Pericytes from human non-small cell lung carcinomas: An attractive target for anti-angiogenic therapy Rebecca G. Bagley ⁎ , Cecile Rouleau, Sharon D. Morgenbesser, William Weber, Brian P. Cook, Srinivas Shankara, Stephen L. Madden, Beverly A. Teicher Genzyme Corporation, 5 Mountain Road, Framingham, MA 01701-9322, USA Received 28 October 2005; revised 21 February 2006; accepted 1 March 2006 Available online 19 April 2006 Abstract Anti-angiogenic strategies have largely focused on endothelial cells and progenitors. However, pericytes are also an important component of vasculature. Perivascular cells from normal tissues have been widely reported, yet have not been extensively studied from human tumors. We have investigated pericytes from tumors of patients with lung cancer, the leader of cancer-related deaths in both men and women. Antibodies and magnetic beads were used to isolate cells from non-small cell lung carcinomas (NSCLC). The morphology of the pericytes was distinct with multiple elongated cytoplasmic extensions. Molecular expression of angiogenic genes was quantified by RT- PCR. Flow cytometric analysis shows that NSCLC pericytes express antigens such NG2 and VEGFR1 and present the ganglioside 3G5. The value of pericytes as models of tumor vasculature was demonstrated in cell-culture-based angiogenesis assays such as tube formation and proliferation. Results show that pericytes from some NSCLC but not all were able to maintain tubes networks on Matrigel. Pericyte function can be influenced by angiogenic growth factors or anti-angiogenic agents. Pericytes displayed invasive action against NSCLC clusters in the absence of other cell types. Perivascular cells contribute to the progression of disease and are an attractive target for anti- angiogenic therapy. © 2006 Elsevier Inc. All rights reserved. Keywords: Pericytes; Non-small cell lung carcinoma; Adenocarcinoma; Squamous cell carcinoma; Angiogenesis; Vasculature; Assays; Models Introduction Lung cancer has recently received greater visibility in the field of cancer as the leader of cancer-related deaths for both men and women. The lack of effective treatments illustrates the need for future therapies against novel targets that have previously been overlooked. While anti-angiogenic strategies have supplemented approaches that heretofore have been directed towards cancer cells, the endothelium has been the focus of vascular destabilization. This report details the characteristics and function of perivascular cells or pericytes from malignant tissue. Pericytes are a vital structural component of vasculature and have been identified, isolated, and investigated from multiple tumors surgically resected from patients with NSCLC. Pericytes were first identified in the microvasculature in 1873 (Rouget, 1873). Since then, pericytes have been described as Rouget cells or myofibroblasts. Pericytes are recognized in tissue sections through immunohistochemical staining for desmin or alpha-smooth muscle actin (αSMA) (Nehls and Drenckhahn, 1993). Electron micrographs show that pericytes Microvascular Research 71 (2006) 163 – 174 www.elsevier.com/locate/ymvre Abbreviations: EC, endothelial cells; VEGF, vascular endothelial growth factor; VEGFR, vascular endothelial growth factor receptor; HMVEC, human microvascular endothelial cells; HDF, human dermal fibroblast; bFGF, basic fibroblast growth factor; MSC, mesenchymal stem cell; PDGF/PDGFβ, platelet- derived growth factor; PDGF-R, platelet-derived growth factor receptor; EGF, epidermal growth factor; ANG, angiopoietin; MMP, matrix metalloproteinase; HGF, hepatocellular growth factor; IL-8, interleukin 8; IGF-1, insulin-like growth factor-1; CD54/ICAM, intercellular adhesion molecule-1; αSMA, alpha-smooth muscle actin; CD106/VCAM, vascular cell adhesion molecule; TGF-β, transforming growth factor-beta; NG2, neurite growth proteoglycan2; FC, flow cytometry; SAGE, serial analysis of gene expression. ⁎ Corresponding author. Fax: +1 508 872 9080. E-mail address: Rebecca.Bagley@Genzyme.com (R.G. Bagley). 0026-2862/$ - see front matter © 2006 Elsevier Inc. All rights reserved. doi:10.1016/j.mvr.2006.03.002