Jan K Hennigs1,2,3,4,5, Aiqin Cao1,2,3, Caiyun G Li1,2,3,6, Minyi Shi2,7, Julia Mienert4,5, Kazuya Miyagawa1,2,3, Jakob Körbelin4,5, David P Marciano2,3,7, Pin-I Chen1,2,3, Matthew Roughley1,2,3, Matthew V Elliott1,2,3, Rebecca L Harper1,2,3, Matthew A Bill1,2,3, James Chappell1,2,3,7, Jan-Renier Moonen1,2,3, Isabel Diebold1,2,3, Lingli Wang1,2,3, Michael P Snyder2,7, Marlene Rabinovitch2,3. 1. Vera Moulton Wall Center for Pulmonary Vascular Disease (J.K.H., A.C., C.G.L., K.M., P.-I.C., M.R., M.V.E., R.L.H., M.A.B., J.C., J.-R.M., I.D., L.W.), Stanford University School of Medicine. 2. Cardiovascular Institute (J.K.H., A.C., C.G.L., M.S., K.M., D.P.M., P.-I.C., M.R., M.V.E., R.L.H., M.A.B., J.C., J.-R.M., I.D., L.W., M.P.S., M.R.), Stanford University School of Medicine. 3. Department of Pediatrics (J.K.H., A.C., C.G.L., K.M., D.P.M., P.-I.C., M.R., M.V.E., R.L.H., M.A.B., J.C., J.-R.M., I.D., L.W., M.R.), Stanford University School of Medicine. 4. Department of Pneumology and Center for Pulmonary Arterial Hypertension Hamburg (J.K.H., J.M., J.K.), University Medical Center Hamburg-Eppendorf, Germany. 5. II. Department of Medicine (J.K.H., J.M., J.K.), University Medical Center Hamburg-Eppendorf, Germany. 6. Department of Radiation Oncology (C.G.L.), Stanford University School of Medicine. 7. Department of Genetics (M.S., D.P.M., J.C., M.P.S.), Stanford University School of Medicine.
Abstract
RATIONALE: In pulmonary arterial hypertension (PAH), endothelial dysfunction and obliterative vascular disease are associated with DNA damage and impaired signaling of BMPR2 (bone morphogenetic protein type 2 receptor) via two downstream transcription factors, PPARγ (peroxisome proliferator-activated receptor gamma), and p53. OBJECTIVE: We investigated the vasculoprotective and regenerative potential of a newly identified PPARγ-p53 transcription factor complex in the pulmonary endothelium. METHODS AND RESULTS: In this study, we identified a pharmacologically inducible vasculoprotective mechanism in pulmonary arterial and lung MV (microvascular) endothelial cells in response to DNA damage and oxidant stress regulated in part by a BMPR2 dependent transcription factor complex between PPARγ and p53. Chromatin immunoprecipitation sequencing and RNA-sequencing established an inducible PPARγ-p53 mediated regenerative program regulating 19 genes involved in lung endothelial cell survival, angiogenesis and DNA repair including, EPHA2 (ephrin type-A receptor 2), FHL2 (four and a half LIM domains protein 2), JAG1 (jagged 1), SULF2 (extracellular sulfatase Sulf-2), and TIGAR (TP53-inducible glycolysis and apoptosis regulator). Expression of these genes was partially impaired when the PPARγ-p53 complex was pharmacologically disrupted or when BMPR2 was reduced in pulmonary artery endothelial cells (PAECs) subjected to oxidative stress. In endothelial cell-specific Bmpr2-knockout mice unable to stabilize p53 in endothelial cells under oxidative stress, Nutlin-3 rescued endothelial p53 and PPARγ-p53 complex formation and induced target genes, such as APLN (apelin) and JAG1, to regenerate pulmonary microvessels and reverse pulmonary hypertension. In PAECs from BMPR2 mutant PAH patients, pharmacological induction of p53 and PPARγ-p53 genes repaired damaged DNA utilizing genes from the nucleotide excision repair pathway without provoking PAEC apoptosis. CONCLUSIONS: We identified a novel therapeutic strategy that activates a vasculoprotective gene regulation program in PAECs downstream of dysfunctional BMPR2 to rehabilitate PAH PAECs, regenerate pulmonary microvessels, and reverse disease. Our studies pave the way for p53-based vasculoregenerative therapies for PAH by extending the therapeutic focus to PAEC dysfunction and to DNA damage associated with PAH progression.
RATIONALE: In pulmonary arterial hypertension (PAH), endothelial dysfunction and obliterative vascular disease are associated with DNA damage and impaired signaling of BMPR2 (bone morphogenetic protein type 2 receptor) via two downstream transcription factors, PPARγ (peroxisome proliferator-activated receptor gamma), and p53. OBJECTIVE: We investigated the vasculoprotective and regenerative potential of a newly identified PPARγ-p53 transcription factor complex in the pulmonary endothelium. METHODS AND RESULTS: In this study, we identified a pharmacologically inducible vasculoprotective mechanism in pulmonary arterial and lung MV (microvascular) endothelial cells in response to DNA damage and oxidant stress regulated in part by a BMPR2 dependent transcription factor complex between PPARγ and p53. Chromatin immunoprecipitation sequencing and RNA-sequencing established an inducible PPARγ-p53 mediated regenerative program regulating 19 genes involved in lung endothelial cell survival, angiogenesis and DNA repair including, EPHA2 (ephrin type-A receptor 2), FHL2 (four and a half LIM domains protein 2), JAG1 (jagged 1), SULF2 (extracellular sulfatase Sulf-2), and TIGAR (TP53-inducible glycolysis and apoptosis regulator). Expression of these genes was partially impaired when the PPARγ-p53 complex was pharmacologically disrupted or when BMPR2 was reduced in pulmonary artery endothelial cells (PAECs) subjected to oxidative stress. In endothelial cell-specific Bmpr2-knockout mice unable to stabilize p53 in endothelial cells under oxidative stress, Nutlin-3 rescued endothelial p53 and PPARγ-p53 complex formation and induced target genes, such as APLN (apelin) and JAG1, to regenerate pulmonary microvessels and reverse pulmonary hypertension. In PAECs from BMPR2 mutant PAH patients, pharmacological induction of p53 and PPARγ-p53 genes repaired damaged DNA utilizing genes from the nucleotide excision repair pathway without provoking PAEC apoptosis. CONCLUSIONS: We identified a novel therapeutic strategy that activates a vasculoprotective gene regulation program in PAECs downstream of dysfunctional BMPR2 to rehabilitate PAH PAECs, regenerate pulmonary microvessels, and reverse disease. Our studies pave the way for p53-based vasculoregenerative therapies for PAH by extending the therapeutic focus to PAEC dysfunction and to DNA damage associated with PAH progression.
Authors: Christopher J Rhodes; Hogune Im; Aiqin Cao; Jan K Hennigs; Lingli Wang; Silin Sa; Pin-I Chen; Nils P Nickel; Kazuya Miyagawa; Rachel K Hopper; Nancy F Tojais; Caiyun G Li; Mingxia Gu; Edda Spiekerkoetter; Zhaoying Xian; Rui Chen; Mingming Zhao; Mark Kaschwich; Patricia A Del Rosario; Daniel Bernstein; Roham T Zamanian; Joseph C Wu; Michael P Snyder; Marlene Rabinovitch Journal: Am J Respir Crit Care Med Date: 2015-08-01 Impact factor: 21.405
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Authors: Yingyao Zhou; Bin Zhou; Lars Pache; Max Chang; Alireza Hadj Khodabakhshi; Olga Tanaseichuk; Christopher Benner; Sumit K Chanda Journal: Nat Commun Date: 2019-04-03 Impact factor: 14.919
Authors: Kylie M Drake; Benjamin J Dunmore; Lauren N McNelly; Nicholas W Morrell; Micheala A Aldred Journal: Am J Respir Cell Mol Biol Date: 2013-09 Impact factor: 6.914