The evolutionarily conserved Hippo signaling pathway is a key regulator of stem cell self-renewal, differentiation, and organ size. While alterations in Hippo signaling are causally linked to uncontrolled cell growth and a broad range of malignancies, genetic mutations in the Hippo pathway are uncommon and it is unclear how the tumor suppressor function of the Hippo pathway is disrupted in human cancers. Here, we report a novel epigenetic mechanism of Hippo inactivation in the context of hepatocellular carcinoma (HCC). We identify a member of the microrchidia (MORC) protein family, MORC2, as an inhibitor of the Hippo pathway by controlling upstream Hippo regulators, neurofibromatosis 2 (NF2) and kidney and brain protein (KIBRA). Mechanistically, MORC2 forms a complex with DNA methyltransferase 3A (DNMT3A) at the promoters of NF2 and KIBRA, leading to their DNA hyper-methylation and transcriptional repression. As a result, NF2 and KIBRA are crucial targets of MORC2 to regulate confluence-induced activation of Hippo signaling and contact inhibition of cell growth under both physiological and pathological conditions. The MORC2-NF2/KIBRA axis is critical for maintaining self-renewal, sorafenib resistance, and oncogenicity of HCC cells in vitro and in nude mice. Furthermore, MORC2 expression is elevated in HCC tissues, associated with stem-like properties of cancer cells, and disease progression in patients. Collectively, MORC2 promotes cancer stemness and tumorigenesis by facilitating DNA methylation-dependent silencing of Hippo signaling and could be a potential molecular target for cancer therapeutics.

The immune system is crucial in the development and advancement of cancerous tumors, particularly in head and neck squamous cell carcinoma (HNSC). This study aimed to identify immune-related gene signatures (IRGs) for predicting the prognosis of HNSC. Transcriptome data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO), in addition to immune gene data from ImmPort, were examined. Using Cox-LASSO screening, nine IRGs were identified, and patients were classified into high- and low-risk cohorts based on risk scores. Differential expression, survival analysis, Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), single-sample GSEA (ssGSEA), CIBERSORT, and drug sensitivity analyses were performed between the cohorts. The high-risk cohorts exhibited lower immune scores and survival rates, while the low-risk cohorts exhibited higher immune scores and better outcomes. Cox regression identified CD19, CD79A, CTLA4, ICOS, and LAT as protective genes and CHGB, DKK1, PDGFA, and PTX3 as risk genes. Based on these nine genes, we established a nomogram prediction model to further assess patient prognosis. These findings highlight the prognostic value of IRGs in HNSC, offering the potential for personalized treatment strategies based on immune risk profiles.

The dynamic interplay between neoplastic cells and the host has been increasingly recognized as important players in the pathogenesis of cancer cachexia, a syndrome affecting ~50–80% of cancer patients with various incidences of different types of malignancies. Despite its prevalence, a comprehensive understanding of cancer cachexia progression, with a holistic view at the cross-organismal, cellular and molecular levels, remains elusive. In this review, we undertake an in-depth exploration of the relevant target organs and their regulatory roles in cancer cachexia, with a particular focus on macroenvironmental interactions via various organismal crosstalk axes. Moreover, we highlight how systemic metabolic remodeling, a hallmark of cancer cachexia, plays essential roles in modulating the inflammatory responses of immune and stromal cells in the tumor microenvironment (TME). These cellular responses, in turn, disrupt energy metabolism in distant organs and perturb organismal homeostasis by secreting a variety of mediators that activate specific signaling pathways, thereby fostering a vicious cycle that exacerbates cancer cachexia. We comprehensively summarize these complex cellular and molecular networks that constitute reciprocally regulatory dynamics between systemic metabolic reprogramming and inflammatory cascades. Notably, targeting the multifaceted interplay of organismal metabolic remodeling and cancer-associated inflammation holds great promise for clinical translation, as illustrated by a series of innovative therapeutic strategies and ongoing clinical trials aimed at mitigating cachexia in cancer patients.

Acute activation of mTORC1 by amino acids (AAs) is pivotal for growth regulation, yet it remains unclear how the intracellular nutrient-sensing machinery might be rewired by environmental cues to execute distinct functions. Here we report that, despite nutrient insufficiency, cancer-intrinsic AA-sensing mTORC1 signalling is hijacked by inflammatory cytokines in the tumour microenvironment (TME). ZBTB5 translates inflammatory signals to restore mTORC1 pathway via disrupting the GATOR1 complex. Mechanistically, inflammatory cues promote phosphorylation of ZBTB5-S127, thereby recruiting the Cullin3ZBTB5 E3 ubiquitin ligase to degrade NPRL2 within GATOR1 and reactivate mTORC1 signalling. Consequently, tumoural AA uptake is boosted to exacerbate nutrient restriction and death of CD8+ T cells, leading to immunoevasion, tumour progression and inferior response to immune-checkpoint inhibitors. As such, blocking ZBTB5-pS127 ameliorates primary and acquired resistance to checkpoint blockade. Thus, targeting aberrant nutrient-sensing via the ZBTB5-pS127-mTORC1 axis represents a proof-of-concept strategy to sensitize cancer immunotherapy by alleviating AA restriction in the TME.

Artificial intelligence (AI) has become an integral force in the clinical landscape of gastrointestinal (GI) oncology. Recent advances in model architectures ranging from traditional machine learning and convolutional neural networks (CNNs) to transformer-based foundational models and graph neural networks (GNNs) have enabled the extraction of complex features from diverse data modalities, including endoscopic images, radiology, pathology whole-slide images, and multi-omics profiles. In this review, AI models are systematically classified into supervised learning, unsupervised clustering, multimodal fusion, and interpretable modeling. The advantages of each model are delineated in unravelling tumor heterogeneity, anatomical characteristics, and treatment-relevant biomarkers. Furthermore, three types of clinical application are emphasized: (1) early screening and lesion localization via segmentation or anomaly detection; (2) molecular subtyping and patient stratification for diagnosis with risk assessment; (3) therapy guidance through response prediction and personalized treatment planning. We also discuss major challenges on the application of AI in integration of heterogeneous clinical data, model generalizability across centers, and the interpretability of predictions. Collectively, this review highlights the transformative potential of AI in better understanding tumor biology and its clinical value in advancing personalized medicine for GI cancer patients.

Tumor vaccines hold promise in inhibiting tumor metastasis and recurrence by stimulating the immune system to target and eliminate heterogeneous tumors. However, the development of optimal strategies for in situ tumor vaccines and the establishment of reliable methods to ensure their efficacy remain challenges. In this study, we engineered a folate-modified antigen-trapping nanoprobe (AGO@FA-lip). This nanoprobe initially acquires tumor antigens and double-stranded DNA (dsDNA) by inducing immunogenic cell death in tumor cells through the DNA-damaging agent oxaliplatin (OXA). Nano-aluminum hydroxide (nano-Al(OH)3) effectively captures and aggregates the released antigens, facilitating their recognition and presentation by dendritic cells. The combination of exogenous cyclic GMP-AMP (cGAMP) and dsDNA synergistically amplifies the activation of STING signaling pathways, leading to a remodeling of the tumor immune microenvironment and an enhancement of the anti-tumor immune response. In ovarian tumor models, AGO@FA-lip-mediated chemoimmunotherapy significantly inhibited tumor growth, metastasis, and recurrence, ultimately prolonging survival in mice. Transcriptomic analysis revealed extensive immune activation, particularly in pathways associated with antigen processing and presentation, T cell differentiation, and TNF inflammatory signaling. The design concept of AGO@FA-lip provides a clinically scalable strategy for developing in situ tumor vaccines to inhibit the metastasis and recurrence of ovarian cancer.

The impact of different mechanical ventilation modes on pulmonary outcomes following laparoscopic surgery in the Trendelenburg position remains unclear. This study aimed to compare the effects of two common ventilation modes on postoperative pulmonary complications (PPCs) in elderly patients undergoing such procedures. Elderly patients scheduled for laparoscopic surgery in the Trendelenburg position were randomly allocated to receive either pressure-controlled ventilation (PCV) or volume-controlled ventilation (VCV). Both groups were managed with a lung-protective ventilation strategy. The primary outcome was the incidence of PPCs within the first three postoperative days. Airway pressures, details enabling the calculation of respiratory system dynamic compliance (Cdyn) and arterial blood gas levels were also recorded at predetermined intraoperative time points: before anesthesia induction (T0); 10 min after tracheal intubation in the supine position without pneumoperitoneum (T1); 30 min (T2) and 60 min (T3) after establishing pneumoperitoneum and the Trendelenburg position; and at the end of surgery after returning to the supine position (T4). Compared with the VCV group (32.1%), the PCV group exhibited a significantly lower incidence of PPCs (13.0%; χ2 = 5.758, P = 0.016) (RR = 0.403, 95% CI: 0.183-0.888). Furthermore, patients managed with PCV exhibited significantly lower intraoperative airway pressures-including peak airway pressure (Ppeak), plateau pressure (Pplat), and driving pressure (ΔP)-as well as reduced dead space fraction (VD/VT) and arterial partial pressure of carbon dioxide (PaCO₂). Cdyn was higher in the PCV group. In elderly patients undergoing laparoscopic surgery in the Trendelenburg position, pressure-controlled ventilation was shown to improve Cdyn and was associated with a lower composite rate of postoperative pulmonary complications than volume-controlled ventilation. Whether these physiological advantages translate into clinically meaningful benefit requires confirmation in larger studies.

Background: Visceral fat plays a central role in cardiometabolic risk among people with type 2 diabetes mellitus (T2DM), yet its assessment in routine clinical practice remains largely dependent on imaging techniques or indirect anthropometric measures. Identifying accessible blood-based markers that reflect visceral adiposity may facilitate improved phenotyping in this population. This study aimed to investigate whether circulating coiled-coil domain–containing protein 3 (CCDC3) reflects visceral fat accumulation in adults with T2DM. Methods: Public RNA-sequencing datasets and human adipose tissue samples were analyzed to identify CCDC3 as a visceral fat–enriched secretory gene. In this cross-sectional study of 160 adults with T2DM undergoing dual-energy X-ray absorptiometry, plasma CCDC3 was measured by ELISA. Associations between plasma CCDC3 and visceral fat area (VFA) were examined using multivariable regression. Logistic regression models for abdominal obesity (VFA ≥ 100 cm2), with and without CCDC3, were evaluated using receiver operating characteristic (ROC) analysis, calibration curves, decision curve analysis (DCA), and Shapley additive explanations (SHAP). Results: Circulating CCDC3 levels were positively associated with VFA (β = 3.11, p < 0.001), independent of demographic and metabolic factors. Incorporating CCDC3 into the baseline model significantly improved discrimination of abdominal obesity (AUC 0.820 vs. 0.663; p = 0.009). Calibration curves and DCA supported better model fit and higher net clinical benefit with CCDC3. SHAP analysis showed that CCDC3 contributed the greatest incremental importance beyond waist circumference, sex, and age. Conclusions: Circulating CCDC3 may serve as a blood-based biomarker reflecting visceral adiposity in adults with T2DM and provides complementary information beyond traditional anthropometric measures.

The gut mycobiome is increasingly linked to host physiology, yet functional insights are dominated by yeasts, leaving the roles of filamentous fungi largely unexplored. Here, we identify Mucor racemosus as a gut-resident symbiotic filamentous fungus that establishes intestinal colonization via both hyphal and spore forms. M. racemosus confers radioprotection through an integrated metabolic program coupling direct host support with microbiota-mediated amplification. The fungus produces L-glutamate, L-aspartate, and DL-lysine, which are transferred to the host to enhance DNA damage repair in radiation-exposed intestinal epithelial cells. In parallel, M. racemosus-derived methylthioadenosine remodels the gut bacterial community by enriching Limosilactobacillus reuteri and reprogramming sulfur metabolism to generate radioprotective methionine. This coordinated fungal-bacterial metabolic axis reinforces intestinal radiotolerance. Consistent with this mechanism, dietary administration of M. racemosus-fermented cheese confers radioprotection in vivo. Together, these findings establish gut filamentous fungi as active metabolic organizers of host-microbiota interactions.

Pancreatic β-cell failure in diabetes is driven by chronic inflammation, yet how metabolic stress determines pro-inflammatory cell fate remains unclear. Here, we report that sublethal oxidative stress activates a β-cell-enriched epigenetic switch that licenses intrinsic inflammation. We identify a β-cell-enriched vulnerability wherein oxidative stress disrupts mitochondrial NAD⁺/acetyl-CoA flux, skewing the nuclear equilibrium between the deacetylase SIRT1 and acetyltransferases p300/CBP. This metabolic-epigenetic imbalance induces hyperacetylation of the alarmin HMGB1 at K96/K128-a modification remarkably prominent in β-cells compared to macrophages or hepatocytes. This site-specific acetylation acts as a molecular gate for HMGB1 nucleocytoplasmic translocation, triggering TLR/RAGE-mediated inflammation. Simultaneously, we discover that oxidative stress co-opts the mechanosensitive Hippo pathway, which sequesters YAP to transcriptionally repress SIRT1, thereby forming a functionally integrated signalling axis that exacerbates HMGB1 acetylation. Therapeutically, reconstruction of mitochondrial retrograde signalling via NAD⁺ supplementation (e.g., NMN) or dual inhibition of mitochondrial ROS and Hippo signalling restored acetylation homeostasis and suppressed HMGB1-driven inflammation. Notably, this combinatorial targeting demonstrates greater efficacy than either intervention alone (~73% reduction in inflammatory markers) in mitigating β-cell failure across murine and porcine models. Further validation in non-human primates was specifically implemented to address the unique translational gap of rodent models and available human single-cell datasets: by leveraging human-like islet anatomy and systemic physiological microenvironment, we verified the pharmacodynamic robustness and in vivo feasibility of this strategy in a clinically recapitulative large-animal setting, rather than merely confirming cross-species molecular concordance. Our work unveils a stress-responsive signalling network in which metabolic and mechanical cues are integrated at the epigenetic level to control the inflammatory fate of β-cells, providing a new mechanistic framework for diabetic pathogenesis and a rationale for combinatorial therapeutic intervention. Oxidative stress triggers mitochondrial dysfunction in pancreatic β-cells, depleting NAD⁺ and accumulating acetyl-CoA. This metabolic crisis skews the SIRT1/p300 balance, inducing β-cell-enriched hyperacetylation of HMGB1 at K96/K128-a molecular switch for its nucleocytoplasmic translocation, which activates TLR/RAGE-mediated intrinsic inflammation. Concurrent Hippo pathway activation further exacerbates this process by repressing SIRT1, forming an integrated signaling axis. To ensure reproducibility, we define a minimal validation workflow using β-TC-6 cells or primary islets under standardized sublethal stress. The central mechanism can be verified by monitoring HMGB1 K96/K128 acetylation and nucleocytoplasmic translocation, while the inflammatory cascade can be effectively blunted through modular rescue approaches, including NAD+ supplementation (NMN), mitochondrial ROS inhibition or hippo pathway inhibition.

The kinase MAPKAPK2 regulates cell survival, proliferation, and death, and is upregulated in colorectal carcinoma (CRC) where it is associated with tumor growth and progression. However, how it regulates tumor progression in conjunction with other signaling pathways, such as MEK/ERK, remains elusive. Solid tumors are often subjected to metabolic stress, notably glucose deprivation. Here, we demonstrate that MAPKAPK2 protein levels in CRC regulate cell fate decision during stress conditions, such as glucose deprivation and therapeutic treatment. While MAPKAPK2 expression is a limiting factor for CRC growth in vitro, depleting MAPKAPK2 or inhibiting its activity pharmacologically provides a survival advantage to CRC cells under glucose limiting conditions. Subjecting CRC cells to low glucose resulted in an ERK1/2-mediated decline in MAPKAPK2 to promote survival. Additionally, cells with reduced MAPKAPK2 activity were less sensitive to trametinib under glucose limiting conditions. Utilizing transcriptomic profiling, we found that glucose deprivation and MAPKAPK2 depletion activate pathways associated with survival during metabolic stress. This relationship was also observed in CRC patients (TCGA), where tumors with low MAPKAPK2 expression had higher ERK1/2 activation and upregulated stress-induced pathways, leading to poor survival. Finally, MAPKAPK2 modulated growth of CRC organoids, subcutaneous tumors, and patient-derived xenografts (PDX), and reduced MAPKAPK2 levels decreased efficacy of trametinib, in vitro and in vivo. Overall, this study identifies an interrelationship between MEK/ERK and p38/MAPKAPK2 signaling pathways during glucose deprivation to support cell survival and features MAPKAPK2 loss as a possible mechanism leading to reduced efficacy of trametinib-based anticancer therapy and poor patient outcomes in CRC.

Tissue invasion is an initiating step of the cancer metastatic cascade. Unraveling the mechanisms underlying intracellular signaling pathway rewiring that activates downstream transcriptional machinery to drive invasiveness could help identify improved strategies to prevent and treat metastasis. Through an unbiased genome-wide CRISPR screen in a mouse model of gastric adenocarcinoma (GAC), an E3 ubiquitin ligase, tripartite motif-containing protein 49 (TRIM49), was identified as a potent suppressor of cancer invasiveness. In two thirds of GAC, TRIM49 expression was downregulated in invading cancer cells, in which TRIM49 deficiency correlated with deeper tumor infiltration and lymph node metastasis and was indicative of shorter overall patient survival. In multiple orthotopic GAC mouse models, TRIM49-deficient cancer cells were highly infiltrative, leading to multiorgan metastasis. Mechanistically, galectin-3, a putative regulator of cancer invasion, was stabilized in TRIM49-deficient cancer, largely because of the failure to undergo TRIM49-mediated polyubiquitination and proteasomal degradation. Consequently, galectin-3 assembled a complex with EGR1, thereby regulating transcriptional activities of a proinvasive gene module. As the galectin-3/EGR1 complex acted as a key node relaying proinvasive signaling, its disruption using GB1107, an oral galectin-3 inhibitor, suppressed tissue infiltration and metastasis of patient-derived xenografts. Taken together, a proinvasive galectin-3/EGR1 transcriptional complex was exploited by TRIM49-deficient GAC to fuel tissue invasion, representing an Achilles' heel that is potentially targetable to prevent metastasis.Significance: A proinvasion galectin-3/EGR1 transcriptional complex is a therapeutic vulnerability in the highly invasive TRIM49-deficient gastric adenocarcinoma, which can be disrupted by the oral galectin-3 inhibitor GB1107 to prevent cancer spreading.

Cardiovascular-kidney-metabolic (CKM) syndrome and cardiometabolic multimorbidity (CMM) carry substantial mortality, yet whether a composite index integrating insulin resistance and frailty shows stronger associations with advanced CKM syndrome, CMM, and mortality than conventional metabolic markers remains unclear. This study evaluated the associations of the triglyceride-glucose-frailty index (TyG-FI) with CKM, CMM, and all-cause and cardiovascular mortality, and explored the role of baseline estimated glomerular filtration rate (eGFR). The analysis included 11,228 adults aged 20–79 years from the National Health and Nutrition Examination Survey 2001–2018. TyG-FI was calculated as the TyG index multiplied by the frailty index. Survey-weighted logistic and Cox proportional-hazards models were used to estimate odds and hazard ratios, with sequential adjustment for demographic, socioeconomic, and behavioral confounders. Restricted cubic splines examined non-linear relationships. Overall model performance was assessed using receiver operating characteristic curves with DeLong’s test, time-dependent AUC, Harrell’s C-index, calibration, and decision curve analysis. Exploratory mediation analyses quantified the proportion of mortality associations accounted for by baseline eGFR. Robustness was verified through multiple sensitivity and subgroup analyses. Over a median follow-up of 90 months (1,220 all-cause and 379 cardiovascular deaths), higher TyG-FI quartiles were associated with graded decreases in survival. Compared with the lowest TyG-FI quartile, the highest quartile yielded markedly elevated odds for advanced CKM syndrome (OR = 4.57, 95

Background: Type 2 Long QT Syndrome (LQT 2 ), caused by KCNH 2 mutations, is associated with an increased risk of sudden cardiac death (SCD). Clinical severity varies considerably among carriers. We investigated a family harboring the KCNH 2 p.Y427H variant, comprising a symptomatic proband (nine cardiac events), an asymptomatic carrier mother, and a mutation-negative father. Hypothesis: We hypothesized that patient-specific induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) can recapitulate the observed clinical phenotype discordance in vitro. Methods: Whole-exome sequencing identified the variant. Peripheral blood mononuclear cells (PBMCs) from all three individuals were reprogrammed into iPSCs and differentiated into cardiomyocytes. Electrophysiological assessment included patch-clamp recordings of I Kr and I Ca,L , measurements of action potential duration APD 90 and APD 50 under pacing, and evaluation of early afterdepolarizations (EADs) and triggered activities (TAs) under spontaneous conditions. Nifedipine, an L-type calcium channel blocker, was used to evaluate arrhythmia suppression. Results: I Kr was markedly reduced in the proband-derived iPSC-CMs (0.26 [0.19–0.29] pA/pF) and mother-derived iPSC-CMs (0.29 [0.27–0.30]) compared to the father-derived iPSC-CMs (1.65 [1.23–2.11], P = 0.001). APD 90 was significantly prolonged in the proband-derived iPSC-CMs (824 [691–893] ms) versus the mother-derived iPSC-CMs (559 [491–574] ms, P = 0.010) and father-derived iPSC-CMs (397 [331–403] ms, P < 0.001). Proband-derived iPSC-CMs exhibited a higher incidence of spontaneous EADs and TAs. Notably, I Ca,L was upregulated in the proband-derived iPSC-CMs (–18.4 [–19.6 to –13.0] pA/pF) compared to the mother-derived iPSC-CMs (–7.23 [–9.5 to –5.9], P = 0.002) and father-derived iPSC-CMs (–7.49 [–8.66 to –5.91], P = 0.001). Nifedipine significantly suppressed EADs and TAs in the proband-derived iPSC-CMs. Conclusion: The KCNH 2 p.Y427H variant induces I Kr loss-of-function. Divergent calcium current responses among family members may reflect a compensatory mechanism modulating phenotypic severity. Patient-specific iPSC-CMs reproduce this variability, providing a platform for mechanistic insights, personalized risk assessment, and therapy in LQT 2 .

Nicotinamide adenine dinucleotide (NAD+) plays an important role in tumor progression, but its role in non-small cell lung cancer with brain metastasis (NSCLC BM) remains unclear. Herein, we investigated NAD+ biosynthesis targeting as a new therapeutic strategy for NSCLC BM. Therapeutic activity of nicotinamide phosphoribosyl transferase (NAMPT) inhibitors was evaluated in mouse models of NSCLC BM and using various assays such as NAD+ quantitation, cell viability, and apoptosis assays. To explore impact on downstream signaling, RNA sequencing was used in NAMPT inhibitor-treated and control cells, followed by validation with genetic knockdown, western blot and qRT-PCR. Expression of NAMPT and downstream proteins in human NSCLC BM and its association with patient prognosis were examined. Finally, combination of NAMPT inhibitor and cisplatin was tested in vivo. Systemic treatment with NAMPT inhibitors demonstrated intracranial activity in an NSCLC BM model. NAMPT inhibitors decreased cellular NAD levels and suppressed proliferation and invasion, and induced apoptosis in NSCLC cells. Supplementation with NAD+ precursor NMN rescued these NAMPT inhibitor effects. Mechanistically, disruption of NAMPT-mediated NAD+ biosynthesis suppressed TGF-β1/Smads/RAB26 signaling, leading to inhibition of NSCLC cells. Expression of NAMPT/TGF-β1/Smads/RAB26 axis proteins was upregulated in NSCLC BM tissues and correlated with poor prognosis. Combining NAMPT inhibitors with cisplatin further extended the survival of NSCLC BM-bearing mice. Targeting NAD+ biosynthesis provides a new therapeutic strategy for NSCLC BM and can be effectively combined with cisplatin. Our studies identified the TGF-β1/Smads/RAB26 signaling downstream of NAMPT, which was targeted by NAMPT inhibition to mediate anti-cancer effects.

Baseline characteristics associated with pathological response

Flow diagram of trial enrollment and analysis

Neoadjuvant and adjuvant treatment-related adverse events

Bioengineered tracheas have shown considerable potential in tracheal injury repair; however, their practical value is limited by challenges in tracheal cartilage regeneration, and postoperative tracheal stenosis remains a common issue. Here, inspired by the 3-layer structure of the trachea and the multi-segmental characteristics of its cartilage, a multilayered bioengineered tracheal scaffold (named Sd@d-ECM/BMSCs/SilMA) with a microgroove structure is designed in this study to repair tracheal defects. In this design, the microgrooved surface of the methacrylated silk fibroin (SilMA) hydrogel provides spatial guidance for the directional growth of bone marrow mesenchymal stem cells (BMSCs) and enhances their adhesion and proliferation. The extracellular matrix of the decellularized cartilage scaffold offers the necessary microenvironment and mechanical support for BMSCs to differentiate into cartilage. Under the influence of the dual-layer structure (inner and outer), the middle-layer BMSCs can undergo stable chondrogenic differentiation without any inducing agents. Sd@d-ECM/BMSCs/SilMA effectively promotes tracheal cartilage formation in a rabbit defect model, reduces the incidence of tracheal stenosis, and substantially improves respiratory function. Sd@d-ECM/BMSCs/SilMA not only confirms the successful construction of microgroove structures on the surface of the SilMA hydrogel and the effective loading of BMSCs but also demonstrates significant experimental value in tracheal cartilage repair and regenerative medicine.