OBJECTIVE: The development of precision therapeutics for systemic sclerosis (SSc) has been hindered by the lack of models that accurately mimic the disease in vitro. This study was undertaken to design and test a self-assembled skin equivalent (saSE) system that recapitulates the cross-talk between macrophages and fibroblasts in cutaneous fibrosis. METHODS: SSc-derived dermal fibroblasts (SScDFs) and normal dermal fibroblasts (NDFs) were cultured with CD14+ monocytes from SSc patients or healthy controls to allow de novo stroma formation. Monocyte donor-matched plasma was introduced at week 3 prior to seeding keratinocytes to produce saSE with a stratified epithelium. Tissue was characterized by immunohistochemical staining, atomic force microscopy, enzyme-linked immunosorbent assay, and quantitative reverse transcriptase-polymerase chain reaction. RESULTS: Stroma synthesized de novo from NDFs and SScDFs supported a fully stratified epithelium to form saSE. A thicker and stiffer dermis was generated by saSE with SScDFs, and more interleukin-6 and transforming growth factor β (TGFβ) was secreted by saSE with SScDFs compared to saSE with NDFs, regardless of the inclusion of monocytes. Tissue with SSc monocytes and plasma had amplified dermal thickness and stiffness relative to control tissue. Viable CD163+ macrophages were found within the stroma of saSE 5 weeks after seeding. Additionally, SSc saSE contained greater numbers of CD163+ and CD206+ macrophages compared to control saSE. TGFβ blockade inhibited stromal stiffness to a greater extent in SSc saSE compared to control saSE. CONCLUSION: These data suggest reciprocal activation between macrophages and fibroblasts that increases tissue thickness and stiffness, which is dependent in part on TGFβ activation. The saSE system may serve as a platform for preclinical therapeutic testing and for molecular characterization of SSc skin pathology through recapitulation of the interactions between macrophages and fibroblasts.
OBJECTIVE: The development of precision therapeutics for systemic sclerosis (SSc) has been hindered by the lack of models that accurately mimic the disease in vitro. This study was undertaken to design and test a self-assembled skin equivalent (saSE) system that recapitulates the cross-talk between macrophages and fibroblasts in cutaneous fibrosis. METHODS: SSc-derived dermal fibroblasts (SScDFs) and normal dermal fibroblasts (NDFs) were cultured with CD14+ monocytes from SSc patients or healthy controls to allow de novo stroma formation. Monocyte donor-matched plasma was introduced at week 3 prior to seeding keratinocytes to produce saSE with a stratified epithelium. Tissue was characterized by immunohistochemical staining, atomic force microscopy, enzyme-linked immunosorbent assay, and quantitative reverse transcriptase-polymerase chain reaction. RESULTS: Stroma synthesized de novo from NDFs and SScDFs supported a fully stratified epithelium to form saSE. A thicker and stiffer dermis was generated by saSE with SScDFs, and more interleukin-6 and transforming growth factor β (TGFβ) was secreted by saSE with SScDFs compared to saSE with NDFs, regardless of the inclusion of monocytes. Tissue with SSc monocytes and plasma had amplified dermal thickness and stiffness relative to control tissue. Viable CD163+ macrophages were found within the stroma of saSE 5 weeks after seeding. Additionally, SSc saSE contained greater numbers of CD163+ and CD206+ macrophages compared to control saSE. TGFβ blockade inhibited stromal stiffness to a greater extent in SSc saSE compared to control saSE. CONCLUSION: These data suggest reciprocal activation between macrophages and fibroblasts that increases tissue thickness and stiffness, which is dependent in part on TGFβ activation. The saSE system may serve as a platform for preclinical therapeutic testing and for molecular characterization of SSc skin pathology through recapitulation of the interactions between macrophages and fibroblasts.
Authors: Loubna Chadli; Britt Sotthewes; Kejie Li; Stefan N Andersen; Ellen Cahir-McFarland; Marc Cheung; Patrick Cullen; Annemarie Dorjée; Jeska K de Vries-Bouwstra; Tom W J Huizinga; David F Fischer; Jeroen DeGroot; Joanne L Viney; Timothy S Zheng; Jamil Aarbiou; Agnes Gardet Journal: Sci Rep Date: 2019-03-14 Impact factor: 4.379
Authors: Jennifer L Sargent; Zhenghui Li; Antonios O Aliprantis; Matthew Greenblatt; Raphael Lemaire; Ming-Hua Wu; Jun Wei; Jaclyn Taroni; Adam Harris; Kristen B Long; Chelsea Burgwin; Carol M Artlett; Elizabeth P Blankenhorn; Robert Lafyatis; John Varga; Stephen H Clark; Michael L Whitfield Journal: Arthritis Rheumatol Date: 2016-08 Impact factor: 10.995
Authors: Alexander V Misharin; Luisa Morales-Nebreda; Paul A Reyfman; Carla M Cuda; James M Walter; Alexandra C McQuattie-Pimentel; Ching-I Chen; Kishore R Anekalla; Nikita Joshi; Kinola J N Williams; Hiam Abdala-Valencia; Tyrone J Yacoub; Monica Chi; Stephen Chiu; Francisco J Gonzalez-Gonzalez; Khalilah Gates; Anna P Lam; Trevor T Nicholson; Philip J Homan; Saul Soberanes; Salina Dominguez; Vince K Morgan; Rana Saber; Alexander Shaffer; Monique Hinchcliff; Stacy A Marshall; Ankit Bharat; Sergejs Berdnikovs; Sangeeta M Bhorade; Elizabeth T Bartom; Richard I Morimoto; William E Balch; Jacob I Sznajder; Navdeep S Chandel; Gökhan M Mutlu; Manu Jain; Cara J Gottardi; Benjamin D Singer; Karen M Ridge; Neda Bagheri; Ali Shilatifard; G R Scott Budinger; Harris Perlman Journal: J Exp Med Date: 2017-07-10 Impact factor: 14.307
Authors: Mareike Lehmann; Lara Buhl; Hani N Alsafadi; Stephan Klee; Sarah Hermann; Kathrin Mutze; Chiharu Ota; Michael Lindner; Jürgen Behr; Anne Hilgendorff; Darcy E Wagner; Melanie Königshoff Journal: Respir Res Date: 2018-09-15