IL-1β-positive macrophages may contribute to inflammatory reprogramming of tumor cells and support pro-tumor features of IL-1R1+iCAFs in hepatocellular carcinoma: preliminary pharmacological and multi-omics evidence

Hepatocellular carcinoma (HCC) is one of the most lethal malignancies and severely threatens human health. Inflammatory and immunoregulatory signals co‑exist within the HCC tumor microenvironment (TME), impairing tissue repair and cytotoxic immune responses. Tumor‑associated macrophages(TAMs) exhibit substantial heterogeneity and exert critical disease‑related functions; nevertheless, their diversity, functional states and regulatory roles within the TME remain incompletely characterized. We performed deep‑depth single‑cell RNA sequencing on 37 tumor and paired paracancerous tissue samples collected from 20 patients with HCC. Patients were stratified into immune‑excluded-like subtype(IES) and immune‑infiltrated-like subtype(IIS) according to immune cell infiltration proportions in the TME. Published spatial transcriptomic datasets were integrated with our in‑house multiplex immunofluorescence and single‑cell RNA‑seq data to identify TAM and cancer‑associated fibroblast(CAF) subsets potentially linked to tumor progression and immunotherapy response. In‑vitro cellular assays provided preliminary pharmacological evidence for candidate signaling axes. Single‑cell profiling uncovered two distinct immune subtypes of HCC defined by TME cellular composition. Compared with IIS, the TME of IES was predominantly infiltrated by macrophages and stromal cells, accompanied by reduced infiltration of T‑ and B‑lymphocytes. Within the IES group, we identified an IL‑1β+ macrophage(Mac) subset with prominent pro‑inflammatory signatures. Multi‑omics analyses suggest potential crosstalk between IL‑1β+Mac and tumor cells via the PGE2‑IL‑1β axis, which might favor tumor progression. Our preliminary in‑vitro pharmacological observations further hint that IL‑1β+Mac‑derived signals could support pro‑tumor properties of inflammation‑associated CAF (iCAFs) through the IL‑1β/IL‑1R1 axis. We additionally characterized an inflammatory fibroblast subset, IL‑1R1+iCAFs, whose molecular features are associated with impaired T‑cell cytotoxicity and putative immunotherapy resistance. Our multi‑omics data highlight the putative role of IL‑1β+ Mac in shaping a pro‑tumor TME in HCC. Preliminary pharmacological evidence points to the therapeutic potential of targeting the IL‑1R1+iCAFs subpopulation to improve immunotherapy outcomes, although in‑vivo functional validation is still required.