دورية أكاديمية
An integrated multiomic and quantitative label-free microscopy-based approach to study pro-fibrotic signalling in ex vivo human precision-cut lung slices.
| العنوان: | An integrated multiomic and quantitative label-free microscopy-based approach to study pro-fibrotic signalling in ex vivo human precision-cut lung slices. |
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| المؤلفون: | Khan MM; European Molecular Biology Laboratory, Heidelberg, Germany.; Discovery Biology, Cellzome GmbH, GSK, Heidelberg, Germany.; Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany., Poeckel D; Discovery Biology, Cellzome GmbH, GSK, Heidelberg, Germany., Halavatyi A; European Molecular Biology Laboratory, Heidelberg, Germany.; Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany., Zukowska-Kasprzyk J; European Molecular Biology Laboratory, Heidelberg, Germany., Stein F; European Molecular Biology Laboratory, Heidelberg, Germany., Vappiani J; Discovery Biology, Cellzome GmbH, GSK, Heidelberg, Germany., Sevin DC; Discovery Biology, Cellzome GmbH, GSK, Heidelberg, Germany., Tischer C; European Molecular Biology Laboratory, Heidelberg, Germany., Zinn N; Discovery Biology, Cellzome GmbH, GSK, Heidelberg, Germany., Eley JD; Novel Human Genetics Research Unit, GSK, Stevenage, UK., Gudmann NS; Nordic Bioscience, Herlev, Denmark., Muley T; Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany.; Biobank Thoraxklinik, University Hospital Heidelberg, Heidelberg, Germany., Winter H; Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany.; Biobank Thoraxklinik, University Hospital Heidelberg, Heidelberg, Germany., Fisher AJ; Newcastle University Translational and Clinical Research Institute and Institute of Transplantation, Newcastle upon Tyne Hospitals, Newcastle upon Tyne, UK., Nanthakumar CB; Novel Human Genetics Research Unit, GSK, Stevenage, UK., Bergamini G; Discovery Biology, Cellzome GmbH, GSK, Heidelberg, Germany.; G. Bergamini and R. Pepperkok contributed equally to this article as lead authors and supervised the work., Pepperkok R; European Molecular Biology Laboratory, Heidelberg, Germany pepperko@embl.de.; Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), Heidelberg, Germany.; G. Bergamini and R. Pepperkok contributed equally to this article as lead authors and supervised the work. |
| المصدر: | The European respiratory journal [Eur Respir J] 2021 Jul 01; Vol. 58 (1). Date of Electronic Publication: 2021 Jul 01 (Print Publication: 2021). |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: European Respiratory Society Country of Publication: England NLM ID: 8803460 Publication Model: Electronic-Print Cited Medium: Internet ISSN: 1399-3003 (Electronic) Linking ISSN: 09031936 NLM ISO Abbreviation: Eur Respir J Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Sheffield, United Kingdom : European Respiratory Society Original Publication: Copenhagen : Published jointly by the Society and Munksgaard, 1988- |
| مواضيع طبية MeSH: | Microscopy* , Pulmonary Fibrosis*/pathology, Fibrosis ; Humans ; Lung/pathology ; Proteomics ; Transforming Growth Factor beta1 |
| مستخلص: | Fibrosis can affect any organ, resulting in the loss of tissue architecture and function with often life-threatening consequences. Pathologically, fibrosis is characterised by the expansion of connective tissue due to excessive deposition of extracellular matrix (ECM) proteins, including the fibrillar forms of collagen. A significant limitation for discovering cures for fibrosis is the availability of suitable human models and techniques to quantify mature fibrillar collagen deposition as close as possible to human physiological conditions.Here we have extensively characterised an ex vivo cultured human lung tissue-derived, precision-cut lung slices (hPCLS) model using label-free second harmonic generation (SHG) light microscopy to quantify fibrillar collagen deposition and mass spectrometry-based techniques to obtain a proteomic and metabolomic fingerprint of hPCLS in ex vivo culture.We demonstrate that hPCLS are viable and metabolically active, with mesenchymal, epithelial, endothelial and immune cell types surviving for at least 2 weeks in ex vivo culture. Analysis of hPCLS-conditioned supernatants showed a strong induction of pulmonary fibrosis-related ECM proteins upon transforming growth factor-β1 (TGF-β1) stimulation. This upregulation of ECM proteins was not translated into an increased deposition of fibrillar collagen. In support of this observation, we revealed the presence of a pro-ECM degradation activity in our ex vivo cultures of hPCLS, inhibition of which by a metalloproteinase inhibitor resulted in increased collagen deposition in response to TGF-β1 stimulation.Together the data show that an integrated approach of measuring soluble pro-fibrotic markers alongside quantitative SHG-based analysis of fibrillar collagen is a valuable tool for studying pro-fibrotic signalling and testing anti-fibrotic agents. Competing Interests: Conflict of interest: M.M. Khan has nothing to disclose. Conflict of interest: D. Poeckel has nothing to disclose. Conflict of interest: A. Halavatyi has nothing to disclose. Conflict of interest: J. Zukowska-Kasprzyk has nothing to disclose. Conflict of interest: F. Stein has nothing to disclose. Conflict of interest: J. Vappiani has nothing to disclose. Conflict of interest: D.C. Sevin has nothing to disclose. Conflict of interest: C. Tischer has nothing to disclose. Conflict of interest: N. Zinn has nothing to disclose. Conflict of interest: J.D. Eley has nothing to disclose. Conflict of interest: N.S. Gudmann has nothing to disclose. Conflict of interest: T. Muley has nothing to disclose. Conflict of interest: H. Winter has nothing to disclose. Conflict of interest: A.J. Fisher reports grants from GlaxoSmithKline, during the conduct of the study, and grants from Pfizer, Nuformix and Genentech, outside the submitted work. Conflict of interest: C.B. Nanthakumar has nothing to disclose. Conflict of interest: G. Bergamini has nothing to disclose. Conflict of interest: R. Pepperkok has nothing to disclose. (Copyright ©ERS 2021.) |
| التعليقات: | Comment in: Eur Respir J. 2021 Jul 1;58(1):. (PMID: 34215663) |
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| المشرفين على المادة: | 0 (Transforming Growth Factor beta1) |
| تواريخ الأحداث: | Date Created: 20201228 Date Completed: 20210809 Latest Revision: 20210809 |
| رمز التحديث: | 20221213 |
| مُعرف محوري في PubMed: | PMC8318569 |
| DOI: | 10.1183/13993003.00221-2020 |
| PMID: | 33361096 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1399-3003 |
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| DOI: | 10.1183/13993003.00221-2020 |