RATIONALE: In patients with pulmonary alveolar proteinosis (PAP) syndrome, disruption of granulocyte/macrophage colony-stimulating factor (GM-CSF) signaling is associated with pathogenic surfactant accumulation from impaired clearance in alveolar macrophages. OBJECTIVES: The aim of this study was to overcome these barriers by using monocyte-derived induced pluripotent stem (iPS) cells to recapitulate disease-specific and normal macrophages. METHODS: We created iPS cells from two children with hereditary PAP (hPAP) caused by recessive CSF2RA(R217X) mutations and three normal people, differentiated them into macrophages (hPAP-iPS-Mφs and NL-iPS-Mφs, respectively), and evaluated macrophage functions with and without gene-correction to restore GM-CSF signaling in hPAP-iPS-Mφs. MEASUREMENTS AND MAIN RESULTS: Both hPAP and normal iPS cells had human embryonic stem cell-like morphology, expressed pluripotency markers, formed teratomas in vivo, had a normal karyotype, retained and expressed mutant or normal CSF2RA genes, respectively, and could be differentiated into macrophages with the typical morphology and phenotypic markers. Compared with normal, hPAP-iPS-Mφs had impaired GM-CSF receptor signaling and reduced GM-CSF-dependent gene expression, GM-CSF- but not M-CSF-dependent cell proliferation, surfactant clearance, and proinflammatory cytokine secretion. Restoration of GM-CSF receptor signaling corrected the surfactant clearance abnormality in hPAP-iPS-Mφs. CONCLUSIONS: We used patient-specific iPS cells to accurately reproduce the molecular and cellular defects of alveolar macrophages that drive the pathogenesis of PAP in more than 90% of patients. These results demonstrate the critical role of GM-CSF signaling in surfactant homeostasis and PAP pathogenesis in humans and have therapeutic implications for hPAP.
RATIONALE: In patients with pulmonary alveolar proteinosis (PAP) syndrome, disruption of granulocyte/macrophage colony-stimulating factor (GM-CSF) signaling is associated with pathogenic surfactant accumulation from impaired clearance in alveolar macrophages. OBJECTIVES: The aim of this study was to overcome these barriers by using monocyte-derived induced pluripotent stem (iPS) cells to recapitulate disease-specific and normal macrophages. METHODS: We created iPS cells from two children with hereditary PAP (hPAP) caused by recessive CSF2RA(R217X) mutations and three normal people, differentiated them into macrophages (hPAP-iPS-Mφs and NL-iPS-Mφs, respectively), and evaluated macrophage functions with and without gene-correction to restore GM-CSF signaling in hPAP-iPS-Mφs. MEASUREMENTS AND MAIN RESULTS: Both hPAP and normal iPS cells had human embryonic stem cell-like morphology, expressed pluripotency markers, formed teratomas in vivo, had a normal karyotype, retained and expressed mutant or normal CSF2RA genes, respectively, and could be differentiated into macrophages with the typical morphology and phenotypic markers. Compared with normal, hPAP-iPS-Mφs had impaired GM-CSF receptor signaling and reduced GM-CSF-dependent gene expression, GM-CSF- but not M-CSF-dependent cell proliferation, surfactant clearance, and proinflammatory cytokine secretion. Restoration of GM-CSF receptor signaling corrected the surfactant clearance abnormality in hPAP-iPS-Mφs. CONCLUSIONS: We used patient-specific iPS cells to accurately reproduce the molecular and cellular defects of alveolar macrophages that drive the pathogenesis of PAP in more than 90% of patients. These results demonstrate the critical role of GM-CSF signaling in surfactant homeostasis and PAP pathogenesis in humans and have therapeutic implications for hPAP.
Authors: Fangjun Jia; Kitchener D Wilson; Ning Sun; Deepak M Gupta; Mei Huang; Zongjin Li; Nicholas J Panetta; Zhi Ying Chen; Robert C Robbins; Mark A Kay; Michael T Longaker; Joseph C Wu Journal: Nat Methods Date: 2010-02-07 Impact factor: 28.547
Authors: Veronika Kleff; Ursula R Sorg; Carsten Bury; Takuji Suzuki; Ina Rattmann; Moran Jerabek-Willemsen; Christopher Poremba; Michael Flasshove; Bertram Opalka; Bruce Trapnell; Uta Dirksen; Thomas Moritz Journal: Mol Ther Date: 2008-03-04 Impact factor: 11.454
Authors: Junying Yu; Kejin Hu; Kim Smuga-Otto; Shulan Tian; Ron Stewart; Igor I Slukvin; James A Thomson Journal: Science Date: 2009-03-26 Impact factor: 47.728
Authors: Takuro Sakagami; Kanji Uchida; Takuji Suzuki; Brenna C Carey; Robert E Wood; Susan E Wert; Jeffrey A Whitsett; Bruce C Trapnell; Maurizio Luisetti Journal: N Engl J Med Date: 2009-12-31 Impact factor: 91.245
Authors: Mary Jane Thomassen; Barbara P Barna; Achut G Malur; Tracey L Bonfield; Carol F Farver; Anagha Malur; Heidi Dalrymple; Mani S Kavuru; Maria Febbraio Journal: J Lipid Res Date: 2007-09-11 Impact factor: 5.922
Authors: In-Hyun Park; Natasha Arora; Hongguang Huo; Nimet Maherali; Tim Ahfeldt; Akiko Shimamura; M William Lensch; Chad Cowan; Konrad Hochedlinger; George Q Daley Journal: Cell Date: 2008-08-07 Impact factor: 41.582
Authors: Takuji Suzuki; Takuro Sakagami; Bruce K Rubin; Lawrence M Nogee; Robert E Wood; Sarah L Zimmerman; Teresa Smolarek; Megan K Dishop; Susan E Wert; Jeffrey A Whitsett; Gregory Grabowski; Brenna C Carey; Carrie Stevens; Johannes C M van der Loo; Bruce C Trapnell Journal: J Exp Med Date: 2008-10-27 Impact factor: 14.307
Authors: Margarita Martinez-Moczygemba; Minh L Doan; Okan Elidemir; Leland L Fan; Sau Wai Cheung; Jonathan T Lei; James P Moore; Ghamartaj Tavana; Lora R Lewis; Yiming Zhu; Donna M Muzny; Richard A Gibbs; David P Huston Journal: J Exp Med Date: 2008-10-27 Impact factor: 14.307
Authors: Hong Ji; Xue Zhang; Sunghee Oh; Christopher N Mayhew; Ashley Ulm; Hari K Somineni; Mark Ericksen; James M Wells; Gurjit K Khurana Hershey Journal: J Allergy Clin Immunol Date: 2014-10-14 Impact factor: 10.793
Authors: Miriam Hetzel; Takuji Suzuki; Anna Rafiei Hashtchin; Paritha Arumugam; Brenna Carey; Marc Schwabbauer; Alexandra Kuhn; Johann Meyer; Axel Schambach; Johannes Van Der Loo; Thomas Moritz; Bruce C Trapnell; Nico Lachmann Journal: Hum Gene Ther Methods Date: 2017-08-30 Impact factor: 2.396