Efferocytosis is a critical physiological process in which phagocytes clear apoptotic cells to maintain tissue homeostasis. However, within the tumour microenvironment (TME), this process is systematically hijacked by tumour cells, transforming it into a key pathological mechanism that drives immunosuppression, tumour progression and therapeutic resistance. This review systematically elucidates the central role of metabolic reprogramming in this functional reversal, emphasising that efferocytosis is essentially an immunometabolic intersection process precisely regulated by metabolism. By releasing various metabolites such as ATP, lactate, adenosine and sphingosine-1-phosphate (S1P), apoptotic tumour cells not only recruit tumour-associated macrophages (TAMs) but also metabolically pre-program their functions, inducing polarisation towards a pro-tumourigenic M2-like phenotype. During the recognition stage, tumour cells exploit metabolic abnormalities, such as glycosylation and lipid oxidation, to modify surface 'eat-me/don't-eat-me' signals, thereby hijacking macrophage recognition and engulfment programs. Upon completion of engulfment, systemic reprogramming of amino acid, lipid and glucose metabolism occurs within macrophages. These metabolic alterations synergistically lock their immunosuppressive phenotype and establish a metabolic symbiosis between the tumour and stromal cells. Based on these mechanisms, this review further explores translational strategies targeting the efferocytic-metabolic axis, aiming to reprogram the immunosuppressive efferocytosis into immune-activating events to overcome TME-mediated immunosuppression and enhance current therapeutic efficacy. By deeply dissecting the metabolic regulatory networks of efferocytosis, we aim to pave new directions for cancer immunotherapy, achieving a paradigm shift from 'metabolic hijacking' to 'metabolic interventional therapy'.

Acute leukaemia is a highly aggressive malignancy with significant unmet therapeutic needs, partly due to epigenetic dysregulation. Here, we uncover deoxynucleotidyl transferase terminal-interacting protein 1 (DNTTIP1) within the mitotic deacetylase complex (MiDAC) as a previously unrecognised epigenetic regulator crucial for leukaemic cell survival and elucidate its mechanistic and translational significance. Using cellular, biochemical, and genetic perturbations, coupled with validation in multiple in vivo leukaemia mouse models, we characterised DNTTIP1's role in acute leukaemia. An integrated multi-omics analysis incorporating RNA-seq, cleavage under targets and tagmentation (CUT&Tag) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) revealed that DNTTIP1 recruits histone deacetylase 1/2 (HDAC1/2) to silence BCL2-modifying factor (BMF) and drive leukaemogenesis, validated by chromatin immunoprecipitation quantitative PCR (ChIP-qPCR). Drug synergy assays identify poly(ADP-ribose) polymerase (PARP)/HDAC/BCL2 inhibitor combinatorial efficacy. DNTTIP1 depletion impaired MiDAC recruitment in acute leukaemia, leading to histone H3 lysine 27 (H3K27) hyperacetylation at the BMF promoter and reactivating this effector. Upregulated BMF disrupted BCL2-mediated survival, triggering coordinated autophagy and apoptosis. Combined HDAC1/2 and BCL2 inhibition exerts synergistic anti-leukaemic effects, a therapeutic strategy currently under clinical evaluation. Further, PARP inhibition profoundly enhanced this synergy by impairing DNA damage repair, unveiling a novel triple-combination strategy. Our work defines the DNTTIP1‒HDAC1/2‒BMF axis as a pivotal epigenetic vulnerability in acute leukaemia and provides preclinical rationale for targeting this axis. These findings offer a validated biological framework for advancing this targeted combination therapy into clinical trials. DNTTIP1 is overexpressed in acute leukaemia and associated with poor prognosis. DNTTIP1 acts as a scaffold for the MiDAC complex, recruiting HDAC1/2 to silence BMF and inhibit leukaemic cell death. Pharmacological disruption of the DNTTIP1-HDAC1/2-BMF axis impairs leukaemogenesis.

DNA methylation and chromatin accessibility are pivotal epigenetic regulators of gene expression and cellular identity, with significant implications in tumorigenesis and progression. Current single-cell multi-omics methods are limited in throughput and sensitivity, hindering comprehensive biomarker discovery. We developed single-cell split-pool ligation-based multi-omics sequencing technology (SpliCOOL-seq), a high-throughput single-cell sequencing technology that simultaneously profiles whole-genome DNA methylation and chromatin accessibility in thousands of cells. By integrating in situ GpC methylation, universal Tn5 tagmentation, and split-pool combinatorial barcoding, SpliCOOL-seq achieves enhanced sensitivity and scalability. SpliCOOL-seq accurately distinguished lung cancer cell types based on genetic and multiple epigenetic modalities and revealed that the two DNA methyltransferase (DNMT) inhibitors, 5-Azacitidine and Decitabine, both cause large-scale demethylation but in distinct patterns. Applied to primary lung adenocarcinoma, SpliCOOL-seq identified tumour subclones within the tumour lesion and uncovered novel DNA methylation biomarkers (e.g., FAM124B, SFN, OR7E47P) associated with patient survival. Additionally, we demonstrated accelerated epigenetic ageing and mitotic activity in tumour subclones, providing new insights into tumorigenesis. SpliCOOL-seq achieves parallel profiling of whole-genome DNA methylation and chromatin accessibility in the same individual cells in a high-throughput manner and is hopefully used to illustrate regulatory interactions under different cell states. SpliCOOL-seq enables high-resolution, multi-modal epigenetic profiling at single-cell resolution, offering a powerful platform for discovering cancer biomarkers. Its application reveals novel therapeutic targets and early-diagnostic markers, underscoring its potential in precision oncology. SpliCOOL-seq achieves high-throughput single-cell co-profiling of DNA methylation and chromatin accessibility. DNMT inhibitors caused cancer cell demethylation with divergent patterns. SpliCOOL-seq enables the discovery of genes related to LUAD tumorigenesis. Ageing and LUAD tumorigenesis may share similar epigenetic alterations.

The prognostic relevance of HLA class I (HLA-I)-mediated immunity in cancer immunotherapy remains unclear. We introduce deltaHED, a novel metric that quantifies evolutionary divergence between germline and tumour-acquired HLA-I alleles, integrating both inherited and somatic immunogenetic variation. Using whole-exome sequencing, we analysed deltaHED across three independent cohorts: 164 patients with recurrent/metastatic nasopharyngeal carcinoma (RM/NPC) from the POLARIS-02 trial (PD-1 monotherapy), 88 melanoma patients receiving PD-1 monotherapy, and 477 esophageal squamous cell carcinoma (ESCC) patients from the JUPITER-06 trial (PD-1 plus chemotherapy vs. chemotherapy alone). High deltaHED was significantly associated with increased tumour mutational burden and neoantigen load (p<.001), but predicted worse progression-free survival (PFS) and overall survival (OS) in patients receiving PD-1 blockade across all three cancers. In ESCC, this association was observed only in the immunotherapy arm, not in patients treated with chemotherapy alone. High deltaHED also correlated with increased mutations in antigen-processing and T-cell receptor pathways. These findings establish deltaHED as a clinically relevant biomarker of immune divergence with potential to improve patient stratification and guide personalised immunotherapy strategies.

Background and RationaleSynthetic lethality (SL)‐based strategies hold significant promise for overcoming therapeutic resistance, a critical bottleneck in cancer treatment where cancer cells evade anticancer therapies, leading to diminished efficacy or treatment failure. The core of SL lies in exploiting tumour‐specific vulnerabilities: drug‐resistant cells often acquire unique genetic defects or compensatory adaptive responses, and SL strategies selectively target genes or pathways dependent on these vulnerabilities to induce specific cell death, thereby reversing resistance.Content and FocusThis review systematically elaborates on SL mechanisms and the multi‐faceted nature of tumour drug resistance, then focuses on how SL counteracts resistant phenotypes by leveraging resistant cells’ vulnerabilities. We further delineate SL applications in preclinical resistance models, highlight representative SL‐related drugs and predictive biomarkers and critically analyse challenges in clinical translation.ConclusionBy integrating mechanistic insights, preclinical validation and translational perspectives, this review aims to provide novel insights for precision therapy and a foundational reference to advance SL strategies in overcoming tumour resistance and facilitating their clinical implementation.Key pointsSL‐based strategies exploit tumour‐specific vulnerabilities in drug‐resistant cells to induce selective cell death and overcome therapeutic resistance.This review dissects SL mechanisms, diverse drivers of tumour drug resistance and how SL counteracts resistant phenotypes via these vulnerabilities.It summarises clinical translational applications of SL from preclinical studies to trials, approvals and emerging targets, and discusses future precision therapy.

BackgroundDFNB16, the second most common genetic cause of hearing loss, is caused by mutations of the STRC gene encoding stereocilin, a protein essential for the effective functioning of outer hair cells (OHCs) as cochlear amplifiers. Strc−/− mice, which lack stereocilin, display severe to profound deafness and constitute a relevant preclinical model for DFNB16.MethodsUsing Strc−/− mice, we developed a gene therapy strategy based on the use of dual AAV9‐PHP.eB vectors to deliver the full‐length Strc cDNA. Therapeutic efficacy was assessed by evaluating stereocilin expression, OHC bundle architecture, and their attachment to the tectorial membrane, together with functional recovery using distortion product otoacoustic emissions (DPOAEs), auditory brainstem responses (ABR) measurements and Go/No‐Go behavioral testing with psychometric analysis.ResultsDual‐AAV–mediated Strc gene delivery restored stereocilin expression, OHC bundle architecture and their attachment to the tectorial membrane, leading to the recovery of cochlear amplification and hearing to near normal thresholds, as confirmed by distortion product otoacoustic emission (DPOAE) and auditory brainstem response measurements. Behavioural assessment showed that treated Strc−/− mice regained normal frequency discrimination, indicating a restoration of higher‐order auditory processing, up to 100 days post‐treatment.ConclusionThese findings provide the first proof‐of‐principle that peripheral gene therapy can restore OHC function, cochlear amplification and central auditory perception in a DFNB16 model.Key pointsDual AAV‐mediated gene delivery restored peripheral hearing in a DFNB16 preclinical mouse model.The same treatment also restored central auditory processing.AAV‐mediated gene therapy represents a promising curative strategy for DFNB16.These results reinforce the translational potential for treating human genetic deafness.

BackgroundBRD7 has been confirmed to be lowly expressed in nasopharyngeal carcinoma (NPC) tissues and exerts tumour suppressive roles. However, the molecular mechanism of the downregulation of BRD7 expression and whether the strategy of activating BRD7 expression plays anti‐tumour effects still needs to be clarified.MethodsMethylation‐specific polymerase chain reaction (PCR) was used to identify the methylation levels of BRD7 promoter. In vitro experiments were used to evaluate the effects of BRD7‐targeted demethylation system on the malignant progression of NPC cells. Chromatin immunoprecipitation (ChIP)‐qPCR experiment was employed to examine the regulatory mechanisms underlying the demethylation system. Xenograft tumour models were used to assess impact of this demethylation system on tumour growth in vivo and the anti‐tumour effects of the lentivirus‐mediated demethylation system in NPC.ResultsThere was hypermethylation modification in BRD7 promoter, which was negatively correlated with BRD7 expression. Next, we constructed a LentiCRISPRv2/dCas9‐TET1CD‐sgRNAs system targeting specific methylation sites of BRD7 promoter based on five sgRNAs, and confirmed that all five sgRNA‐guided CRISPR/dCas9 systems could activate BRD7 and inhibit cell proliferation to varying degrees, among which sgRNA2&sgRNA5 were the most significant. Further, we constructed NPC cell lines stably transfected with LentiCRISPRv2/dCas9‐TET1CD‐sgRNA2&5, and confirmed that both sgRNA2&sgRNA5 could promote the transcriptional activation by reducing its methylation, and inhibit the cell proliferation, migration, invasion and tumour growth in vivo of NPC, and the combination of them has a more significant demethylation, transcriptional activation and anti‐tumour effect. In addition, BRD7 had hypermethylation modification in its promoter and decreased expression in NPC tissues, and both of them were negatively correlated, making it a potential diagnostic marker for NPC diagnosis.ConclusionsThe hypermethylation modification of BRD7 is an important mechanism leading to the inactivation of BRD7, and targeting demethylation of BRD7 inhibits the malignant progression of NPC, which might be a promising targeted therapeutic approach for treating NPC.

BackgroudPersistent infection with high‐risk human papillomavirus type 16 (HPV16) is a principal etiological factor in cervical cancer. Nevertheless, the molecular events linking HPV16‐associated lesion progression to malignant transformation remain insufficiently characterized, particularly those involving vesicular trafficking and autophagy regulation.MethodsProteomic analysis was conducted across five stages of HPV16‐associated cervical lesion progression to identify differentially expressed proteins. The expression of vesicle‐associated membrane protein 7 (VAMP7) was validated in cervical tissue specimens and cellular models. Gain‐ and loss‐of‐function approaches were employed to assess the effects of VAMP7 on cellular proliferation, migration, invasion, and apoptosis. Autophagic activity was evaluated by LC3 lipidation, autophagosome accumulation, and analysis of SNARE complexrelated proteins. The in vivo effects of VAMP7 were examined using xenograft tumor models.ResultsVAMP7 demonstrated dynamic expression changes during cervical lesion progression, characterized by decreased expression in HPV16‐positive non‐malignant tissues and a gradual increase with disease severity, reaching the highest levels in advanced cervical cancer. Functionally, VAMP7 enhanced proliferation, migration, and invasion while inhibiting apoptosis in cervical cancer cells, whereas distinct effects were observed in non‐tumor cervical epithelial cells. Mechanistically, VAMP7 regulated autophagic flux through modulation of SNARE‐mediated vesicle fusion, resulting in altered autophagosome accumulation and autophagy‐related signaling. In xenograft models, VAMP7 overexpression significantly promoted tumor growth and increased the expression of autophagy‐associated markers.ConclusionThese data indicate that dysregulation of VAMP7‐mediated autophagy contributes to cervical carcinogenesis in an HPV16‐associated context. VAMP7 may represent a potential therapeutic target for the treatment of cervical cancer.Key pointsVAMP7 displays dynamic expression changes during HPV16‐associated cervical lesion progression.VAMP7 promotes malignant phenotypes of cervical cancer cells by regulating autophagic flux via SNARE‐mediated vesicle fusion.Dysregulated VAMP7‐mediated autophagy contributes to cervical carcinogenesis in an HPV16‐associated context.

BackgroundNeoadjuvant anti‐programmed cell death 1 (PD‐1) immunochemotherapy has shown promising efficiency in the treatment of early‐stage non‐small‐cell lung cancer (NSCLC), but it has not consistently yielded durable responses. Biomarkers for the prediction of efficacy are warranted.MethodsWe performed shotgun metagenomic and plasma/faecal metabolomic studies in 44 NSCLC patients who underwent neoadjuvant tislelizumab plus platinum‐based doublet chemotherapy. Samples were collected at baseline and before surgical resection, and the major pathologic response (MPR) was evaluated.ResultsMPR patients showed a significantly higher gut‐microbial alpha diversity, an enrichment of Ruminococcaceae, Lachnospiraceae and Clostridiales species, and an increased plasma level of tryptophan metabolites at baseline. On the contrary, non‐MPR patients were characterized by enrichment of Prevotella species in faecal samples and higher plasma levels of linoleic acid metabolites. A high predictive accuracy was achieved using a small panel of differential microbial (Clostridium sp. M62/1 and Eisenbergiella tayi) or metabolomic features (linoleic acid, oxindole‐3‐acetic acid and quinolinic acid) with AUCs > .85.ConclusionsThe baseline characteristics of the gut microbiota and plasma metabolites could provide early predictions of the response to neoadjuvant anti‐PD‐1 immunochemotherapy.Trial registrationNCT05244837.Key pointsBaseline metagenomic and metabolomic signatures were significantly associated with the major pathologic response of neoadjuvant anti‐PD‐1 immunochemotherapy.Integrated microbial model (consists of Clostridium sp. M62/1 and Eisenbergiella tayi) and metabolomic model (consists of linoleic acid, oxindole‐3‐acetic acid and quinolinic acid) could provide early predictions of the response.