HCC Cancer Research Articles

Current status and perspectives of TMIGD2-targeted therapy in thyroid cancer

In recent studies, TMIGD2 has found that it may play an important role in a new therapeutic target in the process of accurate tumor treatment. High expression of TMIGD2 in HCC and glioma cancer can be used as a potential therapeutic target, and some studies showed that the expression level of TMIGD2 in thyroid cancer tissue is also higher than that in normal thyroid tissue, and its expression level is correlated with the malignancy and poor prognosis of tumors. These more cutting-edge findings suggest that TMIGD2 may be used as a potential target for the treatment of thyroid cancer for precise treatment. This article will introduce the current situation and development trend of TMIGD2 in the clinical precision treatment of thyroid cancer.

Comprehensive analysis of paraspeckle-associated gene modules unveils prognostic signatures and immunological relevance in multi-cancers

BackgroundHepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths worldwide, characterized by high rates of angiogenesis and immune evasion. Paraspeckle genes, involved in gene regulation and RNA metabolism, have recently been linked to tumor progression. This study aims to elucidate the relationship between paraspeckle genes and HCC prognosis, focusing on SFPQ, DDX39B, and UBAP2.MethodsWe analyzed HCC (LIHC) and prostate cancer (PRAD) samples from the TCGA database to explore the correlation between paraspeckle genes and angiogenesis. We conducted unsupervised clustering, risk scoring, and survival analysis to identify distinct patient groups and their clinical outcomes. Gene expression data were used to perform differential analysis and Gene Ontology (GO) enrichment.ResultsOur analysis identified significant correlations between paraspeckle genes and angiogenesis across multiple cancer types. Elevated expression levels of SFPQ, DDX39B, and UBAP2 were associated with poor prognosis in HCC patients, and all of them has statistical significance. Unsupervised clustering of HCC samples based on paraspeckle gene expression revealed two distinct clusters, with high-risk patients exhibiting stronger immune suppression and tumor immune evasion. GO enrichment highlighted critical pathways related to angiogenesis and immune regulation. Additionally, a risk scoring model based on these genes effectively distinguished high-risk and low-risk patient groups, providing valuable prognostic insights.ConclusionThis study demonstrates that SFPQ, DDX39B, and UBAP2 are significantly associated with poor prognosis in HCC, likely due to their roles in promoting angiogenesis and immune suppression. These findings highlight the potential of paraspeckle genes as prognostic biomarkers and therapeutic targets, offering new avenues for personalized treatment strategies in HCC. Further research into their functional mechanisms and clinical applicability is crucial for advancing HCC treatment and improving patient outcomes.

A comprehensive analysis of portal vein thrombosis in malignancy.

e24042 Background: Portal vein thrombosis (PVT) is a rare venous thromboembolism, is often linked to cirrhosis, infection, and malignancy. Advancements in imaging have noted an increasing incidence of PVT, particularly in solid cancers. Treating PVT with malignancy is challenging due to lack of data, poor recanalization, and substantial bleeding risk with anticoagulation. This study aims to enhance understanding of malignancy-related PVT, anticoagulation trends, and assess prognosis. Methods: We conducted a multicenter, observational retrospective analysis of patients diagnosed with PVT (ICD code 452,181) from 2013 to 2023. Inclusion criteria: age > 18 and acute portal vein thrombosis. Exclusion criteria: presence of tumor thrombosis, and thrombus in other splanchnic vessels (e.g., mesenteric vein). Data entry was performed on a study-specific REDCap database. Patients with concomitant malignancy were identified and further assessed for cancer type, anticoagulation use, bleeding outcomes, and prognosis. Proportion analysis using Z test was done to compare outcomes between groups. Results: In our study of 583 patients, 236 (40.48%) had concurrent malignancy, predominantly male (144, 61.01%),and female (92, 38.98%) mean age 65. Common malignancies included HCC (28.38%), pancreatic (24.15%), colo-rectal (11.86%), cholangiocarcinoma (8.47%), myeloproliferative neoplasm, prostate, and gynecological cancers each (2.96%), and other cancers (18.22%). 41% had metastasis to liver. Cirrhosis was present in 40%, particularly in HCC patients (89%). Of the total,163 (69%) received anticoagulation; 26.38% showed improvement or re-canalization within a year. The primary anticoagulants were Direct Oral Anticoagulants (DOACs) (57%), with an increasing trend in the last 5 years (69.44%), followed by Low-Molecular-Weight Heparin (LMWH) (19%) and warfarin (14%). Most patients initiated on heparin infusion (9.81%) transitioned to hospice care. Bleeding events in patients receiving DOAC versus LMWH + Warfarin were 36.55% and 33.33%, respectively (p = 0.69). Patients without anticoagulation demonstrated a higher one-year mortality compared to those with anticoagulation (83.56% vs. 63.19%, p value < 0.05). Overall,malignancy-associated PVT had a significantly higher case fatality rate than non-malignancy cases, regardless of anticoagulation (81.35% vs. 32.85%, p < 0.05). Conclusions: There is a significant association between malignancy and PVT, particularly in HCC and pancreatic cancer. The increased mortality in malignancy-related PVT emphasizes the critical role of anticoagulation, which has demonstrated efficacy in improving outcomes, including one-year mortality. Although DOACs have seen increased use in the last 5 years, bleeding events have remained comparable. Further research is warranted to explore patient selection for optimal anticoagulation strategies.

Abstract CT097: IMM27M, a humanized Fc-engineered anti CTLA-4 antibody, in patients with advanced solid tumors: A phase I dose-escalation study

Abstract Background: IMM27M is a humanized Fc-engineered IgG1 CTLA-4 monoclonal antibody with enhanced ADCC. In pre-clinical models, IMM27M could enhance immune responses and promote Treg depletion. The pre-clinical results showed IMM27M induced a significantly stronger anti-tumor activity than ipilimumab and resulted in complete tumor remission even at a low dose. Methods: This study is an open-label, multi-center, phase I dose-escalation study to evaluate the safety, tolerability, maximum tolerated dose/recommended dose for expansion, PK and anti-tumor activity in patients with advanced solid tumors. The study was designed with an accelerated titration followed by a standard 3+3 design. IMM27M (0.1, 0.3, 1.0, 2.0, 3.0, 5.0, 7.5, 10.0 mg/kg) was administered as monotherapy Q3W. Results: As of 3 Nov 2023, the dose has been escalated to 7.5 mg/kg. 25 patients (20 females, 5 males) were enrolled and treated (1 at 0.1 mg/kg, 3 at 0.3 mg/kg, 3 at 1.0 mg/kg, 4 at 2.0 mg/kg, 4 at 3.0 mg/kg, 7 at 5.0 mg/kg, 3 at 7.5 mg/kg), including 13 patients with breast cancer (10 HR+ mBC), 4 patients with melanoma, 3 patients with RCC, each 1 patient with HCC, NSCLC and ovarian cancer, respectively. Median age was 51 years (range 31-72). 92.0% of the patients previously received ≥ 2 lines of systemic therapies and 52.0% received anti PD-1/PD-L1 treatment. Treatment-related adverse events (TRAEs) occurred in 24 patients (96.0%) by the data cutoff. Most TRAEs were grade 1 or 2. The most common TRAEs (≥ 30%) of all grades were anaemia (56.0%), lymphocyte count decreased (48.0%), aspartate aminotransferase increased (40.0%), hypoalbuminaemia (40.0%), decreased appetite (40.0%). Grade ≥3 TRAEs occurred in 11 patients (44.0%). Grade ≥3 TRAEs (≥10%) were lymphocyte count decreased (16.0%) and anaemia (12.0%). Treatment related SAE occurred in 8 patients (32.0%). No DLT was observed. One TRAE (Grade 3 immune related enteritis) led to treatment discontinuation. No TEAE leading to death was reported. Recommended Phase 2 Dose (RP2D) was 5 mg/kg. In 25 response evaluable patients, 2 patients had confirmed PR: 1 patient with mBC (HR+/HER2+, IO naïve, 6L previous treatments) at 3.0 mg/kg and response durable for 7 months; 1 patient with mBC (HR+/HER-, IO naïve, 4L previous treatments) at 5.0 mg/kg and response durable for 3 months by data cut-off. In addition, 9 patients had BOR SD and 4 out of 9 were previously treated with IO. Another 2 out of 8 patients with HR+ mBC had BOR SD with over 10% decreased tumor burden. IMM27M in vivo exposure increased with dose. The T1/2 mean value range in the 3-7.5mg/kg dose group was 8.2-11.5 days. Conclusions: IMM27M monotherapy in general was well-tolerated at the dose levels evaluated and preliminarily single agent activity was seen in patients with heavily pretreated advanced solid tumors. The dose escalation study of IMM27M in advanced solid tumors is ongoing.Clinical trial information: NCT05235438. Citation Format: Shusen Wang, Qiufan Zheng, Quanli Gao, Qingyuan Zhang, Shikai Wu, Zhihua Li, Enxiao Li, Qiying Lu, Frank Gan, Wenzhi Tian. IMM27M, a humanized Fc-engineered anti CTLA-4 antibody, in patients with advanced solid tumors: A phase I dose-escalation study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT097.

Abstract 4557: Overexpressed CPAP promotes cancer stemness and enhances chemo-resistance in HCC

Abstract Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and is highly resistant to chemotherapy and targeted therapy. It was reported that HCC cancer stem cells (CSCs) are involved in chemo-drug resistance by regulating multidrug resistance (MDR) gene expression, such as ABC transporters. Our previous reports indicated that CPAP, a centrosomal protein, can promote HCC progression by up-regulating NF-κB and STAT3 transcriptional activities. Here, we demonstrated that overexpressed CPAP can increase cancer stemness features and CSCs-derived sorafenib resistance in HCC by up-regulating the transcriptional activities of NF-κB and β-catenin. Importantly, CPAP stably expressed HCC spheroids are significantly resistance to sorafenib in vitro and in vivo. Our results suggested that CD24 and ABCG2 are two potential downstream effectors in CPAP overexpressed HCC. CPAP can transcriptionally up-regulate CD24 and ABCG2 expression via Wnt/β-catenin and NF-κB signaling pathways, respectively; high CPAP/ABCG2/CD24 expression levels are negatively correlated with disease-specific survival rates in HCC patients. Combined treatment with sorafenib and Wnt/β-catenin pathway inhibitor PRI-724 significantly inhibited CPAP-overexpressing Hep3B spheroids-derived tumor growth in a xenograft mouse model. Our study highlights the oncogenic role of CPAP in promoting CSCs-induced sorafenib resistance in HCC and provides insights into potential therapeutic strategies. Citation Format: Liang-Yi Hung. Overexpressed CPAP promotes cancer stemness and enhances chemo-resistance in HCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4557.

Circ_HMGCS1 modulates hepatocellular carcinoma chemoresistance via miR-338-5p/IL-7 pathway.

Hepatocellular cancer is one of the most serious types of cancer in the world, with high incidence and mortality rates. Most HCC patients with long-term chemotherapy develop chemoresistance, leading to a poor prognosis. However, the underlying mechanism of circRNAs in HCC chemoresistance remains unclear. Our research found that circ_0072391(circ_HMGCS1) expression was significantly upregulated in cisplatin-resistant HCC cells. The silence of circ_HMGCS1 attenuated the cisplatin resistance in HCC. Results showed that circ_HMGCS1 regulated the expression of miR-338-5p via acting as microRNA sponges. Further study confirmed that miR-338-5p regulated the expression of IL-7. IL-7 could remodel the immune system by improving T-cell function and antagonising the immunosuppressive network. IL-7 is an ideal target used to enhance the function of the immune system. circ_HMGCS1 exerts its oncogenic function through the miR-338-5p/IL-7 pathway. Inhibition of circ_HMGCS1/miR-338-5p/IL-7 could effectively attenuate the chemoresistance of HCC. IL-7 might be a promising immunotherapy target for HCC cancer treatment.

Fatty acid binding protein 5 is a novel therapeutic target for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is an aggressive subtype of liver cancer and is one of the most common cancers with high mortality worldwide. Reprogrammed lipid metabolism plays crucial roles in HCC cancer cell survival, growth, and evolution. Emerging evidence suggests the importance of fatty acid binding proteins (FABPs) in contribution to cancer progression and metastasis; however, how these FABPs are dysregulated in cancer cells, especially in HCC, and the roles of FABPs in cancer progression have not been well defined. To understand the genetic alterations and expression of FABPs and their associated cancer hallmarks and oncogenes in contributing to cancer malignancies. We used The Cancer Genome Atlas datasets of pan cancer and liver hepatocellular carcinoma (LIHC) as well as patient cohorts with other cancer types in this study. We investigated genetic alterations of FABPs in various cancer types. mRNA expression was used to determine if FABPs are abnormally expressed in tumor tissues compared to non-tumor controls and to investigate whether their expression correlates with patient clinical outcome, enriched cancer hallmarks and oncogenes previously reported for patients with HCC. We determined the protein levels of FABP5 and its correlated genes in two HCC cell lines and assessed the potential of FABP5 inhibition in treating HCC cells. We discovered that a gene cluster including five FABP family members (FABP4, FABP5, FABP8, FABP9 and FABP12) is frequently co-amplified in cancer. Amplification, in fact, is the most common genetic alteration for FABPs, leading to overexpression of FABPs. FABP5 showed the greatest differential mRNA expression comparing tumor with non-tumor tissues. High FABP5 expression correlates well with worse patient outcomes (P < 0.05). FABP5 expression highly correlates with enrichment of G2M checkpoint (r = 0.33, P = 1.1e-10), TP53 signaling pathway (r = 0.22, P = 1.7e-5) and many genes in the gene sets such as CDK1 (r = 0.56, P = 0), CDK4 (r = 0.49, P = 0), and TP53 (r = 0.22, P = 1.6e-5). Furthermore, FABP5 also correlates well with two co-expressed oncogenes PLK1 and BIRC5 in pan cancer especially in LIHC patients (r = 0.58, P = 0; r = 0.58, P = 0; respectively). FABP5high Huh7 cells also expressed higher protein levels of p53, BIRC5, CDK1, CDK2, and CDK4 than FABP5low HepG2 cells. FABP5 inhibition more potently inhibited the tumor cell growth in Huh7 cells than in HepG2 cells. We discovered that FABP5 gene is frequently amplified in cancer, especially in HCC, leading to its significant elevated expression in HCC. Its high expression correlates well with worse patient outcome, enriched cancer hallmarks and oncogenes in HCC. FABP5 inhibition impaired the cell viability of FABP5high Huh7 cells. All these support that FABP5 is a novel therapeutic target for treating FABP5high HCC.

In-Depth Proteome Profiling of Small Extracellular Vesicles Isolated from Cancer Cell Lines and Patient Serum.

Extracellular vesicle (EV) secretion has been observed in many types of both normal and tumor cells. EVs contain a variety of distinctive cargoes, allowing tumor-derived serum proteins in EVs to act as a minimally invasive method for clinical monitoring. We have undertaken a comprehensive study of the protein content of the EVs from several cancer cell lines using direct data-independent analysis. Several thousand proteins were detected, including many classic EV markers such as CD9, CD81, CD63, TSG101, and Syndecan-1, among others. We detected many distinctive cancer-specific proteins, including several known markers used in cancer detection and monitoring. We further studied the protein content of EVs from patient serum for both normal controls and pancreatic cancer and hepatocellular carcinoma. The EVs for these studies have been isolated by various methods for comparison, including ultracentrifugation and CD9 immunoaffinity column. Typically, 500-1000 proteins were identified, where most of them overlapped with the EV proteins identified from the cell lines studied. We were able to identify many of the cell-line EV protein markers in the serum EVs, in addition to the large numbers of proteins specific to pancreatic and HCC cancers.

Trends of liver cell carcinoma mortality in the United States by demographics and geography

BackgroundLiver cancer is a complex disease that presents many challenges in its diagnosis, treatment, and prevention. It's mortality rate in the United States is a significant and warrants attention. ObjectiveTo assess the trend of mortality rate due to HCC in the US from 1999 to 2020 by demographic groups for differences in trend of mortality. MethodsWe used the CDC wonder database to collect mortality rate data due to HCC as a multiple cause of death in the US from 1999 to 2020 by sex, race, age, and state of residence. The SEER Joinpoint program was used to calculate trends, defined as average annual percent change (AAPC) and to identify disparities between groups. All age-adjusted rates (AAMR) are reported per 100,000. ResultsFrom 1999 to 2020, we found that women observed an uptrend (AAPC1.6%) and men observed a slightly higher uptrend in mortality (AAPC 1.8%). In addition, AI/AN population had a significant uptrend (AAPC 2.3%). The AAPI population observed a downtrend (AAPC -2.6%). The Black or African American population observed an uptrend (AAPC 1.8%) The white population also observed an uptrend (AAPC 2.2%). In the 2010 to 2020 time period, Mississippi had the lowest AAMR of any state with 15.2, while Hawaii had with the highest with 38.8. ConclusionThis investigation assesses mortality rates and trends due to HCC cancer in the US and found significant differences in mortality rates and mortality rate trends due to HCC by demographic status in the US. Addressing the disparities in HCC incidence and mortality by race, ethnicity, state, and region, as well as improving access to screening, surveillance, and effective treatments, can reduce the burden of HCC and improve outcomes for patients.

LncTUG1 promotes hepatocellular carcinoma immune evasion via upregulating PD-L1 expression

HCC is one of the most common malignant tumors worldwide. Although traditional treatment methods have been improved in recent years, the survival rate of HCC patients has not been significantly improved. Immunotherapy has shown extremely high clinical value in a variety of tumors. In this study, we found that TUG1 could regulate the expression of PD-L1 through JAK2/STAT3 to mediate immunosuppression. Here, The expression of TUG1 and PD-L1 in HCC tissues was evaluated through analysis of databases and verified in HCC tissue and HCC cancer cells by qRT-PCR. The effect of TUG1 on tumor immune escape was detected by coculture, and cell viability was detected with a CCK8 assay. The results demonstrated that TUG1 was closely associated with anticancer immunity. TUG1 and PD-L1 were highly expressed in HCC tissues and HCC cancer cells, and high expression of TUG1 and PD-L1 was related to the poor prognosis of HCC patients. In addition, knocking down TUG1 expression could reduce PD-L1 expression and enhance the cancer cell-killing capability of T cells. Downregulating TUG1 expression could also decrease the mRNA and protein expression of JAK2 and STAT3. To sum up, TUG1 and PD-L1 are overexpressed in patients with liver cancer and are related to the poor prognosis of these patients. Silencing TUG1 expression reduced the mRNA and protein expression of PD-L1 by affecting the JAK2/STAT3 pathway.

OCTN2 enhances PGC-1α-mediated fatty acid oxidation and OXPHOS to support stemness in hepatocellular carcinoma.

The Metabolic reprogramming of tumor cells plays a vital role in the progression of hepatocellular carcinoma. Organic cation/carnitine transporter 2 (OCTN2), a sodium-ion dependent carnitine transporter and a sodium-ion independent tetraethylammonium (TEA) transporter, has been reported to contribute tumor malignancies and metabolic dysregulation in renal and esophageal carcinoma. However, the role of lipid metabolism deregulation mediated by OCTN2 in HCC cells has not been clarified. Bioinformatics analyses and immunohistochemistry assay were employed to identify OCTN2 expression in HCC tissues. The correlation between OCTN2 expression and prognosis was elucidated through K-M survival analysis. The expression and function of OCTN2 were examined via the assays of western blotting, sphere formation, cell proliferation, migration and invasion. The mechanism of OCTN2-mediated HCC malignancies was investigated through RNA-seq and metabolomic analyses. Furthermore, xenograft tumor models based on HCC cells with different OCTN2 expression levels were conducted to analyze the tumorigenic and targetable role of OCTN2 in vivo. We found that gradually focused OCTN2 was significantly upregulated in HCC and tightly associated with poor prognosis. Additionally, OCTN2 upregulation promoted HCC cells proliferation and migration in vitro and augmented the growth and metastasis of HCC. Moreover, OCTN2 promoted the cancer stem-like properties of HCC by increasing fatty acid oxidation and oxidative phosphorylation. Mechanistically, PGC-1α signaling participated in the HCC cancer stem-like properties mediated by OCTN2 overexpression, which is confirmed by in vitro and in vivo analyses. Furthermore, OCTN2 upregulation may be transcriptionally activated by YY1 in HCC. Particularly, treatment with mildronate, an inhibitor of OCTN2, showed a therapeutic influence on HCC in vitro and in vivo. Our findings demonstrate that OCTN2 plays a critical metabolic role in HCC cancer stemness maintenance and HCC progression, providing evidence for OCTN2 as a promising target for HCC therapy.

Transarterial Radioembolization for Hepatic Malignancies in North and South America

AbstractTransarterial radioembolization (TARE) with yttrium 90 is increasingly utilized for the treatment of hepatic neoplasms, whether primary (particularly hepatocellular carcinoma [HCC]) or metastatic (particularly colorectal). Extensive data and practical experience have led to a better understanding of its most appropriate usage and optimal techniques, particularly regarding dosimetry. Different clinical contexts and technical parameters allow its use with either palliative or curative intent. Improved patient outcomes have led to its inclusion in management guidelines of HCC and colorectal cancer. While available in multiple centers in the United States and Canada, its availability in South America is still very limited. The objective of this article is to review available treatment platforms, indications, techniques, recent advances, and clinical results, as well as briefly explore the disparities in availability throughout the United States.

Uncovering LAG-3 related tumor immunology in renal cell carcinoma and pan-cancer evaluation.

e16517 Background: LAG-3 (lymphocyte activation gene 3) is of interest as a target molecule for next-generation immune checkpoint inhibitors in several solid tumors, including renal cell carcinoma (RCC). The aim of this study is uncovering the tumor immune microenvironment (TIME) of the LAG-3-dominant RCC. Methods: Firstly, we performed an automated single-cell count for immunolabelled LAG-3, TIM-3 and TIGIT and TIME evaluation panel molecules (CD3, CD8, CD39, PD1, PDL1, Ctla4, CD68, CD163, CD47, Ki67, CD73, IDO1, GLUT1, CD34 and D2-40) to 105 primary ccRCC tumor samples and created the new classification model. The TIME and clinical value of LAG-3-dominant RCC were evaluated. Secondly, The TIME features of LAG-3 dominant RCC in metastatic sites was evaluated in 47 metastatic samples. Lastly, mRNA seq data from the public cohort and clinical trial data (RCC, n = 1144, other solid cancers, n = 3691) was analyzed with the digital cytometry and evaluated whether the TIME features of LAG-3 dominant RCC are RCC-specific or applicable to other solid tumors. Results: In our cohort and our validated cohort (n = 96), the LAG-3 group had significantly shorter OS (p = 0.037 and p = 0.003). The LAG-3 high expression RCC also showed shorter OS in the TCGA RCC cohort (p = 0.016). As the result of TIME evaluation, LAG-3 dominant RCCs had statistically high level of CD8, high CD39+CD8+, and high M2 macrophages. In metastasis samples, LAG-3 dominant RCCs preserved high level of CD8, high CD39+CD8+. In public mRNA seq analysis, LAG-3 dominant RCCs showed high level of CD8, high CD8+Tregs and high M2 macrophages. High Tregs in LAG-3 dominant tumor were seen in melanoma, HCC and prostate cancer. High M2 macrophage TIME in LAG-3 dominant tumor was unique to RCC, not seen in other solid cancer. Conclusions: LAG-3 dominant RCC had poor prognosis and could exhibit a unique, immunosuppressive TIME.

Treatment with Increasing Amounts and for Various Durations of theAnticancer Agent Curcumol’s Effect on Stimulation of the Clearanceof HEP3b Xenograft Tumors and Decreasing of Jak3/Stat3 Signalingand Phosphorylation.

Human Hepatocellular Carcinoma is the second leading cause of cancer deaths, and it is undoubtedly easy to be infectedwith a destructive lifestyle. However, humans’ immune systems will be effective for cancer treatment if there is nosuppression of T cells brought by Jak3/Stat3 molecular activities. Fortunately, Curcumol has been proven effective forseveral cancer treatments and might also be helpful in HCC cancer treatment. This paper tends to prove that treatmentwith increasing amounts and for various durations of the anticancer agent Curcumol will stimulate the clearance ofHEP3b xenograft tumors and decrease Jak3/Stat3 signaling and phosphorylation in the in vivo conditions. STAT3phosphorylation and STAT3 mediated transcription will be measured by western blot analysis and STAT3 reporter assay.HEP3b killing will be measured by MTT assay. In the experiment, the Positive control is Taxol, and the negative controlis PBS/DMSO. Each experiment will be repeated five times with different concentrations and various treatment durationof Curcumol extractives, while all the experiments will be done in vivo. .

Extrachromosomal circular MiR-17-92 amplicon promotes HCC.

Extrachromosomal circular DNAs (eccDNAs) are prevalent in cancer genomes and emerge as a class of crucial yet less characterized oncogenic drivers. However, the structure, composition, genome-wide frequency, and contribution of eccDNAs in HCC, one of the most fatal and prevalent cancers, remain unexplored. In this study, we provide a comprehensive characterization of eccDNAs in human HCC and demonstrate an oncogenic role of microRNA (miRNA)-17-92-containing eccDNAs in tumor progression. Using the circle-sequencing method, we identify and characterize more than 230,000 eccDNAs from 4 paired samples of HCC tumor and adjacent nontumor liver tissues. EccDNAs are highly enriched in HCC tumors, preferentially originate from certain chromosomal hotspots, and are correlated with differential gene expression. Particularly, a series of eccDNAs carrying the miRNA-17-92 cluster are validated by outward PCR and Sanger sequencing. Quantitative PCR analyses reveal that miRNA-17-92-containing eccDNAs, along with the expression of their corresponding miRNAs, are elevated in HCC tumors and associated with poor outcomes and the age of HCC patients. More intriguingly, exogenous expression of artificial DNA circles harboring the miR-17-92 cluster, which is synthesized by the ligase-assisted minicircle accumulation method, can significantly accelerate HCC cell proliferation and migration. These findings delineate the genome-wide eccDNAs profiling of HCC and highlight the functional significance of miRNA-containing eccDNAs in tumorigenesis, providing insight into HCC pathogenesis and cancer therapy, as well as eccDNA and miRNA biology.

Abstract 2671: Imipridones ONC201 and ONC206 reduce expression of neogenin and EZH1/2 which correlate with synergy following their combination with EZH1/2 or HDAC inhibitors in treatment of DMG and other tumors

Abstract Imipridones are a family of small molecule compounds including ONC201, ONC206 and ONC212 with anti-cancer activity mediated in part through activation of the integrated stress response, induction of TRAIL and its receptor, DR5, and activation of mitochondrial caseinolytic protease ClpP with consequent impairment of oxidative phosphorylation. Early clinical data have indicated that ONC201 provides clinical benefit in a subset of patients with histone H3K27M-mutated diffuse midline glioma (DMG). The H3K27M mutation prevents the function of EZH2 in methylating H3K27 on the mutated protein. This phenomenon implicates H3K27M and EZH2 in the mechanism of the anti-cancer effect of ONC201. EZH1 is a homolog of EZH2 and forms an alternative for EZH2 in assembling the PRC2 complex. We investigated the effects of ONC201, ONC206 or ONC212 on EZH1/2 by treating DMG cells and a panel of other solid tumor cells including GBM, DMG, CRC, PDAC, SCLC, prostate cancer, HCC and breast cancer cells with ONC201, ONC206 or ONC212. We observed that imipridones inhibit expression of both EZH1 and EZH2 in these tumors. RNA-seq analysis and RT-PCR showed no regulation of EZH1/2 at the level of transcription after ONC201 treatment. Linear regression revealed a correlation between extent of EZH1/2 inhibition and extent of cell viability suppression by ONC201 thereby providing further rationale for the combination of ONC201 and EZH1/2 inhibitors or HDAC inhibitors. We combined ONC201 or ONC206 or ONC212 plus a dual EZH1/2 inhibitor or the triple combination of ONC201, EZH2i and HDACi in DMG, GBM, prostate cancer and SCLC cells for 72 hours, performed a CellTiterGlo assay and observed synergies. The effective combination index (CI) of ONC201 plus tazemetostat ranges 0.04-0.64 in SU-DIPG-25 and 0.55-0.84 in SU-DIPG-13 DMG cells. The effective CI of ONC206 plus tazemetostat and panobinostat ranges 0.27-0.77 in U251 GBM and 0.11-0.71 in SU-DIPG-13 DMG cells. The effective CI of ONC201 plus dual EZH1/2 inhibitor, valemetostat, ranges 0.44-0.89 in H1048 SCLC cells, 0.14-0.90 in LNCaP prostate cancer cells, and 0.13-0.79 in SNB19 GBM cells. The effective CI of ONC212 plus valemetostat ranges 0.33-0.83 in 22Rv1. The synergy in inducing apoptosis was demonstrated by immunoblotting of cleaved-PARP. We also observed that ONC201 or ONC206 inhibit neogenin which is associated with invasion of DMG in SU-DIPG-13 cells. We conclude that ONC201 inhibits EZH1/2 in tumor cells and also neogenin in DMG cells. Inhibition of these proteins may play roles in the anti-cancer effect of ONC201. The synergies between ONC201 and EZH1/2 or HDAC inhibitors provide clues for developing novel therapy for the mentioned tumors. Overexpression of EZH2 is in process to elucidate the role of EZH2 in the mechanism of the anti-cancer effect of ONC201. Citation Format: Yiqun Zhang, Wafik S. El-Deiry. Imipridones ONC201 and ONC206 reduce expression of neogenin and EZH1/2 which correlate with synergy following their combination with EZH1/2 or HDAC inhibitors in treatment of DMG and other tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2671.

Hepatocellular Carcinoma: Aetiology and Mechanisms of Development. A Literature Review

Liver cancer remains a global challenge of healthcare, with the incidence growing worldwide. According to various authors, over 1 million patients will be diagnosed with liver cancer each year by 2025. The molecular pathogenesis of HCC varies with respect to genotoxic lesions and aetiologies. Although our understanding of the HCC pathophysiology and drivers tends to improve, it is still distant from translation into clinical practice. About 25 % of HCC cases are associated with variant mutations. HCC pathophysiology is a complex multi-step process. The interaction of various factors underlies the early stages of malignant hepatocyte transformation towards the development of HCC. Overall, about 20–25 % of HCC patients have at least one potential driver mutation. Obesity should also be noted as being associated with a higher risk of HCC and various other cancers. Despite many issues in the HCC pathogenesis being already known, the unresolved questions remain. Modern molecular genetic diagnostics and animal modelling of malignant tumours are expanding our horizons of knowledge in this field.

Outcomes of adolescent and young patients with hepatocellular carcinoma after curative liver resection: a retrospective study

BackgroundThe risk of HCC is documented to be age-related. The outcomes of young HCC patients on postoperative prognosis are not well understood. The study aims to compare the characteristic differences between adolescent and young (AYA) and non-AYA HCC patients.MethodsWe performed a retrospective analysis of the clinical and pathological findings and the survival of 243 HCC patients who underwent operations between 2007 and 2018.ResultsThe AYA group had a higher AFP level and a higher prevalence of family history of HCC or other cancers than the non-AYA group (P < 0.01 and P < 0.05). AYA patients had more unfavorable pathological characteristics including bigger lesion size, microvascular invasion, portal vein invasion, and hepatic capsule invasion. They also had a more unfavorable Edmondson grade and less tumor capsule formation (P < 0.01). Age was an independent predictor of survival in HCC patients. AYA patients had poorer disease-free and overall survival than non-AYA patients did (P < 0.01). Patients under 30 years old had an even poorer disease-free survival than those aged 30–40 (P = 0.047).ConclusionsAYA patients exhibited a higher recurrence rate and disease-related death rate with more unfavorable pathological characteristics. Enhanced follow-up for young HCC patients should be applied.

LncRNA SNHG5 promotes the proliferation and cancer stem cell-like properties of HCC by regulating UPF1 and Wnt-signaling pathway

The role of long noncoding RNA (lncRNAs) had been demonstrated in different types of cancer, including hepatocellular carcinoma. This study was intended to investigate the role of lncRNA small nucleolar RNA host gene 5 (SNHG5) in HCC proliferation and the liver CSC-like properties. Through functional experiments, we determined that knockdown of SNHG5 repressed HCC cell proliferation and CSC-like properties, while over-expression of SNHG5 promoted cell growth. At the same time, CSC markers (CD44, CD133, and ALDH1) and related transcription factors (OCT4, SOX2, and NANOG) were downregulated when SNHG5 was knocked down. Mechanically, RNA immunoprecipitation (RIP) and RNA pulldown assay showed that SNHG5 regulated the proliferation and CSC-like properties of HCC by binding UPF1. Further investigations showed that expression of critical components of Wnt/β-catenin pathway (β-catenin, TCF4, c-myc, cyclinD1, and c-Jun) were upregulated with depletion of UPF1 in liver CSCs, which were downregulated with depletion of SNHG5. After use of the inhibitor of Wnt/β-catenin pathway, the formation of liver CSCs sphere decreased. Taken together, SNHG5 plays a critical role to promote HCC cell proliferation and cancer stem cell-like properties via UPF1 and Wnt/β-catenin pathway.

Identification of the Tumor Immune Microenvironment and Therapeutic Biomarkers by a Novel Molecular Subtype Based on Aging-Related Genes in Hepatocellular Carcinoma.

BackgroundHepatocellular carcinoma (HCC) is one of the most prevalent malignant tumors with poor prognosis. Increasing evidence has revealed that immune cells and checkpoints in the tumor microenvironment (TME) and aging are associated with the prognosis of HCC. However, the association between aging and the tumor immune microenvironment (TIME) in HCC is still unclear.MethodsRNA expression profiles and clinical data concerning HCC were downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Based on differentially expressed aging-related genes (DEAGs), unsupervised clustering was used to identify a novel molecular subtype in HCC. The features of immune cell infiltration and checkpoints were further explored through CIBERSORTx. Enrichment analysis and both univariate and multivariate Cox analyses were conducted to construct a 3-gene model for predicting prognosis and chemosensitivity. Finally, the mRNA and protein expression levels of the 3 genes were verified in HCC and other cancers through database searches and experiments.ResultsEleven differentially expressed AGs (GHR, APOC3, FOXM1, PON1, TOP2A, FEN1, HELLS, BUB1B, PPARGC1A, PRKDC, and H2AFX) correlated with the prognosis of HCC were used to divide HCC into two subtypes in which the prognosis was different. In cluster 2, which had a poorer prognosis, the infiltration of naive B cells and monocytes was lower in the TCGA and GEO cohorts, while the infiltration of M0 macrophages was higher. In addition, the TCGA cohort indicated that the microenvironment of cluster 2 had more immunosuppression through immune checkpoints. Enrichment analysis suggested that the MYC and E2F targets were positively associated with cluster 2 in the TCGA and GEO cohorts. Additionally, 3 genes (HMGCS2, SLC22A1, and G6PD) were screened to construct the prognostic model through univariate/multivariate Cox analysis. Then, the model was validated through the TCGA validation set and GEO dataset (GSE54236). Cox analysis indicated that the risk score was an independent prognostic factor and that patients in the high-risk group were sensitive to multiple targeted drugs (sorafenib, gemcitabine, rapamycin, etc.). Finally, significantly differential expression of the 3 genes was detected across cancers.ConclusionWe systematically described the immune differences in the TME between the molecular subtypes based on AGs and constructed a novel three-gene signature to predict prognosis and chemosensitivity in patients with HCC.