Effect of Prepro-Calcitonin Gene-Related Peptide–Expressing Endothelial Progenitor Cells on Pulmonary Hypertension Qiang Zhao, MD, Zixiong Liu, MD, Zhe.

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Effect of Prepro-Calcitonin Gene-Related Peptide–Expressing Endothelial Progenitor Cells on Pulmonary Hypertension Qiang Zhao, MD, Zixiong Liu, MD, Zhe Wang, MD, Cheng Yang, MD, Jun Liu, MD, Jun Lu, MD The Annals of Thoracic Surgery Volume 84, Issue 2, Pages 544-552 (August 2007) DOI: 10.1016/j.athoracsur.2007.03.067 Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Identification of endothelial progenitor cells derived from human peripheral blood mononuclear cells. Endothelial progenitor cells exhibited spindle-shaped or cobblestone-like morphology on day 7 (A). Adherent cells took up 1,1′-dioctadecyl-3,3,3,′3′-tetramethylindocarbocyanine–labeled acetylated low-density lipoprotein from the medium (B), stained positive for Ulex europaeus agglutinin-lectin (C), and exhibited double positivity by merging B and C in the same field (D). The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Influences of endothelial progenitor cell (EPC) transplantation or calcitonin gene-related peptide (CGRP) gene-transduced endothelial progenitor cell transplantation on total pulmonary vascular resistance (TPR; A) and mean pulmonary arterial pressure (mPAP; B) in rats after shunt operation. * indicates significantly different from control group; # indicates significantly different from shunt group; + indicates significantly different from shunt + EPC group. The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Time course of calcitonin gene-related peptide (CGRP) secretion into the culture medium from calcitonin gene-related peptide gene–transduced endothelial progenitor cells (EPC) and endothelial progenitor cells without transfection. The period of calcitonin gene-related peptide overproduction in transfected endothelial progenitor cells lasted for more than 3 weeks, with its peak appearing on day 5. The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Changes in lung calcitonin gene-related peptide (CGRP) concentrations in control rats, shunt rats, and shunt rats treated with endothelial progenitor cells (EPC) or calcitonin gene-related peptide gene–transduced endothelial progenitor cells. * indicates significantly different from control group; # indicates significantly different from shunt group. The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Distribution of intravenously administered endothelial progenitor cells in lung tissue. Green fluorescence protein–expressing human endothelial progenitor cells (green) were detected in rat pulmonary vascular beds (stained red by rhodamine-B isothiocyanate–conjugated anti-rat CD31). The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 Changes in small pulmonary artery wall thickness in control rats, shunt rats, and shunt rats treated with endothelial progenitor cells (EPC) or calcitonin gene-related peptide (CGRP) gene–transduced endothelial progenitor cells. * indicates significantly different from control group; # indicates significantly different from shunt group; + indicates significantly different from shunt + EPC group. The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 7 Representative ultrastructure of pulmonary arteries under a transmission electron microscope. (A) Control. (B) Shunt. (C) Shunt + endothelial progenitor cells. (D) Shunt + calcitonin gene-related peptide gene–transduced endothelial progenitor cells. (Scale bar = 2 μm.) The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 8 Kaplan–Meier survival chart of shunt rats, shunt rats treated with endothelial progenitor cells (EPC) or calcitonin gene-related peptide (CGRP) gene–transduced endothelial progenitor cells. Survival was observed from the time of cell injection. The survival of rats treated with calcitonin gene-related peptide gene–transduced endothelial progenitor cells is significantly greater than that of shunt rats, whereas there is no difference between shunt rats and shunt rats treated only with endothelial progenitor cells. The Annals of Thoracic Surgery 2007 84, 544-552DOI: (10.1016/j.athoracsur.2007.03.067) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions