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Respir Res. 2018 Apr 19;19(1):67. doi: 10.1186/s12931-018-0768-1.

FK506-binding protein 10 (FKBP10) regulates lung fibroblast migration via collagen VI synthesis.

Author information

1
Comprehensive Pneumology Center, Ludwig-Maximilians-Universität and Helmholtz Zentrum Munich, Max-Lebsche-Platz 31, 81377, Munich, Germany.
2
Member of the German Center of Lung Research (DZL), Munich, Germany.
3
Asklepios Fachkliniken Munich-Gauting, Munich, Germany.
4
Thoraxchirurgisches Zentrum, Klinik für Allgemeine-, Viszeral-, Transplantations-, Gefäß- und Thoraxchirurgie, Klinikum Großhadern, Ludwig-Maximilians-Universität, Munich, Germany.
5
Medizinische Klinik und Poliklinik V, Klinikum der Ludwig-Maximilians-Universität, Munich, Germany.
6
Colorado Anschutz Medical Campus, Pulmonary and Critical Care Medicine University, Denver, Colorado, USA.
7
Comprehensive Pneumology Center, Ludwig-Maximilians-Universität and Helmholtz Zentrum Munich, Max-Lebsche-Platz 31, 81377, Munich, Germany. staab-weijnitz@helmholtz-muenchen.de.
8
Member of the German Center of Lung Research (DZL), Munich, Germany. staab-weijnitz@helmholtz-muenchen.de.

Abstract

BACKGROUND:

In idiopathic pulmonary fibrosis (IPF), fibroblasts gain a more migratory phenotype and excessively secrete extracellular matrix (ECM), ultimately leading to alveolar scarring and progressive dyspnea. Here, we analyzed the effects of deficiency of FK506-binding protein 10 (FKBP10), a potential IPF drug target, on primary human lung fibroblast (phLF) adhesion and migration.

METHODS:

Using siRNA, FKBP10 expression was inhibited in phLF in absence or presence of 2ng/ml transforming growth factor-β1 (TGF-β1) and 0.1mM 2-phosphoascorbate. Effects on cell adhesion and migration were monitored by an immunofluorescence (IF)-based attachment assay, a conventional scratch assay, and single cell tracking by time-lapse microscopy. Effects on expression of key players in adhesion dynamics and migration were analyzed by qPCR and Western Blot. Colocalization was evaluated by IF microscopy and by proximity ligation assays.

RESULTS:

FKBP10 knockdown significantly attenuated adhesion and migration of phLF. Expression of collagen VI was decreased, while expression of key components of the focal adhesion complex was mostly upregulated. The effects on migration were 2-phosphoascorbate-dependent, suggesting collagen synthesis as the underlying mechanism. FKBP10 colocalized with collagen VI and coating culture dishes with collagen VI, and to a lesser extent with collagen I, abolished the effect of FKBP10 deficiency on migration.

CONCLUSIONS:

These findings show, to our knowledge for the first time, that FKBP10 interacts with collagen VI and that deficiency of FKBP10 reduces phLF migration mainly by downregulation of collagen VI synthesis. The results strengthen FKBP10 as an important intracellular regulator of ECM remodeling and support the concept of FKBP10 as drug target in IPF.

KEYWORDS:

FKBP10; FKBP65; collagen VI; fibroblast; fibulin; focal adhesion; lung fibrosis; migration

PMID:
29673351
PMCID:
PMC5909279
DOI:
10.1186/s12931-018-0768-1
[Indexed for MEDLINE]
Free PMC Article

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