Sialylation, a dynamic post-translational modification catalyzed by sialyltransferases and counterbalanced by neuraminidases, entails the attachment of sialic acids to the terminal residues of glycoproteins and glycolipids. This modification profoundly influences diverse biological processes, including early embryogenesis, neurodevelopment, maintenance of stem cell pluripotency, and oncogenic transformation. In cancer, aberrant sialylation manifests as altered linkage patterns and dysregulated expression of sialylated glycans, which directly drive malignant behaviors such as uncontrolled proliferation, enhanced adhesion and invasion, immune evasion, and therapy resistance. Deciphering the underlying molecular mechanisms is therefore crucial for advancing our understanding of tumor biology. In this review, we systematically summarize recent advances in the study of sialylation in cancer, with a focus on the biological functions of distinct sialyltransferases and neuraminidases. We further discuss the diagnostic, prognostic, and therapeutic implications of targeting sialylation, highlighting its emerging potential as a promising avenue for cancer treatment.

BACKGROUND:The precise causal relationship between alterations in the gut microbiota, microbiota-derived metabolites, and the development of diffuse large B-cell lymphoma (DLBCL) remains unclear. OBJECTIVES:To investigate the potential causal relationships between gut microbiota, microbiota-derived metabolites, and DLBCL. MATERIAL AND METHODS:Genetic data on gut microbiota were obtained from the MiBioGen consortium, while data on microbiota-derived metabolites were sourced from the TwinsUK and KORA studies.Statistics for DLBCL were retrieved from FinnGen. Mendelian randomization (MR) analysis was performed, with inverse-variance weighting (IVW) used as the primary analytical method. Sensitivity analyses included Cochran's Q test, the MR-Egger intercept test, and MR-PRESSO. Reverse MR analysis was conducted to assess potential bidirectional causal relationships between gut microbiota and DLBCL. Bayesian weighted MR (BWMR) was applied for additional validation to enhance the robustness of the findings. RESULTS:Among 196 gut microbial taxa analyzed, Bilophila (odds ratio (OR) = 1.777, 95% confidence interval (95% CI): 1.053-3.000, p = 0.031) was associated with an increased risk of DLBCL. In contrast, Alistipes (OR = 0.521, 95% CI: 0.311-0.873, p = 0.013) and Ruminococcaceae UCG011 (OR = 0.749, 95% CI: 0.574-0.978, p = 0.034) were associated with a reduced risk. Reverse MR analysis demonstrated a positive association between DLBCL risk and the abundance of Anaerofilum (OR = 1.087, 95% CI: 1.008-1.173, p = 0.031). Negative associations were observed between DLBCL risk and the abundance of Deltaproteobacteria (OR = 0.959, 95% CI: 0.922-0.997, p = 0.037), Desulfovibrionales (OR = 0.959, 95% CI: 0.922-0.998, p = 0.041), Oxalobacteraceae (OR = 0.914, 95% CI: 0.843-0.992, p = 0.031), and Oxalobacter (OR = 0.909, 95% CI: 0.837-0.988, p = 0.024). Analysis of microbiota-derived metabolites identified a causal association between indolepropionate (OR = 0.296, 95% CI: 0.131-0.669, p = 0.003) and reduced DLBCL risk, whereas 7-alpha-hydroxy-3-oxo-4-cholestenoate (7-HOCA) (OR = 9.561, 95% CI: 1.426-64.088, p = 0.020) was associated with an increased risk. No evidence of directional pleiotropy or heterogeneity was detected. CONCLUSIONS:This MR study provides evidence that specific gut microbial taxa and microbiota-derived metabolites may causally influence the risk of DLBCL.

The standard of care for advanced cervical cancer includes chemotherapy, antiangiogenic, and/or immune checkpoint blockade regimens. Although effective, it leads to pleiotropic side effects. Deescalation chemotherapy together with immunotargeted therapies has been proven effective and less toxic in other cancers. In this study, we conducted a multicenter, single-arm, phase II study of first-line deescalated platinum-based chemotherapy plus anlotinib and penpulimab, followed by maintenance therapy solely with anlotinib and penpulimab in patients with PD-L1-positive, persistent, recurrent, or metastatic cervical cancer. Of 32 efficacy-evaluable patients, 30 (93.8%, 95% confidence interval, 79.2%-99.2%) had an investigator-confirmed objective response. Single-nucleus RNA sequencing implied enhanced chemotaxis and proliferative activity of tumor-infiltrating T cells, and activated germinal center B cells portended optimal treatment response. Patients with a high tertiary lymphoid structure-to-tumor area ratio exhibited better survival. Our findings lay the groundwork for the feasibility of first-line de-escalated chemotherapy plus anlotinib and penpulimab in patients with metastatic, persistent, or recurrent cervical cancer. SIGNIFICANCE:We recruited 34 patients with advanced cervical cancer receiving two cycles of platinum-based chemotherapy plus anlotinib and penpulimab, followed by maintenance therapy solely with anlotinib and penpulimab, and showed safety and efficacy of this deescalation regimen. This work highlights the potential for personalized treatment strategies and feasibility of reduced-toxicity regimens.

This study aimed to identify essential genes driving lung adenocarcinoma (LUAD) progression by integrating CRISPR-Cas9 dependency data from the Cancer Dependency Map (DepMap) portal with transcriptomic profiles from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Through a machine learning-based screening pipeline, we identified GINS complex subunit 1(GINS1) as a pivotal oncogenic driver in LUAD. Multi-omics analysis and in-house clinical validation confirmed that GINS1 is significantly upregulated at both mRNA and protein levels, serving as an independent prognostic marker. Functional assays demonstrated that GINS1 knockdown markedly inhibited LUAD cell proliferation and migration while inducing G1-phase arrest and apoptosis. Notably, GINS1 depletion sensitized tumor cells to ferroptosis, evidenced by increased reactive oxygen species (ROS) accumulation and a significant reduction in the Half Maximal Inhibitory Concentration (IC50) of the ferroptosis inducer Imidazole Ketone Erastin (IKE). Mechanistically, GINS1 promotes an immunosuppressive microenvironment by driving M2 macrophage polarization via the C-C Motif Chemokine Ligand 2(CCL2) axis and fostering T-cell exhaustion and immune exclusion. Clinically, high GINS1 expression predicted unfavorable responses to immune checkpoint blockade (ICB) therapy across multiple independent cohorts. In conclusion, our study identifies GINS1 as a central regulator of tumor cell survival and immune evasion, highlighting its potential as a prognostic biomarker and a promising therapeutic target for precision oncology in LUAD.

IntroductionEvidence indicates that Chinese patent medicines can significantly increase bone mass in patients with osteoporosis and alleviate symptoms associated with low bone density. Although the therapeutic effects of these two drugs have been compared both directly and indirectly, no economic-related studies currently exist. Therefore, this study aims to assess the cost-effectiveness of Xianling Gubao Capsules compared to Jintiange Capsules and non-treatment for postmenopausal osteoporosis from the perspective of Chinese healthcare providers.MethodsA Markov microsimulation model was employed to estimate the cost-effectiveness of the Xianling Gubao capsule and the Jintiange capsule in a hypothetical cohort of postmenopausal osteoporotic women aged 55 to 74 years with no prior history of fractures, over a treatment period of 6 months. Model parameters, including transition probabilities and costs, were derived from Chinese sources. Efficacy data for the treatments were obtained from two network meta-analyses. Outcomes were expressed as incremental costs per quality-adjusted life-year (QALY) gained. Sensitivity analyses were performed to ensure the robustness of the findings, with a cost-effectiveness threshold established at three times the Gross Domestic Product (GDP) per capita in China ($38,223) per QALY.ResultCompared to the control group that did not receive drug treatment, the preventive therapy using Chinese patent medicine significantly increased bone mineral density and reduced the probability of fractures across all age groups in the intervention group. The incremental cost-effectiveness ratios (ICERs) for the Jintiange capsule compared to the Xianling Gubao capsule ranged from $11,955 per QALY at age 55 to $9,711 per QALY at age 74, indicating that the cost-effectiveness of the Jintiange capsule improved consistently with age. Sensitivity analyses confirmed the robustness of the results across all parameter variations, with the annual cost of the Jintiange capsule identified as the most sensitive factor.ConclusionFrom the perspective of Chinese healthcare providers, preventive therapy using Chinese patent medicine, when compared to a control group that did not receive drug treatment, resulted in increased bone mineral density and a reduced probability of fractures across all age levels in the intervention group. Additionally, the Jintiange capsule appears to be a cost-effective treatment option for postmenopausal women with osteoporosis.

Burkitt lymphoma (BL) is characterized by the translocation and dysregulation of the proto-oncogene MYC. Owing to its high aggressiveness, the clinical therapeutics for BL remain unmet. In view of this pathological feature, molecular glue degraders have been widely explored as a targeted therapeutic strategy. This study reports a first-in-class, orally bioavailable dual-target molecular glue degrader that co-degrades GSPT1 and CK1α and demonstrates potent anti-tumor activity in BL. MYC hyperactivity imposes a non-oncogene addiction to GSPT1, its depletion of which collapses protein synthesis and selectively eradicates MYC-addicted tumor cells. Furthermore, down-regulation of CK1α attenuates MYC expression, implying that co-targeting GSPT1 and CK1α may yield superior therapeutic efficacy. To identify the desired molecular glue, we designed a targeted library focused on cereblon (CRBN) and successfully identified the molecular glue degrader, INNO-235. We found that CRBN functions as the requisite E3 ubiquitin ligase for INNO-235, enabling nanomolar-level, proteasome-dependent degradation of both CK1α and GSPT1. Notably, no impact on IKZF1 or IKZF3 was observed. Additionally, INNO-235 demonstrated minimal toxicity in healthy volunteer-derived peripheral blood mononuclear cells, indicating a potentially favorable therapeutic window. In a systematic evaluation, INNO-235 exhibited potent anti-proliferative activity in BL cell lines, including those with TP53 mutations, as shown by nanomolar IC50 values. At the cellular phenotypic level, INNO-235 treatment significantly induced apoptosis and caused cell cycle arrest at the G0/G1 phase. In a Daudi cell-derived xenograft mouse model, orally administered INNO-235 exhibited dose-dependent tumor growth inhibition without observable significant changes in mouse body weight. Notably, compared to INNO-220 (a CK1α single-target molecular glue degrader) and MRT-2359 (a GSPT1 single-target molecular glue degrader), INNO-235 demonstrated superior anti-tumor activity and overall survival. To elucidate INNO-235's mechanism of action, we performed RNA-seq analysis in INNO-235 treated Daudi cells. The results revealed significant enrichment of differentially expressed genes involved in endoplasmic reticulum protein processing, implicating the activation of the integrated stress response (ISR). Subsequent validation confirmed dose-dependent upregulation of key ISR effector (p-eIF2α, ATF4, CHOP) and pro-apoptotic PUMA, concurrent with downregulation of anti-apoptotic BCL-2. These findings indicated that INNO-235 induced p53-independent apoptosis in TP53-mutated BL cells through ISR pathway activation. In conclusion, INNO-235 represents a first-in-class GSPT1/CK1α dual-target molecular glue degrader with significant therapeutic potential for lymphomas. By inducing both p53-dependent and TP53-independent apoptosis, INNO-235 offers a promising therapeutic strategy for TP53-mutated BL and other MYC-driven malignancies with limited treatment options.

Objective This systematic review examines recent pharmacoeconomic literature on denosumab'cost-effectiveness for bone metastasis treatment,providing evidence-based insights to guide healthcare policy decisions. Methods A comprehensive literature search was performed across Cochrane,PubMed,EMBASE(Ovid),CNKI,and Wanfang databases to identify original articles published between 2017 and 2023.Key words consisted of bone metastases,denosumab,and cost-effectiveness in the search strategy.The methodological quality of the included studies was assessed utilizing the revised Consolidated Health Economic Evaluation Reporting Standards(CHEERS 2022).Data was extracted regarding methodological characteristics and cost-effectiveness analyses. Results A total of 111 studies were retrieved,of which 6 met the inclusion criteria.All included studies were based on clinical trials and published literature data and exhibited high methodological quality.Up to 83%(5 out of 6)of comparisons demonstrated that denosumab was more cost-effective or dominant compared to zoledronic acid.The adjusted incremental cost-effectiveness ratios varied substantially by tumor type,ranging from CZK 436,339.09 to USD 136,234 per skeletal-related event avoided and from CZK 61,580.95 to USD 118,392.11 per quality-adjusted life year gained. Conclusions The majority of the included studies support denosumab as a more cost-effective treatment option for bone metastases in solid tumors compared to zoledronic acid.The application of CHEER(2022)enhances the reliability of pharmacoeconomic evaluations.

Dysregulation of amino acid metabolism is recognized to have a substantial influence on tumorigenesis and the modulation of tumor microenvironment. However, the role of amino acid metabolism-related genes in diffuse large B-cell lymphoma (DLBCL) remains undefined. Therefore, we aimed to explore the influence of amino acid metabolism-related genes in DLBCL using bioinformatics approaches. Consensus clustering demonstrated that the reprogramming of amino acid metabolism has prognostic value in DLBCL. Subsequently, we developed a risk model using LASSO-Cox regression analysis to accurately predict DLBCL prognosis and identified kynureninase (KYNU) as a potentially valuable biomarker. Analysis of immune infiltration was conducted to examine the correlation between risk scores and immune profiles. Furthermore, RT-qPCR showed that the KYNU mRNA levels were upregulated in OCI-LY1, OCI-LY3, and OCI-LY10 DLBCL cells compared with normal CD19+B lymphocytes. Cell proliferation assays and flow cytometry analysis showed that inhibition of KYNU expression reduced cell proliferation and induced apoptosis of DLBCL cells. Overall, we demonstrated the significant impact of amino acid metabolism on DLBCL. Our findings may help improve the assessment of disease prognosis and provide potential therapeutic strategies for DLBCL.

Over 95% of primary central nervous system lymphoma (PCNSL) cases are histologically classified as diffuse large B-cell lymphoma (DLBCL), but present markedly poorer prognosis than systemic DLBCL. This discrepancy may stem from its high molecular heterogeneity, immune privilege, inhibitory tumor microenvironment (TME) and dilemma in drug delivery. Current understanding in the tumor ecosystem of PCNSL remains limited. This study employed single-cell RNA sequencing (scRNA-seq) and spatial transcriptomic (ST) analyses to decode tumor microenvironmental features of PCNSL. We performed scRNA-seq of PCNSL tumor samples and reactive lymphoid hyperplasia samples, and additionally conducted ST on PCNSL tumor samples. Unsupervised clustering of B cells identified 4 malignant B cell (mBc) subpopulations with apparently transcriptional and spatial discrepancy. Pseudotime trajectory analysis revealed that mBc3 represented the origin, which evolved into mBc4 and mBc1 and ultimately differentiated into mBc2. To assess the clinical relevance of these subsets, we applied BayesPrism for deconvolution of bulk RNA-seq data from DLBCL cohorts, presenting mBc2 subset was significantly correlated to poor prognosis (p< 0.001). Kyoto Encyclopedia of Genes and Genomes pathway analysis and scMetabolism analyses indicated that mBc2 exhibited a strong oxidative phosphorylation (OXPHOS) metabolic signature. ST analyses of tumor specimens further confirmed the overlap of mBc2 localization and the regions with elevated OXPHOS activity. Based on its high OXPHOS characteristics, we further explored its molecular expression features in an attempt to identify potential therapeutic targets. COX7B, a structural subunit of mitochondrial respiratory chain complex IV, was identified as an mBc2 marker through Venn analysis, specifically overexpressed in the mBc2 subset, the terminal stage of developmental trajectory. In vitroassays demonstrated that COX7B expression is upregulated in activated B-cell-like DLBCL cell lines, and its knockdown significantly reduced cell viability and induced apoptosis, supporting its potential as a therapeutic target. The previous studies have shown that clonally expanded CD8+T cells in PCNSL undergo a dynamic transition from a pre-exhausted state to an exhausted state, exhibiting more pronounced T cell exhaustion features compared to systemic DLBCL, suggesting the presence of an immunosuppressive TME in PCNSL. However, the specific mechanisms driving T cell exhaustion and immune suppression remain incompletely understood. Recognizing the critical role of T cells in the TME and in determining therapeutic response, we further investigated the immunosuppressive landscape of T cells and regulatory mechanisms mediated by tumor cells. T cell cluster analysis demonstrated a substantial enrichment of exhausted T cells (exhTc) and regulatory T cells (TregCD4) in PCNSL. Key exhausted molecules (TIGIT, HAVCR2/TIM-3, LAG3, CTLA4, and PDCD1/PD-1) were notably upregulated. ST confirmed high expression of HAVCR2/TIM-3 in T-cell-enriched regions. Importantly, the ligands for HAVCR2/TIM-3, CTLA4 and CEACAM1 were found to be upregulated in malignant B cells, with CD80 (CTLA4 ligands) particularly enriched in B-cell regions of spatial slices. These findings suggested active involvement of mBc in shaping the immunosuppressive TME. To explore intercellular signaling, we used CellChat to construct the communication network within the PCNSL microenvironment. mBc subsets showed stronger interactions with exhTc and TregCD4 compared to other TME cell types, primarily mediated by the macrophage migration inhibitory factor (MIF) signaling pathway. CD74+CXCR4 and CD74+CD44 were identified as key ligand-receptor pairs in this pathway. Analysis of spatial slices revealed elevated expression of CXCR4 and CD74 in tumor regions, further indicating widespread activation of the MIF pathway. These findings suggest that mBc subsets mediate the immunosuppression of exhTc and TregCD4 via MIF signaling. In conclusion, this study reveals the intratumoral heterogeneity and immunosuppressive microenvironment of PCNSL, identifies the high-risk COX7B⁺ mBc2 subset, and indicates COX7B as a potential therapeutic target. Additionally, we clarify the key role of MIF signaling in immune suppression, offering new directions for targeted and immune therapies in PCNSL.

The modest reduction in casein kinase 1 alpha (CK1α) by lenalidomide contributes to its clinical effectiveness in treating del(5q) myelodysplastic syndrome. However, the mechanism by which CK1α impacts lymphoma survival remains inadequately defined. We developed INNO-220, a CRBN-dependent CK1α degrader, by leveraging cytokine expression profiling in T cells. Unlike lenalidomide, INNO-220 is a highly selective and potent degrader of CK1α without affecting IKZF1/3. Screening across lymphoma cell lines revealed that cells harboring wild-type p53 and exhibiting constitutive NF-κB signaling were particularly sensitive to CK1α degradation yet resistant to Bruton tyrosine kinase inhibitors. Moreover, INNO-220 suppresses NF-κB signaling and activates p53 pathway, leading to complete inhibition of lymphoma tumor growth in vivo. Mechanistically, INNO-220 disrupts the assembly and function of the CARD11/BCL10/MALT1 complex, thereby inhibiting NF-κB signaling in stimulated T cells and lymphoma cells that harbor an activating mutation in CARD11. Moreover, we observed that activation of wild-type p53 upon INNO-220 treatment was sufficient to induce potent cancer cell death even in the absence of constitutive NF-κB activity. In summary, our findings introduce a selective CK1α degrader as a novel therapeutic approach for lymphoma, providing both mechanistic insights and a potential patient selection strategy in treating lymphoma and possibly other cancers.

Upon viral infection, retinoic acid-inducible gene-I (RIG-I)-like receptors (RLRs) detect viral RNA to initiate antiviral innate immune response, which is mediated by the mitochondrial adaptor protein VISA virus-induced signaling adaptor; also known as mitochondiral antiviral-signaling protein (MAVS). The stability and activity of VISA are tightly regulated by various posttranslational modifications, among which polyubiquitination plays important roles. Various E3 ubiquitin ligases, including atrophin interacting protein 4 (AIP4), mediate polyubiquitination of VISA and result in its degradation. However, how polyubiquitination of VISA is regulated remains unclear. Here, we uncovered a dual function for proprotein convertase subtilisin/kexin type 9 (PCSK9), a key enzyme in cholesterol homeostasis and a well-known therapeutic target in cardiovascular diseases, modulating host responses to RNA viruses both extracellularly and intracellularly. Secreted PCSK9 inhibited sendai virus (SeV) and vesicular stomatitis virus (VSV) infection, while the intracellular PCSK9 potentiated RLRs-mediated interferons (IFNs) induction by stabilizing VISA on mitochondria. Viral infection induced the translocation of PCSK9 to mitochondria where it competed with AIP4 for VISA, thereby inhibiting its polyubiquitination and degradation. Consequently, overexpression of PCSK9 enhanced VISA-mediated innate immune response against RNA viral infection, whereas its deficiency had the opposite effects and resulted in more robust replication of the virus. Pcsk9-/- mice produced lower levels of type I IFNs and proinflammatory cytokines, rendering the increased sensitivity to VSV and influenza A virus infection. Altogether, our findings uncovered an important and unexpected role of PCSK9 in virus-host interaction and contribute to the understanding of the sophisticated mechanism governing the proper and efficient immune response to viral infection.

BACKGROUND:Cervical cancer (CC) is a leading cause of cancer-related deaths in women, and understanding the tumor immune microenvironment is crucial for identifying novel biomarkers and therapeutic targets. While T cells have been extensively studied in oncology, the role of B cells in CC remains poorly understood. METHODS:In this study, we generated and integrated multi-omics data, including single-cell RNA sequencing, single-nucleus RNA sequencing, spatial transcriptomics, bulk RNA sequencing, and multiplex immunofluorescence to investigate the composition and transcriptomic states of B cells in CC. RESULTS:Initially, we analyzed plasma cells, classifying them into IgA+ and IgG+ subtypes. We found that the proportion of IgA+ plasma cells was lower in cancer samples, whereas IgG+ plasma cells were higher. IgA+ plasma cells exhibited protein folding and B cell activation characteristics, while IgG+ plasma cells were associated with apoptosis and immune suppression. Subsequently, we focused on two specific plasma cell populations, MANF_PC and HSPA1B_PC. MANF_PC, which were predominantly enriched in normal tissues, were linked to enhanced antibody synthesis and better prognosis, while HSPA1B_PC, primarily found in cancerous tissues, were associated with apoptotic pathways and poor clinical outcomes. Importantly, HSPA1B_PC are associated with tertiary lymphoid structures (TLS), where they show significant spatial colocalization with immunosuppressive T cell subsets, including Tregs and Th17 cells within TLS. CONCLUSIONS:We identified distinct plasma cell subsets in CC and found that HSPA1B_PC are enriched in tumors, associated with immunosuppressive T cells in TLS. These findings suggest that HSPA1B_PC contribute to an immunosuppressive microenvironment in CC, highlighting their potential as therapeutic targets.

Sialylation is a terminal modification of cell glycosylation, including α-2,3-, α-2,6-, and α-2,8-forms, which exert a dominant role in cell recognition and immune regulation. Among them, α-2,6-sialylation and α-2,6-sialyltransferases have shown promising potential in the study of tumor mechanisms and treatment, but their function remains unknown in diffuse large B-cell lymphoma (DLBCL). This study aims to explore the function and underlying mechanism of α-2,6-sialylation in DLBCL. We first detected the expression of α-2,6-sialylation in lymph node tissues from DLBCL patients and reactive hyperplasia cases with informed consent. DLBCL tissues exhibited higher levels of α-2,6-sialylation compared to the control cases. Survival analysis revealed that DLBCL patients with elevated α-2,6-sialylation levels showed poorer clinical outcomes (P=0.0003). Within the sialyltransferase family, elevated expression of the α-2,6-sialyltransferase ST6GALNAC4 was found to be associated with a poor prognosis in DLBCL patients (P<0.05). The above findings prompted us to investigate the biological function of ST6GALNAC4 in DLBCL. Knockdown of ST6GALNAC4 declined cell proliferation and induced cell cycle arrest in the G0/G1 phase. Conversely, overexpression of the gene facilitatedcell proliferation. Moreover, the oncogenic role of ST6GALNAC4 was confirmed in DLBCL xenograft models. Both ST6GALNAC4 deficiency and sialyltransferase inhibitor-treated groups exhibited significantly delayed tumor growth compared to the control group. To explore the underlying mechanism of ST6GALNAC4 in DLBCL tumorigenesis, we performed mass spectrometry to identify interacting proteins of ST6GALNAC4. Since sialic acids are typically found at glycan termini on the cell surface, we primarily focused on cell membrane proteins as potential targets of ST6GALNAC4. Among them, BMPR-1B showed a significant positive correlation with ST6GALNAC4 (P<0.001). The correlation was also validated in DLBCL tissues. BMPR-1B, a member of the TGF-β superfamily, is implicated in tumorigenesis and progression. Co-immunoprecipitation confirmed the interaction between ST6GALNAC4 and BMPR-1B, and ST6GALNAC4 knockdown significantly reduced the protein expression of BMPR-1B in DLBCL cell lines. Notably, knockdown of ST6GALNAC4 led to an enrichment of the SMAD signaling pathway, as indicated by KEGG analysis. Furthermore, in vitro and in vivo experiments verified that inhibition of BMPR-1B could inhibit cell growth. Therefore, BMPR-1B was identified as a downstream target of ST6GALNAC4. We further elucidated the molecular mechanism by which ST6GALNAC4 regulates the expression of BMPR-1B through sialylation. Lectin immunoprecipitation demonstrated that ST6GALNAC4 mediated the sialylation of BMPR-1B. A followed cycloheximide chase experiment was carried out to show that ST6GALNAC4 knockdown decreased the half-life of BMPR-1B protein. We then used site mutations to determine the important modification sites on BMPR-1B. The potential site of BMPR-1B sialylation was predicted by the NetOGlyc web, where Thr24, Thr109, and Ser20 displayed potent potential to be sialylated. We mutated these sites respectively and examined the sialylation level, and found that both Thr109 and Thr24 mutations dramatically reduced the sialylation level of BMPR-1B. Importantly, ST6GALNAC4 significantly enhanced the sialylation levels of wild-type and Thr109-mutant BMPR-1B, but not Thr24-mutant BMPR-1B. This suggested that ST6GALNAC4 specifically mediated sialylation at Thr24. We then confirmed that the Thr24 mutation affected BMPR-1B half-life compared to wild-type BMPR-1B. Furthermore, we validated that ST6GALNAC4 knockdown decreased levels of pSMAD1/5/8, while overexpression of ST6GALNAC4 had the opposite effect. Importantly, overexpression of BMPR-1B could partially reverse the effects of ST6GALNAC4, showing that the oncogenic effects of ST6GALNAC4 were mediated by BMPR-1B. Taken together, these findings suggest that ST6GALNAC4 mediates BMPR-1B sialylation at Thr24, affecting its protein stability and ultimately regulating the SMAD signaling pathway in DLBCL. In conclusion, our results revealed that ST6GALNAC4 induced α-2,6-sialylation of BMPR-1B, which promoted malignant progression of DLBCL. This underscores the potential of ST6GALNAC4 as a novel prognostic biomarker and therapeutic target for DLBCL.

Cardiovascular and renal complications of type 2 diabetes are the main causes of death in diabetic patients. Clinical studies have found that polyethylene glycol loxenatide (PEG-Loxe), a GLP-1 analog widely used to treat type 2 diabetes, boosts renal and cardiac functions in diabetic patients. However, its mechanism of action remains to be elucidated. Using injury models of HK-2 human renal proximal tubular epithelial cells and H9C2 rat myocardial cells, as well as db/db mouse models of type 2 diabetes, this study assessed the protective effects of PEG-Loxe on T2DM mice kidneys and hearts and revealed their mechanisms of action. PEG-Loxe treatment significantly reduced the contents of serum creatinine, urea nitrogen, and 24 h urine protein, alleviated glomerular injury and inflammatory reaction, markedly elevated cardiac left ventricular ejection fraction (LVEF) and fractional shortening (LVFS) levels, diminished pathological injuries in cardiac tissues, and improved renal and cardiac functions in db/db mice. In addition, PEG-Loxe considerably decreased the GRP78 mRNA and protein expressions of GRP78, p-eIF2α, ATF4, and CHOP in the kidneys of T2DM mice, inhibited GRP78/PERK/eIF2α pathway-related proteins in HK-2 cells cultured in high glucose concentrations, subdued renal endoplasmic reticulum stress, and eased renal injury in T2DM mice. PEG-Loxe also obstructed the TLR4/NF-κB inflammatory pathway and myocardial apoptosis and mitigated cardiac trauma in T2DM by reducing TLR4, MyD88, and p-NF-κBp65 protein expressions in cardiac tissues. The H9C2 cell experiment further confirmed PEG-Loxe’s ability to protect the cardiovascular system of T2DM patients by inhibiting the TLR4/NF-κB inflammatory pathway and lessening LDH and CK-MB levels. We showed that PEG-Loxe could decrease renal stress response and improve renal injury in T2DM by inhibiting endoplasmic reticulum stress via the GRP78/PERK/eIF2α pathway. Additionally, PEG-Loxe could hinder the TLR4/NF-κB inflammatory pathway and myocardial apoptosis and boost cardiac function, thus exerting protective effects on the cardiovascular system in T2DM.

Phosphoribosyl pyrophosphate synthase 2 (PRPS2), a key enzyme in pentose phosphate pathway (PPP), catalyzes the conversion of ribulose-5-phosphate to adenosine triphosphate to produce phosphoribosyl pyrophosphate. The metabolites of the PPP are critical raw materials for the anabolism of fatty acids and nucleic acids in cells. Herein, we sought to unpack the specific regulatory mechanism underlying PRPS2's modulation of metabolism reprogramming in diffuse large B-cell lymphoma (DLBCL), expecting to propose novel diagnostic approaches and personalized therapeutic strategies. We first performed untargeted metabolomics on peripheral blood specimens from 60 newly diagnosed DLBCL patients and 60 healthy volunteers with informed consent and identified the PPP was significantly enriched in DLBCL patients. The transcriptome sequencing data of DLBCL patients in the GEO database (GSE56315) further verified above results. Survival analysis suggested that high expression of PRPS2 a key gene in the PPP, was significantly associated with poor prognosis in DLBCL patients. We next observed the upregulation of PRPS2 mRNA and protein levels in DLBCL cells, which was further confirmed in a cohort of newly diagnosed DLBCL patients. To further explore the biological functions of PRPS2, knockdown and overexpression models of PRPS2 were performed. PRPS2 knockdown significantly impaired cell proliferation and induced cell cycle arrest in G0/G1 phase. Moreover, PRPS2 knockdown markedly triggered cell apoptosis and dysregulation of apoptotic proteins. On the contrary, PRPS2 overexpression resulted in reduced apoptosis. To validation of PRPS2's biological function in vivo, xenograft DLBCL mice model was established and we found PRPS2 knockdown significantly reduced DLBCL tumor growth in mice and decreased Ki67 expression. Animal experiments were performed in accordance with the principles of the Institutional Animal Care. To evaluate the potential roles of PRPS2 in DLBCL, assay for transposase accessible chromatin with high-throughput sequencing (ATAC-seq) and unique molecular identifier-RNA sequencing (UMI RNA-seq) were performed on cells with stable PRPS2 knockdown. GSEA enrichment analysis showed significant enrichment in fatty acid metabolic pathway. Furthermore, PRPS2 expression was found to be positively related to triglyceride and cholesterol levels in peripheral blood serum of DLBCL patients by correlation analysis. Absolute quantitative lipidomics revealed reduced triglyceride level in PRPS2 knockdown cells, further confirming abnormalities in fatty acid metabolism. And we experimentally verified that PRPS2 knockdown significantly reduced intracellular triglyceride and lipid droplet contents. Moreover, orlistat, a fatty acid synthesis inhibitor, inhibited DLBCL cell proliferation in a dose- and time-dependent manner. PRPS2 knockdown increased orlistat's inhibition of cell proliferation, while orlistat treatment counteracted PRPS2 overexpression's effect on cell proliferation. The above results support that PRPS2-mediated lipid accumulation promotes DLBCL cell growth. Next, we explore the underlying mechanism by which PRPS2 regulates fatty acid metabolism. Combining ATAC-seq data with UMI RNA-seq data, we observed increased transcription of ACOX1 in the promoter region after PRPS2 knockdown. In PRPS2 knockdown cells, ACOX1 mRNA and protein expression elevated, as well as an increase in the rate of fatty acid β-oxidation. Further investigation was carried out to determine how PRPS2 regulates ACOX1 expression. According to hTFtarget data, PU.1 had the highest peak value in the promoter region when searching for transcription factors that regulate ACOX1 expression. Evidence was provided by chromatin immunoprecipitation (ChIP)-qPCR confirming the enhanced binding of PU.1 to the ACOX1 promoter region after PRPS2 knockdown. Correlation analysis revealed that the mRNA expression level of PU.1 was negatively correlated with that of PRPS2. Our study confirmed that PRPS2 knockdown decreased PU.1 expression and increased PU.1 phosphorylation, which could promote DNA binding and cofactor recruitment. Our study demonstrates that the suppression of PRPS2 could inhibit the development and progression of DLBCL by regulating the PU.1/ACOX1 axis to reprogram fatty acid metabolism, which provides a promising therapeutic insight into DLBCL treatment.

Pancreatic cancer is a devastating malignancy with a high mortality rate, poor prognosis, and limited treatment options. The tumor microenvironment (TME) plays a crucial role in tumor progression and therapy resistance. Multiple subpopulations of cancer-associated fibroblasts (CAFs) within the TME can switch between different states, exhibiting both antitumorigenic and protumorigenic functions in pancreatic cancer. It seems that targeting fibroblast-related proteins and other stromal components is an appealing approach to combat pancreatic cancer. This study employed single-cell transcriptome sequencing to identify MME (Membrane Metalloendopeptidase)-expressing CAFs in pancreatic cancer. Systematic screening was conducted based on tumor differentiation, lymph node metastasis, and T-stage parameters to identify and confirm the existence of a subpopulation of fibroblasts termed MME+CAFs. Subsequent analyses included temporal studies, exploration of intercellular communication patterns focusing on the hypoxia signaling pathway, and investigation of MME+CAF functions in the pancreatic cancer microenvironment. The pathway enrichment analysis and clinical relevance revealed a strong association between high MME expression and glycolysis, hypoxia markers, and pro-cancer inflammatory pathways. The role of MME+CAFs was validated through in vivo and in vitro experiments, including high-throughput drug screening to evaluate potential targeted therapeutic strategies. Single-cell transcriptome sequencing revealed tumor-associated fibroblasts with high MME expression, termed MME+CAF, exhibiting a unique end-stage differentiation function in the TME. MME+CAF involvement in the hypoxia signaling pathway suggested the potential effects on pancreatic cancer progression through intercellular communication. High MME expression was associated with increased glycolysis, hypoxia markers (VEGF), and pro-cancer inflammatory pathways in pancreatic cancer patients, correlating with lower survival rates, advanced disease stage, and higher oncogene mutation rates. Animal experiments confirmed that elevated MME expression in CAFs increases tumor burden, promotes an immunosuppressive microenvironment, and enhances resistance to chemotherapy and immunotherapy. The developed MME+CAF inhibitor IOX2 (a specific prolyl hydroxylase-2 (PHD2) inhibitor), combined with AG (Paclitaxel + Gemcitabine) and anti-PD1 therapy, demonstrated promising antitumor effects, offering a translational strategy for targeting MME in CAFs of pancreatic cancer. The study findings highlighted the significant role of MME+CAF in pancreatic cancer progression by shaping the TME and influencing key pathways. Targeting MME presented a promising strategy to combat the disease, with potential implications for therapeutic interventions aimed at disrupting MME+CAF functions and enhancing the efficacy of pancreatic cancer treatments.

BACKGROUND:Prostate cancer's complex interplay with the immune microenvironment prompted an investigation into immune-related pathogenic mechanisms and potential therapeutic targets. METHODS:Within the GSE176031 data set, Seurat meticulously dissected single-cell profiles from radical prostatectomy patients. Leveraging CellMarker and SingleR cell identities were precisely annotated. Then, monocle traced pseudotime trajectories, illuminating cellular paths, complemented by CellChat's insights into intricate intercellular communications. Furthermore, mendelian randomization (MR) robustly substantiated causal associations within prostate cancer contexts. RESULTS:Employing single-cell analysis on intraoperative tumor and normal tissue, we identified 15 distinct cell types, notably observing a significant T cell reduction in tumor samples. Intercellular communication analysis revealed multiple pathways between epithelial cells and T cells, highlighting interleukin (IL)-IL7R-IL2RG interactions. IL7R, crucial in T cell apoptosis, showed differential expression across T cell development stages. Patients with IL7 amplification had poorer outcomes (p < 0.05), supported by MR in two cohorts (ieu-b-4809 cohort: odds ratio [OR] = 1.005, p = 0.002, 95% confidence interval [CI] [1.002-1.008]; ebi-a-GCST90018905: OR = 1.063, p = 0.032, 95% CI [1.005-1.125]), confirming IL7 as a prostate cancer risk factor. CONCLUSIONS:These findings suggest T cell depletion via IL7-IL7R signaling may drive prostate cancer progression, offering novel therapeutic insights.

Almonertinib, a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, is highly selective for EGFR-activating mutations as well as the EGFR T790M mutation in patients with advanced non-small cell lung cancer (NSCLC). However, the development of resistance inevitably occurs and poses a major obstacle to the clinical efficacy of almonertinib. Therefore, a clear understanding of the mechanism is of great significance to overcome drug resistance to almonertinib in the future. In this study, NCI-H1975 cell lines resistant to almonertinib (NCI-H1975 AR) were developed by concentration-increasing induction and were employed for clarification of underlying mechanisms of acquired resistance. Through RNA-seq analysis, the HIF-1 and TGF-β signaling pathways were significantly enriched by gene set enrichment analysis. Lipocalin-2 (LCN2), as the core node in these two signaling pathways, were found to be positively correlated to almonertinib-resistance in NSCLC cells. The function of LCN2 in the drug resistance of almonertinib was investigated through knockdown and overexpression assays in vitro and in vivo. Moreover, matrix metalloproteinases-9 (MMP-9) was further identified as a critical downstream effector of LCN2 signaling, which is regulated via the LCN2-MMP-9 axis. Pharmacological inhibition of MMP-9 could overcome resistance to almonertinib, as evidenced in both in vitro and in vivo models. Our findings suggest that LCN2 was a crucial regulator for conferring almonertinib-resistance in NSCLC and demonstrate the potential utility of targeting the LCN2-MMP-9 axis for clinical treatment of almonertinib-resistant lung adenocarcinoma.

Abstract Objective To evaluate the safety and efficacy of the granisetron transdermal delivery system (GTDS) combined with Dexamethasone for preventing chemotherapy-induced nausea and vomiting (CINV) in patients receiving Capecitabine plus Oxaliplatin (CapeOX) therapy. Design Open-label, prospective, multi-center phase II trial. Setting Three institutions. Participants Fifty-four patients scheduled to receive CapeOX chemotherapy. Interventions Participants received GTDS (3.1 mg applied to the upper arm 48 h before chemotherapy, replaced on day 5, and discarded on day 12) and Dexamethasone. Main outcome measures The primary endpoint was the complete control rate of CINV. Secondary endpoints included the duration of delayed complete control, complete control rate in the acute phase, safety, and quality of life. Results The complete control rate for delayed CINV over the entire period (25–480 h) was 72.7% (95% CI 0.57–0.88). The duration of delayed complete control was 17.2 ± 4.5 days, with 51.5% of patients experiencing no nausea during the delayed phase. The complete control rate in the acute phase was 81.8% (95% CI 0.69–0.95). No serious adverse events related to the antiemetic regimen were reported. Conclusion Prolonged administration of GTDS is safe and effective for preventing CINV in patients with gastrointestinal malignancies treated with CapeOX. Trial Registration ClinicalTrials.gov registry (NCT05325190); registered on October 10, 2021.

Introduction: The human gastrointestinal tract contains approximately one hundred trillion microorganisms that provide numerous benefits, including energy absorption, protection against pathogens, and regulation of the immune system. Alterations in the gut microbiota, known as dysbiosis, can disrupt these normal physiological processes and have been associated with adverse conditions such as inflammatory states, immune dysregulation, and even malignancies. Studies have shown that the gut microbiota of patients with diffuse large B-cell lymphoma (DLBCL) differs significantly from that of healthy individuals, and changes in the abundance of specific gut microbiota contributes to the prognosis of DLBCL patients. Certain microbiota could regulate the development of DLBCL through their metabolites. However, these case-control studies are susceptible to confounding factors such as age, environment, lifestyle, other diseases, and treatments, which may bias the results. Herein, we employ Mendelian randomization (MR) to investigate the causal relationships between gut microbiota, derived metabolites, and DLBCL. Methods: Gut microbiota data were sourced from the MiBioGen consortium, metabolite information from the TwinsUK and KORA studies, and DLBCL-related data from FinnGen. The single nucleotide polymorphisms (SNPs) were used to serve as instrumental variables (IVs). The primary method for our MR analysis was inverse-variance weighting, enhanced by MR-Egger regression, weighted median, simple mode and weighted mode methods to investigate the causal impacts of the gut microbiome and derived metabolites on DLBCL. We also employed reverse MR analysis to explore the causal effect of DLBCL on gut microbiota. The results underwent verification through a series of sensitivity analyses, including Cochran's Q test, the MR-Egger regression test, and the MR-PRESSO test. Additionally, Bayesian Mendelian randomization (BWMR) was used to further substantiate the findings. Result: Our results indicated that three genus and two microbiome-derived metabolites were causally related to DLBCL. Additionally, reverse MR analysis revealed that DLBCL had a causal relationship with one class, one order, one family, and two genus. Specifically, Alistipes (P=0.013) and Ruminococcaceae UCG011 (P=0.034) were associated with a reduced risk of DLBCL, while Bilophila (P=0.031) was linked to an increased risk. Further reverse MR analysis indicated that DLBCL contributed to a reduction in the abundance of the class Deltaproteobacteria (P=0.037), order Desulfovibrionales (P=0.041), family Oxalobacteraceae (P=0.031), and genus Oxalobacter (P=0.024), along with an increase in the abundance of genus Anaerofilum (P=0.031). The study also found that indolepropionate (P=0.003) decreased the risk of DLBCL, while 7-alpha-hydroxy-3-oxo-4-cholestenoate (P=0.02) increased the risk. Comprehensive sensitivity analyses, including Cochran's Q test, MR-Egger regression test, and MR-PRESSO test, did not identify any outliers, underscoring the reliability of our findings. Moreover, a leave-one-out analysis confirmed that no single SNP disproportionately influenced the MR estimates, further validating the robustness and reliability of our results. Conclusion: Our results provide evidence supporting a potential causal relationship between gut microbiota, derived metabolites, and DLBCL, thus opening new avenues for exploring the pathogenesis of DLBCL and its potential applications in diagnosis and treatment. Keywords: Mendelian randomization; gut microbiota; metabolites; diffuse large B-cell lymphoma; SNPs. Disclosures: No relevant conflicts of interest to declare. Category: Aggressive Lymphomas: Clinical and Epidemiological