scRNA-Seq Identifies IL-1 Responsive Cell Subsets in the Skin Injury-induced Inflammatory Response

Authors

  • Kayla Harpold Indiana University School of Medicine
  • Hong-Ming Zhou Indiana University School of Medicine
  • Radomir Slominski Indiana University School of Medicine
  • Leroy Seymour Indiana University School of Medicine
  • Maria Bell Indiana University School of Medicine
  • Priya Dave Life-Health Science Internship Program, IUPUI
  • Joseph Atumonye Indiana University School of Medicine
  • William Wright III Life-Health Science Internship Program, IUPUI
  • Avery Dawes Indiana University School of Medicine
  • Brad Griesenauer Indiana University School of Medicine
  • Sophie Paczesny Indiana University School of Medicine
  • Mark Kaplan Indiana University School of Medicine
  • Dan Spandau Indiana University School of Medicine
  • Yunglong Liu Indiana University School of Medicine
  • Xiaoling Xuei Indiana University School of Medicine
  • Hongyu Gao Indiana University School of Medicine
  • Aki Hoki Indiana University School of Medicine
  • Matthew Turner Indiana University School of Medicine

DOI:

https://doi.org/10.18060/24623

Abstract

Inflammation is an integral aspect of skin wound healing; however, the mechanisms that regulate inflammatory cascades in this context are not well defined. To better understand how skin inflammation impacts wound healing, we developed an ex vivo skin culture system to model key aspects of the inflammatory phase of wound healing. In this model, a defined set of proinflammatory cytokines and chemokines, mirroring those produced in wounds in vivo, are produced when mouse or human skin biopsies are cultured ex vivo.  We refer to this pattern of cytokine and chemokine induction as the skin injury-induced inflammatory response. Previous studies in our laboratory demonstrated this response is initiated by the cytokine, interleukin 1 alpha (IL-1α). To understand the cellular sources and targets of IL-1α during the skin injury-induced inflammatory response, skin biopsies from mouse tail skin were cultured ex vivo for 8 hours followed by processing for single cell RNA sequencing (scRNAseq). Using bioinformatic software, R, and the package, Seurat, analysis of scRNAseq data from this experiment identified 22 distinct cell population clusters. While no populations exhibited significant expression of Il1a transcripts, multiple cell populations expressed Il1r1 transcripts, which encodes the ligand-specific subunit of the IL-1 receptor.  Notably, fibroblast, endothelial cell and stromal cell clusters were characterized by expression of Il1r1 and the skin injury-induced inflammatory response transcripts Il6, Cxcl1 and/or Csf3. Furthermore, Reactome Pathway Analysis suggested the Il-1 signaling axis was activated in these cell populations. This information provides a basis for future studies to understand how IL-1 signaling in fibroblasts, endothelial cells and stromal cells impacts wound healing in vivo, which could in turn lead to novel therapeutic approaches to clinically relevant outcomes.  

Author Biographies

Hong-Ming Zhou, Indiana University School of Medicine

Departments of Dermatology

Radomir Slominski, Indiana University School of Medicine

Departments of Dermatology

Leroy Seymour, Indiana University School of Medicine

Departments of Dermatology

Avery Dawes, Indiana University School of Medicine

Departments of Dermatology

Brad Griesenauer, Indiana University School of Medicine

Microbiology & Immunology

Melvin and Bren Simon Cancer Center

Sophie Paczesny, Indiana University School of Medicine

Microbiology & Immunology

Melvin and Bren Simon Cancer Center

Mark Kaplan, Indiana University School of Medicine

Microbiology & Immunology

Biochemistry & Molecular Biology

Dan Spandau, Indiana University School of Medicine

Departments of Dermatology

Biochemistry & Molecular Biology

Yunglong Liu, Indiana University School of Medicine

Center for Computation Biology and Bioinformatics

Medical & Molecular Genetics

Biostatistics

Informatics and Computing

Xiaoling Xuei, Indiana University School of Medicine

Medical & Molecular Genetics

Hongyu Gao, Indiana University School of Medicine

Medical & Molecular Genetics

Aki Hoki, Indiana University School of Medicine

Pediatrics

Matthew Turner, Indiana University School of Medicine

Departments of Dermatology

Microbiology & Immunology

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Published

2020-12-15

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Abstracts