Endothelial cell processes: proliferation

Transcription

1 Exploring the Link Between Malignancy and Pulmonary Arterial Hypertension The "cancer paradigm" in pulmonary hypertension Rubin M. Tuder, M. D. Program in Translational Lung Program Division of Pulmonary Sciences and Critical Care Medicine, Department of Medicine University of Colorado, Denver School of Medicine Pulmonary Vascular Research Institute 2nd International Conference Neonatal and Childhood Pulmonary Vascular Disease, UCSF 29 Dysregulated Angiogenesis PH between inflammation and cancer Voelkel et al. Chest 21 Metabolic Shift Endothelial cell processes: proliferation Early Lesions: Plexiform Lesions: a b Concentric Lesions: Genetic Selection TGF-beta Cool et al. AJP, 1999 Abnormal growth factor release MIB-1 1

2 Can EC pathology discriminate between mild vs severe pulmonary hypertension? Plexiform Lesion Concentric lesion Angiomatoid Lesion Mild/Moderate NO NO NO Severe YES All forms YES All forms YES All forms Endothelial cell dysfunction in severe pulmonary hypertension Drug induced: Phen-fen Aminorex Idiopathic Collagen Vascular Diseases: CREST, Lupus Erythematosus Familial Liver cirrhosis Endothelial cell dysfunction Schistosomiasis Secondary (2ry) PH HIV-related High left-to-right heart malformations Three-dimensional organization of intima and plexiform lesions: 1-15 um 5-7 cm 3 /1 sections/29 segments (7,) Plexiform Intima fibrosis Plexiform lesions Intima lesions Where do pulmonary vascular lesions occur? Number of pulmonary Arteries (17 orders) 1 million Elastic (orders 17-1): 3, Muscular (orders 9-5): 8, Precapillary (orders 4-1; 25 um): 7 million Yaginuma, G-Y et al. Thorax 45:586, 199 2

3 Clonal Growth of Endothelial cells in Random X-chromosome inactivation Monoclonal expansion in PPH B Paternal Maternal Endothelial cells Lee et al: JCI,98 Tuder et al, AJRCCM, 99 Gene silencing Germline/Somatic mutations DNA Pol δ hmlh hmsh2 WT Translational STOP -1-2 gaagg NNNNNNNNNNN gcctg hmsh2 Intron 5 (A 26 ) TGF-β RII Exon 3 (A 1 ) BAX Exon 3 (G 8 ) dysregulation Normal EC Conceptual framework for severe PAH: selection of apoptosis resistance Trigger agent Dysfunctional EC NO-PGI 2, serotonin EC Selection (action of growth factors/genes/virus) Apoptosis resistance PAP Shear Stress Viral factors Drugs Mutations Hypoxia/HIF EC death (?apoptosis) Clonal Growth Neoplastic-like Tuder et al. Eur Resp J, 21 Approach to Inhibit KDR: SU 5416 (Sugen) KDR SU 5416 SU 5416: Highly lipophilic kinase inhibitor Group of 3-substituted indolin-2- ones Localizes to the ATP binding site of RTK IC5=1.4 um (2uM for PDGFr and 1 um for EGFr) Used for anti-angiogenic therapies for Kaposi s Sarcoma and Renal cell Carcinoma 3

4 How to best model RV failure: SU5416+CH rat model SPH Time: Unknown Time: 3 weeks mpap 5-75 mmhg mpap mmhg H U M A N D R A T Taraseviciene- Stewart et al, Faseb J 21 Role of EC APOPTOSIS in the genesis of severe PH SU5416+CH and MCT Models SU5416 causes increased EC death in chronically SU5416+CH hypoxic rats Chronic hypoxia Chronic hypoxia + SU5416 Active Caspase 3/Vessel CH CH+SU5416 D7 D14 D21 D42 Stewart et al. Faseb Journal, 21 Activated caspase 3 % caspase positive/perimeter Monocrotaline p<.1 p<.1 Con D14 D21 D3 Zaiman A et al. AJRCCM, 27 PAP (mmhg) Role of EC APOPTOSIS in the genesis of severe PH SU5416+CH and Monocrotaline Models N=12 CH CH+SU5416 Z-Asp+CH+SU5416 Control SU5416+CH N=14 Chronic Hypoxia N=5 SU5416 Z -Asp -CH 2 Stewart L. Faseb Journal 21 RSVP mmhg Monocrotaline P <.3 P < 5 Control MCT MCT +Z-Asp Duncan Stewart Caspase inhibition prevents severe pulmonary hypertension and endothelial cell proliferation induced by chronic hypoxia+su5416 Factor VIII SMA Activated caspase 3 4

5 BMPs Inhibits Apoptosis in human PAEC post serum withdrawal Serum withdrawal % of Annexin V positive cells FCS * SF BMP-2 BMP-7 Duncan Stewart, Circ Res Normal EC Conceptual framework for severe PAH: selection of apoptosis resistance Trigger agent Dysfunctional EC NO-PGI 2, serotonin EC Selection (action of growth factors/genes/virus) Apoptosis resistance PAP Shear Stress Viral factors Drugs Mutations Hypoxia/HIF EC death (?apoptosis) Clonal Growth Neoplastic-like Tuder et al. Eur Resp J, 21 Hypoxia facilitates genetic instability (if apoptosis resistant) Genome changes Clonal heterogeneity Clonal selection Apoptosis resistance Tumor aggressiveness Tumor progression Normoxia Angiogenic switch: the HIF/VEGF paradigm HIF-PAH :HIF 1α and 1β-PAH VEGF-PAH Proteome changes Protein degradation po2 (mmhg) Vaupel P. The Oncologist, 26 Semenza G. Nature Cancer Rev, 3 5

6 Hyperproliferative apoptosis-resistant endothelial cells in idiopathic pulmonary arterial hypertension Increased proliferation Masri et al, Am J Physiol, 27 Decreased apoptosis Endothelial cell energetic switch: the HIF/VEGF paradigm in cultured human cells Increased angiogenic tube formation Control Fijalkowska et al. Submitted. 29 Ctl -ECs -ECs energetic switch: Decreased mitochondria PH Endothelial Cells: Shift to a Glycolytic Pathway Standardized uptake values (SUV) of FDG-PET +NO Xu et al. PNAS, 27 Fijalkowska, Xu et al, Submitted. 29 Xu W et al. Alterations of Cellular Bioenergetics in Pulmonary Artery Endothelial Cells. PNAS 27 6

7 PH Endothelial Cells: Shift to a Glycolytic Pathway HIF-1a Normal -ECs energetic switch: decreased mitochondria MnSOD NO HIF-1a Oxidative stress, Gene microarray, Models Expression studies In vivo imagining HIF-1α target: CAIX Carbonic anhydrase IX RCC: +ctl Resistance to apoptosis Decreased mitochondria: Energetic Shift Somatic Mutations: clonal expansion Growth factors Excessive cell growth Fijalkowska, Xu et al, Submitted. 29 How to best model RV failure: SU5416+CH rat model SPH Time: Unknown Time: 3 weeks mpap 5-75 mmhg mpap mmhg EVIDENCE OF HYPOXIA IN SU5416+CH (3 weeks) SU+CH CH+Vehicle Hydroxyprobe H U M A N D R A T v v v Taraseviciene- Stewart et al, Faseb J 21 7

8 2-metoxy-estrodial (2ME) in PH 2-metoxy-estrodial (2ME) in PH: Prevention SU5416+CH: Panzem SU5416+CH: Vehicle NFkB, Stat3,HIF-1α Teresa LaVallee EntreMed PANZEM: Treatment SU5416+CH: Panzem SU5416+CH: Vehicle Conceptual framework for severe PAH: selection of apoptosis resistance EC Selection (action of growth factors/genes/virus) Apoptosis resistance Clonal Growth Normal EC Dysfunctional EC NO-PGI 2, serotonin EC death (?apoptosis) Treatment: days Trigger agent PAP Shear Stress Viral factors Drugs Neoplastic-like Tuder et al. Eur Resp J, 21 8

9 PH Endothelial Cells: Shift to a Glycolytic Pathway HIF-1a targets Acknowledgments: Hopkins Lab: Iwona Fijalkowska, Lijie Zhen, and Amy Richter Hopkins: Ari Zaiman, and Gregg Semenza Univ. Colorado: Laima Stewart VCU: Norbert Voelkel Cleveland Clinic: Serpil Erzurum, Weiling Xu, Suzy Comhair, Raed Dweik, Kewal Asosingh Pelicano et al. Oncogene 95 EntreMed: Teresa LaValle p16 beta-catenin MDR survivin MDR EGFR-3 9