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Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages

Patient material from rare diseases such as very early-onset inflammatory bowel disease (VEO-IBD) is often limited. The use of patient-derived induced pluripotent stem cells (iPSCs) for disease modeling is a promising approach to investigate disease pathomechanisms and therapeutic strategies. We suc...

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Autores principales: Hoffmann, Dirk, Sens, Johanna, Brennig, Sebastian, Brand, Daniel, Philipp, Friederike, Vollmer Barbosa, Philippe, Kuehle, Johannes, Steinemann, Doris, Lenz, Daniela, Buchegger, Theresa, Morgan, Michael, Falk, Christine S., Klein, Christoph, Lachmann, Nico, Schambach, Axel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003874/
https://www.ncbi.nlm.nih.gov/pubmed/33804706
http://dx.doi.org/10.3390/jpm11030221
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author Hoffmann, Dirk
Sens, Johanna
Brennig, Sebastian
Brand, Daniel
Philipp, Friederike
Vollmer Barbosa, Philippe
Kuehle, Johannes
Steinemann, Doris
Lenz, Daniela
Buchegger, Theresa
Morgan, Michael
Falk, Christine S.
Klein, Christoph
Lachmann, Nico
Schambach, Axel
author_facet Hoffmann, Dirk
Sens, Johanna
Brennig, Sebastian
Brand, Daniel
Philipp, Friederike
Vollmer Barbosa, Philippe
Kuehle, Johannes
Steinemann, Doris
Lenz, Daniela
Buchegger, Theresa
Morgan, Michael
Falk, Christine S.
Klein, Christoph
Lachmann, Nico
Schambach, Axel
author_sort Hoffmann, Dirk
collection PubMed
description Patient material from rare diseases such as very early-onset inflammatory bowel disease (VEO-IBD) is often limited. The use of patient-derived induced pluripotent stem cells (iPSCs) for disease modeling is a promising approach to investigate disease pathomechanisms and therapeutic strategies. We successfully developed VEO-IBD patient-derived iPSC lines harboring a mutation in the IL-10 receptor β-chain (IL-10RB) associated with defective IL-10 signaling. To characterize the disease phenotype, healthy control and VEO-IBD iPSCs were differentiated into macrophages. IL-10 stimulation induced characteristic signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) downstream signaling and anti-inflammatory regulation of lipopolysaccharide (LPS)-mediated cytokine secretion in healthy control iPSC-derived macrophages. In contrast, IL-10 stimulation of macrophages derived from patient iPSCs did not result in STAT3 phosphorylation and subsequent SOCS3 expression, recapitulating the phenotype of cells from patients with IL-10RB deficiency. In line with this, LPS-induced cytokine secretion (e.g., IL-6 and tumor necrosis factor-α (TNF-α)) could not be downregulated by exogenous IL-10 stimulation in VEO-IBD iPSC-derived macrophages. Correction of the IL-10RB defect via lentiviral gene therapy or genome editing in the adeno-associated virus integration site 1 (AAVS1) safe harbor locus led to reconstitution of the anti-inflammatory response. Corrected cells showed IL-10RB expression, IL-10-inducible phosphorylation of STAT3, and subsequent SOCS3 expression. Furthermore, LPS-mediated TNF-α secretion could be modulated by IL-10 stimulation in gene-edited VEO-IBD iPSC-derived macrophages. Our established disease models provide the opportunity to identify and validate new curative molecular therapies and to investigate phenotypes and consequences of additional individual IL-10 signaling pathway-dependent VEO-IBD mutations.
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spelling pubmed-80038742021-03-28 Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages Hoffmann, Dirk Sens, Johanna Brennig, Sebastian Brand, Daniel Philipp, Friederike Vollmer Barbosa, Philippe Kuehle, Johannes Steinemann, Doris Lenz, Daniela Buchegger, Theresa Morgan, Michael Falk, Christine S. Klein, Christoph Lachmann, Nico Schambach, Axel J Pers Med Article Patient material from rare diseases such as very early-onset inflammatory bowel disease (VEO-IBD) is often limited. The use of patient-derived induced pluripotent stem cells (iPSCs) for disease modeling is a promising approach to investigate disease pathomechanisms and therapeutic strategies. We successfully developed VEO-IBD patient-derived iPSC lines harboring a mutation in the IL-10 receptor β-chain (IL-10RB) associated with defective IL-10 signaling. To characterize the disease phenotype, healthy control and VEO-IBD iPSCs were differentiated into macrophages. IL-10 stimulation induced characteristic signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) downstream signaling and anti-inflammatory regulation of lipopolysaccharide (LPS)-mediated cytokine secretion in healthy control iPSC-derived macrophages. In contrast, IL-10 stimulation of macrophages derived from patient iPSCs did not result in STAT3 phosphorylation and subsequent SOCS3 expression, recapitulating the phenotype of cells from patients with IL-10RB deficiency. In line with this, LPS-induced cytokine secretion (e.g., IL-6 and tumor necrosis factor-α (TNF-α)) could not be downregulated by exogenous IL-10 stimulation in VEO-IBD iPSC-derived macrophages. Correction of the IL-10RB defect via lentiviral gene therapy or genome editing in the adeno-associated virus integration site 1 (AAVS1) safe harbor locus led to reconstitution of the anti-inflammatory response. Corrected cells showed IL-10RB expression, IL-10-inducible phosphorylation of STAT3, and subsequent SOCS3 expression. Furthermore, LPS-mediated TNF-α secretion could be modulated by IL-10 stimulation in gene-edited VEO-IBD iPSC-derived macrophages. Our established disease models provide the opportunity to identify and validate new curative molecular therapies and to investigate phenotypes and consequences of additional individual IL-10 signaling pathway-dependent VEO-IBD mutations. MDPI 2021-03-20 /pmc/articles/PMC8003874/ /pubmed/33804706 http://dx.doi.org/10.3390/jpm11030221 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Hoffmann, Dirk
Sens, Johanna
Brennig, Sebastian
Brand, Daniel
Philipp, Friederike
Vollmer Barbosa, Philippe
Kuehle, Johannes
Steinemann, Doris
Lenz, Daniela
Buchegger, Theresa
Morgan, Michael
Falk, Christine S.
Klein, Christoph
Lachmann, Nico
Schambach, Axel
Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages
title Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages
title_full Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages
title_fullStr Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages
title_full_unstemmed Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages
title_short Genetic Correction of IL-10RB Deficiency Reconstitutes Anti-Inflammatory Regulation in iPSC-Derived Macrophages
title_sort genetic correction of il-10rb deficiency reconstitutes anti-inflammatory regulation in ipsc-derived macrophages
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003874/
https://www.ncbi.nlm.nih.gov/pubmed/33804706
http://dx.doi.org/10.3390/jpm11030221
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