Analyzing immune response to engineered hydrogels by hierarchical clustering of inflammatory cell subsets

• Type of Material: Article
• Authors: Marc A Fernandez-Yague, Emory University; Lauren A Hymel, Emory University; Claire E Olingy, Emory University; Claire McClain, Emory University; Molly E Ogle, Emory University; José R García, Georgia Institute of Technology; Dustin Minshew, Emory University; Sofiya Vyshnya, Emory University; Hong Seo Lim, Emory University; Peng Qiu, Emory University; Andrés J García, Georgia Institute of Technology; Edward Botchwey, Emory University
• Language: English
• Date: 2022-02-01
• Publisher: AAAS
• Publication Version: Final Published Version
• Copyright Statement: © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1126/sciadv.abd8056
• Title of Journal or Parent Work: Science Advances
• Volume: 8
• Issue: 8
• Start Page: eabd8056
• End Page: eabd8056
• Grant/Funding Information: We acknowledge funding from the National Institutes of Health, the National Institute of Arthritis and Musculoskeletal and Skin Diseases, and National Institute of Dental and Craniofacial Research under awards R01 AR062920 (A.J.G.), R01 AR062368 (A.J.G.), R01 EB024322 (A.J.G.), R56 AR071708 (E.A.B.), and R56 DE029703 (E.A.B. and A.J.G.). M.A.F.-Y. was funded by a Marie Skłodowska-Curie Individual Fellowship (grant no. 898737). L.A.H. was funded by a National Science Foundation Graduate Research Fellowship (grant no. DGE-1650044). C.M. was supported by the NIGMS-sponsored Cell and Tissue Engineering NIH Biotechnology Training Grant (grant no. T32 GM-008433). C.E.O. was funded by American Heart Association Predoctoral Fellowship (grant no. 15PRE25090024).
• Keywords: Cytokines; Biocompatible Materials; Oligopeptides; Immunity; Proteomics; Hydrogels; Cluster Analysis; Vascular Endothelial Growth Factor A
• Research Categories: Engineering, Biomedical; Engineering, Mechanical

Abstract

Understanding the immune response to hydrogel implantation is critical for the design of immunomodulatory biomaterials. To study the progression of inflammation around poly(ethylene glycol) hydrogels presenting Arg-Gly-Asp (RGD) peptides and vascular endothelial growth factor, we used temporal analysis of high-dimensional flow cytometry data paired with intravital imaging, immunohistochemistry, and multiplexed proteomic profiling. RGD-presenting hydrogels created a reparative microenvironment promoting CD206+ cellular infiltration and revascularization in wounded dorsal skin tissue. Unbiased clustering algorithms (SPADE) revealed significant phenotypic transition shifts as a function of the cell-adhesion hydrogel properties. SPADE identified an intermediate macrophage subset functionally regulating in vivo cytokine secretion that was preferentially recruited for RGD-presenting hydrogels, whereas dendritic cell subsets were preferentially recruited to RDG-presenting hydrogels. Last, RGD-presenting hydrogels controlled macrophage functional cytokine secretion to direct polarization and vascularization. Our studies show that unbiased clustering of single-cell data provides unbiased insights into the underlying immune response to engineered materials.