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Angiogenesis array data for “Identification of a pro-angiogenic functional role for FSP1-positive fibroblast subtype in wound healing”

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modified on 2019-08-16, 09:48

Abstract


Fibrosis accompanying wound healing can drive the failure of many different organs. Activated fibroblasts are the principal determinants of post-injury pathological fibrosis along with physiological repair, making them a difficult therapeutic target. Although activated fibroblasts are phenotypically heterogeneous, they are not recognized as distinct functional entities. Using mice that express GFP under the FSP1 or αSMA promoter, we characterized two non-overlapping fibroblast subtypes from mouse hearts after myocardial infarction. Here, we report the identification of FSP1-GFP+ cells as a non-pericyte, non-hematopoietic fibroblast subpopulation with a predominant pro-angiogenic role, characterized by in vitro phenotypic/cellular/ultrastructural studies and in vivo granulation tissue formation assays combined with transcriptomics and proteomics. This work identifies a fibroblast subtype that is functionally distinct from the pro-fibrotic αSMA-expressing myofibroblast subtype. Our study has the potential to shift our focus towards viewing fibroblasts as molecularly and functionally heterogeneous and provides a paradigm to approach treatment for organ fibrosis.

Angiogenesis protein array

The Proteome Profiler Mouse Angiogenesis Array Kit (# ARY015; R & D) was used to assess the relative levels of 53 mouse angiogenesis-related proteins in 300 µg of FSP1+ and αSMA+ cell lysates according to manufacturer’s instructions. Angiogenesis Array is a membrane-based sandwich immunoassay. The cells were cultured for 72 h in 0.5% FBS prior to protein collection. Briefly, a cocktail of biotinylated detection antibodies was mixed with cell lysates. The array membrane was then treated with the cell lysate and antibody mixture overnight at 4 °C, washed, and then incubated with streptavidin-HRP. The array membrane is spotted with capture antibodies to specific target proteins. The captured proteins were visualized using chemiluminescence.



Funding

Elucidating fibroblast heterogeneity as a pathway to target organ fibrosis

National Institute of General Medical Sciences

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Thermosensitive injectable polymer-based stem cell therapy for limb ischemia

National Institute of Biomedical Imaging and Bioengineering

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17SDG33630187 American Heart Association Career Development Grant