Research Article

Regulation of mitochondrial fragmentation in microvascular endothelial cells isolated from the SU5416/hypoxia model of pulmonary arterial hypertension

Published Online:https://doi.org/10.1152/ajplung.00396.2018

Pulmonary arterial hypertension (PAH) is a morbid disease characterized by progressive right ventricle (RV) failure due to elevated pulmonary artery pressures (PAP). In PAH, histologically complex vaso-occlusive lesions in the pulmonary vasculature contribute to elevated PAP. However, the mechanisms underlying dysfunction of the microvascular endothelial cells (MVECs) that comprise a significant portion of these lesions are not well understood. We recently showed that MVECs isolated from the Sugen/hypoxia (SuHx) rat experimental model of PAH (SuHx-MVECs) exhibit increases in migration/proliferation, mitochondrial reactive oxygen species (ROS; mtROS) production, intracellular calcium levels ([Ca2+]i), and mitochondrial fragmentation. Furthermore, quenching mtROS with the targeted antioxidant MitoQ attenuated basal [Ca2+]i, migration and proliferation; however, whether increased mtROS-induced [Ca2+]i entry affected mitochondrial morphology was not clear. In this study, we sought to better understand the relationship between increased ROS, [Ca2+]i, and mitochondrial morphology in SuHx-MVECs. We measured changes in mitochondrial morphology at baseline and following inhibition of mtROS, with the targeted antioxidant MitoQ, or transient receptor potential vanilloid-4 (TRPV4) channels, which we previously showed were responsible for mtROS-induced increases in [Ca2+]i in SuHx-MVECs. Quenching mtROS or inhibiting TRPV4 attenuated fragmentation in SuHx-MVECs. Conversely, inducing mtROS production in MVECs from normoxic rats (N-MVECs) increased fragmentation. Ca2+ entry induced by the TRPV4 agonist GSK1017920A was significantly increased in SuHx-MVECs and was attenuated with MitoQ treatment, indicating that mtROS contributes to increased TRPV4 activity in SuHx-MVECs. Basal and maximal respiration were depressed in SuHx-MVECs, and inhibiting mtROS, but not TRPV4, improved respiration in these cells. Collectively, our data show that, in SuHx-MVECs, mtROS production promotes TRPV4-mediated increases in [Ca2+]i, mitochondrial fission, and decreased mitochondrial respiration. These results suggest an important role for mtROS in driving MVEC dysfunction in PAH.

REFERENCES

  • 1. Adapala RK, Talasila PK, Bratz IN, Zhang DX, Suzuki M, Meszaros JG, Thodeti CK. PKCα mediates acetylcholine-induced activation of TRPV4-dependent calcium influx in endothelial cells. Am J Physiol Heart Circ Physiol 301: H757–H765, 2011. doi:10.1152/ajpheart.00142.2011.
    LinkISIGoogle Scholar
  • 2. Aggarwal S, Gross CM, Sharma S, Fineman JR, Black SM. Reactive oxygen species in pulmonary vascular remodeling. Compr Physiol 3: 1011–1034, 2013. doi:10.1002/cphy.c120024.
    CrossrefPubMedISIGoogle Scholar
  • 3. Al-Mehdi AB, Pastukh VM, Swiger BM, Reed DJ, Patel MR, Bardwell GC, Pastukh VV, Alexeyev MF, Gillespie MN. Perinuclear mitochondrial clustering creates an oxidant-rich nuclear domain required for hypoxia-induced transcription. Sci Signal 5: ra47, 2012. doi:10.1126/scisignal.2002712.
    CrossrefPubMedISIGoogle Scholar
  • 4. Alvarez DF, King JA, Weber D, Addison E, Liedtke W, Townsley MI. Transient receptor potential vanilloid 4-mediated disruption of the alveolar septal barrier: a novel mechanism of acute lung injury. Circ Res 99: 988–995, 2006. doi:10.1161/01.RES.0000247065.11756.19.
    CrossrefPubMedISIGoogle Scholar
  • 5. Archer SL. Mitochondrial dynamics—mitochondrial fission and fusion in human diseases. N Engl J Med 369: 2236–2251, 2013. doi:10.1056/NEJMra1215233.
    CrossrefPubMedISIGoogle Scholar
  • 6. Archer SL, Gomberg-Maitland M, Maitland ML, Rich S, Garcia JG, Weir EK. Mitochondrial metabolism, redox signaling, and fusion: a mitochondria-ROS-HIF-1α-Kv1.5 O2-sensing pathway at the intersection of pulmonary hypertension and cancer. Am J Physiol Heart Circ Physiol 294: H570–H578, 2008. doi:10.1152/ajpheart.01324.2007.
    LinkISIGoogle Scholar
  • 7. Archer SL, Huang J, Henry T, Peterson D, Weir EK. A redox-based O2 sensor in rat pulmonary vasculature. Circ Res 73: 1100–1112, 1993. doi:10.1161/01.RES.73.6.1100.
    CrossrefPubMedISIGoogle Scholar
  • 8. Bagur R, Hajnóczky G. Intracellular Ca2+ sensing: its role in calcium homeostasis and signaling. Mol Cell 66: 780–788, 2017. doi:10.1016/j.molcel.2017.05.028.
    CrossrefPubMedISIGoogle Scholar
  • 9. Balakrishna S, Song W, Achanta S, Doran SF, Liu B, Kaelberer MM, Yu Z, Sui A, Cheung M, Leishman E, Eidam HS, Ye G, Willette RN, Thorneloe KS, Bradshaw HB, Matalon S, Jordt SE. TRPV4 inhibition counteracts edema and inflammation and improves pulmonary function and oxygen saturation in chemically induced acute lung injury. Am J Physiol Lung Cell Mol Physiol 307: L158–L172, 2014. doi:10.1152/ajplung.00065.2014.
    LinkISIGoogle Scholar
  • 10. Bell EL, Klimova TA, Eisenbart J, Moraes CT, Murphy MP, Budinger GR, Chandel NS. The Qo site of the mitochondrial complex III is required for the transduction of hypoxic signaling via reactive oxygen species production. J Cell Biol 177: 1029–1036, 2007. doi:10.1083/jcb.200609074.
    CrossrefPubMedISIGoogle Scholar
  • 11. Bonnet S, Michelakis ED, Porter CJ, Andrade-Navarro MA, Thébaud B, Bonnet S, Haromy A, Harry G, Moudgil R, McMurtry MS, Weir EK, Archer SL. An abnormal mitochondrial-hypoxia inducible factor-1alpha-Kv channel pathway disrupts oxygen sensing and triggers pulmonary arterial hypertension in fawn hooded rats: similarities to human pulmonary arterial hypertension. Circulation 113: 2630–2641, 2006. doi:10.1161/CIRCULATIONAHA.105.609008.
    CrossrefPubMedISIGoogle Scholar
  • 12. Bowers R, Cool C, Murphy RC, Tuder RM, Hopken MW, Flores SC, Voelkel NF. Oxidative stress in severe pulmonary hypertension. Am J Respir Crit Care Med 169: 764–769, 2004. doi:10.1164/rccm.200301-147OC.
    CrossrefPubMedISIGoogle Scholar
  • 13. Cereghetti GM, Stangherlin A, Martins de Brito O, Chang CR, Blackstone C, Bernardi P, Scorrano L. Dephosphorylation by calcineurin regulates translocation of Drp1 to mitochondria. Proc Natl Acad Sci USA 105: 15803–15808, 2008. doi:10.1073/pnas.0808249105.
    CrossrefPubMedISIGoogle Scholar
  • 14. Cho B, Choi SY, Cho HM, Kim HJ, Sun W. Physiological and pathological significance of dynamin-related protein 1 (drp1)-dependent mitochondrial fission in the nervous system. Exp Neurobiol 22: 149–157, 2013. doi:10.5607/en.2013.22.3.149.
    CrossrefPubMedGoogle Scholar
  • 15. Coelho-Santos V, Socodato R, Portugal C, Leitão RA, Rito M, Barbosa M, Couraud PO, Romero IA, Weksler B, Minshall RD, Fontes-Ribeiro C, Summavielle T, Relvas JB, Silva AP. Methylphenidate-triggered ROS generation promotes caveolae-mediated transcytosis via Rac1 signaling and c-Src-dependent caveolin-1 phosphorylation in human brain endothelial cells. Cell Mol Life Sci 73: 4701–4716, 2016. doi:10.1007/s00018-016-2301-3.
    CrossrefPubMedISIGoogle Scholar
  • 16. Contreras L, Drago I, Zampese E, Pozzan T. Mitochondria: the calcium connection. Biochim Biophys Acta 1797: 607–618, 2010. doi:10.1016/j.bbabio.2010.05.005.
    CrossrefPubMedISIGoogle Scholar
  • 17. Diebold LP, Gil HJ, Gao P, Martinez CA, Weinberg SE, Chandel NS. Mitochondrial complex III is necessary for endothelial cell proliferation during angiogenesis. Nat Metab 1: 158–171, 2019. doi:10.1038/s42255-018-0011-x.
    CrossrefPubMedGoogle Scholar
  • 18. Duong HT, Comhair SA, Aldred MA, Mavrakis L, Savasky BM, Erzurum SC, Asosingh K. Pulmonary artery endothelium resident endothelial colony-forming cells in pulmonary arterial hypertension. Pulm Circ 1: 475–486, 2011. doi:10.4103/2045-8932.93547.
    CrossrefPubMedGoogle Scholar
  • 19. Evrard SM, Lecce L, Michelis KC, Nomura-Kitabayashi A, Pandey G, Purushothaman KR, d’Escamard V, Li JR, Hadri L, Fujitani K, Moreno PR, Benard L, Rimmele P, Cohain A, Mecham B, Randolph GJ, Nabel EG, Hajjar R, Fuster V, Boehm M, Kovacic JC. Endothelial to mesenchymal transition is common in atherosclerotic lesions and is associated with plaque instability. Nat Commun 7: 11853, 2016. [Erratum in Nat Commun 8: 14710, 2017.] doi:10.1038/ncomms11853.
    CrossrefPubMedISIGoogle Scholar
  • 20. Fiorio Pla A, Ong HL, Cheng KT, Brossa A, Bussolati B, Lockwich T, Paria B, Munaron L, Ambudkar IS. TRPV4 mediates tumor-derived endothelial cell migration via arachidonic acid-activated actin remodeling. Oncogene 31: 200–212, 2012. doi:10.1038/onc.2011.231.
    CrossrefPubMedISIGoogle Scholar
  • 21. Gandre-Babbe S, van der Bliek AM. The novel tail-anchored membrane protein Mff controls mitochondrial and peroxisomal fission in mammalian cells. Mol Biol Cell 19: 2402–2412, 2008. doi:10.1091/mbc.e07-12-1287.
    CrossrefPubMedISIGoogle Scholar
  • 22. Giedt RJ, Yang C, Zweier JL, Matzavinos A, Alevriadou BR. Mitochondrial fission in endothelial cells after simulated ischemia/reperfusion: role of nitric oxide and reactive oxygen species. Free Radic Biol Med 52: 348–356, 2012. doi:10.1016/j.freeradbiomed.2011.10.491.
    CrossrefPubMedISIGoogle Scholar
  • 23. Goedicke-Fritz S, Kaistha A, Kacik M, Markert S, Hofmeister A, Busch C, Bänfer S, Jacob R, Grgic I, Hoyer J. Evidence for functional and dynamic microcompartmentation of Cav-1/TRPV4/K(Ca) in caveolae of endothelial cells. Eur J Cell Biol 94: 391–400, 2015. doi:10.1016/j.ejcb.2015.06.002.
    CrossrefPubMedISIGoogle Scholar
  • 24. Goldenberg NM, Ravindran K, Kuebler WM. TRPV4: physiological role and therapeutic potential in respiratory diseases. Naunyn Schmiedebergs Arch Pharmacol 388: 421–436, 2015. doi:10.1007/s00210-014-1058-1.
    CrossrefPubMedISIGoogle Scholar
  • 25. Goldenberg NM, Wang L, Ranke H, Liedtke W, Tabuchi A, Kuebler WM. TRPV4 is required for hypoxic pulmonary vasoconstriction. Anesthesiology 122: 1338–1348, 2015. doi:10.1097/ALN.0000000000000647.
    CrossrefPubMedISIGoogle Scholar
  • 26. Guerra F, Guaragnella N, Arbini AA, Bucci C, Giannattasio S, Moro L. Mitochondrial dysfunction: a novel potential driver of epithelial-to-mesenchymal transition in cancer. Front Oncol 7: 295, 2017. doi:10.3389/fonc.2017.00295.
    CrossrefPubMedISIGoogle Scholar
  • 27. Gusarova GA, Trejo HE, Dada LA, Briva A, Welch LC, Hamanaka RB, Mutlu GM, Chandel NS, Prakriya M, Sznajder JI. Hypoxia leads to Na,K-ATPase downregulation via Ca2+ release-activated Ca2+ channels and AMPK activation. Mol Cell Biol 31: 3546–3556, 2011. doi:10.1128/MCB.05114-11.
    CrossrefPubMedISIGoogle Scholar
  • 28. Han XJ, Lu YF, Li SA, Kaitsuka T, Sato Y, Tomizawa K, Nairn AC, Takei K, Matsui H, Matsushita M. CaM kinase I alpha-induced phosphorylation of Drp1 regulates mitochondrial morphology. J Cell Biol 182: 573–585, 2008. doi:10.1083/jcb.200802164.
    CrossrefPubMedISIGoogle Scholar
  • 29. Huetsch JC, Jiang H, Larrain C, Shimoda LA. The Na+/H+ exchanger contributes to increased smooth muscle proliferation and migration in a rat model of pulmonary arterial hypertension. Physiol Rep 4: e12729, 2016. doi:10.14814/phy2.12729.
    CrossrefPubMedISIGoogle Scholar
  • 30. Humbert M, Montani D, Perros F, Dorfmüller P, Adnot S, Eddahibi S. Endothelial cell dysfunction and cross talk between endothelium and smooth muscle cells in pulmonary arterial hypertension. Vascul Pharmacol 49: 113–118, 2008. doi:10.1016/j.vph.2008.06.003.
    CrossrefPubMedISIGoogle Scholar
  • 31. Jeyaraju DV, Cisbani G, Pellegrini L. Calcium regulation of mitochondria motility and morphology. Biochim Biophys Acta 1787: 1363–1373, 2009. doi:10.1016/j.bbabio.2008.12.005.
    CrossrefPubMedISIGoogle Scholar
  • 32. Jezek J, Cooper KF, Strich R. Reactive oxygen species and mitochondrial dynamics: the yin and yang of mitochondrial dysfunction and cancer progression. Antioxidants (Basel) 7: E13, 2018. doi:10.3390/antiox7010013.
    CrossrefPubMedISIGoogle Scholar
  • 33. Lanna A, Dustin ML. Mitochondrial fusion fuels T cell memory. Cell Res 26: 969–970, 2016. doi:10.1038/cr.2016.94.
    CrossrefPubMedISIGoogle Scholar
  • 34. Le A, Lane AN, Hamaker M, Bose S, Gouw A, Barbi J, Tsukamoto T, Rojas CJ, Slusher BS, Zhang H, Zimmerman LJ, Liebler DC, Slebos RJ, Lorkiewicz PK, Higashi RM, Fan TW, Dang CV. Glucose-independent glutamine metabolism via TCA cycling for proliferation and survival in B cells. Cell Metab 15: 110–121, 2012. doi:10.1016/j.cmet.2011.12.009.
    CrossrefPubMedISIGoogle Scholar
  • 35. Mammoto T, Muyleart M, Konduri GG, Mammoto A. Twist1 in hypoxia-induced pulmonary hypertension through transforming growth factor-β-Smad signaling. Am J Respir Cell Mol Biol 58: 194–207, 2018. doi:10.1165/rcmb.2016-0323OC.
    CrossrefPubMedISIGoogle Scholar
  • 36. Mandegar M, Fung YC, Huang W, Remillard CV, Rubin LJ, Yuan JX. Cellular and molecular mechanisms of pulmonary vascular remodeling: role in the development of pulmonary hypertension. Microvasc Res 68: 75–103, 2004. doi:10.1016/j.mvr.2004.06.001.
    CrossrefPubMedISIGoogle Scholar
  • 37. Marsboom G, Toth PT, Ryan JJ, Hong Z, Wu X, Fang YH, Thenappan T, Piao L, Zhang HJ, Pogoriler J, Chen Y, Morrow E, Weir EK, Rehman J, Archer SL. Dynamin-related protein 1-mediated mitochondrial mitotic fission permits hyperproliferation of vascular smooth muscle cells and offers a novel therapeutic target in pulmonary hypertension. Circ Res 110: 1484–1497, 2012. doi:10.1161/CIRCRESAHA.111.263848.
    CrossrefPubMedISIGoogle Scholar
  • 38. McLelland GL, Goiran T, Yi W, Dorval G, Chen CX, Lauinger ND, Krahn AI, Valimehr S, Rakovic A, Rouiller I, Durcan TM, Trempe JF, Fon EA. Mfn2 ubiquitination by PINK1/parkin gates the p97-dependent release of ER from mitochondria to drive mitophagy. eLife 7: e32866, 2018. doi:10.7554/eLife.32866.
    CrossrefPubMedISIGoogle Scholar
  • 39. Oka M, Homma N, Taraseviciene-Stewart L, Morris KG, Kraskauskas D, Burns N, Voelkel NF, McMurtry IF. Rho kinase-mediated vasoconstriction is important in severe occlusive pulmonary arterial hypertension in rats. Circ Res 100: 923–929, 2007. doi:10.1161/01.RES.0000261658.12024.18.
    CrossrefPubMedISIGoogle Scholar
  • 40. Orlova DY, Zimmerman N, Meehan S, Meehan C, Waters J, Ghosn EE, Filatenkov A, Kolyagin GA, Gernez Y, Tsuda S, Moore W, Moss RB, Herzenberg LA, Walther G. Earth Mover’s Distance (EMD): a true metric for comparing biomarker expression levels in cell populations. PLoS One 11: e0151859, 2016. doi:10.1371/journal.pone.0151859.
    CrossrefPubMedISIGoogle Scholar
  • 41. Ouellet M, Guillebaud G, Gervais V, Lupien St-Pierre D, Germain M. A novel algorithm identifies stress-induced alterations in mitochondrial connectivity and inner membrane structure from confocal images. PLOS Comput Biol 13: e1005612, 2017. doi:10.1371/journal.pcbi.1005612.
    CrossrefPubMedISIGoogle Scholar
  • 42. Palmer CS, Osellame LD, Laine D, Koutsopoulos OS, Frazier AE, Ryan MT. MiD49 and MiD51, new components of the mitochondrial fission machinery. EMBO Rep 12: 565–573, 2011. doi:10.1038/embor.2011.54.
    CrossrefPubMedISIGoogle Scholar
  • 43. Peng T-I, Jou M-J. Oxidative stress caused by mitochondrial calcium overload. Ann N Y Acad Sci 1201: 183–188, 2010. doi:10.1111/j.1749-6632.2010.05634.x.
    CrossrefPubMedISIGoogle Scholar
  • 44. Picard M, Shirihai OS, Gentil BJ, Burelle Y. Mitochondrial morphology transitions and functions: implications for retrograde signaling? Am J Physiol Regul Integr Comp Physiol 304: R393–R406, 2013. doi:10.1152/ajpregu.00584.2012.
    LinkISIGoogle Scholar
  • 45. R Development Core Team. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing, 2018. https://www.r-project.org.
    Google Scholar
  • 46. Rabinovitch M. Molecular pathogenesis of pulmonary arterial hypertension. J Clin Invest 122: 4306–4313, 2012. doi:10.1172/JCI60658.
    CrossrefPubMedISIGoogle Scholar
  • 47. Rambold AS, Kostelecky B, Elia N, Lippincott-Schwartz J. Tubular network formation protects mitochondria from autophagosomal degradation during nutrient starvation. Proc Natl Acad Sci USA 108: 10190–10195, 2011. doi:10.1073/pnas.1107402108.
    CrossrefPubMedISIGoogle Scholar
  • 48. Rosca MG, Vazquez EJ, Chen Q, Kerner J, Kern TS, Hoppel CL. Oxidation of fatty acids is the source of increased mitochondrial reactive oxygen species production in kidney cortical tubules in early diabetes. Diabetes 61: 2074–2083, 2012. doi:10.2337/db11-1437.
    CrossrefPubMedISIGoogle Scholar
  • 49. Ryan J, Dasgupta A, Huston J, Chen KH, Archer SL. Mitochondrial dynamics in pulmonary arterial hypertension. J Mol Med (Berl) 93: 229–242, 2015. doi:10.1007/s00109-015-1263-5.
    CrossrefPubMedISIGoogle Scholar
  • 50. Sanders SP, Zweier JL, Kuppusamy P, Harrison SJ, Bassett DJ, Gabrielson EW, Sylvester JT. Hyperoxic sheep pulmonary microvascular endothelial cells generate free radicals via mitochondrial electron transport. J Clin Invest 91: 46–52, 1993. doi:10.1172/JCI116198.
    CrossrefPubMedISIGoogle Scholar
  • 51. Scheraga RG, Abraham S, Niese KA, Southern BD, Grove LM, Hite RD, McDonald C, Hamilton TA, Olman MA. TRPV4 mechanosensitive ion channel regulates lipopolysaccharide-stimulated macrophage phagocytosis. J Immunol 196: 428–436, 2016. doi:10.4049/jimmunol.1501688.
    CrossrefPubMedISIGoogle Scholar
  • 52. Seifert EL, Estey C, Xuan JY, Harper ME. Electron transport chain-dependent and -independent mechanisms of mitochondrial H2O2 emission during long-chain fatty acid oxidation. J Biol Chem 285: 5748–5758, 2010. doi:10.1074/jbc.M109.026203.
    CrossrefPubMedISIGoogle Scholar
  • 53. Suresh K, Servinsky L, Jiang H, Bigham Z, Yun X, Kliment C, Huetsch J, Damarla M, Shimoda LA. Reactive oxygen species induced Ca2+ influx via TRPV4 and microvascular endothelial dysfunction in the SU5416/hypoxia model of pulmonary arterial hypertension. Am J Physiol Lung Cell Mol Physiol 314: L893–L907, 2018. doi:10.1152/ajplung.00430.2017.
    LinkISIGoogle Scholar
  • 54. Suresh K, Servinsky L, Reyes J, Baksh S, Undem C, Caterina M, Pearse DB, Shimoda LA. Hydrogen peroxide-induced calcium influx in lung microvascular endothelial cells involves TRPV4. Am J Physiol Lung Cell Mol Physiol 309: L1467–L1477, 2015. doi:10.1152/ajplung.00275.2015.
    LinkISIGoogle Scholar
  • 55. Suresh K, Servinsky L, Reyes J, Undem C, Zaldumbide J, Rentsendorj O, Modekurty S, Dodd-O JM, Scott A, Pearse DB, Shimoda LA. CD36 mediates H2O2-induced calcium influx in lung microvascular endothelial cells. Am J Physiol Lung Cell Mol Physiol 312: L143–L153, 2017. doi:10.1152/ajplung.00361.2016.
    LinkISIGoogle Scholar
  • 57. Suzuki T, Carrier EJ, Talati MH, Rathinasabapathy A, Chen X, Nishimura R, Tada Y, Tatsumi K, West J. Isolation and characterization of endothelial-to-mesenchymal transition cells in pulmonary arterial hypertension. Am J Physiol Lung Cell Mol Physiol 314: L118–L126, 2018. doi:10.1152/ajplung.00296.2017.
    LinkISIGoogle Scholar
  • 58. Szabadkai G, Simoni AM, Bianchi K, De Stefani D, Leo S, Wieckowski MR, Rizzuto R. Mitochondrial dynamics and Ca2+ signaling. Biochim Biophys Acta 1763: 442–449, 2006. doi:10.1016/j.bbamcr.2006.04.002.
    CrossrefPubMedISIGoogle Scholar
  • 59. Szarka N, Pabbidi MR, Amrein K, Czeiter E, Berta G, Pohoczky K, Helyes Z, Ungvari Z, Koller A, Buki A, Toth P. Traumatic brain injury impairs myogenic constriction of cerebral arteries: role of mitochondria-derived H2O2 and TRPV4-dependent activation of BKCa channels. J Neurotrauma. In press. doi:10.1089/neu.2017.5056.
    CrossrefPubMedISIGoogle Scholar
  • 60. Tang H, Babicheva A, McDermott KM, Gu Y, Ayon RJ, Song S, Wang Z, Gupta A, Zhou T, Sun X, Dash S, Wang Z, Balistrieri A, Zheng Q, Cordery AG, Desai AA, Rischard F, Khalpey Z, Wang J, Black SM, Garcia JGN, Makino A, Yuan JX-J. Endothelial HIF-2α contributes to severe pulmonary hypertension due to endothelial-to-mesenchymal transition. Am J Physiol Lung Cell Mol Physiol 314: L256–L275, 2018. doi:10.1152/ajplung.00096.2017.
    LinkISIGoogle Scholar
  • 61. Tang H, Yamamura A, Yamamura H, Song S, Fraidenburg DR, Chen J, Gu Y, Pohl NM, Zhou T, Jiménez-Pérez L, Ayon RJ, Desai AA, Goltzman D, Rischard F, Khalpey Z, Black SM, Garcia JGN, Makino A, Yuan JXJ. Pathogenic role of calcium-sensing receptors in the development and progression of pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol 310: L846–L859, 2016. doi:10.1152/ajplung.00050.2016.
    LinkISIGoogle Scholar
  • 62. Taraseviciene-Stewart L, Kasahara Y, Alger L, Hirth P, Mc Mahon G, Waltenberger J, Voelkel NF, Tuder RM. Inhibition of the VEGF receptor 2 combined with chronic hypoxia causes cell death-dependent pulmonary endothelial cell proliferation and severe pulmonary hypertension. FASEB J 15: 427–438, 2001. doi:10.1096/fj.00-0343com.
    CrossrefPubMedISIGoogle Scholar
  • 63. Thorneloe KS, Cheung M, Bao W, Alsaid H, Lenhard S, Jian MY, Costell M, Maniscalco-Hauk K, Krawiec JA, Olzinski A, Gordon E, Lozinskaya I, Elefante L, Qin P, Matasic DS, James C, Tunstead J, Donovan B, Kallal L, Waszkiewicz A, Vaidya K, Davenport EA, Larkin J, Burgert M, Casillas LN, Marquis RW, Ye G, Eidam HS, Goodman KB, Toomey JR, Roethke TJ, Jucker BM, Schnackenberg CG, Townsley MI, Lepore JJ, Willette RN. An orally active TRPV4 channel blocker prevents and resolves pulmonary edema induced by heart failure. Sci Transl Med 4: 159ra148, 2012. doi:10.1126/scitranslmed.3004276.
    CrossrefPubMedISIGoogle Scholar
  • 64. Tian L, Neuber-Hess M, Mewburn J, Dasgupta A, Dunham-Snary K, Wu D, Chen KH, Hong Z, Sharp WW, Kutty S, Archer SL. Ischemia-induced Drp1 and Fis1-mediated mitochondrial fission and right ventricular dysfunction in pulmonary hypertension. J Mol Med (Berl) 95: 381–393, 2017. doi:10.1007/s00109-017-1522-8.
    CrossrefPubMedISIGoogle Scholar
  • 65. Touyz RM. Reactive oxygen species as mediators of calcium signaling by angiotensin II: implications in vascular physiology and pathophysiology. Antioxid Redox Signal 7: 1302–1314, 2005. doi:10.1089/ars.2005.7.1302.
    CrossrefPubMedISIGoogle Scholar
  • 66. Tsushima K, Bugger H, Wende AR, Soto J, Jenson GA, Tor AR, McGlauflin R, Kenny HC, Zhang Y, Souvenir R, Hu XX, Sloan CL, Pereira RO, Lira VA, Spitzer KW, Sharp TL, Shoghi KI, Sparagna GC, Rog-Zielinska EA, Kohl P, Khalimonchuk O, Schaffer JE, Abel ED. Mitochondrial reactive oxygen species in lipotoxic hearts induce post-translational modifications of AKAP121, DRP1, and OPA1 that promote mitochondrial fission. Circ Res 122: 58–73, 2018. doi:10.1161/CIRCRESAHA.117.311307.
    CrossrefPubMedISIGoogle Scholar
  • 67. Wakabayashi J, Zhang Z, Wakabayashi N, Tamura Y, Fukaya M, Kensler TW, Iijima M, Sesaki H. The dynamin-related GTPase Drp1 is required for embryonic and brain development in mice. J Cell Biol 186: 805–816, 2009. doi:10.1083/jcb.200903065.
    CrossrefPubMedISIGoogle Scholar
  • 68. Wang W, Wang Y, Long J, Wang J, Haudek SB, Overbeek P, Chang BH, Schumacker PT, Danesh FR. Mitochondrial fission triggered by hyperglycemia is mediated by ROCK1 activation in podocytes and endothelial cells. Cell Metab 15: 186–200, 2012. doi:10.1016/j.cmet.2012.01.009.
    CrossrefPubMedISIGoogle Scholar
  • 69. Wegierski T, Lewandrowski U, Müller B, Sickmann A, Walz G. Tyrosine phosphorylation modulates the activity of TRPV4 in response to defined stimuli. J Biol Chem 284: 2923–2933, 2009. doi:10.1074/jbc.M805357200.
    CrossrefPubMedISIGoogle Scholar
  • 70. Weinberg F, Hamanaka R, Wheaton WW, Weinberg S, Joseph J, Lopez M, Kalyanaraman B, Mutlu GM, Budinger GR, Chandel NS. Mitochondrial metabolism and ROS generation are essential for Kras-mediated tumorigenicity. Proc Natl Acad Sci USA 107: 8788–8793, 2010. doi:10.1073/pnas.1003428107.
    CrossrefPubMedISIGoogle Scholar
  • 71. Xia Y, Fu Z, Hu J, Huang C, Paudel O, Cai S, Liedtke W, Sham JS. TRPV4 channel contributes to serotonin-induced pulmonary vasoconstriction and the enhanced vascular reactivity in chronic hypoxic pulmonary hypertension. Am J Physiol Cell Physiol 305: C704–C715, 2013. doi:10.1152/ajpcell.00099.2013.
    LinkISIGoogle Scholar
  • 72. Xie Q, Wu Q, Horbinski CM, Flavahan WA, Yang K, Zhou W, Dombrowski SM, Huang Z, Fang X, Shi Y, Ferguson AN, Kashatus DF, Bao S, Rich JN. Mitochondrial control by DRP1 in brain tumor initiating cells. Nat Neurosci 18: 501–510, 2015. doi:10.1038/nn.3960.
    CrossrefPubMedISIGoogle Scholar
  • 73. Xiong J, Kawagishi H, Yan Y, Liu J, Wells QS, Edmunds LR, Fergusson MM, Yu ZX, Rovira II, Brittain EL, Wolfgang MJ, Jurczak MJ, Fessel JP, Finkel T. A metabolic basis for endothelial-to-mesenchymal transition. Mol Cell 69: 689–698.e7, 2018. doi:10.1016/j.molcel.2018.01.010.
    CrossrefPubMedISIGoogle Scholar
  • 74. Xu H, Zhao H, Tian W, Yoshida K, Roullet JB, Cohen DM. Regulation of a transient receptor potential (TRP) channel by tyrosine phosphorylation. SRC family kinase-dependent tyrosine phosphorylation of TRPV4 on TYR-253 mediates its response to hypotonic stress. J Biol Chem 278: 11520–11527, 2003. doi:10.1074/jbc.M211061200.
    CrossrefPubMedISIGoogle Scholar
  • 75. Xu W, Erzurum SC. Endothelial cell energy metabolism, proliferation, and apoptosis in pulmonary hypertension. Compr Physiol 1: 357–372, 2011.
    PubMedISIGoogle Scholar
  • 76. Xu W, Kaneko FT, Zheng S, Comhair SAA, Janocha AJ, Goggans T, Thunnissen FBJM, Farver C, Hazen SL, Jennings C, Dweik RA, Arroliga AC, Erzurum SC. Increased arginase II and decreased NO synthesis in endothelial cells of patients with pulmonary arterial hypertension. FASEB J 18: 1746–1748, 2004. doi:10.1096/fj.04-2317fje.
    CrossrefPubMedISIGoogle Scholar
  • 77. Xu W, Koeck T, Lara AR, Neumann D, DiFilippo FP, Koo M, Janocha AJ, Masri FA, Arroliga AC, Jennings C, Dweik RA, Tuder RM, Stuehr DJ, Erzurum SC. Alterations of cellular bioenergetics in pulmonary artery endothelial cells. Proc Natl Acad Sci USA 104: 1342–1347, 2007. doi:10.1073/pnas.0605080104.
    CrossrefPubMedISIGoogle Scholar
  • 78. Yu T, Robotham JL, Yoon Y. Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology. Proc Natl Acad Sci USA 103: 2653–2658, 2006. doi:10.1073/pnas.0511154103.
    CrossrefPubMedISIGoogle Scholar
  • 79. Zecchin A, Kalucka J, Dubois C, Carmeliet P. How endothelial cells adapt their metabolism to form vessels in tumors. Front Immunol 8: 1750, 2017. doi:10.3389/fimmu.2017.01750.
    CrossrefPubMedISIGoogle Scholar