Impaired Angiogenesis in Aminopeptidase N-null Mice. Roberto Rangel, Ph.D. Arap & Pasqualini Laboratory
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1 Impaired Angiogenesis in Aminopeptidase N-null Mice Roberto Rangel, Ph.D. Arap & Pasqualini Laboratory
2 Profiling Cell Surface Markers: Aminopeptidase N (CD13) Tumor cell - endothelium cell adhesion and extravasation Peptides mimic ligands Peptides mimic surface molecules on endothelial cells Target Cell Cell - cell interactions anchorage-dependent cell growth Peptides mimic adhesion receptors on tumor cells
3 Organ Vascular Diversity Yields a ZIP Code System Kolonin et al. Curr. Opin. Chem. Biol. 21
4 Design & Construction of the APN-null Mouse Wild type allele 11 kb Targeting vector 3 probe IRES-LacZ PGK-Neo DTA Mutant allele 8 kb IRES-LacZ PGK-Neo 3 probe Kb +/+ +/- -/- Kb M +/+ +/- -/ Rangel et al. PNAS 27
5 Expression Analysis of APN-null Mice APN +/+ APN +/- APN -/- Kb 12. Brain Kidney M +/+ +/- -/- +/+ +/- -/- A B C Brain 1..5 D E F.1 Kidney G3PDH Rangel et al. PNAS 27
6 Immunohistochemical Analysis of APN & CD31 Pancreas Liver Spleen Ovary Intestine APN +/+ APN +/+ APN +/- APN -/- APN -/- CD13 CD31 Rangel et al. PNAS 27
7 Non-invasive Phenotypic Studies in APN-null Mice LOCOMOTOR ACTIVITY PAIN REFLEXES Percent DISTANCE IN CENTRAL REGION Seconds LATENCY TO ENTER CENTRAL REGION ACTIVITY CENTRAL REGION Percent Seconds THERMAL PLATE APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- MUSCLE GRIP STRENGTH BLOOD PRESSURE.125 FORELIMB.1 HINDLIMB 125 SYSTOLIC 6 PULSE Kilograms Kilograms mm Hg bpm APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- Rangel et al. PNAS 27
8 Comprehensive In-cage Monitoring of APN-null Mice 1.2 RESPIRATORY EXCHANGE RATIO 3 O 2 UPTAKE 3 CO2 PRODUCTION.5 HEAT GENERATION 1.4 Arbitrary Units ml/kg/h 2 1 ml/kg/h 2 1 Kcal/Kg/h APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- Grams FOOD CONSUMPTION WATER CONSUMPTION 4 3 Mililiter 2 1 Seconds AMBULATION Seconds REARING ACTIVITY APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- APN +/+ APN -/- Rangel et al. PNAS 27
9 Failed Retinal Neovascularization in APN-null Mice in Hypoxia 75% O 2 Angiogenesis P1 P7 P12 P17 P21 Neovascular nuclei/section Blood Vessel Count APN -/- APN +/- APN +/+ Rangel et al. PNAS 27 APN +/+ APN +/- APN -/-
10 Impaired Angiogenesis Upon Growth Factor Stimulation in APN-null Mice 6 5 Hemoglobin mg/ml APN +/+ APN -/- APN +/+ APN -/- Rangel et al. PNAS 27
11 Growth Inhibition of Tumor Xenografts in APN-null Mice 35 3 Tumor volumen (mm3) APN +/+ APN -/- APN +/+ APN -/- Rangel et al. (Unpublished)
12 Conclusions 1. Despite the broad range of CD13 functions, APN-null mice exhibit no developmental, fertility, behavioral or physiological p abnormalities. 2. APN-deficient mice display a severely impaired dose-dependent dependent angiogenic response to oxygen-induced retinopathy and angiogenic factors in vivo. 3. APN-null mice may be useful reagents in other medical fields such as malignant, cardiovascular, immunological, i or infectious disease.
13 The People Principal Investigators Wadih Arap, M.D., Ph.D. Renata Pasqualini, Ph.D. Staff Faculty Emmanuel Dias, Ph.D. Ricardo Giordano, Ph.D. Erkki Koivunen,, Ph.D. Graduate & Medical Students Dawn Christianson, Ph.D. candidate Danielle Conley, Ph.D. student Katja Karjalainen,, M.Sc., Ph.D. student Alessandro Lee, M.Sc., Ph.D. student Michael Ozawa, M.D./Ph.D. candidate Tracey Smith, Ph.D. student Jessica Sun, M.D., Ph.D. student Postdoctoral Fellows Marina Cardó-Vila Vila,, Ph.D. Liliana Guzman-Rojas, Ph.D. Jeasun Kim, Ph.D. Jami Mandelin,, Ph.D. Diana Nunes,, Ph.D. Roberto Rangel, Ph.D. Masanori Sato, M.D. Glauco Souza, Ph.D. Fernanda Staquicini,, Ph.D. Wouter Driessen,, Ph.D. Research Technicians Natalie Hernandez, B.S. Connie Sun, M.Sc. Diana Quian,, B.S.
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