Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis
Abstract Background Exaggerated fibroblast proliferation is a well‐known feature in idiopathic pulmonary fibrosis (IPF) which may be – in part – due to insufficient autophagy, a lysosome dependent cellular surveillance pathway. Bcl2‐associated athanogene 3 (BAG3) is a pivotal co‐chaperone of the aut...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-07-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.935 |
| _version_ | 1811221663263490048 |
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| author | Shashipavan Chillappagari Julian Schwarz Vidyasagar Kesireddy Jessica Knoell Martina Korfei Konrad Hoetzenecker M. Lienhard Schmitz Christian Behl Saverio Bellusci Andreas Guenther Poornima Mahavadi |
| author_facet | Shashipavan Chillappagari Julian Schwarz Vidyasagar Kesireddy Jessica Knoell Martina Korfei Konrad Hoetzenecker M. Lienhard Schmitz Christian Behl Saverio Bellusci Andreas Guenther Poornima Mahavadi |
| author_sort | Shashipavan Chillappagari |
| collection | DOAJ |
| description | Abstract Background Exaggerated fibroblast proliferation is a well‐known feature in idiopathic pulmonary fibrosis (IPF) which may be – in part – due to insufficient autophagy, a lysosome dependent cellular surveillance pathway. Bcl2‐associated athanogene 3 (BAG3) is a pivotal co‐chaperone of the autophagy pathway. Here, we studied whether therapeutic modulation of BAG3‐mediated autophagy can rescue insufficient autophagy and impact IPF fibroblast proliferation. Methods Primary interstitial fibroblasts or precision cut lung slices (PCLS) of IPF lungs were treated with (1) the antifibrotic drug pirfenidone (Pirf), (2) the demethylating agent 5‐azacytidine (Aza), (3) the BAG3 modulator cantharidin (Ctd). Autophagy flux was measured following pretreatment with the autophagy inhibitors or by GFP‐RFP‐LC3B transfection followed by drug treatments. Proliferation was measured by 5‐bromo‐2′‐deoxyuridine assay. BAG3, filamin C (FLNC), proliferating‐cell‐nuclear‐antigen (PCNA), collagen1A1 (COL1A1) and autophagy proteins were assessed by immunoblotting or immunofluorescence. Loss of function experiments were performed by siRNA mediated knockdown of BAG3. Results In comparison with healthy donors, increased BAG3 protein was observed in IPF lung homogenates and IPF fibroblasts. In addition, the substrate of BAG3‐mediated autophagy, FLNC, was increased in IPF fibroblasts, implying insufficient activation of BAG3‐dependent autophagy. Therapeutic modulation of this pathway using Aza and Ctd alone or in combination with the IPF therapy drug Pirf rescued the insufficient BAG3‐mediated autophagy and decreased fibroblast proliferation. Such effects were observed upon therapeutic modulation of BAG3 but not upon knock down of BAG3 per se in IPF fibroblasts. Similarly, PCLS of IPF patients showed a significant decrease in collagen deposition in response to these drugs, either alone or in a more potent form in combination with Pirf. Conclusions Our study reveals that repurposing drugs that modulate autophagy regulating proteins render therapeutic benefits in IPF. Fine tuning of this pathway may hence signify a promising therapeutic strategy to ameliorate antifibrotic properties and augment the efficacy of current IPF therapy. |
| first_indexed | 2024-04-12T08:03:31Z |
| format | Article |
| id | doaj.art-6f9ebcc8bb3a4739be193fb80c667671 |
| institution | Directory Open Access Journal |
| issn | 2001-1326 |
| language | English |
| last_indexed | 2024-04-12T08:03:31Z |
| publishDate | 2022-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Clinical and Translational Medicine |
| spelling | doaj.art-6f9ebcc8bb3a4739be193fb80c6676712022-12-22T03:41:14ZengWileyClinical and Translational Medicine2001-13262022-07-01127n/an/a10.1002/ctm2.935Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosisShashipavan Chillappagari0Julian Schwarz1Vidyasagar Kesireddy2Jessica Knoell3Martina Korfei4Konrad Hoetzenecker5M. Lienhard Schmitz6Christian Behl7Saverio Bellusci8Andreas Guenther9Poornima Mahavadi10Department of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Thoracic Surgery Vienna General Hospital Vienna AustriaUniversities of Giessen and Marburg Lung Center (UGMLC) Member of the German Centre for Lung Research (DZL) Giessen Hessen GermanyInstitute of Pathobiochemistry, The Autophagy Lab University Medical Center Johannes Gutenberg University Mainz GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyDepartment of Internal Medicine Justus‐Liebig University (JLU) Giessen Giessen Hessen GermanyAbstract Background Exaggerated fibroblast proliferation is a well‐known feature in idiopathic pulmonary fibrosis (IPF) which may be – in part – due to insufficient autophagy, a lysosome dependent cellular surveillance pathway. Bcl2‐associated athanogene 3 (BAG3) is a pivotal co‐chaperone of the autophagy pathway. Here, we studied whether therapeutic modulation of BAG3‐mediated autophagy can rescue insufficient autophagy and impact IPF fibroblast proliferation. Methods Primary interstitial fibroblasts or precision cut lung slices (PCLS) of IPF lungs were treated with (1) the antifibrotic drug pirfenidone (Pirf), (2) the demethylating agent 5‐azacytidine (Aza), (3) the BAG3 modulator cantharidin (Ctd). Autophagy flux was measured following pretreatment with the autophagy inhibitors or by GFP‐RFP‐LC3B transfection followed by drug treatments. Proliferation was measured by 5‐bromo‐2′‐deoxyuridine assay. BAG3, filamin C (FLNC), proliferating‐cell‐nuclear‐antigen (PCNA), collagen1A1 (COL1A1) and autophagy proteins were assessed by immunoblotting or immunofluorescence. Loss of function experiments were performed by siRNA mediated knockdown of BAG3. Results In comparison with healthy donors, increased BAG3 protein was observed in IPF lung homogenates and IPF fibroblasts. In addition, the substrate of BAG3‐mediated autophagy, FLNC, was increased in IPF fibroblasts, implying insufficient activation of BAG3‐dependent autophagy. Therapeutic modulation of this pathway using Aza and Ctd alone or in combination with the IPF therapy drug Pirf rescued the insufficient BAG3‐mediated autophagy and decreased fibroblast proliferation. Such effects were observed upon therapeutic modulation of BAG3 but not upon knock down of BAG3 per se in IPF fibroblasts. Similarly, PCLS of IPF patients showed a significant decrease in collagen deposition in response to these drugs, either alone or in a more potent form in combination with Pirf. Conclusions Our study reveals that repurposing drugs that modulate autophagy regulating proteins render therapeutic benefits in IPF. Fine tuning of this pathway may hence signify a promising therapeutic strategy to ameliorate antifibrotic properties and augment the efficacy of current IPF therapy.https://doi.org/10.1002/ctm2.9355‐azacytidineautophagyBAG3cantharidinfibroblastsfilamin C |
| spellingShingle | Shashipavan Chillappagari Julian Schwarz Vidyasagar Kesireddy Jessica Knoell Martina Korfei Konrad Hoetzenecker M. Lienhard Schmitz Christian Behl Saverio Bellusci Andreas Guenther Poornima Mahavadi Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis Clinical and Translational Medicine 5‐azacytidine autophagy BAG3 cantharidin fibroblasts filamin C |
| title | Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis |
| title_full | Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis |
| title_fullStr | Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis |
| title_full_unstemmed | Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis |
| title_short | Therapeutic induction of Bcl2‐associated athanogene 3‐mediated autophagy in idiopathic pulmonary fibrosis |
| title_sort | therapeutic induction of bcl2 associated athanogene 3 mediated autophagy in idiopathic pulmonary fibrosis |
| topic | 5‐azacytidine autophagy BAG3 cantharidin fibroblasts filamin C |
| url | https://doi.org/10.1002/ctm2.935 |
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