title: Activation of the Wnt/planar cell polarity pathway is required for pericyte recruitment during pulmonary angiogenesis.
abstract: Pericytes are perivascular cells localized to capillaries that promote vessel maturation, and their absence can contribute to vessel loss. Whether impaired endothelial-pericyte interaction contributes to small vessel loss in pulmonary arterial hypertension (PAH) is unclear. Using 3G5-specific, immunoglobulin G-coated magnetic beads, we isolated pericytes from the lungs of healthy subjects and PAH patients, followed by lineage validation. PAH pericytes seeded with healthy pulmonary microvascular endothelial cells failed to associate with endothelial tubes, resulting in smaller vascular networks compared to those with healthy pericytes. After the demonstration of abnormal polarization toward endothelium via live-imaging and wound-healing studies, we screened PAH pericytes for abnormalities in the Wnt/planar cell polarity (PCP) pathway, which has been shown to regulate cell motility and polarity in the pulmonary vasculature. PAH pericytes had reduced expression of frizzled 7 (Fzd7) and cdc42, genes crucial for Wnt/PCP activation. With simultaneous knockdown of Fzd7 and cdc42 in healthy pericytes in vitro and in a murine model of angiogenesis, motility and polarization toward pulmonary microvascular endothelial cells were reduced, whereas with restoration of both genes in PAH pericytes, endothelial-pericyte association was improved, with larger vascular networks. These studies suggest that the motility and polarity of pericytes during pulmonary angiogenesis are regulated by Wnt/PCP activation, which can be targeted to prevent vessel loss in PAH.
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Stanford University, Stanford, California; Stanford Cardiovascular Institute, Stanford University, Stanford, California.
authors.forename: Ke
authors.initials: K
authors.lastname: Yuan
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Stanford University, Stanford, California; Stanford Cardiovascular Institute, Stanford University, Stanford, California.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Orcholski
authors.affiliation: Division of Endocrinology, Gerontology & Metabolism, Stanford University, Stanford, California.
authors.forename: Cristina
authors.initials: C
authors.lastname: Panaroni
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Stanford University, Stanford, California; Stanford Cardiovascular Institute, Stanford University, Stanford, California.
authors.forename: Eric M
authors.initials: EM
authors.lastname: Shuffle
authors.affiliation: Stanford Cardiovascular Institute, Stanford University, Stanford, California; VA Palo Alto Health Care System, Stanford University, Stanford, California; Department of Cardiothoracic Surgery, Stanford University, Stanford, California.
authors.forename: Ngan F
authors.initials: NF
authors.lastname: Huang
authors.affiliation: Stanford Cardiovascular Institute, Stanford University, Stanford, California; VA Palo Alto Health Care System, Stanford University, Stanford, California.
authors.forename: Xinguo
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Stanford Cardiovascular Institute, Stanford University, Stanford, California; VA Palo Alto Health Care System, Stanford University, Stanford, California.
authors.forename: Wen
authors.initials: W
authors.lastname: Tian
authors.affiliation: Department of Pathology, Vera Moulton Wall Center for Pulmonary Vascular Disease, Stanford University School of Medicine, Stanford, California.
authors.forename: Eszter K
authors.initials: EK
authors.lastname: Vladar
authors.affiliation: Department of Pediatrics, Stanford University School of Medicine, Stanford, California.
authors.forename: Lingli
authors.initials: L
authors.lastname: Wang
authors.affiliation: Stanford Cardiovascular Institute, Stanford University, Stanford, California; VA Palo Alto Health Care System, Stanford University, Stanford, California.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Nicolls
authors.affiliation: Division of Endocrinology, Gerontology & Metabolism, Stanford University, Stanford, California.
authors.forename: Joy Y
authors.initials: JY
authors.lastname: Wu
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Stanford University, Stanford, California; Stanford Cardiovascular Institute, Stanford University, Stanford, California. Electronic address: vdejesus@stanford.edu.
authors.forename: Vinicio A
authors.initials: VA
authors.lastname: de Jesus Perez
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D002465:Cell Movement; D016764:Cell Polarity; D002648:Child; D018920:Coculture Techniques; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000166:cytology; D005260:Female; D051157:Frizzled Receptors; D055785:Gene Knockdown Techniques; D006801:Humans; D006976:Hypertension, Pulmonary / Q000378:metabolism / Q000503:physiopathology*; D007074:Immunoglobulin G / Q000737:chemistry; D008168:Lung / Q000098:blood supply / Q000503:physiopathology*; D008280:Magnetics; D008297:Male; D051379:Mice; D016513:Mice, SCID; D008833:Microcirculation; D008875:Middle Aged; D009389:Neovascularization, Pathologic; D020286:Pericytes / Q000166:cytology*; D034741:RNA, Small Interfering / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction; D051153:Wnt Proteins / Q000378:metabolism*; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism
chemical_list: C581598:Cdc42 protein, mouse; D051157:Frizzled Receptors; C492041:Fzd7 protein, mouse; D007074:Immunoglobulin G; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled; D051153:Wnt Proteins; D020764:cdc42 GTP-Binding Protein