Recently profound therapeutic efficacy of intratracheally tr

Recently, profound therapeutic efficacy of intratracheally transplanted bone marrow-derived Mφ has been demonstrated in two murine disease models of herPAP, indicating a therapeutic potential also of iPSC-derived Mφ (Happle et al., 2014; Suzuki et al., 2014a). In this context genetically repaired human herPAP patient-specific iPSCs lines already have been generated, and functional Mφ derived from these lines potentially represent an autologous cell source for herPAP therapy (Lachmann et al., 2014; Suzuki et al., 2014b). Moreover, in a murine disease model of adenosine deaminase deficiency, therapeutic benefit has been demonstrated following the intrapulmonary administration of iPSC-derived AM-like cells with a characteristic F4/80+ CD11c+, SIGLEC-F+ surface phenotype and an AM-typical gene-expression pattern (Litvack et al., 2016). On this background pulmonary macrophage transfer of autologous gene-corrected iPSC-Mφ may represent an innovative therapeutic option for other disease entities associated with malfunction of AMs such as chronic obstructive pulmonary disease (Hodge et al., 2003), bronchopulmonary dysplasia (Blackwell et al., 2011), or cystic fibrosis (Bruscia et al., 2009; Di et al., 2006). Moreover, site-specific transplantation of iPSC-Mφ into organs such as the liver or the buy Manumycin A may turn out to be relevant also for other diseases associated with Mφ malfunction such as heme oxygenase-1 deficiency (Kovtunovych et al., 2014) or adrenoleukodystrophy (Okamura et al., 2009). In summary, we here describe a highly robust and efficient protocol for the generation of murine iPSC-Mφ, which phenotypically and functionally show high similarity to their primary Mφ counterparts. Moreover, we employed this protocol to obtain disease-specific iPSC-Mφ from herPAP mice and recapitulated in vitro the hallmark Mφ phenotype of this disorder. Thus, this study introduces a unique tool for the study of Mφ and iPSC-Mφ biology in healthy and disease contexts, provides an in vitro disease model to study CSF2RB-deficient herPAP, and may eventually contribute a platform upon which to develop advanced therapeutic approaches in herPAP or other severe Mφ-related disease entities.
Experimental Procedures
Author Contributions
Acknowledgments The authors thank M. Ballmaier and C. Struckmann (Cell Sorting Facility, Hannover Medical School) for scientific support and D. Lüttge and T. Buchegger for experimental and technical support. In addition, we would like to thank Elena Lopez-Rodriguez for performing immunofluorescence staining of lung sections. This work was supported by grants from the Else Kröner-Fresenius-Stiftung, the Deutsche Forschungsgemeinschaft (Cluster of Excellence REBIRTH, Exc62/1, grants MO 886/6-1, LA 3680/2-1, and the SFB738), the Bundesministerium für Bildung und Forschung (BMBF, project PidNet), and the German Center for Lung Research and Hannover Medical School internal programs (Hochschulinterne Leistungsförderung [HiLF] and Young Academy).