64c34db83828069d4a9a762d310b9645 pmc_33924264.pdf 965668770424a854f87b49bb7ea0922d8b2fc69e pmc_33924264.pdf 626d420857a7405cf921f5451c41f4a473d26caba6cbb286d5592fd3aa63b4f5 pmc_33924264.pdf Title: Crohn’s Disease Increases the Mesothelial Properties of Adipocyte Progenitors in the Creeping Fat Subject: Our understanding of the interplay between human adipose tissue and the immune system is limited. The mesothelium, an immunologically active structure, emerged as a source of visceral adipose tissue. After investigating the mesothelial properties of human visceral and subcutaneous adipose tissue and their progenitors, we explored whether the dysfunctional obese and Crohn’s disease environments influence the mesothelial/mesenchymal properties of their adipocyte precursors, as well as their ability to mount an immune response. Using a tandem transcriptomic/proteomic approach, we evaluated the mesothelial and mesenchymal expression profiles in adipose tissue, both in subjects covering a wide range of body-mass indexes and in Crohn’s disease patients. We also isolated adipose tissue precursors (adipose-derived stem cells, ASCs) to assess their mesothelial/mesenchymal properties, as well as their antigen-presenting features. Human visceral tissue presented a mesothelial phenotype not detected in the subcutaneous fat. Only ASCs from mesenteric adipose tissue, named creeping fat, had a significantly higher expression of the hallmark mesothelial genes mesothelin (MSLN) and Wilms’ tumor suppressor gene 1 (WT1), supporting a mesothelial nature of these cells. Both lean and Crohn’s disease visceral ASCs expressed equivalent surface percentages of the antigen-presenting molecules human leucocyte antigen—DR isotype (HLA-DR) and CD86. However, lean-derived ASCs were predominantly HLA-DR dim, whereas in Crohn’s disease, the HLA-DR bright subpopulation was increased 3.2-fold. Importantly, the mesothelial-enriched Crohn’s disease precursors activated CD4+ T-lymphocytes. Our study evidences a mesothelial signature in the creeping fat of Crohn’s disease patients and its progenitor cells, the latter being able to present antigens and orchestrate an immune response. Keywords: Crohn’s disease; adipose tissue; mesothelium; adipose-derived stem cells Author: Ana Madeira, Carolina Serena, Miriam Ejarque, Elsa Maymó-Masip, Monica Millan, M. Carmen Navarro-Ruiz, Rocío Guzmán-Ruiz, María M. Malagón, Eloy Espin, Marc Martí, Margarita Menacho, Ana Megía, Joan Vendrell and Sonia Fernández-Veledo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Apr 21 08:29:26 2021 CEST ModDate: Wed Apr 21 08:33:28 2021 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2466013 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PRJKGQ+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 KORLVE+VnURWPalladioL Type 1 Custom yes yes no 15 0 ZNZRVT+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 NPMMTL+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 ROQDDT+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 36 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 39 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 61 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 65 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 69 0 WABLSD+SFRM1000 Type 1 Custom yes yes no 81 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 97 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 101 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 105 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 109 0 COGLGI+CMSY10 Type 1 Builtin yes yes no 121 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 129 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 133 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 137 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 141 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 157 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 161 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 165 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 169 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 185 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 189 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 193 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 197 0 OQIVQT+EURM10 Type 1 Builtin yes yes no 209 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 217 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 221 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 225 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 229 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 245 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 249 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 253 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 257 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 03:19:59 CEST RepresentationInformation: pmc_33924264.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:09:52 CEST Size: 2466013 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 542 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: A Mesothelial Gene Signature Distinguishes Visceral Adipose Tissue and Its Precursors from Subcutaneous Adipose Tissue Destination: subsection.2.1 Item: Title: Creeping Fat Precursor Cells Have Mesothelial-Enriched Properties Destination: subsection.2.2 Item: Title: Creeping Fat-Derived ASCs Have an Altered Immunophenotypic Profile and Act as Antigen-Presenting Cells Destination: subsection.2.3 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Study Subjects Destination: subsection.4.1 Item: Title: Adipose Stem Cell Isolation and Cell Culture Destination: subsection.4.2 Item: Title: Immunophenotyping of ASCs Destination: subsection.4.3 Item: Title: Gene Expression Analysis Destination: subsection.4.4 Item: Title: Proteomics Destination: subsection.4.5 Item: Title: Antigen Presentation Assay between ASCs and Autologous CD4+ T-lymphocytes Destination: subsection.4.6 Item: Title: Statistical Analysis Destination: subsection.4.7 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Crohn’s Disease Increases the Mesothelial Properties of Adipocyte Progenitors in the Creeping Fat Author: Ana Madeira, Carolina Serena, Miriam Ejarque, Elsa Maymó-Masip, Monica Millan, M. Carmen Navarro-Ruiz, Rocío Guzmán-Ruiz, María M. Malagón, Eloy Espin, Marc Martí, Margarita Menacho, Ana Megía, Joan Vendrell and Sonia Fernández-Veledo Subject: Our understanding of the interplay between human adipose tissue and the immune system is limited. The mesothelium, an immunologically active structure, emerged as a source of visceral adipose tissue. After investigating the mesothelial properties of human visceral and subcutaneous adipose tissue and their progenitors, we explored whether the dysfunctional obese and Crohn’s disease environments influence the mesothelial/mesenchymal properties of their adipocyte precursors, as well as their ability to mount an immune response. Using a tandem transcriptomic/proteomic approach, we evaluated the mesothelial and mesenchymal expression profiles in adipose tissue, both in subjects covering a wide range of body-mass indexes and in Crohn’s disease patients. We also isolated adipose tissue precursors (adipose-derived stem cells, ASCs) to assess their mesothelial/mesenchymal properties, as well as their antigen-presenting features. Human visceral tissue presented a mesothelial phenotype not detected in the subcutaneous fat. Only ASCs from mesenteric adipose tissue, named creeping fat, had a significantly higher expression of the hallmark mesothelial genes mesothelin (MSLN) and Wilms’ tumor suppressor gene 1 (WT1), supporting a mesothelial nature of these cells. Both lean and Crohn’s disease visceral ASCs expressed equivalent surface percentages of the antigen-presenting molecules human leucocyte antigen DR isotype (HLA-DR) and CD86. However, lean-derived ASCs were predominantly HLA-DR dim, whereas in Crohn’s disease, the HLA-DR bright subpopulation was increased 3.2-fold. Importantly, the mesothelial-enriched Crohn’s disease precursors activated CD4+ T-lymphocytes. Our study evidences a mesothelial signature in the creeping fat of Crohn’s disease patients and its progenitor cells, the latter being able to present antigens and orchestrate an immune response. 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