title: Increased Wnt5a in squamous cell lung carcinoma inhibits endothelial cell motility.
abstract: Angiogenesis is important both in normal tissue function and disease and represents a key target in lung cancer (LC) therapy. Unfortunately, the two main subtypes of non-small-cell lung cancers (NSCLC) namely, adenocarcinoma (AC) and squamous cell carcinoma (SCC) respond differently to anti-angiogenic e.g. anti-vascular endothelial growth factor (VEGF)-A treatment with life-threatening side effects, often pulmonary hemorrhage in SCC. The mechanisms behind such adverse reactions are still largely unknown, although peroxisome proliferator activator receptor (PPAR) gamma as well as Wnt-s have been named as molecular regulators of the process. As the Wnt microenvironments in NSCLC subtypes are drastically different, we hypothesized that the particularly high levels of non-canonical Wnt5a in SCC might be responsible for alterations in blood vessel growth and result in serious adverse reactions.
  
  
  
  PPARgamma, VEGF-A, Wnt5a, miR-27b and miR-200b levels were determined in resected adenocarcinoma and squamous cell carcinoma samples by qRT-PCR and TaqMan microRNA assay. The role of PPARgamma in VEGF-A expression, and the role of Wnts in overall regulation was investigated using PPARgamma knock-out mice, cancer cell lines and fully human, in vitro 3 dimensional (3D), distal lung tissue aggregates. PPARgamma mRNA and protein levels were tested by qRT-PCR and immunohistochemistry, respectively. PPARgamma activity was measured by a PPRE reporter system. The tissue engineered lung tissues expressing basal level and lentivirally delivered VEGF-A were treated with recombinant Wnts, chemical Wnt pathway modifiers, and were subjected to PPARgamma agonist and antagonist treatment.
  
  
  
  PPARgamma down-regulation and VEGF-A up-regulation are characteristic to both AC and SCC. Increased VEGF-A levels are under direct control of PPARgamma. PPARgamma levels and activity, however, are under Wnt control. Imbalance of both canonical (in AC) and non-canonical (in SCC) Wnts leads to PPARgamma down-regulation. While canonical Wnts down-regulate PPARgamma directly, non-canonical Wnt5a increases miR27b that is known regulator of PPARgamma.
  
  
  
  During carcinogenesis the Wnt microenvironment alters, which can downregulate PPARgamma leading to increased VEGF-A expression. Differences in the Wnt microenvironment in AC and SCC of NSCLC lead to PPARgamma decrease via mechanisms that differentially alter endothelial cell motility and branching which in turn can influence therapeutic response.
authors.affiliation: Department of Pharmaceutical Biotechnology, School of Pharmacy, University of Pecs, 2 Rokus Str, Pecs, 7624, Hungary.|János Szentágothai Research Centre, University of Pécs, 20 Ifjúság Str, Pecs, 7622, Hungary.|Humeltis Ltd, János Szentágothai Research Center, University of Pécs, 20 Ifjúság Str, Pécs, 7622, Hungary.
authors.forename: J
authors.initials: J
authors.lastname: Rapp
authors.affiliation: Department of Pharmaceutical Biotechnology, School of Pharmacy, University of Pecs, 2 Rokus Str, Pecs, 7624, Hungary.|János Szentágothai Research Centre, University of Pécs, 20 Ifjúság Str, Pecs, 7622, Hungary.|Humeltis Ltd, János Szentágothai Research Center, University of Pécs, 20 Ifjúság Str, Pécs, 7622, Hungary.
authors.forename: E
authors.initials: E
authors.lastname: Kiss
authors.affiliation: Medical Microbiology and Immunity, University of Pécs, 12 Szigeti Str, Pécs, 7624, Hungary.|Humeltis Ltd, János Szentágothai Research Center, University of Pécs, 20 Ifjúság Str, Pécs, 7622, Hungary.
authors.forename: M
authors.initials: M
authors.lastname: Meggyes
authors.affiliation: Biophysics, University of Pécs, 12 Szigeti Str, Pécs, 7624, Hungary.|János Szentágothai Research Centre, University of Pécs, 20 Ifjúság Str, Pecs, 7622, Hungary.
authors.forename: E
authors.initials: E
authors.lastname: Szabo-Meleg
authors.affiliation: Department of Pharmaceutical Biotechnology, School of Pharmacy, University of Pecs, 2 Rokus Str, Pecs, 7624, Hungary.|János Szentágothai Research Centre, University of Pécs, 20 Ifjúság Str, Pecs, 7622, Hungary.|Humeltis Ltd, János Szentágothai Research Center, University of Pécs, 20 Ifjúság Str, Pécs, 7622, Hungary.
authors.forename: D
authors.initials: D
authors.lastname: Feller
authors.affiliation: Pathology, University of Pécs, 12 Szigeti Str, Pécs, 7624, Hungary.
authors.forename: G
authors.initials: G
authors.lastname: Smuk
authors.affiliation: Pathology, University of Pécs, 12 Szigeti Str, Pécs, 7624, Hungary.
authors.forename: T
authors.initials: T
authors.lastname: Laszlo
authors.affiliation: Internal Medicine, Pulmonology, University of Pécs, 2 Rakoczi Str, Pécs, 7623, Hungary.
authors.forename: V
authors.initials: V
authors.lastname: Sarosi
authors.affiliation: Operational Medicine, University of Pécs, 12 Szigeti Str, Pécs, 7624, Hungary.|Department of Surgery, Thoracic Surgery Unit, Petz A Hospital, 2-4 Vasvari Str, Győr, 9023, Hungary.
authors.forename: T F
authors.initials: TF
authors.lastname: Molnar
authors.affiliation: Department of Pharmaceutical Biotechnology, School of Pharmacy, University of Pecs, 2 Rokus Str, Pecs, 7624, Hungary.|János Szentágothai Research Centre, University of Pécs, 20 Ifjúság Str, Pecs, 7622, Hungary.
authors.forename: K
authors.initials: K
authors.lastname: Kvell
authors.affiliation: Department of Pharmaceutical Biotechnology, School of Pharmacy, University of Pecs, 2 Rokus Str, Pecs, 7624, Hungary. pongracz.e.judit@pte.hu.|János Szentágothai Research Centre, University of Pécs, 20 Ifjúság Str, Pecs, 7622, Hungary. pongracz.e.judit@pte.hu.|Humeltis Ltd, János Szentágothai Research Center, University of Pécs, 20 Ifjúság Str, Pécs, 7622, Hungary. pongracz.e.judit@pte.hu.
authors.forename: J E
authors.initials: JE
authors.lastname: Pongracz
mesh_terms: D000230:Adenocarcinoma / Q000098:blood supply / Q000378:metabolism / Q000473:pathology; D000818:Animals; D014408:Biomarkers, Tumor; D002289:Carcinoma, Non-Small-Cell Lung / Q000098:blood supply / Q000378:metabolism / Q000473:pathology; D002294:Carcinoma, Squamous Cell / Q000098:blood supply / Q000378:metabolism / Q000473:pathology*; D002465:Cell Movement; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008175:Lung Neoplasms / Q000098:blood supply / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000378:metabolism / Q000473:pathology; D047495:PPAR gamma / Q000502:physiology*; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D000071818:Wnt-5a Protein / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs; D047495:PPAR gamma; D042461:Vascular Endothelial Growth Factor A; D000071818:Wnt-5a Protein
keywords: Angiogenesis; Lung cancer; NSCLC; PPARgamma; Wnt