ADAMTS12 promotes fibrosis by restructuring extracellular matrix to enable activation of injury-responsive fibroblasts
Konrad Hoeft, … , Sikander Hayat, Rafael Kramann
Konrad Hoeft, … , Sikander Hayat, Rafael Kramann
Published September 17, 2024
Citation Information: J Clin Invest. 2024;134(18):e170246. https://doi.org/10.1172/JCI170246.
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Research Article Cardiology Nephrology

ADAMTS12 promotes fibrosis by restructuring extracellular matrix to enable activation of injury-responsive fibroblasts

Abstract

Fibrosis represents the uncontrolled replacement of parenchymal tissue with extracellular matrix (ECM) produced by myofibroblasts. While genetic fate-tracing and single-cell RNA-Seq technologies have helped elucidate fibroblast heterogeneity and ontogeny beyond fibroblast to myofibroblast differentiation, newly identified fibroblast populations remain ill defined, with respect to both the molecular cues driving their differentiation and their subsequent role in fibrosis. Using an unbiased approach, we identified the metalloprotease ADAMTS12 as a fibroblast-specific gene that is strongly upregulated during active fibrogenesis in humans and mice. Functional in vivo KO studies in mice confirmed that Adamts12 was critical during fibrogenesis in both heart and kidney. Mechanistically, using a combination of spatial transcriptomics and expression of catalytically active or inactive ADAMTS12, we demonstrated that the active protease of ADAMTS12 shaped ECM composition and cleaved hemicentin 1 (HMCN1) to enable the activation and migration of a distinct injury-responsive fibroblast subset defined by aberrant high JAK/STAT signaling.

Authors

Konrad Hoeft, Lars Koch, Susanne Ziegler, Ling Zhang, Steffen Luetke, Maria C. Tanzer, Debashish Mohanta, David Schumacher, Felix Schreibing, Qingqing Long, Hyojin Kim, Barbara M. Klinkhammer, Carla Schikarski, Sidrah Maryam, Mathijs Baens, Juliane Hermann, Sarah Krieg, Fabian Peisker, Laura De Laporte, Gideon J.L. Schaefer, Sylvia Menzel, Joachim Jankowski, Benjamin D. Humphreys, Adam Wahida, Rebekka K. Schneider, Matthias Versele, Peter Boor, Matthias Mann, Gerhard Sengle, Sikander Hayat, Rafael Kramann

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Figure 6

Rescue of ADAMTS12 KO by overexpression of catalytically active or inactive ADAMTS12.

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Rescue of ADAMTS12 KO by overexpression of catalytically active or inact...
(A) Sequencing of ADAMTS12 expression plasmids with a WT active (Act) or a mutated inactive (Inact) catalytic domain. Seq, original ADAMTS12 sequence; Act, active catalytic domain; Inact, inactive catalytic domain. (B) Western blot for HA, tubulin, and GFP in ADAMTS12-KO, active or inactive ADAMTS12-expressing PDGFRβ+ cells. (C) Trajectory maps and quantification of the average migration speed of ADAMTS12-KO and active and inactive ADAMTS12-expressing PDGFRβ+ cells (n = 16 per group, pKO vs. Act< 0.0001, PAct vs. Inact = 0.0002, by ordinary 1-way ANOVA). Results were reproducible in 3 independent experiments. (D) PROGENy pathway analysis based on the DEGs shown in Supplemental Figure 6, G–I. ActvsKO, catalytically active ADAMTS12 expression versus ADAMTS12-KO; ActvsInact, catalytically active versus inactive ADAMTS12 expression; InactvsKO, catalytically inactive ADAMTS12 expression versus ADAMTS12-KO. (E) Top enriched biological process GO terms based on the top upregulated genes shown in Supplemental Figure 6, G–I. Comparisons are described in D (for abbreviations, see Supplemental Table 14). (F) ADAMTS12 active versus KO signature (Act vs. KO) in a scRNA-Seq framework of murine cardiac fibroblasts in heart failure. (G) ADAMTS12 active versus inactive signature (Act vs. Inact) in the above dataset. ***P < 0.001 and ****P < 0.0001. For C, F, and G, a 1-way ANOVA with Tukey’s post hoc test was performed.