J Pers Med. 2021 Mar 20;11(3):221. doi: 10.3390/jpm11030221.

Dirk Hoffmann ^1 2, Johanna Sens ^1 2, Sebastian Brennig ^1 2, Daniel Brand ^1 2, Friederike Philipp ^1 2, Philippe Vollmer Barbosa ^1 2 3, Johannes Kuehle ^1 2, Doris Steinemann ⁴, Daniela Lenz ^1 2, Theresa Buchegger ^1 2, Michael Morgan ^1 2, Christine S Falk ⁵, Christoph Klein ⁶, Nico Lachmann ^1 2, Axel Schambach ^1 2 7

Author information

• ¹Institute of Experimental Hematology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
• ²REBIRTH Research Center for Translational Regenerative Medicine, Hannover Medical School, 30625 Hannover, Germany.
• ³Fraunhofer Institute for Toxicology and Experimental Medicine, 30625 Hannover, Germany.
• ⁴Institute of Cell and Molecular Pathology, Hannover Medical School, 30625 Hannover, Germany.
• ⁵Transplant Immunology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
• ⁶Department of Pediatrics, Dr. von Hauner Children's Hospital, University Hospital, Ludwig Maximilian University Munich, 80337 Munich, Germany.
• ⁷Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.

Abstract

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.