αAzithromycin has enhanced effects on lung fibroblasts from idiopathic pulmonary fibrosis (IPF) patients compared to controls
Identifieur interne : 000006 ( Main/Exploration ); précédent : 000005; suivant : 000007αAzithromycin has enhanced effects on lung fibroblasts from idiopathic pulmonary fibrosis (IPF) patients compared to controls
Auteurs : Kristina Krempaska [Suisse] ; Sandra Barnowski [Suisse] ; Jacopo Gavini [Suisse] ; Nina Hobi [Suisse] ; Simone Ebener [Suisse] ; Cedric Simillion [Suisse] ; Andrea Stokes [Suisse] ; Ronja Schliep [Allemagne] ; Lars Knudsen [Allemagne] ; Thomas K. Geiser [Suisse] ; Manuela Funke-Chambour [Suisse]Source :
- Respiratory Research [ 1465-9921 ] ; 2020.
Abstract
Idiopathic pulmonary fibrosis (IPF) is a chronic fatal lung disease without a cure and new drug strategies are urgently needed. Differences in behavior between diseased and healthy cells are well known and drug response can be different between cells isolated from IPF patients and controls. The macrolide Azithromycin (AZT) has anti-inflammatory and immunomodulatory properties. Recently anti-fibrotic effects have been described. However, the anti-fibrotic effects on primary IPF-fibroblasts (FB) directly compared to control-FB are unknown. We hypothesized that IPF-FB react differently to AZT in terms of anti-fibrotic effects.
Primary normal human lung and IPF-FB were exposed to TGF-β (5 ng/ml), Azithromycin (50 μM) alone or in combination prior to gene expression analysis. Pro-collagen Iα1 secretion was assessed by ELISA and protein expression by western blot (αSMA, Fibronectin, ATP6V1B2, LC3 AB (II/I), p62, Bcl-xL). Microarray analysis was performed to screen involved genes and pathways after Azithromycin treatment in control-FB. Apoptosis and intraluminal lysosomal pH were analyzed by flow cytometry.
AZT significantly reduced collagen secretion in TGF-β treated IPF-FB compared to TGF-β treatment alone, but not in control-FB. Pro-fibrotic gene expression was similarly reduced after AZT treatment in IPF and control-FB. P62 and LC3II/I western blot revealed impaired autophagic flux after AZT in both control and IPF-FB with significant increase of LC3II/I after AZT in control and IPF-FB, indicating enhanced autophagy inhibition. Early apoptosis was significantly higher in TGF-β treated IPF-FB compared to controls after AZT. Microarray analysis of control-FB treated with AZT revealed impaired lysosomal pathways. The ATPase and lysosomal pH regulator ATP6V0D2 was significantly less increased after additional AZT in IPF-FB compared to controls. Lysosomal function was impaired in both IPF and control FB, but pH was significantly more increased in TGF-β treated IPF-FB.
We report different treatment responses after AZT with enhanced anti-fibrotic and pro-apoptotic effects in IPF compared to control-FB. Possibly impaired lysosomal function contributes towards these effects. In summary, different baseline cell phenotype and behavior of IPF and control cells contribute to enhanced anti-fibrotic and pro-apoptotic effects in IPF-FB after AZT treatment and strengthen its role as a new potential anti-fibrotic compound, that should further be evaluated in clinical studies.
Url:
DOI: 10.1186/s12931-020-1275-8
PubMed: 31941499
PubMed Central: 6964061
Affiliations:
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Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>CH-3010 Bern</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0726 5157</institution-id><institution-id institution-id-type="GRID">grid.5734.5</institution-id><institution>Department for BioMedical Research,</institution><institution>University of Bern,</institution></institution-wrap>Bern, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>Bern</wicri:regionArea></affiliation></author><author><name sortKey="Schliep, Ronja" sort="Schliep, Ronja" uniqKey="Schliep R" first="Ronja" last="Schliep">Ronja Schliep</name><affiliation wicri:level="1"><nlm:aff id="Aff8"><institution-wrap><institution-id institution-id-type="ISNI">0000 0000 9529 9877</institution-id><institution-id institution-id-type="GRID">grid.10423.34</institution-id><institution>Institute of Functional and Applied Anatomy, Hannover Medical School,</institution></institution-wrap>Hannover, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Hannover</wicri:regionArea></affiliation></author><author><name sortKey="Knudsen, Lars" sort="Knudsen, Lars" uniqKey="Knudsen L" first="Lars" last="Knudsen">Lars Knudsen</name><affiliation wicri:level="1"><nlm:aff id="Aff8"><institution-wrap><institution-id institution-id-type="ISNI">0000 0000 9529 9877</institution-id><institution-id institution-id-type="GRID">grid.10423.34</institution-id><institution>Institute of Functional and Applied Anatomy, Hannover Medical School,</institution></institution-wrap>Hannover, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Hannover</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff9"><institution-wrap><institution-id institution-id-type="GRID">grid.452624.3</institution-id><institution>Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL),</institution></institution-wrap>Hannover, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Hannover</wicri:regionArea></affiliation></author><author><name sortKey="Geiser, Thomas K" sort="Geiser, Thomas K" uniqKey="Geiser T" first="Thomas K." last="Geiser">Thomas K. Geiser</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0726 5157</institution-id><institution-id institution-id-type="GRID">grid.5734.5</institution-id><institution>Department of Pulmonary Medicine, Inselspital, Bern University Hospital,</institution><institution>University of Bern,</institution></institution-wrap>CH-3010 Bern, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>CH-3010 Bern</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0726 5157</institution-id><institution-id institution-id-type="GRID">grid.5734.5</institution-id><institution>Department for BioMedical Research,</institution><institution>University of Bern,</institution></institution-wrap>Bern, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>Bern</wicri:regionArea></affiliation></author><author><name sortKey="Funke Chambour, Manuela" sort="Funke Chambour, Manuela" uniqKey="Funke Chambour M" first="Manuela" last="Funke-Chambour">Manuela Funke-Chambour</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0726 5157</institution-id><institution-id institution-id-type="GRID">grid.5734.5</institution-id><institution>Department of Pulmonary Medicine, Inselspital, Bern University Hospital,</institution><institution>University of Bern,</institution></institution-wrap>CH-3010 Bern, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>CH-3010 Bern</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0726 5157</institution-id><institution-id institution-id-type="GRID">grid.5734.5</institution-id><institution>Department for BioMedical Research,</institution><institution>University of Bern,</institution></institution-wrap>Bern, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>Bern</wicri:regionArea></affiliation></author></analytic><series><title level="j">Respiratory Research</title><idno type="ISSN">1465-9921</idno><idno type="eISSN">1465-993X</idno><imprint><date when="2020">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p id="Par1">Idiopathic pulmonary fibrosis (IPF) is a chronic fatal lung disease without a cure and new drug strategies are urgently needed. Differences in behavior between diseased and healthy cells are well known and drug response can be different between cells isolated from IPF patients and controls. The macrolide Azithromycin (AZT) has anti-inflammatory and immunomodulatory properties. Recently anti-fibrotic effects have been described. However, the anti-fibrotic effects on primary IPF-fibroblasts (FB) directly compared to control-FB are unknown. We hypothesized that IPF-FB react differently to AZT in terms of anti-fibrotic effects.</p></sec><sec><title>Methods</title><p id="Par2">Primary normal human lung and IPF-FB were exposed to TGF-β (5 ng/ml), Azithromycin (50 μM) alone or in combination prior to gene expression analysis. Pro-collagen Iα1 secretion was assessed by ELISA and protein expression by western blot (αSMA, Fibronectin, ATP6V1B2, LC3 AB (II/I), p62, Bcl-xL). Microarray analysis was performed to screen involved genes and pathways after Azithromycin treatment in control-FB. Apoptosis and intraluminal lysosomal pH were analyzed by flow cytometry.</p></sec><sec><title>Results</title><p id="Par3">AZT significantly reduced collagen secretion in TGF-β treated IPF-FB compared to TGF-β treatment alone, but not in control-FB. Pro-fibrotic gene expression was similarly reduced after AZT treatment in IPF and control-FB. P62 and LC3II/I western blot revealed impaired autophagic flux after AZT in both control and IPF-FB with significant increase of LC3II/I after AZT in control and IPF-FB, indicating enhanced autophagy inhibition. Early apoptosis was significantly higher in TGF-β treated IPF-FB compared to controls after AZT. Microarray analysis of control-FB treated with AZT revealed impaired lysosomal pathways. The ATPase and lysosomal pH regulator ATP6V0D2 was significantly less increased after additional AZT in IPF-FB compared to controls. Lysosomal function was impaired in both IPF and control FB, but pH was significantly more increased in TGF-β treated IPF-FB.</p></sec><sec><title>Conclusion</title><p id="Par4">We report different treatment responses after AZT with enhanced anti-fibrotic and pro-apoptotic effects in IPF compared to control-FB. Possibly impaired lysosomal function contributes towards these effects. In summary, different baseline cell phenotype and behavior of IPF and control cells contribute to enhanced anti-fibrotic and pro-apoptotic effects in IPF-FB after AZT treatment and strengthen its role as a new potential anti-fibrotic compound, that should further be evaluated in clinical studies.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Ley, B" uniqKey="Ley B">B Ley</name></author><author><name sortKey="Collard, Hr" uniqKey="Collard H">HR Collard</name></author><author><name sortKey="King, Te" uniqKey="King T">TE King</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Raghu, G" uniqKey="Raghu G">G Raghu</name></author><author><name sortKey="Collard, Hr" uniqKey="Collard H">HR Collard</name></author><author><name sortKey="Egan, Jj" uniqKey="Egan J">JJ Egan</name></author><author><name sortKey="Martinez, Fj" uniqKey="Martinez F">FJ Martinez</name></author><author><name sortKey="Behr, J" uniqKey="Behr J">J Behr</name></author><author><name sortKey="Brown, Kk" uniqKey="Brown K">KK Brown</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Crestani, Bruno" uniqKey="Crestani B">Bruno Crestani</name></author><author><name sortKey="Huggins, John T" uniqKey="Huggins J">John T Huggins</name></author><author><name sortKey="Kaye, Mitchell" uniqKey="Kaye M">Mitchell Kaye</name></author><author><name sortKey="Costabel, Ulrich" uniqKey="Costabel U">Ulrich Costabel</name></author><author><name sortKey="Glaspole, Ian" uniqKey="Glaspole I">Ian Glaspole</name></author><author><name sortKey="Ogura, Takashi" uniqKey="Ogura T">Takashi Ogura</name></author><author><name sortKey="Song, Jin Woo" uniqKey="Song J">Jin Woo Song</name></author><author><name sortKey="Stansen, Wibke" uniqKey="Stansen W">Wibke Stansen</name></author><author><name sortKey="Quaresma, Manuel" uniqKey="Quaresma M">Manuel Quaresma</name></author><author><name sortKey="Stowasser, Susanne" uniqKey="Stowasser S">Susanne Stowasser</name></author><author><name sortKey="Kreuter, Michael" uniqKey="Kreuter M">Michael Kreuter</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Margaritopoulos, Ga" uniqKey="Margaritopoulos G">GA Margaritopoulos</name></author><author><name sortKey="Trachalaki, A" uniqKey="Trachalaki A">A Trachalaki</name></author><author><name sortKey="Wells, Au" uniqKey="Wells A">AU Wells</name></author><author><name sortKey="Vasarmidi, E" uniqKey="Vasarmidi E">E Vasarmidi</name></author><author><name sortKey="Bibaki, E" uniqKey="Bibaki E">E Bibaki</name></author><author><name sortKey="Papastratigakis, G" uniqKey="Papastratigakis G">G Papastratigakis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Geiser, T" uniqKey="Geiser T">T Geiser</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Phan, Sh" uniqKey="Phan S">SH Phan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Upagupta, C" uniqKey="Upagupta C">C Upagupta</name></author><author><name sortKey="Shimbori, C" uniqKey="Shimbori C">C Shimbori</name></author><author><name sortKey="Alsilmi, R" uniqKey="Alsilmi R">R Alsilmi</name></author><author><name sortKey="Kolb, M" uniqKey="Kolb M">M Kolb</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Biernacka, A" uniqKey="Biernacka A">A Biernacka</name></author><author><name sortKey="Dobaczewski, M" uniqKey="Dobaczewski M">M Dobaczewski</name></author><author><name sortKey="Frangogiannis, Ng" uniqKey="Frangogiannis N">NG Frangogiannis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Araya, J" uniqKey="Araya J">J Araya</name></author><author><name sortKey="Kojima, J" uniqKey="Kojima J">J Kojima</name></author><author><name sortKey="Takasaka, N" uniqKey="Takasaka N">N Takasaka</name></author><author><name sortKey="Ito, S" uniqKey="Ito S">S Ito</name></author><author><name sortKey="Fujii, S" uniqKey="Fujii S">S Fujii</name></author><author><name sortKey="Hara, H" uniqKey="Hara H">H Hara</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sosulski, Meredith L" uniqKey="Sosulski M">Meredith L. 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Abraham</name></author><author><name sortKey="Jessen, Bart A" uniqKey="Jessen B">Bart A. Jessen</name></author></analytic></biblStruct><biblStruct></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Allemagne</li><li>Suisse</li></country><region><li>Canton de Berne</li></region><settlement><li>Berne</li></settlement></list><tree><country name="Suisse"><noRegion><name sortKey="Krempaska, Kristina" sort="Krempaska, Kristina" uniqKey="Krempaska K" first="Kristina" last="Krempaska">Kristina Krempaska</name></noRegion><name sortKey="Barnowski, Sandra" sort="Barnowski, Sandra" uniqKey="Barnowski S" first="Sandra" last="Barnowski">Sandra Barnowski</name><name sortKey="Barnowski, Sandra" sort="Barnowski, Sandra" uniqKey="Barnowski S" first="Sandra" last="Barnowski">Sandra Barnowski</name><name sortKey="Ebener, Simone" sort="Ebener, Simone" uniqKey="Ebener S" first="Simone" last="Ebener">Simone Ebener</name><name sortKey="Ebener, Simone" sort="Ebener, Simone" uniqKey="Ebener S" first="Simone" last="Ebener">Simone Ebener</name><name sortKey="Funke Chambour, Manuela" sort="Funke Chambour, Manuela" uniqKey="Funke Chambour M" first="Manuela" last="Funke-Chambour">Manuela Funke-Chambour</name><name sortKey="Funke Chambour, Manuela" sort="Funke Chambour, Manuela" uniqKey="Funke Chambour M" first="Manuela" last="Funke-Chambour">Manuela Funke-Chambour</name><name sortKey="Gavini, Jacopo" sort="Gavini, Jacopo" uniqKey="Gavini J" first="Jacopo" last="Gavini">Jacopo Gavini</name><name sortKey="Geiser, Thomas K" sort="Geiser, Thomas K" uniqKey="Geiser T" first="Thomas K." last="Geiser">Thomas K. Geiser</name><name sortKey="Geiser, Thomas K" sort="Geiser, Thomas K" uniqKey="Geiser T" first="Thomas K." last="Geiser">Thomas K. Geiser</name><name sortKey="Hobi, Nina" sort="Hobi, Nina" uniqKey="Hobi N" first="Nina" last="Hobi">Nina Hobi</name><name sortKey="Hobi, Nina" sort="Hobi, Nina" uniqKey="Hobi N" first="Nina" last="Hobi">Nina Hobi</name><name sortKey="Krempaska, Kristina" sort="Krempaska, Kristina" uniqKey="Krempaska K" first="Kristina" last="Krempaska">Kristina Krempaska</name><name sortKey="Krempaska, Kristina" sort="Krempaska, Kristina" uniqKey="Krempaska K" first="Kristina" last="Krempaska">Kristina Krempaska</name><name sortKey="Simillion, Cedric" sort="Simillion, Cedric" uniqKey="Simillion C" first="Cedric" last="Simillion">Cedric Simillion</name><name sortKey="Simillion, Cedric" sort="Simillion, Cedric" uniqKey="Simillion C" first="Cedric" last="Simillion">Cedric Simillion</name><name sortKey="Stokes, Andrea" sort="Stokes, Andrea" uniqKey="Stokes A" first="Andrea" last="Stokes">Andrea Stokes</name><name sortKey="Stokes, Andrea" sort="Stokes, Andrea" uniqKey="Stokes A" first="Andrea" last="Stokes">Andrea Stokes</name></country><country name="Allemagne"><noRegion><name sortKey="Schliep, Ronja" sort="Schliep, Ronja" uniqKey="Schliep R" first="Ronja" last="Schliep">Ronja Schliep</name></noRegion><name sortKey="Knudsen, Lars" sort="Knudsen, Lars" uniqKey="Knudsen L" first="Lars" last="Knudsen">Lars Knudsen</name><name sortKey="Knudsen, Lars" sort="Knudsen, Lars" uniqKey="Knudsen L" first="Lars" last="Knudsen">Lars Knudsen</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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