g, Representative immunostaining (brown) for PGC1α in a renal biopsy with chronic kidney disease (CKD). Scale bar, 50 μm. h, Results of scoring PGC1α immunostaining intensity (1, weakest; 4, strongest) in specimens with CKD by blinded operator. Each dot represents a unique specimen. Analysed with Mann–Whitney _U_-test.

Extended Data Figure 9 Evidence for renal-tubular-epithelial-PGC1α-dependent reversible vascular relaxation.

a, Serum creatinine in uninduced (−Dox) versus induced ( + Dox) iNephPGC1α mice (n = 8 mice per group). b, Comparison of serum creatinine with degree of renal PGC1α expression, P < 0.05. c, d, Serial serum creatinines in iNephPGC1α mice versus controls before PGC1α induction (OFF), after 4 weeks of PGC1α induction (ON), and after 4 weeks of washout (OFF), n = 5 per group; *P < 0.05 as calculated using repeated-measures ANOVA. e–g, Comparison of serum creatinine at different time points with renal artery flow in iNephPGC1α mice from d, P < 0.05 when correlation coefficient r = −0.65. h–j, Comparison of resistive index with renal artery flow volume in iNephPGC1α mice from d, P < 0.05 when correlation coefficient r = −0.80. k, Relative renal expression of VEGF and nitric oxide synthases 1 and 3 (n = 6 per group). Analysed by two-way ANOVA with Bonferroni corrections. l, Circulating thyroxine levels in iNephPGC1α mice with and without gene induction (n = 5 per g_r_oup) to rule out Pax8-related thyrotoxicosis driving perfusion differences as previously described45. m, Relative renal expression for VEGF in Pgc1α −/− mice (KO) versus wild-type (WT) littermates (n = 6 per group). Error bars, s.e.m.; NS, not significant.

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

Tran, M., Zsengeller, Z., Berg, A. et al. PGC1α drives NAD biosynthesis linking oxidative metabolism to renal protection.Nature 531, 528–532 (2016). https://doi.org/10.1038/nature17184

Download citation