HCK induces macrophage activation to promote renal inflammation and fibrosis via suppression of autophagy

Abstract

Renal inflammation and fibrosis are the common pathways leading to progressive chronic kidney disease (CKD). We previously identified hematopoietic cell kinase (HCK) as upregulated in human chronic allograft injury promoting kidney fibrosis; however, the cellular source and molecular mechanisms are unclear. Here, using immunostaining and single cell sequencing data, we show that HCK expression is highly enriched in pro-inflammatory macrophages in diseased kidneys. HCK-knockout (KO) or HCK-inhibitor decreases macrophage M1-like pro-inflammatory polarization, proliferation, and migration in RAW264.7 cells and bone marrow-derived macrophages (BMDM). We identify an interaction between HCK and ATG2A and CBL, two autophagy-related proteins, inhibiting autophagy flux in macrophages. In vivo, both global or myeloid cell specific HCK-KO attenuates renal inflammation and fibrosis with reduces macrophage numbers, pro-inflammatory polarization and migration into unilateral ureteral obstruction (UUO) kidneys and unilateral ischemia reperfusion injury (IRI) models. Finally, we developed a selective boron containing HCK inhibitor which can reduce macrophage pro-inflammatory activity, proliferation, and migration in vitro, and attenuate kidney fibrosis in the UUO mice. The current study elucidates mechanisms downstream of HCK regulating macrophage activation and polarization via autophagy in CKD and identifies that selective HCK inhibitors could be potentially developed as a new therapy for renal fibrosis.

Comprehensive analysis of the HCK gene in myeloid neoplasms: Insights into biological functions, prognosis, and response to antineoplastic agents

“… 27 In this present study, the increase in HCK expression in the myeloid compartment was validated in experiments using cell lines and suggests that HCK participates in signal transduction and functions in immune system cells. In this context, HCK has been associated with macrophage activation, 28 neutrophil-mediated autoinflammation, 29 tumor progression through myeloid infiltrates in solid tumors, 30 or even participating in the response to immunotherapy through T cell infiltration. 31 Thus, the role of HCK must be interpreted in a context-dependent manner.…”

Single-cell RNA sequencing of the holothurian regenerating intestine reveals the pluripotency of the coelomic epithelium

“…Mainly because these molecules have been vastly studied across invertebrates, and multiple immune roles have been proposed, including phagocyte recognition and encapsulation (Hanington and Zhang, 2011). A distinct example is that of C10, where the expression of HCK , FER and ITF8 markers suggest this might be a macrophage-like activity (Chen et al, 2023; Dolgachev et al, 2018; Shuttleworth, 2018). Specifically, HCK , a member of Src family kinases (SFK), has been closely related to macrophage activation and polarization (Bhattacharjee et al, 2011; Poh et al, 2015).…”

NLRX1 Prevents M2 Macrophage Polarization and Excessive Renal Fibrosis in Chronic Obstructive Nephropathy

“…It has been well investigated that macrophages react heterogeneously to the kidney microenvironment and assume different metabolic and phenotypic changes during kidney diseases [ 43 , 44 ]. The polarization of macrophages to a pro-fibrotic M2 phenotype is considered to play vital role during the development of renal fibrosis by producing and secreting TGFβ and fuelling a fibrotic kidney microenvironment [ 10 ].…”