Abstract
Hyperoxia is implicated in the pathogenesis of bronchopulmonary dysplasia (BPD), a chronic lung disease of premature infants. High levels of supplemental oxygen can result in microvascular endothelial cell death and may disrupt lung development. In postnatal animals, hyperoxia inhibits expression of vascular endothelial growth factor (VEGF), which is required for normal vascular development. A potential mechanism of oxygen effects on VEGF is induction of p53, a transcription factor that represses VEGF gene transcription. Oxidant DNA damage can increase p53. We used a moderately premature baboon model of hyperoxia to examine p53, oxidant DNA damage, and VEGF expression. Fetal baboons delivered at 140 d of gestation (75% of term) were ventilated with 100% oxygen or oxygen as needed for 6 or 10 d. Lungs from the 10-d 100% oxygen animals had increased nuclear p53, compared with the oxygen as needed animals. The mechanism of increased p53 was probably related to oxidant DNA damage, which was documented by increased oxidized guanine. Dual fluorescent confocal microscopy found increased oxidized guanine in mitochondrial DNA of distal lung epithelial cells. Distal epithelial cell VEGF expression was decreased and p21, another downstream target of p53, was increased in the distal epithelium of the hyperoxic animals. These data show that p53 is induced in hyperoxic fetal lung epithelium and are consistent with p53 repression of VEGF expression in these cells. The findings suggest that oxidant DNA damage may be a mechanism of increased p53 in hyperoxic fetal lung.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- 8-oxoG:
-
8-hydroxy-2′:-deoxyguanosine
- ARDS:
-
acute respiratory distress syndrome
- BPD:
-
bronchopulmonary dysplasia
- HIF-1:
-
hypoxia inducible factor-1
- RPA:
-
ribonuclease protection assay
- VEGF:
-
vascular endothelial growth factor
- VEGFR1:
-
vascular endothelial growth factor receptor-1
References
Crapo JD 1986 Morphologic changes in pulmonary oxygen toxicity. Annu Rev Physiol 48: 721–731
Coalson J 2000 Pathology of chronic lung disease in early infancy. In: Bland R, Coalson JJ (eds) Chronic Lung Disease in Early Infancy. Marcel Dekker, New York, pp 85–124
Bhatt AJ, Pryhuber GS, Huyck H, Watkins RH, Metlay LA, Maniscalco WM 2001 Disrupted pulmonary vasculature and decreased vascular endothelial growth factor, Flt-1, and TIE-2 in human infants dying with bronchopulmonary dysplasia. Am J Respir Crit Care Med 164: 1971–1980
Coalson JJ, Winter VT, Siler-Khodr T, Yoder BA 1999 Neonatal chronic lung disease in extremely immature baboons. Am J Respir Crit Care Med 160: 1333–1346
Maniscalco WM, Watkins RH, Pryhuber GS, Bhatt A, Shea C, Huyck H 2002 Angiogenic factors and alveolar vasculature: development and alterations by injury in very premature baboons. Am J Physiol Lung Cell Mol Physiol 282: L811–L823
Maniscalco WM, Watkins RH, D'Angio CT, Ryan RM 1997 Hyperoxic injury decreases alveolar epithelial cell expression of vascular endothelial growth factor (VEGF) in neonatal rabbit lung. Am J Respir Cell Mol Biol 16: 557–567
Perkett EA, Klekamp JG 1998 Vascular endothelial growth factor expression is decreased in rat lung following exposure to 24 or 48 hours of hyperoxia: implications for endothelial cell survival. Chest 114: 52S–53S
Hosford GE, Olson DM 2003 Effects of hyperoxia on VEGF, its receptors, and HIF-2alpha in the newborn rat lung. Am J Physiol Lung Cell Mol Physiol 285: L161–L168
Thickett DR, Armstrong L, Millar AB 2002 A role for vascular endothelial growth factor in acute and resolving lung injury. Am J Respir Crit Care Med 166: 1332–1337
Tsokos M, Pufe T, Paulsen F, Anders S, Mentlein R 2003 Pulmonary expression of vascular endothelial growth factor in sepsis. Arch Pathol Lab Med 127: 331–335
Carmeliet P, Ferreira V, Breier G, Pollefeyt S, Kieckens L, Gertsenstein M, Fahrig M, Vandenhoeck A, Harpal K, Eberhardt C, Declercq C, Pawling J, Moons L, Collen D, Risau W, Nagy A 1996 Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380: 435–439
Bhatt AJ, Amin SB, Chess PR, Watkins RH, Maniscalco WM 2000 Expression of vascular endothelial growth factor and Flk-1 in developing and glucocorticoid-treated mouse lung. Pediatr Res 47: 606–613
Zeng X, Wert SE, Federici R, Peters KG, Whitsett JA 1998 VEGF enhances pulmonary vasculogenesis and disrupts lung morphogenesis in vivo. Dev Dyn 211: 215–227
Galambos C, Ng YS, Ali A, Noguchi A, Lovejoy S, D'Amore PA, DeMello DE 2002 Defective pulmonary development in the absence of heparin-binding vascular endothelial growth factor isoforms. Am J Respir Cell Mol Biol 27: 194–203
Alon T, Hemo I, Itin A, Pe'er J, Stone J, Keshet E 1995 Vascular endothelial growth factor acts as a survival factor for newly formed retinal vessels and has implications for retinopathy of prematurity. Nature Med 1: 1024–1028
Benjamin LE, Keshet E 1997 Conditional switching of vascular endothelial growth factor (VEGF) expression in tumors: induction of endothelial cell shedding and regression of hemangioblastoma-like vessels by VEGF withdrawal. Proc Natl Acad Sci U S A 94: 8761–8766
Forsythe JA, Jiang BH, Iyer NV, Agani F, Leung SW, Koos RD, Semenza GL 1996 Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol 16: 4604–4613
Tuder RM, Zhen L, Cho CY, Taraseviciene-Stewart L, Kasahara Y, Salvemini D, Voelkel NF, Flores SC 2003 Oxidative stress and apoptosis interact and cause emphysema due to vascular endothelial growth factor receptor blockade. Am J Respir Cell Mol Biol 29: 88–97
Auten RL, Whorton MH, Nicholas Mason S 2002 Blocking neutrophil influx reduces DNA damage in hyperoxia-exposed newborn rat lung. Am J Respir Cell Mol Biol 26: 391–397
Roper JM, Mazzatti DJ, Watkins RH, Maniscalco WM, Keng PC, O'Reilly MA 2004 In vivo exposure to hyperoxia induces DNA damage in a population of alveolar type II epithelial cells. Am J Physiol Lung Cell Mol Physiol 286: L1045–L1054
Bohr VA, Stevnsner T, de Souza-Pinto NC 2002 Mitochondrial DNA repair of oxidative damage in mammalian cells. Gene 286: 127–134
Abid MR, Tsai JC, Spokes KC, Deshpande SS, Irani K, Aird WC 2001 Vascular endothelial growth factor induces manganese-superoxide dismutase expression in endothelial cells by a Rac1-regulated NADPH oxidase-dependent mechanism. FASEB J 15: 2548–2550
Kroll J, Waltenberger J 1998 VEGF-A induces expression of eNOS and iNOS in endothelial cells via VEGF receptor-2 (KDR). Biochem Biophys Res Commun 252: 743–746
O'Reilly MA, Staversky RJ, Stripp BR, Finkelstein JN 1998 Exposure to hyperoxia induces p53 expression in mouse lung epithelium. Am J Respir Cell Mol Biol 18: 43–50
Pal S, Datta K, Mukhopadhyay D 2001 Central role of p53 on regulation of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) expression in mammary carcinoma. Cancer Res 61: 6952–6957
Riccioni T, Cirielli C, Wang X, Passaniti A, Capogrossi MC 1998 Adenovirus-mediated wild-type p53 overexpression inhibits endothelial cell differentiation in vitro and angiogenesis in vivo. Gene Ther 5: 747–754
Zhang L, Yu D, Hu M, Xiong S, Lang A, Ellis LM, Pollock RE 2000 Wild-type p53 suppresses angiogenesis in human leiomyosarcoma and synovial sarcoma by transcriptional suppression of vascular endothelial growth factor expression. Cancer Res 60: 3655–3661
Fujisawa T, Watanabe J, Kamata Y, Hamano M, Hata H, Kuramoto H 2003 Effect of p53 gene transfection on vascular endothelial growth factor expression in endometrial cancer cells. Exp Mol Pathol 74: 276–281
Ravi R, Mookerjee B, Bhujwalla ZM, Sutter CH, Artemov D, Zeng Q, Dillehay LE, Madan A, Semenza GL, Bedi A 2000 Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1alpha. Genes Dev 14: 34–44
Tebar M, Boex JJ, Ten Have-Opbroek AA 2001 Functional overexpression of wild-type p53 correlates with alveolar cell differentiation in the developing human lung. Anat Rec 263: 25–34
Rancourt RC, Keng PC, Helt CE, O'Reilly MA 2001 The role of p21(CIP1/WAF1) in growth of epithelial cells exposed to hyperoxia. Am J Physiol Lung Cell Mol Physiol 280: L617–626
Munshi N, Fernandis AZ, Cherla RP, Park IW, Ganju RK 2002 Lipopolysaccharide-induced apoptosis of endothelial cells and its inhibition by vascular endothelial growth factor. J Immunol 168: 5860–5866
Stromblad S, Becker JC, Yebra M, Brooks PC, Cheresh DA 1996 Suppression of p53 activity and p21WAF1/CIP1 expression by vascular cell integrin alphaVbeta3 during angiogenesis. J Clin Invest 98: 426–433
Coalson JJ, Kuehl TJ, Prihoda TJ, deLemos RA 1988 Diffuse alveolar damage in the evolution of bronchopulmonary dysplasia in the baboon. Pediatr Res 24: 357–366
Coalson JJ, Kuehl TJ, Escobedo MB, Hilliard JL, Smith F, Meredith K, Null DM Jr, Walsh W, Johnson D, Robotham JL 1982 A baboon model of bronchopulmonary dysplasia. II. Pathologic features. Exp Mol Pathol 37: 335–350
Delemos RA, Coalson JJ, Gerstmann DR, Kuehl TJ, Null DM Jr 1987 Oxygen toxicity in the premature baboon with hyaline membrane disease. Am Rev Respir Dis 136: 677–682
Watkins RH, D'Angio CT, Ryan RM, Patel A, Maniscalco WM 1999 Differential expression of VEGF mRNA splice variants in newborn and adult hyperoxic lung injury. Am J Physiol 276: L858–L867
O'Reilly MA, Staversky RJ, Watkins RH, Maniscalco WM 1998 Accumulation of p21(Cip1/WAF1) during hyperoxic lung injury in mice. Am J Respir Cell Mol Biol 19: 777–785
Bruce MC, Honaker CE 1998 Transcriptional regulation of tropoelastin expression in rat lung fibroblasts: changes with age and hyperoxia. Am J Physiol 274: L940–L950
Warner BB, Stuart LA, Papes RA, Wispe JR 1998 Functional and pathological effects of prolonged hyperoxia in neonatal mice. Am J Physiol 275: L110–L117
Compernolle V, Brusselmans K, Acker T, Hoet P, Tjwa M, Beck H, Plaisance S, Dor Y, Keshet E, Lupu F, Nemery B, Dewerchin M, Van Veldhoven P, Plate K, Moons L, Collen D, Carmeliet P 2002 Loss of HIF-2alpha and inhibition of VEGF impair fetal lung maturation, whereas treatment with VEGF prevents fatal respiratory distress in premature mice. Nat Med 8: 702–710
Lassus P, Turanlahti M, Heikkila P, Andersson LC, Nupponen I, Sarnesto A, Andersson S 2001 Pulmonary vascular endothelial growth factor and Flt-1 in fetuses, in acute and chronic lung disease, and in persistent pulmonary hypertension of the newborn. Am J Respir Crit Care Med 164: 1981–1987
Jakkula M, Le Cras TD, Gebb S, Hirth KP, Tuder RM, Voelkel NF, Abman SH 2000 Inhibition of angiogenesis decreases alveolarization in the developing rat lung. Am J Physiol Lung Cell Mol Physiol 279: L600–L607
Gerber HP, McMurtrey A, Kowalski J, Yan M, Keyt BA, Dixit V, Ferrara N 1998 Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3′-kinase/Akt signal transduction pathway. Requirement for Flk-1/KDR activation. J Biol Chem 273: 30336–30343
Carmeliet P, Dor Y, Herbert JM, Fukumura D, Brusselmans K, Dewerchin M, Neeman M, Bono F, Abramovitch R, Maxwell P, Koch CJ, Ratcliffe P, Moons L, Jain RK, Collen D, Keshert E, Keshet E 1998 Role of HIF-1alpha in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis. Nature 394: 485–490
Fontanini G, Boldrini L, Vignati S, Chine S, Basolo F, Silvestri V, Lucchi M, Mussi A, Angeletti CA, Bevilacqua G 1998 Bcl2 and p53 regulate vascular endothelial growth factor (VEGF)-mediated angiogenesis in non-small cell lung carcinoma. Eur J Cancer 34: 718–723
Tai YT, Podar K, Gupta D, Lin B, Young G, Akiyama M, Anderson KC 2002 CD40 activation induces p53-dependent vascular endothelial growth factor secretion in human multiple myeloma cells. Blood 99: 1419–1427
Kosmidou I, Xagorari A, Roussos C, Papapetropoulos A 2001 Reactive oxygen species stimulate VEGF production from C(2)C(12) skeletal myotubes through a PI3K/Akt pathway. Am J Physiol Lung Cell Mol Physiol 280: L585–L592
Dulic V, Kaufmann WK, Wilson SJ, Tlsty TD, Lees E, Harper JW, Elledge SJ, Reed SI 1994 p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest. Cell 76: 1013–1023
Teruyama K, Abe M, Nakano T, Iwasaka-Yagi C, Takahashi S, Yamada S, Sato Y 2001 Role of transcription factor Ets-1 in the apoptosis of human vascular endothelial cells. J Cell Physiol 188: 243–252
Acknowledgements
The authors thank Colleen Shea for excellent technical assistance.
Author information
Authors and Affiliations
Corresponding author
Additional information
This work was supported by NHLBI grants HL63400 (W.M.M.) and HL67392 (M.A.O.).
Rights and permissions
About this article
Cite this article
Maniscalco, W., Watkins, R., Roper, J. et al. Hyperoxic Ventilated Premature Baboons Have Increased p53, Oxidant DNA Damage and Decreased VEGF Expression. Pediatr Res 58, 549–556 (2005). https://doi.org/10.1203/01.pdr.0000176923.79584.f7
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/01.pdr.0000176923.79584.f7
This article is cited by
-
Caffeine is associated with improved alveolarization and angiogenesis in male mice following hyperoxia induced lung injury
BMC Pulmonary Medicine (2019)
-
Early injury of the neonatal lung contributes to premature lung aging: a hypothesis
Molecular and Cellular Pediatrics (2016)
-
Respiratory consequences of prematurity: evolution of a diagnosis and development of a comprehensive approach
Journal of Perinatology (2015)
-
Oxygen differentially affects the hox proteins Hoxb5 and Hoxa5 altering airway branching and lung vascular formation
Journal of Cell Communication and Signaling (2014)
