Targeting metabolic vulnerabilities: REV-ERB agonist SR9009 potentiates sorafenib efficacy in liver cancer.

Objective To establish a new patient-derived xenograft (PDX) model of human liver cancer by inoculating the complex of human primary liver cancer cells and a novel microcarrier (microcarrier 6) into mice with normal immune function. Methods Primary liver cancer cells were isolated and extracted from the fresh human liver cancer tissue of five patients and were then co-cultured with microcarrier 6 to construct a three-dimensional tumor cell culture model in vitro. According to the type of graft, 75 male C57BL/6 mice were divided into cell control group, microcarrier control group, and experimental group (each sample corresponded to three groups, with 15 groups in total and 5 mice in each group). The liver cancer cell-microcarrier complex was implanted into the mice by subcutaneous inoculation, and tumor formation time, tumor formation rate, and histopathological manifestations were observed. The Fisher's exact test was used for comparison of categorical data between two groups. Results As for the liver cancer cells from the five patients, tumor formation was observed in the mice corresponding to three patients. In these three experiments, tumor formation was not observed in the control groups and was only observed in the experimental groups, and 12 of the 15 mice in the experimental groups had successful tumor formation, with a tumor formation rate as high as 80%, which was significantly different from that in the cell control groups and the microcarrier control groups (all P Conclusion This experiment successfully establishes a new PDX model of human liver cancer based on the complex of microcarrier 6 and human primary liver cancer cells in mice with normal immunity. This model can be used to better elucidate the mechanism of the development and progression of liver cancer in the body with normal immunity, and besides, it also provides a new animal model with higher value for the precise treatment of liver cancer.

CPEB4 modulates liver cancer progression by translationally regulating hepcidin expression and sensitivity to ferroptosis.

Sodium Iodide Symporter Is Expressed at the Preneoplastic Stages of Liver Carcinogenesis and in Human Cholangiocarcinoma

MicroRNA-638 induces apoptosis and autophagy in human liver cancer cells by targeting enhancer of zeste homolog 2 (EZH2)

miR-199a-3p increases the anti-tumor activity of palbociclib in liver cancer models

Apoptotic liver cancer cells have important roles in liver tumorigenesis and liver cancer progression. Recent studies have shown that δ‑opioid receptors are highly expressed in human liver and liver cancer cells. The present study aimed to investigate the role of activated δ‑opioid receptors on human liver cancer cell apoptosis and its interrelation with the mitochondria and the protein kinaseC/extracellular‑signal‑regulated kinase (PKC/ERK) signaling pathway. H2O2 was used to induce apoptosis in human liver cancer cells. During apoptosis, mitochondrial transmembrane potentials were observed to decrease, cytochromec expression was found to increase and Bcell lymphoma2 (Bcl‑2) expression decreased. These findings suggested that H2O2‑induced apoptosis was mediated through the mitochondrial pathway. Of note, activated δ‑opioid receptors were observed to inhibit H2O2‑induced apoptosis in human liver cancer cells. Following δ‑opioid receptor activation, the number of apoptotic liver cancer cells decreased, mitochondrial transmembrane potentials were restored, cytoplasmic cytochromec and Bcl‑2‑associatedXprotein expression decreased and Bcl‑2 expression increased. These data suggested that δ‑opioid receptor activation inhibited mitochondria‑mediated apoptosis. In addition, activation of δ‑opioid receptors was observed to increase the expression of PKC and ERK in human liver cancer cells. Furthermore, upon inhibition of the PKC/ERK signaling pathway, the protective effect associated with the δ‑opioid receptor on liver cancer cell apoptosis was inhibited, which was not associated with the status of δ‑opioid receptor activation. These findings suggested that the PKC/ERK signaling pathway has an important role in δ‑opioid receptor‑mediated inhibition of apoptosis in human liver cancer cells.

NRT-YHD_001, a modified antisense miRNA targeting let-7i-5p for liver cancer therapy, acts as a key regulator of macrophage activity within the tumor microenvironment. Through in vitro pharmacology studies, we identified anticancer effects not only on cell growth, proliferation, viability, migration, and invasion but also through the enhancement of macrophage phagocytosis. To evaluate the in vivo efficacy of NRT-YHD_001, we used a Ras-transgenic mouse liver cancer model which develops spontaneous liver cancer mass after 15 ∼ 20 weeks of birth. Based on dose-dependent and injection interval studies, the optimal regimen was determined to be 1 mg/kg administered weekly. We confirmed that weekly intravenous injections of 1mg/kg NRT-YHD_001 showed significantly greater therapeutic efficacy than the sorafenib-treated group. Additionally, xenograft assay using human liver cancer cells in athymic-nude mice showed that weekly injection of NRT-YHD_001 led to improved survival and tumor growth inhibition compared to the untreated group. We performed Absorption, Metabolism, Distribution, and Excretion studies for NRT-YHD_001, stability studies in serum and liver homogenate, and metabolite identification and profiling analysis were completed in mice and cynomolgus monkeys. In maximum tolerated dose (MTD) studies conducted in mice and cynomolgus monkeys, no toxicity of NRT-YHD_001 was observed at doses up to 500 mg/kg administered intravenously. Furthermore, comprehensive evaluations across 4-week repeated dose toxicity and toxicokinetic in mice and monkey, safety pharmacology, and genetic toxicity studies revealed no adverse findings with NRT-YHD_001. For NRT-YHD_001, Drug Substance (DS) and Drug Product (DP) were produced by a global CDMO with experience in U.S. FDA approval of oligonucleotide drugs, and the analysis method was established.These results enabled us to complete the IND-enabling study package for NRT-YHD_001, a liver cancer therapy candidate with FTO analysis completed and intellectual property (IP) fully secured. Regulatory approval submissions to the Korea MFDS and the U.S. FDA are planned for 2025. Citation Format: Suk Woo Nam, Jin Woong Ha, Sang Yean Kim, Min Jeong Na, Soyoung Jeon, Bee Yoo, In Seop Yoon, Hyunmin Lee, Chang Won Park. IND enabling study of NRT-YHD_001, a macrophage immune checkpoint inhibitor in liver cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3209.

Expanding the immunotherapy roadmap for hepatocellular carcinoma.

e15188 Background: Despite success of immunotherapy in a wide range of solid tumors, immune checkpoint inhibitors (ICIs, such as α-PD-1 or α-PD-L1) remain to be ineffective in the treatment of pancreatic adenocarcinoma (PDAC) owing to the very low objective response rate (ORR). Although ICIs have already been approved for the treatment of hepatocellular carcinoma (HCC), the low ORR remains a clinical challenge for the treatment of these two types of highly aggressive malignancies. The oncolytic bacteria product, SGN1, is a genetically engineered attenuated Salmonella strain that overexpresses an L-methioninase, which can specifically target and reduce methionine supply within tumor tissues (Zhou et al, Cell Reports Medicine , 2023, 4, 101070;). In all preclinical models as well as in clinical trials, SGN1 has shown strong activities in inducing oncolysis and suppressing metastasis. Currently, SGN1 is undergoing Phase I/IIa trials in the US, Mainland China, & Taiwan (NCT05103345; NMPA: CTR20233735). Lately, we revealed that SGN1 treatment could elicit a strong elevation of PD-L1 expression and CD8+ T cell infiltration in melanoma model (Zhou et al, JITC, 2024, 12:e008238). We aimed to examine if SGN1 can induce PD-L1 expression, stimulate the infiltration of cytotoxic T cell infiltrations, and the combination application can generate in pancreatic cancer and liver cancer. Methods: Methionine restriction was applied to examine the expression of PD-L1 in cultured PDAC or HCC cells. SGN1 was co-incubated with human PBMC and PD-L1 to examine the combo effects on cultured PDAC or HCC cells. Lastly, tumor suppression effects of combination of SGN1 and α-PD-L1 were evaluated in the growth of pancreatic and liver cancer models. Results: SGN1 can induce strong and significant elevation of PD-L1 expression and CD8+ T cell infiltration in PDAC or HCC tumors. These effects of SGN1 are stronger than that of the bacterial vector alone, indicative of methionine restriction plays a prominent role. In the PDAC and HCC models, sequential use of SGN1 and anti-PD-L1 generated synergistic effect that strongly suppressed the tumor growth of PDAC and HCC, very similar to those effects observed in melanoma model. Conclusions: The genetically engineered oncolytic bacteria agent, SGN1, can induce robust changes in the immune environment and, when used in combination with α-PD-L1, can have strong synergistic effects on restricting the growth of PDAC or HCC tumors. Further clinical trials of such combination applications in these malignancies are warranted.

The increased expression of SIRT1 has recently been identified in numerous human tumors and a possible correlation with c-Myc oncogene has been proposed. However, it remains unclear whether SIRT1 functions as an oncogene or tumor suppressor. We sought to elucidate the role of SIRT1 in liver cancer under the influence of c-Myc and to determine the prognostic significance of SIRT1 and c-Myc expression in human hepatocellular carcinoma. The effect of either over-expression or knock down of SIRT1 on cell proliferation and survival was evaluated in both mouse and human liver cancer cells. Nicotinamide, an inhibitor of SIRT1, was also evaluated for its effects on liver tumorigenesis. The prognostic significance of the immunohistochemical detection of SIRT1 and c-Myc was evaluated in 154 hepatocellular carcinoma patients. SIRT1 and c-Myc regulate each other via a positive feedback loop and act synergistically to promote hepatocellular proliferation in both mice and human liver tumor cells. Tumor growth was significantly inhibited by nicotinamide in vivo and in vitro. In human hepatocellular carcinoma, SIRT1 expression positively correlated with c-Myc, Ki67 and p53 expression, as well as high á-fetoprotein level. Moreover, the expression of SIRT1, c-Myc and p53 were independent prognostic indicators of hepatocellular carcinoma. In conclusion, this study demonstrates that SIRT1 expression supports liver tumorigenesis and is closely correlated with oncogenic c-MYC expression. In addition, both SIRT1 and c-Myc may be useful prognostic indicators of hepatocellular carcinoma and SIRT1 targeted therapy may be beneficial in the treatment of hepatocellular carcinoma.

Revealing anti-PD-1 resistance mechanisms in HCC: A path towards novel combination immunotherapies

Treatment for Hepatocellular Carcinoma in South Asia

Epidemiology of hepatocellular carcinoma in the United States: where are we? Where do we go?

Simple SummaryCurrent immunotherapies based on blockade of immunosuppressive elements provide limited results in liver cancer patients. Here we tested whether combination of this therapy with a vaccine based on the Cold-Inducible RNA Binding Protein (CIRP) would improve its efficacy. Combination of immunotherapy with a CIRP-based vaccine increased vaccine immunogenicity and, when tested in several mouse models of liver cancer, resulted in better therapeutic effects. Despite good immune responses observed in peripheral organs, lymphocytes infiltrating the tumor appeared exhausted, with a weak functional capacity. Finally, by using the same strategy, we prepared a new CIRP-based vaccine containing glypican-3, human antigen commonly found in patients with liver cancer. An equivalent combination enclosing this new vaccine was also highly immunogenic. This suggests that CIRP-based vaccines may enhance the beneficial effects provided by current immunotherapies. However, they should also consider incorporating new elements to overcome limitations observed in tumor lymphocytes.Therapies based on immune checkpoint inhibitors (ICPI) have yielded promising albeit limited results in patients with hepatocellular carcinoma (HCC). Vaccines have been proposed as combination partners to enhance response rates to ICPI. Thus, we analyzed the combined effect of a vaccine based on the TLR4 ligand cold-inducible RNA binding protein (CIRP) plus ICPI. Mice were immunized with vaccines containing ovalbumin linked to CIRP (OVA-CIRP), with or without ICPI, and antigen-specific responses and therapeutic efficacy were tested in subcutaneous and orthotopic mouse models of liver cancer. OVA-CIRP elicited polyepitopic T-cell responses, which were further enhanced when combined with ICPI (anti-PD-1 and anti-CTLA-4). Combination of OVA-CIRP with ICPI enhanced ICPI-induced therapeutic responses when tested in subcutaneous and intrahepatic B16-OVA tumors, as well as in the orthotopic PM299L HCC model. This effect was associated with higher OVA-specific T-cell responses in the periphery, although many tumor-infiltrating lymphocytes still displayed an exhausted phenotype. Finally, a new vaccine containing human glypican-3 linked to CIRP (GPC3-CIRP) induced clear responses in humanized HLA-A2.01 transgenic mice, which increased upon combination with ICPI. Therefore, CIRP-based vaccines may generate anti-tumor immunity to enhance ICPI efficacy in HCC, although blockade of additional checkpoint molecules and immunosuppressive targets should be also considered.

Construction and application of liver cancer models in vitro