PMID- 35834635 OWN - NLM STAT- MEDLINE DCOM- 20220718 LR - 20231105 IS - 2001-1326 (Electronic) IS - 2001-1326 (Linking) VI - 12 IP - 7 DP - 2022 Jul TI - Therapeutic induction of Bcl2-associated athanogene 3-mediated autophagy in idiopathic pulmonary fibrosis. PG - e935 LID - 10.1002/ctm2.935 [doi] LID - e935 AB - 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. CI - (c) 2022 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics. FAU - Chillappagari, Shashipavan AU - Chillappagari S AUID- ORCID: 0000-0002-0080-6552 AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. AD - Department of Biochemistry, Faculty of Medicine, JLU Giessen, Giessen, Hessen, Germany. FAU - Schwarz, Julian AU - Schwarz J AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. FAU - Kesireddy, Vidyasagar AU - Kesireddy V AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. FAU - Knoell, Jessica AU - Knoell J AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. FAU - Korfei, Martina AU - Korfei M AUID- ORCID: 0000-0001-8177-2085 AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. FAU - Hoetzenecker, Konrad AU - Hoetzenecker K AD - Department of Thoracic Surgery, Vienna General Hospital, Vienna, Austria. AD - European IPF Network and European IPF Registry, Giessen, Germany. FAU - Schmitz, M Lienhard AU - Schmitz ML AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. AD - Department of Biochemistry, Faculty of Medicine, JLU Giessen, Giessen, Hessen, Germany. AD - Member of the Cardio-Pulmonary Institute (CPI), JLU Giessen, Giessen, Germany. FAU - Behl, Christian AU - Behl C AD - Institute of Pathobiochemistry, The Autophagy Lab, University Medical Center, Johannes Gutenberg University, Mainz, Germany. FAU - Bellusci, Saverio AU - Bellusci S AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. AD - Member of the Cardio-Pulmonary Institute (CPI), JLU Giessen, Giessen, Germany. FAU - Guenther, Andreas AU - Guenther A AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. AD - European IPF Network and European IPF Registry, Giessen, Germany. AD - Member of the Cardio-Pulmonary Institute (CPI), JLU Giessen, Giessen, Germany. AD - Lung Clinic, Agaplesion Evangelisches Krankenhaus Mittelhessen, Giessen, Germany. FAU - Mahavadi, Poornima AU - Mahavadi P AUID- ORCID: 0000-0002-7580-4738 AD - Department of Internal Medicine, Justus-Liebig University (JLU) Giessen, Giessen, Hessen, Germany. AD - Universities of Giessen and Marburg Lung Center (UGMLC), Member of the German Centre for Lung Research (DZL), Giessen, Hessen, Germany. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Clin Transl Med JT - Clinical and translational medicine JID - 101597971 RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Apoptosis Regulatory Proteins) RN - 0 (BAG3 protein, human) RN - 0 (BCL2 protein, human) RN - 0 (Proto-Oncogene Proteins c-bcl-2) RN - 9007-34-5 (Collagen) SB - IM MH - *Adaptor Proteins, Signal Transducing/genetics/metabolism MH - *Apoptosis Regulatory Proteins/genetics/metabolism MH - *Autophagy/genetics/physiology MH - Collagen/metabolism MH - *Fibroblasts/metabolism MH - Humans MH - *Idiopathic Pulmonary Fibrosis/genetics/metabolism MH - Lung/metabolism MH - Proto-Oncogene Proteins c-bcl-2/metabolism PMC - PMC9282656 OTO - NOTNLM OT - 5-azacytidine OT - BAG3 OT - autophagy OT - cantharidin OT - fibroblasts OT - filamin C OT - idiopathic pulmonary fibrosis OT - pirfenidone COIS- The authors declare no conflicts of interest. EDAT- 2022/07/15 06:00 MHDA- 2022/07/19 06:00 PMCR- 2022/07/14 CRDT- 2022/07/14 14:23 PHST- 2022/05/25 00:00 [revised] PHST- 2021/08/31 00:00 [received] PHST- 2022/06/02 00:00 [accepted] PHST- 2022/07/14 14:23 [entrez] PHST- 2022/07/15 06:00 [pubmed] PHST- 2022/07/19 06:00 [medline] PHST- 2022/07/14 00:00 [pmc-release] AID - CTM2935 [pii] AID - 10.1002/ctm2.935 [doi] PST - ppublish SO - Clin Transl Med. 2022 Jul;12(7):e935. doi: 10.1002/ctm2.935.