Cargando…
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...
Autores principales: | , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1783671792165978112 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8003874 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT hoffmanndirk geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT sensjohanna geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT brennigsebastian geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT branddaniel geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT philippfriederike geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT vollmerbarbosaphilippe geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT kuehlejohannes geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT steinemanndoris geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT lenzdaniela geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT bucheggertheresa geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT morganmichael geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT falkchristines geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT kleinchristoph geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT lachmannnico geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages AT schambachaxel geneticcorrectionofil10rbdeficiencyreconstitutesantiinflammatoryregulationinipscderivedmacrophages |