Microenvironment inflammatory infiltrate drives growth speed and outcome of hepatocellular carcinoma: a prospective clinical study

Abstract

In HCC, tumor microenvironment, heavily influenced by the underlying chronic liver disease, etiology and stage of the tissue damage, affects tumor progression and determines the high heterogeneity of the tumor. Aim of this study was to identify the circulating and tissue components of the microenvironment immune-mediated response affecting the aggressiveness and the ensuing clinical outcome. We analyzed the baseline paired HCC and the surrounding tissue biopsies from a prospective cohort of 132 patients at the first diagnosis of HCC for immunolocalization of PD-1/PD-L1, FoxP3, E-cadherin, CLEC2 and for a panel of 82 microRNA associated with regulation of angiogenesis, cell proliferation, cell signaling, immune control and autophagy. Original microarray data were also explored. Serum samples were analyzed for a panel of 19 cytokines. Data were associated with biochemical data, histopathology and survival. Patients with a more aggressive disease and shorter survival, who we named fast-growing accordingly to the tumor doubling time, at presentation had significantly higher AFP levels, TGF-β1 and Cyphra 21-1 levels. Transcriptomic analysis evidenced a significant downregulation of CLEC2 and upregulation of several metalloproteinases. A marked local upregulation of both PD-1 and PD-L1, a concomitant FoxP3-positive lymphocytic infiltrate, a loss of E-cadherin, gain of epithelial–mesenchymal transition (EMT) phenotype and extreme poor differentiation at histology were also present. Upregulated microRNA in fast-growing HCCs are associated with TGF-β signaling, angiogenesis and inflammation. Our data show that fast HCCs are characterized not only by redundant neo-angiogenesis but also by unique features of distinctively immunosuppressed microenvironment, prominent EMT, and clear-cut activation of TGFβ1 signaling in a general background of long-standing and permanent inflammatory state.

Liver Angiopoietin‐2 Is a Key Predictor of D e N ovo or Recurrent Hepatocellular Cancer After Hepatitis C Virus Direct‐Acting Antivirals

“…). These patients would have had a high probability of developing HCC during the natural history of their liver disease, and, given the intrinsic characteristics of the liver microenvironment, their tumors would have had distinctive clinical and biological aggressiveness. The combination of the clinical and biological risk factors that we have identified gives the unique possibility of selecting the patients at real risk of developing HCC after DAAs, without the need of holding back from treatment patients with cirrhosis at low risk of HCC …”

“…ANGPT2 mRNA levels and neo‐angiogenic transcriptomic signatures were tested by quantitative real‐time PCR, as described . ANGPT2 RNA levels were evaluated in terms of fold change in comparison with surrounding nontumoral tissue.…”

“…In 36 of 49 patients with HCC, frozen tissue (collected during initial liver biopsy in patients with past HCC and during biopsy of newly diagnosed lesions in patients with de novo HCC) was available for testing the presence of the neo-angiogenic transcriptomic signature. (23) This signature was observed among patients with recurrent and de novo HCC, but not in patients with no HCC recurrence (P = 0.0002; Supporting Table S2).…”

“…ANGPT2 mRNA levels and neo-angiogenic transcriptomic signatures were tested by quantitative realtime PCR, as described. (23) ANGPT2 RNA levels were evaluated in terms of fold change in comparison with surrounding nontumoral tissue. iMMUNoHiStocHeMiStry Tumoral and nontumoral liver tissue samples from all patients with HCC were available for programmed death 1 (PD1), programmed death ligand 1 (PDL1), and ANGPT2 immunohistochemical (IHC) analysis, which was performed as described.…”

Functional significance of the platelet immune receptors GPVI and CLEC-2

“…In contrast, the expression of CLEC-2 on cancer cells in hepatocellular carcinoma and gastric cancer is protective. Downregulation of CLEC-2 has been observed on the invasive form of both cancers, and has been linked to increased EMT and tumor aggressiveness (152,153). Therefore, the role of CLEC-2 of in cancer progression and metastasis depends on its expression on cancer cells or platelets.…”