JHEP Reports最新文献

Background & aims: Tumor neoantigens, especially cryptic antigens from non-canonical translation, are vital for cancer immunotherapy. Mass spectrometry (MS)-based de novo sequencing identifies candidates, but unverified immunogenicity and antitumor efficacy limit clinical applicability. This study aimed to identify novel non-canonical neoantigens in hepatocellular carcinoma (HCC) using MS-based de novo sequencing and rigorously validate their immunogenicity and antitumor efficacy.

Methods: Using a C57BL/6 subcutaneous HCC mouse model, immunopeptides were comprehensively profiled via MHC-I immunoprecipitation combined with MS-based de novo sequencing. Identified high-immunogenicity peptides predicted by deep learning were validated using ex vivo ELISpot assays. Endogenous peptide expression was confirmed using parallel reaction monitoring-targeted quantification. The antitumor efficacy of therapeutic peptide vaccines comprising the seven most immunogenic peptides combined with the adjuvant poly(I:C) was evaluated in vivo in the subcutaneous and orthotopic HCC models.

Results: We identified 5,576 immunopeptides, with sequence motifs consistent with prior reports. Remarkably, 95% of deep learning-predicted high-immunogenicity peptides were successfully validated by ELISpot (p <0.05). Parallel reaction monitoring confirmed endogenous expression of these peptides. Most significantly, the peptide vaccines (7 peptides + poly(I:C)) demonstrated potent antitumor efficacy in vivo compared to controls (p <0.05).

Conclusions: MS-based de novo sequencing combined with computational prioritization enables identification of non-canonical, immunogenic neoantigens in HCC. Selected peptides demonstrated endogenous presentation and measurable antitumor activity in preclinical models.

Impact and implications: This study provides robust experimental validation that mass spectrometry-based de novo sequencing effectively identifies novel, highly immunogenic non-canonical neoantigens in hepatocellular carcinoma, overcoming a key limitation of prior predictive methods and opening avenues for exploring this understudied neoantigen class in other cancers. The findings are critical for cancer immunologists and oncologists developing next-generation immunotherapies, demonstrating a viable discovery-to-validation pipeline for novel therapeutic targets. The validated neoantigens and successful peptide vaccine strategy offer a direct pathway towards developing personalized hepatocellular carcinoma immunotherapies, enabling clinicians to adopt similar integrated approaches for patient-specific neoantigen discovery; however, clinical translation beyond this preclinical murine model requires confirmation in human settings due to potential differences in HLA presentation and the tumor microenvironment.

Background & aims: Statins, used for cardiovascular disease (CVD) prevention, may offer hepatoprotective benefits. However, adherence to treatment indications and the associations between statin use, metabolic dysfunction-associated steatotic liver disease (MASLD), and elevated liver stiffness in the general population remain poorly understood.

Methods: This prospective, population-based study included adults aged ≥40 years between 2011 and 2020. We evaluated statin indications for CVD risk using prevailing European Society of Cardiology/European Atherosclerosis Society clinical practice guidelines based on SCORE2 and SCORE2-Older Persons algorithms. We used multivariable regression to examine associations between statins, MASLD, and elevated liver stiffness, adjusting for demographic, socioeconomic and metabolic covariables. We performed dose-response analyses using WHO defined daily dosages.

Results: Of 6,405 eligible individuals, 6,055 participants were included in the analysis (median age 64 years; 56% female); MASLD was present in 32%, elevated liver stiffness in 4.8%, and statin use in 21%. Participants with MASLD had higher predicted 10-year CVD risk compared to participants without MASLD (p <0.001), yet were less likely to use statins: 33% of individuals with MASLD and an indication for statin treatment remained untreated, compared to 19% of those without MASLD (p <0.001). Statin use was associated with lower prevalence of MASLD (adjusted odds ratio 0.76; 95% CI 0.63-0.92) and elevated liver stiffness (adjusted odds ratio 0.65; 95% CI 0.46-0.92) relative to untreated individuals with a statin treatment indication. The highest statin WHO defined daily dosage category was associated with lower prevalence of MASLD (p = 0.033) and elevated liver stiffness (p = 0.035).

Conclusions: Individuals with MASLD are less likely to use statins despite a contemporary guideline-based indication. Statin use is independently associated with lower prevalence of both MASLD and elevated liver stiffness. These findings underscore the need to improve CVD risk management in this population with the potential added benefit of mitigating MASLD.

Impact and implications: In this large prospective, population-based study, statins were underutilized in metabolic dysfunction-associated steatotic liver disease (MASLD) compared to non-MASLD individuals, even though they had the highest cardiovascular risk and met guideline-based treatment criteria. Our findings further demonstrate that individuals who used statins had a lower likelihood of MASLD and elevated liver stiffness compared with statin-eligible individuals who were not treated. Taken together, these results highlight a missed opportunity: optimizing statin use in people with MASLD could strengthen cardiovascular disease prevention while also offering potential benefits for liver health.

Background & aims: As the global prevalence of obesity continues to rise, metabolic dysfunction-associated steatohepatitis (MASH) has emerged as a significant burden on healthcare systems worldwide. Liver sinusoidal endothelial cells (LSECs) are recognized as crucial regulators of hepatic homeostasis. However, their role in the progression of MASH remains poorly understood, particularly given their early capillarization during disease onset. In this study, we sought to investigate the specific role of LSECs in MASH progression and the underlying molecular mechanisms driving their dysfunction.

Methods: We analyzed single-cell RNA sequencing of the liver from advanced MASH mice (from 6 mice/condition) and conducted in vivo experiments using various LSEC-specific transgenic mouse models (n = 3-10 mice/condition). Primary LSECs (n = 3-6 mice/condition) and human umbilical vein endothelial cells (HUVECs) were used for in vitro experiments.

Results: Using single-cell RNA sequencing data from MASH-affected livers, we identified a unique population of biphenotypic cells co-expressing markers of both LSECs and mesenchymal cells. Through lineage tracing in LSEC-specific reporter mice, we demonstrated that these biphenotypic cells arose from LSECs undergoing partial endothelial-mesenchymal transition (EndMT) in MASH (p <0.05-0.0001; n = 3 mice/condition, depending on the parameter). Mechanistically, we found that differentiation of these biphenotypic cells was regulated by TGF-β signaling. Inhibition of TGF-β signaling in LSECs suppressed partial EndMT and attenuated MASH progression (p <0.01-0.0001; n = 10 mice/condition). In addition, we discovered that Notch signaling in LSECs, which is activated in response to MASH, was positively regulated by TGF-β signaling and had a crucial role in both the generation of biphenotypic cells and MASH progression (p <0.01-0.0001; n = 3-10 samples/condition).

Conclusions: Our findings reveal that a pathogenic TGF-β/Notch axis drives MASH progression by inducing partial EndMet in LSECs, thereby establishing LSEC-targeted intervention as a promising therapeutic strategy for this disease.

Impact and implications: This study reveals a novel biphenotypic LSEC population generated via TGF-β-driven partial EndMT, establishing LSEC dysfunction as an active driver of MASH progression. By elucidating the TGF-β/Notch crosstalk in LSEC partial EndMT, our findings provide new mechanistic insights into MASH pathogenesis. Thus, these results highlight TGF-β signaling in LSECs as a promising therapeutic target. Future work should focus on LSEC-specific delivery systems and validation in cohorts of patients with MASH.

Background & aims: Hereditary hemochromatosis is an autosomal recessive disorder of excessive iron accumulation. Early diagnosis enables treatment before organ damage. The C282Y variant in the HFE gene is the main cause, but its penetrance of only 20% limits its utility for population-wide screening. We aimed to identify and quantify novel genetic and non-genetic modifiers of C282Y-related disease from electronic healthcare records, and thereby partly explain its incomplete penetrance.

Methods: We carried out a cohort study on data from 420,543 individuals in the FinnGen project, for whom genotype information and healthcare records were available. We performed both standard and interaction genome-wide association study analyses for hemochromatosis and fitted statistical models including age, sex, 21 million genetic variants, preceding diagnoses, and blood donation history as predictors. Results were validated using data from the UK Biobank.

Results: We identified three novel fine-mapped variants within 4 Mb of the HFE gene. Of these, variant rs181949568 in the CASC15 gene remained significant in the multivariable model (odds ratio 7.25, 95% CI 3.63-28.87, p = 1.96 × 10^-8). We found that donating blood at least twice a year is likely sufficient to reduce the risk of C282Y homozygotes (male risk 0.16, 80% CI 0.13-0.19) to that of C282Y-H63D compound heterozygotes (male risk 0.018, 80% CI 0.015-0.023). Additionally, the S65C variant protects against severe disease (incidence ratio 0.328, 95% CI 0.192-0.562).

Conclusions: We demonstrated that use of large-scale electronic health record data allows for precise quantification of individual-level risk, which we present as risk tables to support clinical practice. Furthermore, our findings suggest that hemochromatosis may be under-recognized in Finland.

Impact and implications: Because the factors influencing the penetrance of the C282Y variant in hemochromatosis remain incompletely understood, a study leveraging newly available large-scale healthcare and genetic data is warranted. We present the findings of our study as an individual-level risk table designed for practicing clinicians, summarizing the combined effects of key variables most frequently observed in the dataset. Our results suggest that asymptomatic individuals who are homozygous for C282Y could significantly reduce their risk of developing hemochromatosis by donating blood just twice a year.

Background & aims: Cholangiocarcinoma (CCA), a heterogeneous malignancy that includes intrahepatic (iCCA), perihilar (pCCA), and distal (dCCA) subtypes, often exhibits perineural invasion (PNI). Despite PNI being associated with postoperative recurrence, evidence on its prognostic impact remains inconsistent. To clarify this, we conducted a systematic review and meta-analysis.

Methods: A comprehensive literature search was conducted across the Medline-PubMed, Embase, and Web of Science databases from their inception through February 27, 2025. We included studies involving patients diagnosed with CCA who underwent surgery and for whom PNI data were available. The primary outcomes were overall survival (OS) and disease-free survival (DFS), extracted from both univariable and multivariable analyses. The study is registered in PROSPERO (CRD42024575458).

Results: Of 1,555 articles screened, 64 retrospective studies comprising 15,543 patients were included. In the univariable analysis, the pooled hazard ratio (HR) for OS from 34 studies (6,084 patients) was 1.92 (95% CI 1.67-2.21, I² = 59.6%). Multivariable-adjusted analysis of 42 studies (10,335 patients) showed an HR of 1.68 (95% CI 1.47-1.90, I² = 47.9%) for patients with PNI. For DFS, the pooled univariable HR in 6,110 patients was 1.70 (95% CI 1.46-1.98, I² = 52.3%), and the multivariable HR was 1.62 (95% CI 1.37-1.92, I² = 70.5%)). Prognostic consistency was observed across anatomical subgroups. Univariable OS HRs were 2.14 for iCCA, 1.69 for pCCA, and 1.66 for dCCA. Corresponding multivariable HRs were 1.74, 1.60, and 1.66, respectively.

Conclusions: PNI is a prognostic factor for both DFS and OS in CCA, regardless of anatomical location. Further research is needed to elucidate its biological mechanisms and potential therapeutic implications.

Impact and implications: This comprehensive review and meta-analysis confirms that perineural invasion (PNI) predicts poorer overall and disease-free survival in cholangiocarcinoma, independent of tumor location. These results highlight the potential of PNI as a reliable histopathological marker to stratify postoperative risk and inform patient management. While most evidence is retrospective and smaller studies may overstate the effect, the consistent findings emphasize the need for large, multicenter prospective studies with standardized pathological assessment. Such efforts could facilitate the integration of PNI into prognostic models and support more individualized treatment strategies for patients with cholangiocarcinoma.

Background & aims: The optimal management and outcomes of patients with HBeAg-negative grey-zone (GZe-) chronic HBV infection remain debatable. We assessed the outcomes and real-life management of GZe-patients and compared them to patients with typical HBeAg-negative chronic infection (CIe-).

Methods: We included all HBeAg-negative patients with baseline HBV DNA ≤20,000 IU/ml or HBV DNA >20,000 IU/ml and ALT <2x the upper limit of normal (ULN). Among patients without treatment indications in year 1, those with persistently normal ALT (≤ULN) and HBV DNA <2,000 IU/ml were defined as typical CIe-, and all others were classified as GZe-. Outcomes included treatment initiation, HBsAg loss, hepatocellular carcinoma (HCC), and liver-related events (LREs: HCC, decompensated cirrhosis, liver transplantation, or liver-related death).

Results: In total, 1,501 patients with a mean follow-up of 6.0 ± 4.6 years were included (GZe-/CIe-baseline characteristics: 811/690; GZe-/CIe-at year 1: 719/677). Compared with CIe-patients, GZe-patients more frequently developed treatment indications after year 1 (year 5: 13.4% vs. 2.2%; log-rank, p <0.001) and were more frequently treated after year 1 (year 5: 37.6% vs. 4.6%; log-rank, p <0.001). GZe-patients, particularly those with GZe-baseline characteristics, were less likely to achieve HBsAg loss (1-/5-year: 0.1/1.0% vs. 1/3%; log-rank, p = 0.012) and more frequently developed HCC (1-/5-year: 1/3% vs. 0%; log-rank, p <0.001) and LREs (1-/5-year: 1/4% vs. 0.1%; log-rank, p <0.001).

Conclusions: GZe-patients represent a large proportion of patients with chronic HBV seen at tertiary centers and meet treatment indications more frequently than CIe-patients. They also have a lower probability of HBsAg loss and higher risks of HCC and LREs despite treatment initiation in most cases; therefore, their optimal management and timing of treatment initiation require further evaluation.

Impact and implications: The outcomes of patients with grey-zone HBeAg-negative chronic HBV infection remain uncertain, hindering their optimal management and timely treatment in clinical practice. Our real-world data from a tertiary HBV center provide valuable insights to guide the management of this patient group. Grey-zone HBeAg-negative patients represent a substantial proportion of the chronic HBV population and require careful monitoring, as they are at increased risk of hepatocellular carcinoma and other liver-related events.