Journal of Experimental & Clinical Cancer Research最新文献

Background: Despite significant therapeutic advances, multiple myeloma (MM) remains incurable in most patients due to frequent tumor relapse. A major contributor to relapse is clonal heterogeneity, where subclones exhibit distinct mechanisms of therapy resistance, along with the presence of drug-resistant dormant cells. Eliminating these distinct populations, which often coexist in the tumor niche, is clinically challenging. Identifying survival mechanisms shared by drug-resistant proliferating and dormant cells holds potential for the simultaneous elimination of different tumor-repopulating clones.

Methods: To identify shared mechanisms of therapeutic resistance, we analyzed clinical databases and drug-resistant myeloma cell lines. We employed pharmacologic approaches to target common candidates identified in our analysis and assessed their impact on tumor progression and survival in preclinical mouse models containing both therapy-resistant and dormant cells.

Results: We identified upregulation of several components of the Notch signaling pathway in both dormant and drug-resistant MM cells, which correlated with poor clinical outcomes in newly diagnosed MM patients. Selective blockade of NOTCH3 with a neutralizing antibody or pan-Notch inhibition with a bone-targeted inhibitor reduced tumor burden and eliminated coexisting dormant and bortezomib-resistant cells in clinically relevant models of MM disease.

Conclusions: Our findings reveal that NOTCH3-dependent survival programs represent a shared vulnerability in both cells refractory to therapy and dormant cells. These programs can be exploited to overcome the diverse mechanisms by which cancer cells evade therapy, potentially preventing disease relapse and extending remission in patients with MM.

Background: Paclitaxel (PTX) serves as a first-line chemotherapeutic agent for the treatment of advanced non-small cell lung cancer (NSCLC). However, the emergence of drug resistance poses a significant threat to patient survival. The serine synthetic pathway (SSP) has been implicated in drug resistance across various cancers and is notably activated in NSCLC. Nevertheless, its role in PTX resistance remains poorly understood.

Methods: In this study, we investigated the influence of the SSP on PTX resistance in NSCLC and explored a novel combination therapeutic strategy involving PTX and anlotinib to reverse NSCLC drug resistance. Specifically, using integrated transcriptomic and metabolomic analyses along with in vitro and in vivo experimental approaches, we aimed to elucidate the regulatory role of activated SSP in PTX resistance and to determine whether the combination of anlotinib and PTX can overcome PTX resistance in NSCLC through modulation of the SSP.

Results: We found that SSP activation drives PTX resistance by promoting the proliferation of PTX-resistant NSCLC cells, increasing the expression and transport function of P-glycoprotein (P-gp), inducing epithelial-to-mesenchymal transition (EMT), and maintaining redox homeostasis. Anlotinib synergizes with PTX by suppressing SSP. This leads to attenuated glycolysis, disruption of the AKT/ERK proliferative signaling pathway, inhibition of P-gp expression and function, reversal of EMT, and redox imbalance, which subsequently elevates reactive oxygen species (ROS) levels and activates the mitochondrial apoptosis pathway, ultimately inducing apoptosis.

Conclusion: Collectively, our study demonstrates that anlotinib combined with PTX, via SSP inhibition, is a promising strategy for overcoming PTX resistance in NSCLC.

Background: Metastatic colorectal cancer (CRC) harboring KRAS mutations presents a major therapeutic challenge due to its aggressive nature, poor prognosis, and resistance to EGFR-targeted therapies. This study aimed to identify novel drivers of metastasis specifically in KRAS-mutant CRC and to elucidate the underlying molecular mechanisms to undercover new therapeutic vulnerabilities.

Methods: We integrated data from clinical databases (TCGA, CPTAC) with experimental validation using human CRC cell lines, a tissue microarray, and two distinct in vivo metastasis models (liver and lung colonization). ZBTB20 expression and function were analyzed by IHC, Western blotting, Transwell assays, and RNA-seq integrated with ChIP-seq data. The mechanism of ZBTB20 regulation was investigated via co-immunoprecipitation, mass spectrometry, truncation analysis, site-directed mutagenesis, and luciferase reporter assays. Statistical significance was determined using Student's t-tests, ANOVA, and survival analysis.

Results: ZBTB20 expression was significantly upregulated with metastatic progression specifically in KRAS-mutant CRC patients and correlated with reduced overall survival. Functionally, ZBTB20 promoted CRC cell migration, invasion, EMT in vitro, and drove metastatic colonization in vivo. Mechanistically, KRAS/ERK signaling directly phosphorylated ZBTB20 at Threonine 138, 142, and 232, a step essential for its nuclear localization and pro-metastatic activity. Integrating transcriptomic and cistromic data, we identified TGFBR2 as a direct transcriptional target of activated ZBTB20. Notably, pharmacological degradation of TGFBR2 with the inhibitor ITD-1 potently abrogated metastatic outgrowth in both liver and lung colonization models.

Conclusions: Our findings delineate a novel KRAS-ERK-ZBTB20-TGFBR2 signaling axis that is a critical driver of metastasis colonization in KRAS-mutant CRC. The robust efficacy of a TGFBR2 degrader in multiple in vivo models validates this axis as a viable therapeutic target, offering a promising strategy to inhibit metastatic progression in patients with this aggressive disease.

Background: Tumor-associated neutrophils and macrophages are key components of the hepatocellular carcinoma (HCC) microenvironment. However, the interplay between them and its contribution to HCC progression remain unclear.

Methods: Bioinformatic analysis of TCGA datasets and clinical HCC samples was used to evaluate neutrophil extracellular trap (NETs) levels and macrophage polarization. Co-culture of neutrophils, macrophages, and HCC cells, along with molecular analysis and in vivo mouse models, were employed to dissect the mechanisms underlying NETs-mediated macrophage reprogramming and tumor progression.

Results: NETs were significantly elevated in HCC patients, particularly in advanced and metastatic stages, which were positively correlated with intrahepatic M2 macrophage infiltration and M2d subset-associated cytokines in blood. In vitro, NETs promoted M2d polarization in the presence of HCC cells via IL-17R/NF-κB signaling activated by IL-17 carried within NETs, which subsequently enhanced angiogenesis, migration, invasion, and epithelial-mesenchymal transition; these effects were partially reversed by IL-17R inhibition. In vivo, NETs-induced M2d polarization accelerated tumor growth, angiogenesis, and metastasis, whereas IL-17R blockade attenuated these pro-tumor effects. Moreover, M2d macrophages indirectly promoted NETs formation by upregulating HCC cell-derived S100A9 through VEGF-NF-κB signaling, establishing a positive feedback loop between neutrophils and macrophages. Furthermore, IL-17 carried by NETs (NETs-IL-17) demonstrated strong predictive value for extrahepatic metastasis in HCC, with an area under the ROC curve (AUC) of 0.89.

Conclusions: A positive feedback loop between neutrophils and macrophages via the NETs-IL-17/VEGF/S100A9 axis accelerates HCC progression and metastasis. More importantly, NETs-IL-17 exhibited potential as an alternative biomarker for predicting extrahepatic metastasis in HCC.

The field of human induced pluripotent stem cell (hiPSC)-derived cell therapies is rapidly advancing, offering a promising "off-the-shelf" approach for treating both solid and hematologic malignancies. Among these, hiPSC-derived Natural Killer (NK) cell therapies have gained significant traction, with several currently in clinical trials and development. NK cell-based immunotherapy has emerged as a safe and effective strategy for patients with advanced leukemia, and ongoing research is focused on optimizing its accessibility, scalability, and efficacy. A key advantage of hiPSC-derived NK cells is their genetic susceptibility, allowing for targeted enhancements in fitness, metabolism, specificity, and cytotoxicity. This overcomes the donor-dependent variability that limits autologous and allogeneic NK cell therapies, which often struggle with expansion and functional consistency. Despite their promise, hiPSC-derived NK cells present unique manufacturing challenges, requiring precise optimization to ensure reproducibility, safety, and clinical-grade scalability. In this review, we will explore what we believe to be the most impactful genetic engineering strategies to enhance hiPSC-derived NK cell function. Additionally, we will also discuss the major hurdles challenging widespread clinical adoption, including licensing constraints, production yield, regulatory ambiguities, and the complexities of multi-step genetic engineering and safety validation. Finally, we will outline the emerging therapeutic pipelines from leading biotech companies, providing a valuable and up-to-date overview of the future landscape of hiPSC-derived NK cell therapy.

Background: Hepatoblastoma (HB) is the most common pediatric liver malignancy with an increasing incidence. However, the functional roles of 3D chromatin organization, epigenetic regulatory factors, and transcriptional reprogramming in HB pathogenesis remain poorly understood.

Methods: ​Integrated multi-omics analyses of HB and matched non-tumor tissues were performed, including Hi-C, H3K27ac CUT&Tag, ATAC-seq, and RNA-seq, to construct high-resolution 3D epigenomic maps and identify genes interacting with HB-specific super-enhancers (SEs). Functional assays of identified targets were conducted in cell lines and animal models. The regulatory mechanisms of SEs and upstream transcription factors (TFs) were investigated using CRISPRi-dCas9, 3C-qPCR, ChIP-qPCR, and luciferase reporter assays.

Results: Comprehensive analysis identified TRIB2 as an HB-specific SE-associated oncogene. Functionally, TRIB2 promoted cell proliferation and accelerated tumor growth both in vitro and in vivo. Patients with high TRIB2 expression exhibited advanced PRETEXT stage and metastasis. Mechanistically, TCF3 directly bound to both the TRIB2-SE and its promoter, promoting TRIB2 overexpression. Moreover, TRIB2 conferred resistance to ferroptosis by disrupting KEAP1-mediated ubiquitination of NRF2, thereby stabilizing NRF2 protein and enhancing antioxidant responses. The TCF3-TRIB2-NRF2 axis showed significant co-expression in HB tissues, effectively distinguished HB from normal liver tissues, and was associated with poorer overall survival.

Conclusions: Our findings reveal that TCF3 and SE mediate TRIB2 overexpression to inhibit ferroptosis via the KEAP1-NRF2 pathway and drive HB pathogenesis, providing potential diagnostic and prognostic markers for HB.

Triple-negative breast cancer (TNBC) is an aggressive and chemotherapy resistant subtype with high metastatic potential and frequent recurrence after standard treatment. While poly (ADP-ribose) polymerase inhibitors (PARPi) show efficacy in TNBCs with DNA repair deficiencies, only ~ 20% of patients respond, underscoring the need for more effective therapies. Mitochondria, as central regulators of cancer cell survival, present a compelling therapeutic target. Here, we introduce a novel gene-chemotherapy combining our mitochondria-targeted luminoptogenetics technology (cmLumiOpto), which directly disrupts mitochondrial membrane potential and induces cancer cell death, with PARPi to enhance TNBC treatment outcomes. To achieve targeted delivery, we conjugated a high-affinity anti-CD276 monoclonal antibody (mAb) that selectively binds human and mouse TNBCs to an exosome-associated adeno-associated virus (mAb-Exo-AAV). In vitro studies confirmed successful transfection, internalization, and functional expression of cmLumiOpto, leading to significantly enhanced cytotoxicity when combined with PARPi. In vivo, the combination therapy achieved a 95-100% reduction in tumor burden, suppressed patient-derived xenograft growth, and inhibited metastasis in four TNBC mouse models. Post-treatment analyses confirmed mitochondrial depolarization, downregulation of DNA replication, cytokine upregulation, and immune cell infiltration in tumor. These findings highlight the potential of mitochondria-targeted gene therapy combined with chemotherapy as a powerful and innovative strategy for TNBC treatment.