The combination of immune checkpoint inhibitors (ICIs) and anti-angiogenic agents represents the standard first-line therapy for patients with unresectable hepatocellular carcinoma (HCC). However, most patients do not derive sustained clinical benefit. Emerging evidence has underscored a strong association between immunotherapy efficacy and the tumor immune microenvironment (TIME) in HCC. Accordingly, this study aimed to characterize the TIME and develop a potential approach for predicting patient survival. We analyzed a cohort of 78 patients with unresectable HCC who received ICIs combined with anti-angiogenic therapy. Four multiplex immunohistochemistry (mIHC) panels were designed to comprehensively evaluate tumor-infiltrating immune cells (TIICs). Digital pathology was applied to raw imaging data to extract TIME features, including positive rates and spatial distributions of TIICs. Machine learning algorithms were then used to construct predictive models based on TIME-associated signatures (TIS). High positive rates of CD103+ and CD103+CD8+ T cells were associated with prolonged overall survival (OS). Conversely, a high positive rate of CD8+PD-L1+ T cells correlated with shorter progression-free survival (PFS). A greater abundance of CD8+ and CD103+ T cells in close proximity to tumor cells was also associated with longer OS. Multivariate Cox models incorporating these cell populations demonstrated that a lower TIS was significantly associated with longer OS and PFS. The composition and spatial distribution of immune cells critically shape the TIME in HCC and influence immunotherapy outcomes. TIS-based models show promise for predicting immunotherapy response in HCC patients, though they require further validation in larger prospective cohorts. These immune cell populations may serve as prognostic biomarkers and potential targets for personalized immunotherapy.

Humoral immunological memory mediated by memory B cells (MBCs) and long-lived plasma cells (LLPCs) is critical for sustained protection following infection or vaccination. LLPCs protect the hosts by secreting protective neutralizing antibodies over extended periods. However, the mechanism regulating their survival and thus the durability of protective antibodies remains unclear. Here, we showed in human and mouse models that intermittent fasting impaired humoral immunological memory by accelerating antibody decay. Fasting selectively depleted LLPCs while sparing MBCs in mice. Mechanistically, this effect was mediated by increased extracellular β-hydroxybutyrate, a ketone body produced during fasting, which acted through the hydroxycarboxylic acid receptor 2 (HCAR2) on plasma cells. Activation of the HCAR2-Gαi-adenylate cyclase-cAMP axis by β-hydroxybutyrate downregulated CXCR4, leading plasma cells to exit their bone marrow niche and undergo apoptosis in the periphery. These findings reveal that fasting-induced metabolic signals regulate humoral immunity duration and suggest that diet and lifestyle could influence vaccine effectiveness.

Mitochondrial dysfunction is a hallmark of aging and a key contributor to age-related diseases including cardiovascular disease. However, molecular pathways that safeguard mitochondrial homeostasis in the aging heart remain poorly understood. Here, we identify MTFR1L as a regulator of mitophagy that binds p-S65-Ub, a key signal amplifying the PINK1/Parkin axis. We find that MTFR1L is enriched in metabolically active tissues, particularly in the heart, where it regulates Parkin signaling. Genetic deletion of Mtfr1l in mice impairs stress-induced mitophagy and Parkin activation, leading to accumulation of damaged mitochondria, increased inflammation and senescence, and accelerated age-related cardiac dysfunction. Strikingly, cardiac expression of MTFR1L progressively decreases along with aging in mice, primates, and humans, coinciding with cardiomyocyte senescence and lipofuscin accumulation. Together, these findings uncover a role for MTFR1L in regulation of the p-S65-Ub/Parkin mitophagy axis and maintenance of mitochondrial homeostasis during cardiac aging and suggest that age-associated loss of MTFR1L may contribute to age-related cardiac dysfunction. Based on these findings, we propose a therapeutic paradigm for the prevention of heart aging by restoring MTFR1L function.

Boron Neutron Capture Therapy (BNCT) is a binary radiotherapy based on the combination of 10B drugs and thermal neutrons. Coordinating 10B with neutron irradiation will optimize the efficacy of BNCT, but further improvements are still required. Here, we develop a delivery system incorporating microneedles (MNs) and a modified boron agent PAMAM-BSH, enabling precise spatiotemporal matching of 10B with neutron irradiation for melanoma BNCT. The PAMAM-BSH is a cationic polymer 10B agent with amphiphilicity, forming nanomicelles in an aqueous solution. It exhibits high boron content and demonstrates good uptake and retention capabilities in cancer cells. When administering PAMAM-BSH-loaded MNs to melanoma-bearing mice, the duration for which the 10B content in tumors exceeds 20 ppm can extend up to 2 h. Notably, the system facilitates the precise administration of 10B into tumor cells, achieving concentrations exceeding 35 ng 10-6 cells. These contribute to an excellent alignment of spatiotemporal matching of 10B with neutron irradiation in BNCT. Meanwhile, the T/N (tumor-to-normal tissue) and T/B (tumor-to-blood) ratios can reach 30 and 100, respectively, which are much higher than the clinical BPA (T/N is 1.1-2.9, T/B is 1.1-3.6). Utilizing the system ultimately demonstrated a significant enhancement in melanoma BNCT, along with improved safety.

Lung adenocarcinoma (LUAD), the most common subtype of lung cancer, is a major contributor to worldwide cancer deaths. Although the slit guidance ligand (SLIT) protein family is implicated in both normal biological processes and disease states, the specific role of SLIT2 in LUAD development remains unclear. To address this gap, the present study integrated bioinformatics analyses and experimental studies to investigate the functional significance and clinical relevance of SLIT2 in LUAD. The present analysis revealed that SLIT2 expression is significantly reduced in LUAD tissues and cell lines, and this decrease is associated with poorer patient outcomes. Functional experiments showed that inhibiting SLIT2 expression enhances LUAD cell growth, migration, invasion and epithelial-mesenchymal transformation of LUAD cells, whereas overexpression of SLIT2 can reverse these carcinogenic effects. Furthermore, it was observed that SLIT2 expression levels correlate with distinct patterns of immune cell infiltration in the tumor environment. These results suggested that targeting SLIT2 pathways could represent a novel therapeutic strategy for LUAD and further research is needed to explore the specific mechanisms through which SLIT2 exerts its tumor-suppressive effects and modulates immune responses. The current study enhances the understanding of LUAD biology and underscores the potential of SLIT2 as a biomarker and therapeutic target in LUAD.

CD47 is a crucial anti-phagocytic signal in regulating macrophage responses and its manipulation offers the therapeutic potential in cancer treatment. However, in many cases, blockade of CD47 by itself is insufficient to activate macrophage effectively, indicating other unidentified phagocytosis-regulating factors to resist the macrophage activity. In this study, a genome-wide human CRISPR-Cas9 library was developed for comprehensive screening of phagocytosis-regulating factors in the context of CD47 blockade. The screening results identified GSTK1 as a potential anti-phagocytic signal counteracting the efficacy of CD47-based phagocytosis. The disruption of GSTK1 significantly increased the phagocytosis rate of cancer cells by macrophages in combination with anti-CD47 antibody. Further mechanism investigation unveiled that GSTK1 blockade increased the membrane exposure of calreticulin in different cancer cells, which might be the primary mechanism driving enhanced macrophage-mediated phagocytosis. To this end, siGSTK1-loaded nanoparticles (siGSTK1-LNPs) were designed to suppress the GSTK1 expression efficiently. The comparable phagocytosis efficacy was also observed when combining siGSTK1-LNPs with anti-CD47 antibody. Above all, GSTK1 blockade was identified as a promising and feasible stimulus for enhancing the effectiveness of anti-CD47 antibody, introducing a novel and effective combination approach in cancer immunotherapy.

The constrained cross-talk between myeloid cells and T cells in the tumor immune microenvironment (TIME) restricts cancer immunotherapy efficacy, whereas the underlying mechanism remains elusive. Parkin, an E3 ubiquitin ligase renowned for mitochondrial quality control, has emerged as a regulator of immune response. Here, we show that both systemic and macrophage-specific ablations of Parkin in mice lead to attenuated tumor progression and prolonged mouse survival. By single-cell RNA-seq and flow cytometry, we demonstrate that Parkin deficiency reshapes the TIME through activating both innate and adaptive immunities to control tumor progression and recurrence. Mechanistically, Parkin activation by AMP-activated protein kinase rather than PTEN-induced kinase 1 mediated major histocompatibility complex I down-regulation on macrophages via Autophagy related 5-dependent autophagy. Furthermore, Parkin deletion synergizes with immune checkpoint blockade treatment and Park2-/- signature aids in predicting the prognosis of patients with solid tumor. Our findings uncover Parkin's involvement in suppressing macrophage antigen presentation for coordinating the cross-talk between macrophages and T cells.

Previous research has demonstrated that Dock2 deficiency results in a reduction in both the quantity and proliferation rate of T cells, thereby heightening the host's vulnerability to various infections. Nevertheless, the impact of DOCK2 on T cell activation remains unexplored. In this study, we employed flow cytometry to assess the activation phenotype of T cells in the peripheral lymphoid tissues of wild-type (Dock2+/+), DOCK2 heterozygous (Dock2+/-) and DOCK2 knockout (Dock2-/-) mice. Our findings revealed that, in comparison to Dock2+/+ mice, Dock2-/- mice exhibited increased expression levels of CD44 and CD69 on CD4+ and/or CD8+ T cells within spleen and mesenteric lymph nodes (MLN). Additionally, there was a significant elevation in the proportions of IFN-γ+/CD4+, IFN-γ+/CD8+ and IL-4+/CD8+ T cells. Furthermore, the percentage of IL-17a+/CD4+ and IL-17a+/CD8+ T cells in the MLN of Dock2-/- mice was higher than that observed in Dock2+/+ mice. These results suggest that Dock2 deficiency induces aberrant T cell activation in peripheral lymphoid tissues. To further investigate the underlying mechanisms of this phenomenon, we conducted transcriptome sequencing on CD8+ T cells collected from all groups of mice. The results indicate that Ccr2 and Ifng are potentially pivotal genes involved in the aberrant activation of T cells in Dock2-/- mice. These findings contribute to elucidating the host defense mechanisms against foreign pathogens and advance our comprehension of the role of cytoskeleton-related proteins in the regulation of cellular immunity.

BackgroundBreast cancer (BC) represents a highly heterogeneous malignancy and continues to be a leading source of death among women worldwide. Enhancing diagnostic and therapeutic approaches necessitates a thorough grasp of the underlying molecular pathways and the identification of dependable biomarkers. Although palmitoyl transferases, particularly ZDHHC9, have been associated with the progression of various cancers, their specific role in BC remains incompletely understood.MethodsIn this investigation, TCGA and GTEx databases were utilized to analyze the expression patterns of ZDHHC9 and to evaluate its prognostic significance. Moreover, the regulatory pathways involving ZDHHC9 were explored via co-expression analysis and differential gene enrichment studies. Insights into ZDHHC9 expression across different cell types and its potential oncogenic pathways were derived from scRNA sequencing analysis. Additionally, immunophenoscore (IPS), EaSIeR and immunotherapy cohorts were utilized to predict immunotherapy responses. The biological significance of ZDHHC9 was verified through in vitro and in vivo experiments.ResultsOur findings revealed that ZDHHC9 is markedly overexpressed in BC, with elevated levels of ZDHHC9 being correlated with poor survival outcomes, suggesting its role as an independent risk factor in BC. Furthermore, high ZDHHC9 expression was found to be associated with multiple immune cell types within BC. Notably, patients exhibiting lower ZDHHC9 expression demonstrated a higher likelihood of benefitting from immunotherapy. Lastly, the vivo and vitro experiments consistently demonstrated that suppression of ZDHHC9 expression could reduce BC cell proliferation.ConclusionsThis study highlights ZDHHC9 as a potential prognostic marker, a regulator of tumor immunity, and a biomarker of therapeutic response in BC, offering a promising avenue for improving BC diagnosis and treatment.

Staphylococcal nuclease and Tudor domain containing 1 (SND1) is an emerging oncoprotein highly expressed in various tumors. Database analyses indicate that SND1 is enriched in tumor-derived exosomes, suggesting its potential role in modulating the tumor microenvironment (TME) via exosomes. Here, we demonstrated that SND1 served as a novel tumor-derived exosome (TEX) marker, influencing macrophage polarization by enriching exosomal membrane proteins. In mice, SND1 enriched in melanoma-derived exosomes promoted lung metastasis, accompanied by increased tumor-associated macrophage (TAM) infiltration. Conversely, SND1-deficient exosomes (ExoSND1-KO) shifted macrophage polarization toward an M1 phenotype, creating an anti-tumor immune microenvironment and inhibiting melanoma lung metastasis. Mechanistically, SND1 promoted ESCRT-dependent CD47 sorting, thereby facilitating its incorporation into melanoma-derived exosomes and allowing them to evade macrophage-mediated phagocytosis through the CD47-SIRPα axis. Consequently, macrophages failed to engulf TEXs or tumor cells. Notably, ExoSND1-KO, lacking CD47, were preferentially phagocytosed by macrophages, triggering M1 reprogramming via exosome-derived dsDNA activation of the cGAS-STING/TBK1/NF-κB pathway. This process led to increased secretion of inflammatory cytokines (IL-1β, IL-6, TNF-α) and activation of type I cell-mediated immunity. Our study suggests that targeting SND1 enrichment in tumor cells could be a promising strategy to inhibit tumor metastasis.

Immune checkpoint inhibitor (ICI) has limited efficacy in the treatment of immune "cold" tumors. Due to insufficient T cell infiltration and heterogeneous programmed death ligand 1 (PD-L1) expression, the ORR is only 5%-8% compared with 30%-40% of "hot" tumors. This article reviews the synergistic mechanism, clinical efficacy and optimization strategy of oncolytic virus (OVs) combined with ICIs in the treatment of refractory malignant tumors. Systematic analysis of mechanistic interactions across tumor types and clinical trial data demonstrates that OVs transform the immunosuppressive microenvironment by inducing immunogenic cell death and activating innate immunity. Concurrently, ICIs enhance adaptive immunity by reversing T-cell exhaustion and expanding T-cell diversity. Clinical trials in melanoma, head and neck cancer and breast cancer showed superior efficacy. The Objective Response Rate (ORR) of combination therapy was 39%-62%, while the ORR of ICI monotherapy was 18%. Treatment heterogeneity is mainly attributed to virus-related factors, including targeting specificity and replication efficiency, tumor characteristics, such as antigen presenting ability and mutation load, and host immune status, including pre-existing antiviral antibodies and microbiome composition. This combined approach represents a paradigm shift in cancer immunotherapy, which effectively transforms immune "cold" tumors into "hot" tumors through the continuous activation of innate and adaptive immune responses. In the future, it is expected to improve the therapeutic effect of treatment-resistant malignant tumors through the integration of immune regulatory molecules, accurate biomarkers to guide the treatment scheme and triple combination strategy by a new generation of engineering viruses.

Four new compounds (1-4), along with 22 known metabolites (5-26), were isolated from the fungus Biscogniauxia sp. 8703. The structures of the new compounds were elucidated based on NMR, MS, and ECD analysis. Compounds 1 and 2 were identified as heliannuol D analogs, which exhibited anti-inflammatory activity by inhibiting NO production in LPS-induced RAW 264.7 cell, with IC50 values of 7.14 and 25.25 μM, respectively.

BackgroundGastric cancer (GC) is a highly aggressive and fatal disease, with limited treatment options. Altered cellular lipid metabolism is a hallmark of cancer that contributes to GC progression. However, the determinants of lipid metabolism in GC and new agents that target lipid metabolic pathways are poorly defined. The aim of this study was to identify potentially effective lead anti-GC compounds by developing and exploring natural marine products. Furthermore, we sought to uncover viable therapeutic targets for GC through the underlying mechanisms of action of the compounds as an anticancer agent.MethodsThe chemical structures of the metabolites produced by the crinoid-derived fungus Penicillium brocae SYSU-CJ17 were elucidated using advanced spectroscopic techniques. These metabolites were screened for their growth-inhibitory effects on GC cell lines. Among them, Penicolinate H (Pen-H) demonstrated the most significant anti-cancer activity. High-throughput RNA sequencing of Pen-H-treated GC samples revealed differentially expressed genes, and transcriptomic data integrated with bioinformatics analyses highlighted the potential pathways and target genes through which Pen-H might exert its anti-cancer effects. Further investigations, including rescue experiments, endogenous affinity pull-down assays, cellular thermal shift assays (CETSA), surface plasmon resonance (SPR) assays, molecular docking, and in vitro analyses, confirmed the interaction between Pen-H and SREBP-1. The feasibility of SREBP-1 as a therapeutic target for GC is supported by single-cell transcriptome analysis, bioinformatics evaluation of GC patient data, and in vitro studies. Additionally, the chemosensitization effect of Pen-H was confirmed by in vivo and in vitro experiments.ResultsOur findings reveal a novel marine-derived compound, Pen-H, which inhibits GC growth and metastasis both in vitro and in vivo by suppressing SREBP-1 mediated lipogenesis. Bioinformatic analysis indicated that SREBP-1 was highly expressed in GC tissues, and high SREBP-1 transcript levels were negatively correlated with prognosis in GC patients. SREBP-1 depletion significantly inhibits the proliferation, migration, and invasion of GC cells. Mechanistic studies have revealed that targeting SREBP-1 by Pen-H significantly reduces de novo fatty acid synthesis and that the anti-GC efficacy of Pen-H is SREBP-1 dependent. Moreover, combination treatment with Pen-H and 5-fluorouracil (5-Fu) resulted in enhanced inhibition of cell proliferation and tumor growth compared to monotherapy.ConclusionTaken together, these findings highlight that SREBP-1 is an effective therapeutic target in GC and that Pen-H is a promising SREBP-1 inhibitor and a candidate for GC treatment.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer associated with severe pain and depression. Neuropeptide VGF (non-acronymic) exhibits robust expression in the pancreas and brain, known for its modulatory roles in metabolic homeostasis, nociception, and depression-like behaviors. Despite elevated VGF expression being linked to poor prognosis in various cancers, its specific role in PDAC remains unexplored. By combining bioinformatic analysis of clinical datasets with experimental validations, we uncover that high VGF expression correlates with improved survival in PDAC patients. Notably, the administration of TLQP-21, a C-terminal peptide derived from VGF, significantly reduces tumor size and enhances the therapeutic efficacy of gemcitabine, resulting in a marked increase in overall survival in an orthotopic mouse model of PDAC. Mechanistically, TLQP-21 suppresses the tumor-promoting effects of tumor-associated macrophages through complement receptors C3aR1 and C1qBP. Additionally, TLQP-21 alleviates depression-like behaviors, allodynia, and muscle wasting in PDAC mice. Collectively, these findings demonstrate the dual efficacy of TLQP-21 in inhibiting tumor growth and mitigating nociceptive and psychiatric symptoms, highlighting the potential of TLQP-21 as a therapeutic option for PDAC.

Resistance to osimertinib, a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), is a significant challenge for patients with advanced EGFR-mutant lung adenocarcinoma (LUAD). Fat mass and obesity-associated protein (FTO), an N6-methyladenosine (m6A) demethylase, plays a critical role in the occurrence, metastasis, and drug resistance of various tumors. We found that high FTO expression was significantly associated with resistance to osimertinib in advanced LUAD patients harboring EGFR mutations. In vitro experiments indicated that FTO enhanced the resistance of LUAD cells to osimertinib by reducing m6A modification. FTO silencing induced G0/G1 phase arrest in resistant cells in an m6A-dependent manner, restoring osimertinib sensitivity. Both in vivo and in vitro studies showed that FB23-2, an FTO inhibitor, synergistically suppressed the growth of resistant cells in combination with osimertinib. Furthermore, LUAD patient-derived organoids maintained histological and genetic consistency with patient tissues, where FTO was highly expressed in osimertinib-resistant organoids. The combination of FB23-2 and osimertinib effectively inhibited the growth of osimertinib-resistant organoids. Overall, this study demonstrates that FTO promotes osimertinib resistance and targeting FTO induces G0/G1 arrest in an m6A-dependent manner. The combination of FTO inhibitor and osimertinib provides a strategy to override osimertinib resistance.

Cancer immunotherapies exhibit impressive efficacy in some cancers but show only limited benefits for refractory hematological malignancies. The complex immune escape mechanisms of hematological cancers remain unclear. Here, we found that P-selectin glycoprotein ligand 1 (PSGL-1) was highly expressed by hematological cancers and negatively correlated with cancer prognosis. PSGL-1 deficiency in tumors suppressed the progression of multiple mouse models of hematological cancer by promoting infiltration of macrophages and their phagocytic activity. Tumor PSGL-1 inhibited the interaction between tumor ICAM-1 and CD11a/CD18 integrin (LFA-1) in macrophages, thereby suppressing prophagocytic signaling downstream of LFA-1. A humanized antibody targeting human PSGL-1 (αhPSGL-1) efficiently triggered macrophage phagocytosis of human hematological malignancies in vitro and slowed cancer progression in vivo. Additionally, PSGL-1 blockade potentiated the efficacy of doxorubicin chemotherapy and anti-CD47 and anti-CD38 antibody therapy. Therefore, PSGL-1 is a previously undescribed phagocytosis checkpoint, and targeting PSGL-1 could be a promising immunotherapy strategy for treating hematological malignancies.

The dysregulation of long non-coding RNAs (lncRNAs) are involved in regulating tumor progression in multiple manner. However, little is known about whether lncRNA is involved in the translation regulation of proteins. Here, we identified that the suppressor of inflammatory macrophage apoptosis lncRNA (SIMALR) was highly expressed in nasopharyngeal carcinoma (NPC) tissues by analyzing the lncRNA microarray. Clinically, the high expression of SIMALR served as an independent predictor for inferior prognosis in NPC patients. SIMALR functioned as an oncogenic lncRNA that promoted the proliferation and metastasis of NPC cells in vitro and in vivo. Mechanistically, SIMALR served as a critical accelerator of protein synthesis by binding to eEF1A2 (eukaryotic translation elongation factor 1 alpha 2), one of the most crucial regulators in the translation machinery of the eukaryotic cells, and enhancing its endogenous GTPase activity. Furthermore, SIMALR mediated the activation of eEF1A2 phosphorylation to accelerate the translation of ITGB4/ITGA6, ultimately promoting the malignant phenotype of NPC cells. In addition, N-acetyltransferase 10 (NAT10) enhanced the stability of SIMALR and caused its overexpression in NPC through the N4-acetylcytidine (ac4C) modification. In sum, our results illustrate SIMALR functions as an accelerator for protein translation and highlight the oncogenic role of NAT10-SIMALR-eEF1A2-ITGB4/6 axis in NPC.