Tubule-derived CCN1 drives renal repair via αvβ5-STAT6-ARG1-dependent reprogramming of macrophages

Macrophages play a critical role in injury and repair following acute kidney injury (AKI), but their regulatory mechanisms remain incompletely understood. Cellular communication network factor 1 (CCN1), a secreted matricellular protein and early biomarker of AKI, may regulate macrophage function during kidney injury. In this study, we first investigated CCN1's interaction with macrophages in a murine model of ischemia-reperfusion (I/R)-induced AKI. The role of CCN1 was further investigated using recombinant protein administration, a renal tubular epithelial cell (RTEC)-specific CCN1 knockdown mouse model via adeno-associated virus, and in vitro studies with bone marrow-derived macrophages (BMDMs). We found that in response to injury, RTECs upregulated and secreted CCN1, which colocalized with infiltrating F4/80⁺ macrophages. RTEC-specific CCN1 knockdown exacerbated renal injury and reduced macrophage infiltration, as confirmed by H&E, KIM-1 and F4/80 staining. Transcriptomic profiling of kidney tissues revealed that CCN1 expression was associated with immune cell infiltration, particularly macrophages, while RNA-seq analysis of BMDMs demonstrated that CCN1 promoted pro-repair arginase1 (Arg1)⁺ macrophage differentiation and activated the STAT6 signaling pathway. BayesPrism deconvolution further confirmed the enrichment of Arg1^hi macrophages following CCN1 treatment. Co-immunoprecipitation coupled with mass spectrometry identified Integrin α_vβ₅ as a direct CCN1-binding partner mediating STAT6/ARG1 activation. Functionally, CCN1-treated macrophages enhanced RTECs' proliferation in vitro and in vivo, an effect abolished by ARG1 inhibition or macrophage depletion. In conclusion, CCN1 regulates macrophages via the α_vβ₅-STAT6-ARG1 axis to promote tubular epithelial proliferation and improve kidney function in I/R-AKI, highlighting a novel tubular-immune communication pathway and potential therapeutic targets for ischemic AKI.