title: Enhanced chondrogenesis of induced pluripotent stem cells from patients with neonatal-onset multisystem inflammatory disease occurs via the caspase 1-independent cAMP/protein kinase A/CREB pathway.
abstract: OBJECTIVE
  Neonatal-onset multisystem inflammatory disease (NOMID) is a dominantly inherited autoinflammatory disease caused by NLRP3 mutations. NOMID pathophysiology is explained by the NLRP3 inflammasome, which produces interleukin-1β (IL-1β). However, epiphyseal overgrowth in NOMID is resistant to anti-IL-1 therapy and may therefore occur independently of the NLRP3 inflammasome. This study was undertaken to investigate the effect of mutated NLRP3 on chondrocytes using induced pluripotent stem cells (iPSCs) from patients with NOMID.
  
  
  METHODS
  We established isogenic iPSCs with wild-type or mutant NLRP3 from 2 NOMID patients with NLRP3 somatic mosaicism. The iPSCs were differentiated into chondrocytes in vitro and in vivo. The phenotypes of chondrocytes with wild-type and mutant NLRP3 were compared, particularly the size of the chondrocyte tissue produced.
  
  
  RESULTS
  Mutant iPSCs produced larger chondrocyte masses than wild-type iPSCs owing to glycosaminoglycan overproduction, which correlated with increased expression of the chondrocyte master regulator SOX9. In addition, in vivo transplantation of mutant cartilaginous pellets into immunodeficient mice caused disorganized endochondral ossification. Enhanced chondrogenesis was independent of caspase 1 and IL-1, and thus the NLRP3 inflammasome. Investigation of the human SOX9 promoter in chondroprogenitor cells revealed that the CREB/ATF-binding site was critical for SOX9 overexpression caused by mutated NLRP3. This was supported by increased levels of cAMP and phosphorylated CREB in mutant chondroprogenitor cells.
  
  
  CONCLUSIONS
  Our findings indicate that the intrinsic hyperplastic capacity of NOMID chondrocytes is dependent on the cAMP/PKA/CREB pathway, independent of the NLRP3 inflammasome.
authors.affiliation: Kyoto University, Kyoto, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Yokoyama
authors.forename: Makoto
authors.initials: M
authors.lastname: Ikeya
authors.forename: Katsutsugu
authors.initials: K
authors.lastname: Umeda
authors.forename: Hirotsugu
authors.initials: H
authors.lastname: Oda
authors.forename: Seishiro
authors.initials: S
authors.lastname: Nodomi
authors.forename: Akira
authors.initials: A
authors.lastname: Nasu
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: Kazushi
authors.initials: K
authors.lastname: Izawa
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Horigome
authors.forename: Toshimasa
authors.initials: T
authors.lastname: Kusaka
authors.forename: Takayuki
authors.initials: T
authors.lastname: Tanaka
authors.forename: Megumu K
authors.initials: MK
authors.lastname: Saito
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yasumi
authors.forename: Ryuta
authors.initials: R
authors.lastname: Nishikomori
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohara
authors.forename: Naoki
authors.initials: N
authors.lastname: Nakayama
authors.forename: Tatsutoshi
authors.initials: T
authors.lastname: Nakahata
authors.forename: Toshio
authors.initials: T
authors.lastname: Heike
authors.forename: Junya
authors.initials: J
authors.lastname: Toguchida
mesh_terms: D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology; D020170:Caspase 1 / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism / Q000473:pathology; D020219:Chondrogenesis / Q000502:physiology*; D056587:Cryopyrin-Associated Periodic Syndromes / Q000235:genetics / Q000473:pathology* / Q000503:physiopathology*; D000242:Cyclic AMP / Q000502:physiology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D058847:Inflammasomes / Q000737:chemistry* / Q000502:physiology; D009154:Mutation / Q000235:genetics; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000473:pathology*; D055758:SOX9 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology
chemical_list: C115163:CREB1 protein, human; D002352:Carrier Proteins; D017362:Cyclic AMP Response Element-Binding Protein; D006025:Glycosaminoglycans; D058847:Inflammasomes; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D020170:Caspase 1