Dynamic fibroblast-immune interactions shape recovery after brain injury [snRNASeq_Tgfb]

Fibroblasts and immune cells coordinate tissue regeneration and necessary scarring after injury. In the brain, fibroblasts are border-enriched cells whose dynamic molecular states and immune interactions after injury remain unclear. We used single nuclear RNA sequencing at multiple timepoints after photothrombotic (PT) injury to test the role of TGFb in regulating fibroblast and macrophage responses to brain injury. We find that inhibition of fibroblast TGFb signaling, or of TGFb activation, impairs profibrotic fibroblast (and macrophage) expansion and function. Overall design: Tissue was collected from wildtype mice, mice with TGFb-blind fibroblasts [Col1a2(creER); Tgfbr2(flox)], or mice treated with ADWA11 (anti-integrin avb8, 0/7dpi). Bulk lesional nuclei (DAPI+) were isolated and sort-purified from mice at 7 or 21 days post injury (dpi) for each condition. 1 male and 1 female mouse were harvested per timepoint/condition and were processed together.