Liver cancer is among the world’s leading causes of cancer-related death, and hepatocellular carcinoma, or HCC, accounts for most primary liver cancers. One reason HCC is difficult to treat is its dependence on a highly adaptable tumor microenvironment. As tumors enlarge, they require oxygen and nutrients that cannot diffuse far through tissue. Cancer cells therefore release signals that encourage nearby blood vessels to sprout and connect with the tumor. This process, called angiogenesis, can also create disordered, leaky vessels that help malignant cells invade surrounding tissue and enter the circulation. Drugs that inhibit angiogenesis have improved outcomes for some patients, but tumors frequently develop resistance by activating alternative vascular signals. The new study, published in Clinical Proteomics, focuses on a molecular layer that may help explain how HCC sustains this vascular supply.

The central protein, LGALS3BP, is a secreted molecule that has been found at elevated levels in several malignant tumors, but its specific role in HCC angiogenesis has been unclear. The researchers first used bioinformatics analyses to examine LGALS3BP expression in liver cancer datasets and its relationship to angiogenesis-related genes and the NOTCH pathway. They then compared messenger RNA and protein levels in HCC tissues and cancer-cell models using quantitative reverse-transcription PCR and Western blotting. They also measured secreted LGALS3BP with enzyme-linked immunosorbent assays. Across these analyses, LGALS3BP was substantially more abundant in HCC tissue and cells than in corresponding noncancerous controls, supporting the idea that it is not merely a passive marker of malignancy but could be an active participant in tumor biology.

To test that possibility, the team reduced LGALS3BP production in HCC cells and measured several behaviors associated with aggressive disease. Loss of the protein diminished cell viability and proliferation, weakened the ability of cells to form colonies and reduced invasion through laboratory membranes in Transwell assays. The researchers also observed lower levels of VEGF-A, a powerful signal that binds receptors on endothelial cells and stimulates their migration, division and organization into new vascular structures. In a co-culture system, cancer cells were grown alongside human umbilical vein endothelial cells. When LGALS3BP was depleted, the endothelial cells formed fewer and less extensive tube-like networks, an in-vitro model of angiogenic activity. Together, the results linked LGALS3BP to both cancer-cell aggressiveness and the capacity of tumor cells to recruit vascular support.

The study then moved upstream to ask how LGALS3BP is regulated. The researchers identified an interaction between LGALS3BP and UGGT1, an enzyme located in the endoplasmic reticulum, the cellular compartment where many newly synthesized proteins are folded and processed. UGGT1, short for UDP-glucose:glycoprotein glucosyltransferase 1, functions as part of the endoplasmic-reticulum quality-control system. It recognizes incompletely folded glycoproteins and adds a glucose residue to specific N-linked carbohydrate chains, allowing the proteins to re-enter a cycle of binding to molecular chaperones. This quality-control process can give a protein additional opportunities to fold correctly before it is transported through the secretory pathway. Because LGALS3BP is secreted, its stability and maturation may be particularly sensitive to such processing.

Using protein-interaction analyses, co-immunoprecipitation and immunofluorescence microscopy, the researchers found evidence that UGGT1 associates with LGALS3BP. They next examined potential N-glycosylation sites—positions where a carbohydrate chain can be attached to an asparagine residue within a characteristic amino-acid sequence. LGALS3BP contains six candidate sites, and the team generated individual mutants in which the relevant asparagine was replaced, as well as a combined mutant lacking all six sites. These experiments showed that UGGT1-mediated glycosylation helped maintain LGALS3BP protein stability. When UGGT1 was reduced, LGALS3BP protein levels and secretion declined; restoring or increasing LGALS3BP could counter some of the downstream effects. The results point to a mechanism in which UGGT1 does not simply alter a signal at the cell surface, but helps preserve the supply of a secreted factor that cancer cells use to remodel their surroundings.

The downstream signaling route involved NOTCH, a communication system that helps neighboring cells coordinate decisions about development, survival and cell identity. NOTCH signaling begins when a ligand on one cell binds a NOTCH receptor on an adjacent cell. Proteolytic cleavage then releases the receptor’s intracellular domain, which travels to the nucleus and partners with DNA-binding proteins to switch target genes on or off. In blood-vessel formation, NOTCH helps regulate the balance between endothelial “tip” cells that lead a new sprout and “stalk” cells that follow and proliferate. Dysregulated NOTCH activity can therefore reshape the architecture and behavior of tumor-associated vessels. In the HCC models, LGALS3BP increased activity of NOTCH-associated molecules and raised VEGF-A expression, while suppressing LGALS3BP weakened this signaling axis. The findings support a UGGT1–LGALS3BP–NOTCH–VEGF-A chain connecting intracellular protein quality control to the formation of new tumor blood vessels.

The most demanding test came in animals bearing xenograft tumors, in which human HCC cells were implanted into mice. Reducing UGGT1 significantly slowed tumor growth and decreased markers of angiogenesis within the tumors. When LGALS3BP was overexpressed, some of the effects of UGGT1 loss were reversed, a result known as partial rescue. Such rescue experiments are important because they help place LGALS3BP downstream of UGGT1 rather than showing only that both proteins happen to be associated with aggressive tumors. Even so, partial rescue does not prove that the pathway is the only route by which UGGT1 influences cancer. UGGT1 participates in broad protein-folding quality control, and altering it could affect many glycoproteins at once. The apparent anti-tumor effect may therefore reflect a combination of LGALS3BP-dependent and independent changes.

The work raises the possibility of attacking liver cancer angiogenesis at a different point from existing VEGF-directed drugs. Directly blocking VEGF-A can starve tumors of vascular support, but cancer cells may compensate by increasing other growth factors or by changing the structure of their vessels. Inhibiting UGGT1 could, in principle, reduce the stability of several pro-tumor glycoproteins, while targeting LGALS3BP might offer a more focused way to interfere with the secreted signal identified in this study. Yet both strategies present challenges. UGGT1 is part of a normal cellular quality-control system, so systemic inhibition could harm healthy secretory tissues or trigger endoplasmic-reticulum stress. LGALS3BP is also expressed in noncancerous contexts, meaning that its role in immunity, tissue repair and other physiological processes will need to be defined before it can be safely targeted. The experiments used cell lines, endothelial co-cultures and xenograft models, which do not fully reproduce the immune system, genetic diversity or treatment history of human HCC.

The researchers describe their findings as evidence for a potential anti-angiogenic target, not as a ready-made therapy. The next steps will include confirming the pathway in larger collections of patient tumors, determining whether UGGT1 or LGALS3BP levels predict vascular activity or treatment response, and developing inhibitors that can distinguish cancer-relevant signaling from essential normal protein processing. It will also be important to test whether disrupting the pathway improves the performance of existing immunotherapies or molecular treatments, and whether tumors can bypass it through alternative angiogenic programs. For now, the study offers a striking mechanistic explanation for how a modification made inside the endoplasmic reticulum may influence events far outside the cancer cell: by stabilizing a glycosylated secreted protein, UGGT1 may help HCC turn on NOTCH and VEGF-A, recruiting the blood vessels that allow the disease to expand.

At the core of this investigation lies the comprehensive analysis of eleven distinct acylation modifications, including diverse modalities such as crotonylation, lactylation, succinylation, and others like benzoylation and butyrylation. These chemical alterations on protein substrates are known to intricately regulate cellular functions, yet their collective impact on HCC progression and prognosis had remained obscure. By generating consensus clusters from patient tumor data, researchers delineated two acylation modification-related subtypes with distinct molecular identities and clinical behaviors.

To unravel the genetic networks underpinning these subtypes, the team employed Weighted Gene Co-Expression Network Analysis (WGCNA). This approach facilitated the detection of gene modules closely correlated with acylation processes, enabling a refined understanding of the transcriptional programs operative within HCC tumors. Subsequently, machine learning algorithms were harnessed to distill these complex genetic profiles into a practical and quantifiable scoring system— the Acylation Modification-Related Gene score (AMRG.score).

This scoring system, comprising 21 rigorously selected key genes, stands as a powerful predictive tool for assessing patient prognosis. Its robustness was validated across multiple independent cohorts beyond the initial discovery set, including diverse datasets such as TCGA-LIHC, LIRI-JP, and several GEO repositories (GSE10143, GSE14520, GSE27150, GSE36376, and GSE76427), as well as a clinical in-house cohort. This extensive validation reinforces the generalizability and clinical relevance of the AMRG.score across heterogeneous patient populations.

Beyond its prognostic capabilities, the AMRG.score revealed profound insights into the intricacies of the tumor microenvironment (TME) in HCC. Patients with elevated scores were found to possess an immunologically active TME characterized by increased infiltration of immune effector cells and heightened expression of immune checkpoint molecules. Such immunological landscapes typically herald enhanced responsiveness to immunotherapies, underscoring the clinical utility of the AMRG.score in stratifying candidates for these treatments.

This study also sheds light on the dynamic interplay between acylation modifications and immunosuppressive mechanisms within HCC. Post-translational modifications like crotonylation and lactylation were implicated in modulating immune evasion pathways, which are pivotal barriers to effective antitumor immune responses. Understanding these modifications at a molecular level paves the way for novel therapeutic strategies that could synergize with existing immunotherapies to overcome resistance.

Furthermore, the integration of multi-omics data underscores the complexity of HCC biology, highlighting how epigenetic and metabolic alterations converge via acylation modifications to influence tumor behavior. This systems-level perspective is critical for developing precision oncology approaches, tailoring interventions based on individual tumor acylation profiles to maximize therapeutic benefit and minimize toxicity.

The identification and functional characterization of the 21 gene signature supporting the AMRG.score offer promising avenues for future research. These genes span diverse biological processes, from metabolic regulation to immune signaling, serving as potential biomarkers and therapeutic targets. Functional validation of these targets could spearhead the design of novel pharmacological agents aimed at modulating acylation-driven pathways.

Importantly, this research underscores the transformative potential of integrating computational biology and clinical oncology. Machine learning not only facilitated the stratification of complex data but also converted biological phenomena into actionable clinical metrics. This approach exemplifies the future of translational research, where data science amplifies the discovery-to-clinic pipeline.

The clinical implications of the AMRG.score extend to patient management paradigms. By predicting both prognosis and immunotherapy sensitivity, this tool empowers oncologists to make informed decisions regarding treatment intensity and modality. Patients with high AMRG.score might benefit from early and aggressive immunotherapeutic interventions, while those with lower scores could be spared unnecessary toxicity from less effective immune-based treatments.

From a broader perspective, the study highlights post-translational acylation as a vital frontier in cancer epigenetics and immunology. As an emerging category of modifications beyond classical phosphorylation and ubiquitination, acylation defines a new layer of regulatory complexity with significant translational promise. This conceptual advance invites the oncology community to revisit molecular mechanisms governing tumor-immune interactions.

The findings also encourage exploration into how acylation modifications might impact other cancer types and treatment contexts. Given the conserved nature of many acylation pathways, it is plausible that similar prognostic and therapeutic paradigms could be extrapolated beyond HCC, potentially revolutionizing personalized medicine across a spectrum of malignancies.

Ultimately, the integration of acylation biology into clinical prognostic frameworks and therapeutic design symbolizes a leap forward in the fight against HCC. This study equips researchers and clinicians with a refined lens to view tumor biology while providing patients with hope for more precise, effective treatment strategies rooted in molecular insight.

As the field progresses, future investigations will undoubtedly delve deeper into the mechanistic underpinnings of acylation-mediated immune modulation and its synergy with emerging immunotherapies, including checkpoint inhibitors and adoptive cell therapies. Combining such knowledge with innovative drug delivery systems could herald a new era of targeted, acylation-informed therapeutics.

In conclusion, this landmark study not only elucidates the prognostic value of acylation-related gene signatures in hepatocellular carcinoma but also bridges fundamental biology with clinical application. Through the creation and validation of the AMRG.score, the research offers a transformative tool capable of guiding personalized treatment and enhancing the efficacy of immunotherapy, marking a seminal contribution to oncology and immunology.

Subject of Research: Post-translational acylation modifications and their impact on immunosuppression and immunotherapy efficacy in hepatocellular carcinoma.

Article Title: Post-translational acylation modulates immunosuppression and immunotherapy efficacy in hepatocellular carcinoma.

Article References:
Li, Y., Bai, S., Hu, J. et al. Post-translational acylation modulates immunosuppression and immunotherapy efficacy in hepatocellular carcinoma. Genes Immun (2025). https://doi.org/10.1038/s41435-025-00362-2

Image Credits: AI Generated

DOI: https://doi.org/10.1038/s41435-025-00362-2