Sanami Takada1, Naotomo Kambe2, Yuri Kawasaki3, Akira Niwa3, Fumiko Honda-Ozaki3, Kazuki Kobayashi3, Mitsujiro Osawa3, Ayako Nagahashi4, Katsunori Semi5, Akitsu Hotta6, Isao Asaka4, Yasuhiro Yamada6, Ryuta Nishikomori7, Toshio Heike7, Hiroyuki Matsue8, Tatsutoshi Nakahata3, Megumu K Saito9. 1. Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan; Department of Dermatology, Chiba University Graduate School of Medicine, Chiba, Japan. 2. Department of Dermatology, Kansai Medical University, Hirakata, Japan. 3. Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan. 4. Department of Fundamental Cell Technology, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan. 5. Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan. 6. Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan; Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan. 7. Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, Japan. 8. Department of Dermatology, Chiba University Graduate School of Medicine, Chiba, Japan; Division of Molecular Immunology, Medical Mycology Research Center, Chiba University, Chiba, Japan. 9. Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan. Electronic address: msaito@cira.kyoto-u.ac.jp.
Abstract
BACKGROUND: Blau syndrome, or early-onset sarcoidosis, is a juvenile-onset systemic granulomatosis associated with a mutation in nucleotide-binding oligomerization domain 2 (NOD2). The underlying mechanisms of Blau syndrome leading to autoinflammation are still unclear, and there is currently no effective specific treatment for Blau syndrome. OBJECTIVES: To elucidate the mechanisms of autoinflammation in patients with Blau syndrome, we sought to clarify the relation between disease-associated mutant NOD2 and the inflammatory response in human samples. METHODS: Blau syndrome-specific induced pluripotent stem cell (iPSC) lines were established. The disease-associated NOD2 mutation of iPSCs was corrected by using a CRISPR-Cas9 system to precisely evaluate the in vitro phenotype of iPSC-derived cells. We also introduced the same NOD2 mutation into a control iPSC line. These isogenic iPSCs were then differentiated into monocytic cell lineages, and the statuses of nuclear factor κB pathway and proinflammatory cytokine secretion were investigated. RESULTS: IFN-γ acted as a priming signal through upregulation of NOD2. In iPSC-derived macrophages with mutant NOD2, IFN-γ treatment induced ligand-independent nuclear factor κB activation and proinflammatory cytokine production. RNA sequencing analysis revealed distinct transcriptional profiles of mutant macrophages both before and after IFN-γ treatment. Patient-derived macrophages demonstrated a similar IFN-γ-dependent inflammatory response. CONCLUSIONS: Our data support the significance of ligand-independent autoinflammation in the pathophysiology of Blau syndrome. Our comprehensive isogenic disease-specific iPSC panel provides a useful platform for probing therapeutic and diagnostic clues for the treatment of patients with Blau syndrome.
BACKGROUND:Blau syndrome, or early-onset sarcoidosis, is a juvenile-onset systemic granulomatosis associated with a mutation in nucleotide-binding oligomerization domain 2 (NOD2). The underlying mechanisms of Blau syndrome leading to autoinflammation are still unclear, and there is currently no effective specific treatment for Blau syndrome. OBJECTIVES: To elucidate the mechanisms of autoinflammation in patients with Blau syndrome, we sought to clarify the relation between disease-associated mutant NOD2 and the inflammatory response in human samples. METHODS:Blau syndrome-specific induced pluripotent stem cell (iPSC) lines were established. The disease-associated NOD2 mutation of iPSCs was corrected by using a CRISPR-Cas9 system to precisely evaluate the in vitro phenotype of iPSC-derived cells. We also introduced the same NOD2 mutation into a control iPSC line. These isogenic iPSCs were then differentiated into monocytic cell lineages, and the statuses of nuclear factor κB pathway and proinflammatory cytokine secretion were investigated. RESULTS: IFN-γ acted as a priming signal through upregulation of NOD2. In iPSC-derived macrophages with mutant NOD2, IFN-γ treatment induced ligand-independent nuclear factor κB activation and proinflammatory cytokine production. RNA sequencing analysis revealed distinct transcriptional profiles of mutant macrophages both before and after IFN-γ treatment. Patient-derived macrophages demonstrated a similar IFN-γ-dependent inflammatory response. CONCLUSIONS: Our data support the significance of ligand-independent autoinflammation in the pathophysiology of Blau syndrome. Our comprehensive isogenic disease-specific iPSC panel provides a useful platform for probing therapeutic and diagnostic clues for the treatment of patients with Blau syndrome.
Authors: Zuzana Parackova; Marketa Bloomfield; Petra Vrabcova; Irena Zentsova; Adam Klocperk; Tomas Milota; Michael Svaton; Jean-Laurent Casanova; Jacinta Bustamante; Eva Fronkova; Anna Sediva Journal: J Clin Immunol Date: 2019-11-23 Impact factor: 8.542