Aim Immune checkpoint blockade (ICB) has improved the treatment landscape of lung squamous cell carcinoma (LUSC), but durable clinical benefit is limited by frequent therapeutic resistance. We aimed to define the microenvironmental mechanisms underlying ICB resistance in LUSC. Methods Surgical specimens from LUSC patients treated with neoadjuvant anti-PD-1 therapy plus chemotherapy were analyzed using integrated single-cell and spatial transcriptomics. Our findings were validated in independent bulk RNA-seq and multiplex immunofluorescence cohorts and functionally tested in fibroblast-specific conditional knockout and humanized mouse models. Results We identified a TMEM158⁺ cancer-associated fibroblast (CAF) subpopulation enriched in non-responders and associated with poor prognosis. Spatial and functional analyses showed that TMEM158⁺ CAFs promoted CD8⁺ T-cell exhaustion. Mechanistically, tumor-derived KLK5 engaged TMEM158, recruited SDCBP, activated the MEK/ERK-ELK1 signaling axis, and induced TNC and SPP1 expression, thereby establishing an immunosuppressive niche. Genetic or functional disruption of TMEM158⁺ CAFs enhanced sensitivity to anti-PD-1 therapy in LUSC. Conclusion TMEM158⁺ CAFs are key stromal mediators of immune evasion and ICB resistance in LUSC and represent a promising biomarker and therapeutic target.
With rapid advances in systemic therapy for hepatocellular carcinoma (HCC), developing evidence-based treatment strategies after first-line therapy failure has become a major clinical challenge. In this context, establishing a standardized consensus for later-line therapy tailored to clinical practice, acknowledging country-level heterogeneity in drug access and treatment patterns, is of significant clinical importance. Grounded in best available evidence and multidisciplinary expert experience, this consensus addresses key issues after failure of targeted therapy, immune checkpoint inhibitors, or combination regimens, including assessment of disease progression, evaluation of resistance, and optimal timing of treatment switching. It proposes unified evaluation criteria and stratified treatment recommendations. Emphasizing evidence-based, individualized decision-making, the consensus balances efficacy with safety. It aims to promote standardized and more consistent later-line decision-making, providing practice-oriented clinical guidance to improve long-term survival and quality of life for patients with advanced HCC. These recommendations integrate regional real-world experience with available evidence and are contextualized in reference to current international guidelines.
Extracellular matrix (ECM) is a key component in the tumor microenvironment (TME). The stiffened ECM not only acts as a barrier to prevent drug delivery but also facilitates tumor progression via activating cell membrane receptors and mechanical sensors, such as Piezo1 ion channels, integrins, and Yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif (TAZ). Accumulating evidence suggests that stiffer ECM and aberrant mechanotransduction contribute to treatment resistance. Targeting ECM stiffness and mechanotransduction pathways and restoring mechanical abnormalities in the TME provide opportunities for the rational design of combination therapies, especially in drug-resistant tumors. Therefore, elucidating how ECM stiffness regulates cancer drug resistance will facilitate the development of new therapeutic strategies for improving patient outcomes. In this review, we summarize the key factors that regulate ECM stiffness during tumor progression. We comprehensively analyze the multi-faceted mechanisms of ECM stiffness in cancer therapy resistance and discuss potential strategies that target ECM components and downstream signaling pathways for improving treatment efficacy. This comprehensive review will enhance the understanding of the functional roles of ECM stiffness and mechanotransduction in cancer drug resistance, providing basic knowledge for exploring novel therapeutic strategies.
Gemcitabine resistance remains a major obstacle in pancreatic cancer therapy. We herein report that O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) and its catalytic enzyme, O-GlcNAc transferase (OGT), are markedly upregulated in gemcitabine-resistant (Gem-R) pancreatic cancer cells, driven by hyperactivated glycolysis. Mechanistically, OGT interacts with and catalyzes the O-GlcNAcylation of BICD2 at serine 192 (S192), which further promotes BICD2 phosphorylation. This dual post-translational modification enhances BICD2 binding to RanBP2 and dynactin (DCTN1) during the G2/M phase, thereby accelerating cell cycle progression and gemcitabine resistance. Genetic depletion of OGT or pharmacological inhibition (OSMI-1) restores gemcitabine sensitivity, whereas the overexpression of OGT or stabilization of O-GlcNAcylation (PugNAc) exacerbates resistance and tumor aggressiveness. In the KPC genetically engineered mouse model of spontaneous pancreatic cancer, the combination of gemcitabine and OSMI-1 exerts an additive effect. Clinically, elevated levels of O-GlcNAcylation, OGT, and O-GlcNAcylated BICD2 correlate with a poor prognosis in patients with pancreatic cancer. Our study unveils the OGT-BICD2 (O-GlcNAcylation/phosphorylation)-RanBP2/DCTN1 axis as a pivotal regulator of gemcitabine resistance and suggests targeting O-GlcNAcylation as a promising therapeutic strategy to overcome chemoresistance in pancreatic cancer.
Recent advances in the field of oncology have increased our understanding of the immune system, its response to malignancies and the immune component of the tumor microenvironment (TME). Indeed, information obtained about tumor biology has become an important strategy in developing treatments for cancer. A plethora of immunotherapy drugs have been approved and have been shown to be efficacious in treating cancers resistant to other therapeutic modalities. However, these drugs produce autoimmunity and non-specific inflammation, and therapeutic resistance can occur. Resistance to immunotherapeutic drugs can be due to: (i) tumor-intrinsic antigens or histocompatibility leukocyte antigens (HLA) loss; (ii) adaptive signaling changes (e.g., phosphoinositide-3-kinase (PI3K)/AKT, mitogen-activated protein kinase (MAPK), Janus kinase (JAK)/signal transducers and activator of transcription (STAT)) and (iii) immunosuppressive TME. Antibody drug conjugate (ADC)-specific issues include target density thresholds, impaired internalization, lysosomal trafficking defects and payload efflux, which could be overcome by rational drug combinations (i.e., immune checkpoint inhibitors (ICI) and virtual reality (VR) blockade, ADC and programmed cell death protein 1 (PD-1) inhibitors, chimeric antigen receptor (CAR) - T cells and interleukin-12 (IL-12)), TME reprogramming (e.g., tumor growth factor-beta (TGFβ) or stimulator of interferon genes (STING) modulators and next-generation constructs (e.g., bispecific antibodies, immune-stimulating ADCs, proteolysis targeting chimera (PROTAC) conjugates). In this review, we provide a cross-platform analysis, with a significant emphasis on ADCs, which has a dual chemical-biological nature that epitomizes the complexity of contemporary drug design.
Gastrointestinal (GI) cancers remain a major cause of cancer-related mortality worldwide, and multidrug resistance (MDR) is a critical barrier to durable therapeutic response. In GI malignancies, MDR arises from multilayered and interdependent factors, including insufficient intracellular drug accumulation, altered drug activation and metabolic detoxification, target rewiring and phenotypic plasticity, apoptosis-autophagy-mediated stress adaptation, and extrinsic protection from the tumor microenvironment, cancer stem cells, metastatic niches, and microbiome-associated mechanisms. Biomimetic delivery systems (BDSs), including cell membrane-coated nanocarriers, extracellular vesicles, and living or virus-mediated platforms, provide biological interface functions that may improve drug circulation, tumor recognition, tissue penetration, membrane interaction, and active trafficking. However, their therapeutic value in MDR GI cancers should not be judged merely by improved delivery efficiency, but by whether they address defined resistance bottlenecks and generate evidence of genuine re-sensitization in resistant disease contexts. Current evidence indicates that many BDSs remain at the level of exposure enhancement or mechanism-aligned sensitization, whereas only a limited subset has demonstrated functional restoration of treatment response in established resistant models. This review organizes current BDS strategies according to their dominant mechanistic functions: enhancing drug accumulation and intratumoral delivery, reversing cellular drug resistance, remodeling resistance-supportive tumor microenvironment, targeting cancer stem cells (CSCs) and metastatic niches, and enabling site-directed intervention using living carriers and living therapeutics. By linking GI cancer-focused MDR biology to function-oriented biomimetic design, this review establishes an evidence-based framework for distinguishing delivery enhancement from true MDR reversal and outlines platform-specific requirements for clinical translation.
AIMS:Non-small cell lung cancer (NSCLC) patients treated with platinum drugs develop chemoresistance. C/EBPβ has alternative translational LAP and LIP isoforms which impact cancer chemoresistance by modulating ABC efflux transporter expression and activity. Differential alternative translation of LAP:LIP reprograms metabolism in murine embryonic fibroblasts; however, little is known in cancer. To target possible metabolic vulnerabilities, we herein investigated whether LAP/LIP rewires NSCLC cell metabolism towards a chemoresistant phenotype. METHODS:LAP- or LIP-overexpressing NSCLC cells were screened for anticancer drug sensitivity, DNA damage and ABC exporter expression and function. Metabolome/lipidome analyses and functional metabolic assays were performed to identify possible chemosensitizing agents. Tumor growth, mass spectrometry imaging and single-cell RNA-sequencing were determined in Hu-CD34+NSG xenografts. RESULTS:LAP induced chemoresistance by increasing ABCB1/ABCC1/ABCC2 levels, activity and oxidative DNA damage. Furthermore, LAP altered metabolome and lipidome composition of plasma membrane and mitochondria, and upregulated HADHA and CPT1A, key enzymes in fatty acid oxidation (FAO). The high metabolic flux through FAO and oxidative phosphorylation increased mitochondrial ATP levels, thereby fueling these ATP-driven multidrug efflux pumps. Conversely, LIP displayed the opposite effect. CPT1A knock-out or catalytically-inactive mutant, FAO inhibition with etomoxir or trimetazidine, surmounted chemoresistance. In LAPhigh chemoresistant immune-xenografts, etomoxir redistributed fatty acids within tumor immune-microenvironment (TIME), metabolically reprogrammed NK cells and enhanced their anti-tumor activity. CONCLUSION:Increased LAP:LIP ratio induced chemoresistance in NSCLC tumors by instigating a FAO-dependence, unveiling a metabolic vulnerability. FAO inhibition emerges as a novel chemosensitization strategy operating via rewiring tumor and TIME metabolism.
Immunotherapeutic strategies for triple-negative breast cancer (TNBC) have yielded only modest response rates, severely limiting patients' clinical outcomes. Clarifying the underlying mechanisms to reverse immunotherapy resistance therefore represents a critical unmet need. In this study, we employed the single-cell RNA sequencing (scRNA-Seq) analysis of human TNBC tissues to identify inositol-trisphosphate 3-kinase B (ITPKB) as a central regulator governing cytotoxic T lymphocyte trafficking into the tumor microenvironment (TME). Ectopic expression of ITPKB robustly enhanced the secretion of C-X-C motif chemokine ligand 9 (CXCL9), a critical chemokine required for effective anti-tumor immune responses. Mechanistically, elevated ITPKB activity accelerated the conversion of inositol 1,4,5-trisphosphate (IP3) to inositol 1,3,4,5-tetrakisphosphate (IP4), thereby dampening IP3-driven endoplasmic reticulum (ER) calcium release. Attenuated intracellular calcium ions (Ca2 +) flux, in turn, promoted the recruitment of signal transducer and activator of transcription 1 (STAT1) and nuclear factor-kappa B (NF-κB) to CXCL9 promoter, driving robust CXCL9 transcription. In addition, utilizing a stepwise virtual screening strategy of 3677 compounds, we identified amentoflavone (AMF) as a novel ITPKB agonist-like compound. AMF treatment could stabilize ITPKB by preventing Trim25-mediated K48-linked ubiquitination and degradation. Notably, in 4T1 and EMT6 syngeneic TNBC murine models, ITPKB-overexpressing or AMF administration exhibited markedly enhanced sensitivity to anti-programmed cell death protein 1 (αPD-1) immune checkpoint blockade. Collectively, these results position ITPKB as a therapeutically tractable molecular switch that remodels the tumor immune landscape via Ca2+-dependent transcriptional rewiring. Targeting ITPKB thus represents a promising strategy to overcome immunotherapy resistance and expand the pool of TNBC patients who may benefit from αPD-1-based immunotherapy.
NK/T-cell lymphoma (NKTL) is a highly aggressive non-Hodgkin's lymphoma characterized by extranodal involvement. Programmed cell death protein 1 (PD-1) monoclonal antibody (anti-PD1) treatment is ineffective in some patients, leading to recurrence or metastasis. The resistance to anti-PD1 treatment remains a major challenge in clinics. To identify the core molecules responsible for anti-PD1 treatment resistance and explore the possible molecular mechanisms behind it, simulating the human immune microenvironment during PD-1 treatment is essential. In this study, human peripheral blood mononuclear cells (PBMCs) were transplanted into immunodeficient mice to reconstitute human immunity in mice. The results showed that after immune reconstitution, human immune cells, especially T cells, remain at a high level (≥90%) for at least 4 weeks, indicating that human PBMCs were successfully reconstituted in immunodeficient mice. Two weeks after PBMCs implantation, human NKTL cells (SNK1, KHYG1 and YT) were subcutaneously inoculated into the right lower abdomen of mice. Then, high-dose anti-PD1 treatment was initiated (about 50 mm3, Sintilimab, twice a week, 10 mg/kg each time). The tumor from the mice was removed (≤than 1500 mm3) to prepare a single-cell suspension and re-implanted into the mice. The above process was repeated 5 times (280 days in total). The tumor growth inhibition (TGI) values of NKTL cells before and after induction (71.89% and 1.08% for KHYG1 cells, 54.37% and 3.79% for SNK1 cells, 51.17% and 6.34% for YT cells, respectively) confirmed the successful construction of anti-hPD1-induced resistance NKTL cells (SNK1-Re, KHYG1-Re and YT-Re). Subsequently, we performed transcriptomic, proteomic, and metabolomic analyses on the above resistant and sensitive cell lines. Through multiple functional experiments and clinical sample verification, we confirmed that the transcriptional repressor Zinc finger BED-type containing 6 (ZBED6) was a key biomarker for anti-PD1 treatment resistance in NKTL. Downregulated ZBED6 creates a thymidine-deficient environment by E2F transcription factor 1 (E2F1)-ribonucleoside-diphosphate reductase subunit M2 (RRM2)/dihydropyrimidine dehydrogenase (DPYD), in which drug-resistant tumor cells scavenge thymidine (TdR) from the tumor microenvironment via solute carrier family 29 member 1 (SLC29A1) to increase nucleotide synthesis, thereby establishing an immunosuppressive niche, resulting in the ineffectiveness of anti-PD1 immunotherapy.
Drug resistance to the androgen receptor (AR) antagonist is a critical obstacle in the clinic for advanced prostate cancers. Especially, AR antagonist treatment-induced neuroendocrine progression represents a lethal and therapy-resistant subtype. Although transcriptional and epigenetic lineage plasticity have been extensively implicated in treatment-induced neuroendocrine progression, the contribution of metabolic adaptation remains incompletely understood. Here, we identified a previously unrecognized metabolic reprogramming mechanism induced by AR antagonists in castration-resistant prostate cancer (CRPC) models. AR antagonist treatment markedly enhanced glycolytic activity and induced glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. Genetic depletion of GAPDH suppressed AR antagonist-induced glycolytic activation, altered transcriptomic and metabolic programs, reduced neuroendocrine-associated marker expression, and inhibited xenograft tumor growth. Mechanistically, GAPDH promoter pulldown coupled with mass spectrometry, siRNA screening, and chromatin immunoprecipitation assays identified myeloid zinc finger-1 (MZF1) as a key transcription factor for Enzalutamide-induced GAPDH gene expression. Pharmacological inhibition of GAPDH using koningic acid (KA) or penta-O-galloyl-β-D-glucopyranose (PGG) significantly suppressed tumor growth and attenuated neuroendocrine-associated molecular programs in CRPC cell-derived xenograft and patient-derived t-NEPC xenograft models. Collectively, our findings identify an AR antagonist-induced MZF1-GAPDH signaling axis that promotes glycolytic activation and neuroendocrine-associated metabolic adaptation during treatment resistance. These results support targeting GAPDH-dependent metabolic reprogramming as a potential therapeutic strategy for treatment-resistant prostate cancer.
AIMS:The global health crisis posed by New Delhi metallo-β-lactamase-producing carbapenem-resistant Enterobacteriaceae (NDM-CRE) and methicillin-resistant Staphylococcus aureus (MRSA) underscores the urgent need for novel antimicrobial agents. Natural antimicrobial peptides (AMPs) often suffer from limited modes of action, narrow antimicrobial spectra, and poor cellular permeability. To address these challenges, we engineered X1-3, a rationally designed AMP that exhibits direct bactericidal activity, efficient cell penetration and synergistic potentiation of β-lactams. METHODS:X1-3 was generated via structure-activity relationship-guided optimization and machine learning-assisted screening, followed by characterization of its physicochemical properties and safety profile. Its antimicrobial and β-lactam synergistic activities were evaluated against clinical NDM-CRE and MRSA isolates using checkerboard assays, time-kill kinetics, and confocal microscopy. Mechanistic studies combined transcriptomic profiling with flow cytometry, gel retardation, and enzymatic assays to elucidate multimodal actions involving membrane disruption, DNA binding, and specific inhibition of NDM-5 and PBP2a. Intracellular penetration and in vivo therapeutic potential were assessed in RAW264.7 macrophage infection model and murine systemic infection model, respectively. RESULTS:X1-3 exhibits high stability and robust activity against NDM-CRE, MRSA, and vancomycin-resistant Enterococcus, with low resistance induction. Additionally, X1-3 shows a robust anti-virulence profile by inhibiting biofilm formation and eradicating mature biofilms in both NDM-CRE and MRSA. Mechanistic studies revealed that X1-3 induces bacterial death primarily through membrane disruption and multiple synergistic mechanisms (DNA binding and metabolic perturbations). As a β-lactam adjuvant, X1-3 enhances antibiotic efficacy by disrupting proton motive force (PMF) to suppress efflux pumps and directly targeting NDM-CRE- and MRSA-associated resistance enzymes, including NDM and penicillin-binding protein 2a (PBP2a). Furthermore, it penetrates host cells via lipid raft-mediated internalization to eradicate intracellular pathogens and attenuate inflammation. In vivo, the combination of X1-3 with β-lactam antibiotics showed promising therapeutic potential with no observable systemic toxicity. Notably, the treatment effectively alleviated the MRSA-infected wound. CONCLUSIONS:Rational structural optimization enables X1-3 to function as both a direct bactericide and a β-lactam adjuvant. This dual strategy significantly improves efficacy against clinical NDM-CRE and MRSA, offering a novel therapeutic paradigm.
Sorafenib resistance remains a major challenge in the treatment of hepatocellular carcinoma (HCC). Through an in vivo CRISPR/Cas9 screen, we identified protein SUMOylation as a key pathway enriched in sorafenib-resistant HCC tumors. SUMO1 expression was significantly upregulated in resistant tumors and cell lines, and its modulation directly influenced sorafenib sensitivity both in vitro and in vivo. Proteomic analysis revealed that SUMO1 overexpression enhanced glycolysis, and metabolic assays confirmed increased extracellular acidification rate (ECAR) and decreased oxygen consumption rate (OCR) in SUMO1-high cells. We further identified PKM2 as a key SUMOylation target, mediated by the E3 ligase TRIM28. The SUMOylation of PKM2 increased its enzymatic activity, promoted aerobic glycolysis, and conferred sorafenib resistance. The inhibition of PKM2 with Compound 3k reversed glycolytic flux and restored sorafenib sensitivity. Clinically, SUMOylated PKM2 was highly expressed in HCC tumors and was associated with the expression of markers of glycolysis and sorafenib resistance in HCC. Our study revealed a novel SUMO1-PKM2 axis that drives glycolysis and sorafenib resistance in HCC, suggesting a potential therapeutic target for overcoming drug resistance.
AIMS:Antimicrobial peptides (AMPs) are promising alternatives to antibiotics; however, their development is often hampered by poor stability, safety concerns, and uncertain resistance risk. Therefore, this study aimed to develop new AMPs with enhanced stability, safety, and low resistance potential against multidrug-resistant bacteria. METHODS:A fluorination-based modification strategy was used to develop new AMPs, and their antimicrobial activity, stability, safety, mechanism, resistance development, and in vivo efficacy and safety were evaluated. RESULTS:A series of fluorinated AMPs was designed and synthesized, and Hrk8 was identified the lead candidate, which exhibited potent activity against multidrug-resistant bacteria, good stability and a favourable safety profile. Hrk8 operated via a multi-mechanism, including rapid membrane disruption, interference with intracellular processes, and species-specific transcriptional responses. In Escherichia coli, Hrk8 exposure was associated with widespread metabolic dysregulation reflecting systemic collapse of cellular homeostasis. In contrast, in methicillin-resistant Staphylococcus aureus, the transcriptional response suggested adaptive envelope remodelling and reduced virulence-associated regulatory programs. Consistent with this combined action, Hrk8 showed a low propensity to develop bacterial resistance and could limit the development of antibiotic resistance, while also exhibiting synergistic or additive antimicrobial activity and eradicating persister cells. Hrk8 achieved efficacy comparable to polymyxin B and vancomycin in mouse models of systemic infection, pneumonia and skin wound infection, but with a more favourable safety profile than polymyxin B. CONCLUSIONS:Fluorination effectively generated developable AMPs, resulting in Hrk8, which exhibited potent, broad-spectrum antimicrobial activity, low resistance potential, and promising therapeutic prospects. This approach provides a practical method to combat drug-resistant infections.
S-palmitoylation is a common and reversible post-translational modification. This study aims to elucidate the functional role and mechanism of S-palmitoylation in lenvatinib resistance and lenvatinib-induced immunogenic cell death (ICD) in hepatocellular carcinoma (HCC). Using small interfering RNA (siRNA) screening in HCC cells, zinc finger DHHC-type palmitoyltransferase 16 (ZDHHC16) was identified as a lenvatinib resistance-associated palmitoyl acyltransferase. ZDHHC16 was abnormally overexpressed in both lenvatinib-resistant tissues and cells. ZDHHC16 knockdown improved the sensitivity of HCC cells to lenvatinib. Furthermore, ZDHHC16 knockdown boosted lenvatinib-induced ICD of HCC cells, characterized by an increase in the release of danger associated molecular patterns (DAMPs), including calreticulin (CRT) exposure on the tumor cell surface, high mobility group box 1 (HMGB1), annexin A1 (ANXA1), and adenosine-5'-triphosphate (ATP), as well as an increase in reactive oxygen species (ROS) production, endoplasmic reticulum (ER) stress, and the secretion of cytotoxic cytokines (tumor necrosis factor (TNF)-α and interferon (IFN)-γ). Mechanistically, ZDHHC16 catalyzed the S-palmitoylation of glutathione peroxidase 2 (GPX2) at C67, which prevented ubiquitination-proteasome degradation of GPX2, thereby enhancing its protein stability. Targeting the ZDHHC16-mediated GPX2 S-palmitoylation improved lenvatinib-induced ICD and antitumor immunity, thereby enhancing the efficacy of lenvatinib. Collectively, our findings reveal that ZDHHC16-induced GPX2 S-palmitoylation plays a crucial role in modulating lenvatinib resistance and lenvatinib-induced ICD in HCC. Targeting this pathway is a promising approach to improve lenvatinib's antitumor activity.
Oxaliplatin-based chemotherapy is the first-line treatment for colorectal cancer (CRC) patients, but its clinical efficacy is hindered by therapeutic resistance. Here, we demonstrate that DNA ligase 1 (LIG1) is highly expressed in CRC and impairs chemosensitivity, highlighting LIG1 as a promising therapeutic target. However, the mechanism by which targeting LIG1 enhances chemosensitivity to oxaliplatin remains incompletely defined, and no highly selective LIG1 inhibitors have been identified to date. In this study, a library of 1220 natural compounds was screened to identify LIG1 inhibitors. As a result, gambogenic acid (GNA) was identified as a selective LIG1-binding candidate followed by cell-based cytotoxicity assays. Subsequently, we show that GNA treatment markedly enhances oxaliplatin chemosensitivity in CRC cells, drug-tolerant persister (DTP) cells, organoids, and in vivo tumor models. Moreover, the combination of GNA with FOLFOX (oxaliplatin + 5-fluorouracil + calcium levofolinate) also improves tumor responsiveness to anti-PD-1 immunotherapy. Mechanistically, GNA combined with oxaliplatin triggers ZBP1-dependent PANoptosis. Further investigations unveil that GNA suppresses DNA repair by targeting LIG1, thereby enhancing the DNA damage response and ultimately promoting PANoptosis and immunogenic cell death (ICD), which establishes a mechanistic link between LIG1 inhibition and PANoptosis induction. Overall, our findings suggest that LIG1 can be explored as a key target for oxaliplatin chemosensitivity in CRC and targeting LIG1 by GNA may be a promising therapeutic strategy for augmenting the sensitivity of CRC cells to oxaliplatin and anti-PD-1 treatment.
The optimal radiation dose for definitive chemoradiotherapy in locally advanced unresectable esophageal squamous cell carcinoma (ESCC) remains controversial, particularly whether dose escalation can overcome treatment resistance. We previously reported that escalation to 60 Gy increased radiation-related toxicity without improving short-term locoregional control versus 50 Gy. Here, we present the long-term outcomes and exploratory analyses of this randomized phase 3 trial. Between 2013 and 2017, 324 patients with stage IIA-IVA ESCC were randomly assigned 1:1 to receive 60 Gy or 50 Gy of conventionally fractionated radiotherapy with concurrent chemotherapy. Pretreatment biopsy specimens were analyzed using NanoString GeoMx digital spatial profiling for spatial transcriptomic and proteomic characterization. The full analysis set included 319 patients (60 Gy, n = 160; 50 Gy, n = 159). After a median follow-up of 99.5 months, no significant differences were observed in locoregional progression-free survival between the 60 Gy and 50 Gy groups, with 5- and 8-year rates of 41.8% and 32.7% versus 43.3% and 36.3%, respectively (HR 1.06, 95% CI 0.80-1.39, p = 0.70). Overall survival, progression-free survival, distant metastasis-free survival, and failure patterns were also comparable between groups. Spatial multi-omics analyses identified distinct baseline tumor microenvironment states associated with outcome. Long survivors showed an immune-activated profile with higher interferon and HLA signaling, and increased CD8+ T cell infiltration, whereas early progressors showed an immunosuppressive stromal profile suggestive of intrinsic resistance to definitive chemoradiotherapy. These findings support 50 Gy as an appropriate standard radiation dose for unselected patients with locally advanced ESCC treated with definitive chemoradiotherapy and do not support routine escalation to 60 Gy. Baseline spatial tumor microenvironment features may help identify resistance-associated states and provide biological context for divergent clinical outcomes.
Tumor microenvironment with high complexity and heterogeneity is one of the hallmarks of tumor and a key driver of treatment resistance. Conventional antitumor therapies, such as chemotherapy and radiotherapy, trigger tumor cell senescence through mechanisms involving DNA damage and oxidative stress. These senescent tumor cells, in turn, promote tumor malignancy via the senescence-associated secretory phenotype (SASP), thereby facilitating treatment resistance. Accumulating evidence indicates that, in addition to tumor cells, various microenvironmental components including endothelial cells, immune cells, and fibroblasts also undergo senescence in response to chemo- and radiotherapeutic stress. This prevalent therapy-induced microenvironmental senescence remodels the tumor microenvironment and fuels treatment resistance, highlighting its potential as a target for combination therapy. In this review, we outline the mechanisms underlying therapy-induced tumor cell senescence, with a particular emphasis on the mechanisms by which therapy-induced senescence of major non-tumoral components within tumor microenvironment mediates therapy resistance. Furthermore, we summarize the current therapeutic strategies targeting therapy-induced senescence, aiming to provide novel insights into the rational combination of senescence-targeted therapies with conventional radiotherapy and chemotherapy for tumor treatment.
AIM:Many patients with non-small cell lung cancer (NSCLC) do not derive clinical benefit from immune checkpoint inhibitors (ICIs), and a reliable method for identifying potential responders is lacking. This study aims to establish an optimized patient-derived organoids (PDOs) culture system and develop a novel organoid-peripheral blood mononuclear cell (PBMC) co-culture platform to evaluate response to ICIs. METHODS:We optimized culture conditions for patient-derived NSCLC organoids by integrating conditional reprogramming techniques. ICIs response was assessed with an organoid-PBMC co-culture system. The accuracy of this ex vivo system was validated by comparing its results with efficacy of ICI therapy in both an in vivo humanized mouse xenograft model and the corresponding patient's clinical response. Moreover, the genomic stability of the organoids during serial passaging was monitored through gene sequencing. RESULTS:An optimal indirect co-culture system was established, culturing organoids with 25% Matrigel. This system consisted of 40 Gy-irradiated Swiss 3T3-J2 fibroblasts in an upper chamber and primary NSCLC cells in a lower chamber, using F-medium (DMEM/F-12 (3:1, v/v) + 10 µM Y-27632) supplemented with 0.5 µM A83-01. This method achieved an 85.59% (95/111) success rate. Using this platform, we evaluated PD-1 antibody efficacy with the organoid-PBMC co-culture system in 70 cases. For in vivo validation, a direct patient-derived organoid xenograft (PDOX) models were established in humanized mice reconstituted with the corresponding patient's PBMCs. Notably, the ex vivo results demonstrated a strong correlation with both the in vivo validation data and patients' actual clinical responses to ICIs, yielding a sensitivity of 100% and a specificity of 62.20%. CONCLUSION:We developed a novel, simple, rapid and reliable organoid-PBMC co-culture system for screening the response of NSCLC patients to ICI therapy.
Therapeutic resistance is a major barrier to durable cancer control in contemporary oncology practice. Despite extensive studies on individual cell death pathways and mitochondrial stress responses, a comprehensive framework describing how mitochondrial organization contributes to the coordination of multiple regulated cell death programs and therapeutic resistance remains insufficiently defined. This review examines resistance as malignant cells evade regulated cell death and adapt to mitochondrial stress. Mitochondria are framed as integrative hubs that link bioenergetics, redox regulation, metabolic flexibility, and stress signaling to apoptotic competence. It also describes how apoptosis connects with other death programs through mitochondrial compartmentalization. Signals from the matrix, inner membrane, and cristae, intermembrane space, and outer membrane influence ferroptosis, necroptosis, mitochondrial permeability transition-driven necrosis, and immunogenic cell death. Stress-response pathways are highlighted as interfaces between mitochondrial dysfunction and fate decisions, including the OMA1-DELE1-heme-regulated inhibitor kinase axis that activates the integrated stress response and ATF4-dependent transcription. Translationally, the review proposes a co-targeting framework that pairs apoptosis-directed therapies, especially BH3 mimetics, with interventions that destabilize mitochondrial homeostasis or tune stress signaling. Examples include electron transport chain inhibitors, integrated stress response modulators, and compartment-targeted strategies that alter cristae remodeling, calcium flux, or cardiolipin oxidation.
