Plecstatin (PST) is a potent anticancer agent in preclinical models, yet its precise mechanism of action and molecular specificity regarding its main targets, plectin and outer dense fiber protein 2 (ODF2), remain incomp...Plecstatin (PST) is a potent anticancer agent in preclinical models, yet its precise mechanism of action and molecular specificity regarding its main targets, plectin and outer dense fiber protein 2 (ODF2), remain incompletely understood. Here, we dissected PST's mode of action using knockouts of plectin (PLEC) and ODF2 in SNU-475 hepatocellular carcinoma (HCC) cells. PST suppressed anchorage-independent growth and impaired 2D and 3D migration in a dose-dependent manner in both wild-type and ODF2-deficient cells, but not in PLEC-deficient cells, establishing plectin as the principal effector of PST's antitumor activity. Proteomic and functional analyses revealed that PST primarily disrupts cytoskeletal remodeling through plectin, while selectively affecting ciliogenesis-related pathways linked to ODF2 loss. Deletion of either protein attenuated PST-induced Ser51 phosphorylation of eIF2α, ATF4/GADD34 induction, and cytochrome c release, indicating cooperative involvement in integrated stress response (ISR). Correlative analysis of patient datasets confirmed associations between PLEC/ODF2 expression and ISR-related gene signatures, supporting the clinical relevance of this pathway. Together, these findings identify plectin as a key target of PST in disrupting cytoskeletal integrity and establish plectin/ODF2 axis in PST-driven stress adaptation in HCC.
Combining chemotherapy with chemosensitizing agents is a common strategy to enhance anticancer efficacy while mitigating treatment-related side effects. This study investigated the potential of dammarenediol II (DM2), a...Combining chemotherapy with chemosensitizing agents is a common strategy to enhance anticancer efficacy while mitigating treatment-related side effects. This study investigated the potential of dammarenediol II (DM2), a ginsenoside precursor, to enhance the anticancer effects of etoposide by downregulating O-linked β-N-acetylglucosamine modification (O-GlcNAcylation) and modulating the Akt signaling pathway in HepG2 human liver cancer cells. The effect of DM2 on O-GlcNAcylation regulation was analyzed using Pharmaco-Net, an artificial intelligence-driven drug screening platform and further validated using O-GlcNAc transferase (OGT) activity assay. DM2 cotreatment enhanced etoposide's anticancer efficacy, which was quantitatively evaluated by viability, Annexin V binding, membrane integrity, and caspase-3/7 activity assays in HepG2 cells. Results showed that DM2 reduced O-GlcNAc levels by directly interacting with OGT, as confirmed through Pharmaco-Net. Cotreatment with 40 μm DM2 and 20 μm etoposide produced synergistic anticancer effects, lowering etoposide's IC for cell viability by 2.29-fold and its EC for caspase-3/7 activity by 3.64-fold. Mechanistically, DM2 dose-dependently suppressed Akt/GSK3β/mTOR signaling. Using the Akt activator SC79, additional experiments confirmed that Akt signaling acts downstream of O-GlcNAcylation regulated by etoposide and DM2. These effects were also observed in multiple human liver cancer cell lines, as well as in A549 lung and Caco-2 colorectal cancer cells. This supports the broader anticancer and Akt-inhibitory potential of DM2. This study is the first to demonstrate that DM2 enhances anticancer synergy by suppressing O-GlcNAcylation and Akt signaling, highlighting its potential as a novel chemotherapy adjuvant.
Brix N, Samaga D, Gehr K
… +11 more, Dankó B, Schumann M, Drexler G, Alnatsha A, Beyer G, Mahajan U, Selmansberger M, Mayerle J, Belka C, Zitzelsberger H, Lauber K
The limiting dilution assay (LDA) is a key method to quantify clonogenic cells with self-renewing capacity in vitro, crucial for preclinical cancer research and therapy response assessment. It estimates the frequency of...The limiting dilution assay (LDA) is a key method to quantify clonogenic cells with self-renewing capacity in vitro, crucial for preclinical cancer research and therapy response assessment. It estimates the frequency of individual clonogenic, stem-like cells within a population based on their ability to form colonies with ≥50 cells at limiting cell numbers. Standard LDA analysis relies on linear, single-hit Poisson models, yet clonogenic growth under single-cell conditions often involves cooperative or competitive dynamics, violating this linearity assumption. Here, we present a modeling framework incorporating non-linear population dynamics into LDA analysis and introduce LDAcoop, an R-based tool for universal quantification of clonogenic cells in LDA formats. Across multiple cancer cell types, we benchmarked LDA against the colony formation assay (CFA) and show that LDA outperforms CFA, especially for patient-derived organoids, suspension cultures, and higher throughput applications. This renders the LDA format particularly suitable for larger-scale pharmacogenomic screening and drug sensitivity testing in complex models. Our results establish LDA and LDAcoop as versatile, scalable tools for robust quantification of clonogenic growth, supporting preclinical drug development and molecular precision oncology research.
Complete surgical resection is essential for oropharyngeal squamous cell carcinoma (OPSCC) therapy, underscoring the need for improved intraoperative margin assessment. To advance in vivo diagnostics for OPSCC, Nanosecon...Complete surgical resection is essential for oropharyngeal squamous cell carcinoma (OPSCC) therapy, underscoring the need for improved intraoperative margin assessment. To advance in vivo diagnostics for OPSCC, Nanosecond infrared laser (NIRL) tissue sampling combined with shotgun lipidomic analysis reveals lipidome differences between OPSCC tissue and adjacent healthy tissue. In this study, ablations were performed on tonsil squamous cell carcinoma in 28 samples from 11 patients using an established chamber setup, and a subset of six samples from three patients with a custom-made laser fiber-coupled applicator, designed for handheld use. Welch's t-test results (p = 0.05, two-fold change) revealed a similar OPSCC lipid profile in seven out of 11 patients. Potential tumor lipid markers were identified as consistently and significantly increased, despite the biological heterogeneity of the samples, underscoring their potential diagnostic value. Tissue ablation with fiber-coupled applicator was successful and the lipidomic analysis was consistent with the chamber setup. While limited to off-line analysis, our approach highlights the potential of fiber-based laser sampling as a rapid and minimally invasive method to obtain tissue material for advanced molecular profiling in surgical and endoscopic settings.
Ojala VK, Ahonen S, Peltola S
… +13 more, Tuohisto-Kokko A, Esparta O, Suominen P, Jokilammi A, Farahani I, Chakroborty D, Dibus N, Boettcher S, Airenne TT, Johnson MS, Eli LD, Elenius K, Kurppa KJ
Receptor tyrosine kinase ERBB4 (HER4) is frequently mutated in human cancer, and ERBB4 mutations have been identified in patients relapsing on targeted therapy. Here, we addressed the functional consequences of recurrent...Receptor tyrosine kinase ERBB4 (HER4) is frequently mutated in human cancer, and ERBB4 mutations have been identified in patients relapsing on targeted therapy. Here, we addressed the functional consequences of recurrent cancer-associated ERBB4 mutations that are located at regions important for receptor activation and/or are paralogous to known oncogenic hotspot mutations in other ERBB genes. Eleven out of 18 analyzed mutations were transforming in cell models, thus suggesting oncogenic potential for more than half of the recurrent ERBB4 mutations. More detailed analyses of the most potent mutations, S303F, E452K, and L798R, showed that they are activating, can co-operate with other ERBB receptors and are sensitive to clinically available second-generation pan-ERBB inhibitors neratinib, afatinib, and dacomitinib. Furthermore, the S303F mutation, together with a previously identified activating ERBB4 mutation, E715K, promoted resistance to third-generation EGFR inhibitor osimertinib in EGFR-mutant lung cancer model in vitro and in vivo. Together, these results are expected to facilitate clinical interpretation of the most recurrent cancer-associated ERBB4 mutations. The findings provide rationale for testing the efficacy of clinically used pan-ERBB inhibitors in patients harboring driver ERBB4 mutations both in the treatment-naïve setting, and upon development of resistance to targeted agents.
Single circulating tumor cell (sCTC) analysis enables the determination of predominant CTC phenotypes and genotypes. We previously demonstrated the feasibility of sCTC detection and genomic characterization in high-grade...Single circulating tumor cell (sCTC) analysis enables the determination of predominant CTC phenotypes and genotypes. We previously demonstrated the feasibility of sCTC detection and genomic characterization in high-grade serous ovarian cancer (HGSOC) by combining immune-magnetic enrichment and image-based sorting, followed by whole-genome amplification (WGA) and next-generation sequencing-based copy number alteration analysis (CNA). Here we aimed to improve our workflow by incorporating HGSOC-specific markers, folate receptor alpha (FRα), and markers to identify epithelial (cytokeratin) and mesenchymal (vimentin) phenotypes for the phenotypic as well as genotypic analysis of sCTCs over the course of treatment in 42 HGSOC patients. We detected a significant reduction of FRα-positive cells (P = 0.0205) and an expansion of cells with a high nuclear staining and no target antigen expression (P = 0.002). Before treatment, sCTCs showed an enrichment in CNAs of Chromosomes 2, 7, and 12, while CNA dynamics of sCTCs suggested a potential selection of distinct CNAs specific to the homologous recombination pathway. sCTCs revealed persistent CNAs in the CDK4 and emerging ones in the ALK oncogene. Notably, primary tumors revealed considerable fractions of shared genomic aberrations.
Copy number variations (CNVs) play a crucial role in cancer diagnostics and prognostics, potentially impacting treatment decisions. Ultra-low-pass whole-genome sequencing (ULP-WGS) has emerged as a promising alternative...Copy number variations (CNVs) play a crucial role in cancer diagnostics and prognostics, potentially impacting treatment decisions. Ultra-low-pass whole-genome sequencing (ULP-WGS) has emerged as a promising alternative to array-based methods for CNV detection, especially in formalin-fixed paraffin-embedded (FFPE) samples. However, sequencing biases and sample heterogeneity necessitate the optimization of CNV detection tools for FFPE sample-derived data. This study evaluates three open-source CNV callers (CNVpytor, ichorCNA, and WisecondorX) using ULP-WGS and compares their performance against a single nucleotide polymorphism (SNP) array. Our results demonstrate that under optimal experimental conditions, ichorCNA and WisecondorX achieved equal detection of true positive results, with reduced false positive results compared to the SNP array. The SNP array detection pattern differed somewhat from that of the CNV callers, while ichorCNA and WisecondorX had the most comparable detection pattern. We highlight the importance of (pre-)analytical parameters such as neoplastic cell content, sequencing coverage, and bin size selection on CNV detection accuracy. Our findings support the adoption of ULP-WGS-based CNV detection as a robust alternative to SNP arrays, with WisecondorX emerging as the most suitable tool for clinical implementation.
Metabolic dysfunction-associated steatohepatitis (MASH) is emerging as a major driver of hepatocellular carcinoma (HCC). Crouchet et al. identify PRDX2 as a key regulator linking oxidative stress, metabolic imbalance, an...Metabolic dysfunction-associated steatohepatitis (MASH) is emerging as a major driver of hepatocellular carcinoma (HCC). Crouchet et al. identify PRDX2 as a key regulator linking oxidative stress, metabolic imbalance, and oncogenic signaling. Across multiple in vivo and in vitro models, PRDX2 inhibition restores metabolic homeostasis, reduces tumor initiation, and selectively impairs HCC cell survival. These findings highlight PRDX2 as a promising biomarker and hepatocyte-directed target for chemoprevention, emphasizing the importance of the interplay between metabolism and liver cancer development.
Monomethyl auristatin E (MMAE) is used as the cytotoxic payload for enfortumab vedotin (EV) in the treatment of locally advanced and metastatic bladder cancer (BC). However, the development of resistance to MMAE in BC is...Monomethyl auristatin E (MMAE) is used as the cytotoxic payload for enfortumab vedotin (EV) in the treatment of locally advanced and metastatic bladder cancer (BC). However, the development of resistance to MMAE in BC is a therapeutic problem. To explore the mechanism of resistance to MMAE in BC, we established MMAE-resistant BC cells (MR-BCs). RNA sequencing analysis showed that the expression of dipeptidyl peptidase 4 (DPP4, also called CD26) increased significantly in MR-BCs compared with parental BC cells. Knock down of DPP4 expression using small interfering RNA inhibited the viability of MR-BCs. In addition, the DPP4 inhibitor sitagliptin suppressed the proliferation, migration, and invasion of BC cells, and cotreatment with MMAE effectively induced cell apoptosis, arrested cells in the GM phase of the cell cycle, increased reactive oxygen species production by inhibiting the AKT pathway, and significantly inhibited the in vivo growth of MMAE-resistant cells. This study provides insights into the use of DPP4 inhibitors as a treatment strategy for MMAE-resistant BC.
Ringborg U, von Braun J, Celis J
… +41 more, Berns A, Baumann M, Albreht T, Abou-Zeid N, Bagnato V, Brandts C, Chen CJ, di Pietro M, Dosanjh M, Dubois T, Eggermont A, Eggert A, Ernberg I, Faithfull S, Förner J, Fröhling S, Heitor M, Hood L, Jiang W, Jönsson B, Kannan R, Leptin M, Li S, Lindgren P, Lowy D, Ma J, Markham A, Nagy P, Oberst S, Parker MI, Rodin D, Ryan K, Schüz J, Sullivan R, Tabernero J, Turkson P, Várhelyi O, Varmus H, Wang C, Weiderpass E, Wilking N
The cancer problem is expanding, particularly in low- and middle-income countries (LMICs). Preventive measures can reduce the incidence by 40-50%, and cure rates have increased during the past decades in a number of canc...The cancer problem is expanding, particularly in low- and middle-income countries (LMICs). Preventive measures can reduce the incidence by 40-50%, and cure rates have increased during the past decades in a number of cancers. However, optimizing prevention programmes and increasing cure rates of cancer remain significant research challenges. The main focus of the conference was on P4 Cancer Medicine (Predictive, Preventive, Personalized and Participatory), a comprehensive strategy encompassing Health-Related Quality of Life (HRQoL) research, aiming to enhance the well-being of patients and individuals at risk. Addressing the cancer problem requires two key elements: translational cancer research and the development of relevant infrastructures. A Comprehensive Cancer Centre (CCC) acts as an innovation hub by integrating high-quality, multidisciplinary therapy and care, with healthcare-dependent prevention, research, and education. The United States has been at the forefront, providing quality-assured CCCs and the Cancer Moonshot for strategic cancer research. The EU has followed with the European Research Council for basic research, the European Innovation Council to boost disruptive innovation, and two EU initiatives on cancer, Europe's Beating Cancer Plan (EBCP) and the Mission on Cancer. The increasing complexity of cancer biology and technologies presents both a research challenge and a healthcare demand. For most patients, a CCC is not available. A critical discussion focused on quality assurance of healthcare outside the catchment area of a CCC and involving patients in clinical research. The strategic deployment of resources to support collective healthcare efforts and research aimed at reducing the cancer problem was discussed with representatives from the United States, EU, Africa, China, India and Taiwan. Analyses of translational cancer research have revealed important gaps in implementing innovations, assessment of clinical effectiveness, HRQoL, outcome and health economics research. The increased release of new anticancer agents over the last 25 years, accompanied by insufficient information on clinical benefits, presents both an economic and ethical problem. Direct healthcare costs have increased due to expenses for anticancer agents for the treatment of patients with incurable diseases. Evidence-based treatment based on HRQoL research is an unmet need. Basic/preclinical research aimed at increasing the cure rate should identify new, broader targets for therapy and develop extended diagnostic technologies for stratifying patients, to inform innovative clinical trials. Present research strategies convert cancer to a chronic disease, a growing burden for the healthcare systems. The increasing complexity of cancer biology and technology, the growing need for translational cancer research, and the demand for supporting infrastructures underscore the importance of international collaborations between CCCs. However, funding for cancer research is not currently aligned to reduce the cancer problem. While public funding for cancer research doubled between 2005 and 2024, the pharmaceutical industry's spending on cancer research increased tenfold. Increasing funding by public and non-profit funding organizations is mandatory. Education is another significant need, but it is currently fragmented and underfunded. The last session of the conference summarized the strategies in a Statement with a strong emphasis on global collaboration addressing the growing cancer burden and pronounced inequalities. Expanding partnerships and fostering innovative, multidisciplinary approaches to cancer prevention, therapeutics/care, as well as research, are not just urgent but essential steps towards reducing incidence, increasing cure rates and enhancing the well-being of cancer patients. Data-driven cancer medicine is currently under development, and modern communication technologies for diagnostics may facilitate interactions across geographical distances. A global cancer research agenda can become a model of solidarity, sustainability, and ethical responsibility.
A hypoxic microenvironment promotes the aggressiveness of lung adenocarcinoma (LUAD) through treatment resistance and generation of new lymphatic vessels (i.e., lymphangiogenesis) favoring metastatic dissemination. Trans...A hypoxic microenvironment promotes the aggressiveness of lung adenocarcinoma (LUAD) through treatment resistance and generation of new lymphatic vessels (i.e., lymphangiogenesis) favoring metastatic dissemination. Transcriptomic analysis of cohorts of LUAD patients highlighted LINC01116, a long noncoding RNA, associated with a bad prognosis, a high rate of recurrence, and induced by hypoxia in tumors. Gain- (overexpression) and loss-of-function (CRISPRi (Clustered Regularly Interspaced Short Palindromic Repeats interference, RNA interference)) approaches performed in LUAD cancer cell lines did not reveal a clear regulatory role for LINC01116 in tumor cells. Analyses of LUAD single-cell RNA sequencing data sets and RNA Fluorescent In Situ Hybridization (RNA-FISH) showed high expression of LINC01116 in lymphatic endothelial cells (LEC) pointing to this transcript as a specific biomarker of tumoral lymphangiogenesis. Efficient knockdown of LINC01116 in LEC in normoxic or hypoxic conditions impacted the proliferation rate under hypoxic stimulation and revealed a gene signature associated with proliferation and hypoxia sensing. Together, our data suggest a role for LINC01116 in pathological lymphangiogenesis of lung tumors.
Balaseviciute U, Huguet-Pradell J, Abril-Fornaguera J
… +15 more, Gris-Oliver A, Rialdi A, Fernández-Martínez E, Montironi C, Del Pozo V, Houghton P, Zanatto L, Mesropian A, Keraite I, Thung S, Armengol C, Sancho-Bru P, Guccione E, Pinyol R, Llovet JM
Hepatoblastoma (HB), the most frequent pediatric liver cancer (2.16 cases/million), has surgery and perioperative chemotherapy as primary treatment, with severe lifelong side effects. This study evaluates the efficacy of...Hepatoblastoma (HB), the most frequent pediatric liver cancer (2.16 cases/million), has surgery and perioperative chemotherapy as primary treatment, with severe lifelong side effects. This study evaluates the efficacy of the Wnt/CTNNB1 inhibitor WNTinib as a potential HB treatment, since CTNNB1 mutations occur in 70-90% of HBs. WNTinib's efficacy was assessed in three animal models (n = 48): (a) patient-derived xenograft (PDX) HB tumors (n = 5 CTNNB1-mutant, n = 1 CTNNB1 wild-type) implanted in NSG mice; (b) PDX-derived TT001- and (c) HepG2-HB cells subcutaneously implanted in Fox1 mice; and in two patient-derived organoids from CTNNB1-mutant HBs. WNTinib delayed tumor growth in n = 4/5 CTNNB1-mutant PDX models and significantly improved survival versus controls (P = 0.03), with no effect in the wild-type model. Further, in the TT001 and HepG2 models, WNTinib reduced tumor growth (P < 0.05 and P = 0.002) and extended survival (P = 0.03 and P = 0.008), respectively. In HB organoids, WNTinib demonstrated greater efficacy than standard-of-care cisplatin (P = 0.009, org-1), and its antitumor effect was further enhanced when combined with chemotherapy (P = 0.01, org-1; P = 0.007, org-22). WNTinib delays tumor progression and increases survival in CTNNB1-mutated HB models, providing rationale to explore its use in human HB.
Potassium channels in brain tissue orchestrate essential cellular processes, including the regulation of membrane potential and neuronal excitability. Among them, large-conductance calcium-activated potassium (BK) channe...Potassium channels in brain tissue orchestrate essential cellular processes, including the regulation of membrane potential and neuronal excitability. Among them, large-conductance calcium-activated potassium (BK) channels play a pivotal role in both normal brain physiology and the pathogenesis of glioblastoma multiforme, a highly aggressive primary brain tumor. Within the central nervous system, BK channels are widely expressed in neurons, astrocytes, and oligodendrocytes, contributing to ion homeostasis and synaptic transmission. In glioblastoma cells, overexpression of BK channels, particularly the glioma-specific gBK variant, facilitates tumor progression by enhancing cell migration, invasion, and therapeutic resistance. Recent evidence highlights the significance of the mitochondrial isoform of the BK channel (mitoBK) in modulating oxidative phosphorylation and reactive oxygen species generation, thereby promoting tumor cell survival under hypoxic and cytotoxic stress. This review summarizes current insights into the role of BK and mitoBK channels in glioblastoma biology, their potential classification as oncochannels, and the emerging pharmacological strategies targeting these channels, emphasizing the translational challenges in developing BK-directed therapies for glioblastoma treatment.
Eller S, Ebner S, Haselrieder C
… +15 more, Günther JK, Drasche A, Strich S, Volani C, Medici A, Nikolajevic A, Deltedesco A, Sigmund JE, Blumer MJ, Hermann M, Vanacker J, Brandacher G, Stefan E, Torres-Quesada O, Troppmair J
The emergence of resistance to mutant BRAF-specific inhibitors (BRAFi) requires novel strategies for melanoma treatment. The progression of these tumors involves metabolic adaptations, which also affect the cellular redo...The emergence of resistance to mutant BRAF-specific inhibitors (BRAFi) requires novel strategies for melanoma treatment. The progression of these tumors involves metabolic adaptations, which also affect the cellular redox status. Previous studies have linked RAF kinase signaling, a key component of the MAPK/ERK pathway involved in cell division and survival, to the suppression of mitochondrial reactive oxygen species (ROS) production, resulting in protection against cell death. In BRAF-transformed cells, we have identified impaired JNK1/2-dependent activation of the mitochondrial prooxidant protein p66Shc as a potential cause. In the present study, we dissected signaling and mitochondrial alterations that characterize the transition from BRAFi responsiveness to resistance in A375 melanoma cells. Insensitivity to BRAFi dabrafenib exposure was associated with reactivation of ERK1/2 phosphorylation, increased JNK1/2 kinase activity, p66ShcS36 phosphorylation, and elevated ROS production. Utilizing high-resolution respirometry (HRR) and transmission electron microscopy (TEM), we show that dabrafenib-resistant cells displayed mitochondrial damage, compensated by increased respiration, leading to high ROS levels. Moreover, dabrafenib-resistant cells (A375D) have more efficient antioxidant systems, which may explain why despite ongoing cell death, net cell growth was observed. Treatment of both parental and resistant cells with phenethyl isothiocyanate (PEITC) increased ROS production but caused substantial cell death only in A375D melanoma cells. This PEITC effect could be demonstrated in two further dabrafenib-resistant cell lines, WM164D and 451LuP. These results suggest that the altered redox status is linked to compromised mitochondria and is associated with the development of BRAFi resistance, rendering cells exquisitely sensitive to the actions of selective ROS-inducing therapeutics.
Mismatch repair-proficient (pMMR) colorectal cancers (CRC) have long been considered nonresponsive to immune checkpoint blockade (ICB), in contrast to their mismatch repair-deficient (dMMR) counterparts. Recent evidence...Mismatch repair-proficient (pMMR) colorectal cancers (CRC) have long been considered nonresponsive to immune checkpoint blockade (ICB), in contrast to their mismatch repair-deficient (dMMR) counterparts. Recent evidence indicates that neoadjuvant immunotherapy can be used to treat pMMR CRC before surgery, potentially reducing postoperative relapse. Tan et al. report results from the NICHE-2 trial, which achieved a 26% response rate in early-stage pMMR colon cancer (CC) patients. Molecular studies show that despite low tumor mutational burden (TMB), responders exhibit higher chromosomal instability (CIN), TP53 mutations, and enrichment of proliferative and cell-cycle signatures, associated with higher density of Ki-67 tumor and CD8 T cells. In contrast, nonresponders display metabolic and stromal reprogramming, enhanced TGF-β signaling, and immune exclusion. Circulating tumor DNA (ctDNA) clearance correlated with pathological response and long-term disease-free survival postsurgery. While the biological and molecular determinants underlying the response rates observed in the NICHE-2 trial remain to be fully elucidated, the work by Tan et al. suggests that biomarker-guided neoadjuvant immunotherapy could represent a valuable strategy to achieve pathological responses in early-stage pMMR CC, despite its clinical relevance requiring further evaluation.
Chordoma, a rare primary bone malignancy, currently lacks effective targeted therapies. Despite surgical resection and adjuvant radiotherapy, prognosis remains poor. Recent preclinical studies have highlighted potential...Chordoma, a rare primary bone malignancy, currently lacks effective targeted therapies. Despite surgical resection and adjuvant radiotherapy, prognosis remains poor. Recent preclinical studies have highlighted potential therapeutic targets, including the transcription factor T-box transcription factor T (TBXT). However, clinical outcomes associated with therapies targeting TBXT remain underexplored or have been modest, warranting further investigation. In this study, we investigated the therapeutic potential of transfer RNA (tRNA) synthetase inhibitors in chordoma treatment. Focused compound screening identified distinct chemotypes targeting human glutamyl-prolyl-tRNA synthetase (EPRS) as being effective in reducing cell viability in chordoma cell lines through a cyclic AMP-dependent transcription factor (ATF4)-mediated stress response rather than through TBXT regulation. Mechanistically significant upregulation of ATF4 and associated stress response genes was identified with consecutive pro-apoptotic DNA damage-inducible transcript 3 protein (DDIT3)-mediated cell death. The prototypic EPRS inhibitor halofuginone demonstrated significant tumour growth inhibition in an in vivo patient-derived xenograft model. These results suggest that targeting metabolic stress pathways via ATF4 activation presents a novel therapeutic approach for chordoma, warranting further clinical investigation.
Breast cancer is known for late recurrences, yet current follow-up lacks radiological or blood-based monitoring for systemic relapse. This study evaluated circulating tumor DNA (ctDNA) monitoring for early detection of s...Breast cancer is known for late recurrences, yet current follow-up lacks radiological or blood-based monitoring for systemic relapse. This study evaluated circulating tumor DNA (ctDNA) monitoring for early detection of systemic relapse after curative treatment. In this case-control study of 70 patients with operable breast cancer (35 with relapse and 35 without relapse), blood samples were collected every 6-12 months during a median 8.3-year follow-up. ctDNA was analyzed by targeted DNA sequencing using Oncomine™ Breast cfDNA Research Assay v2, and results were compared to genetic analysis of tumor and metastasis biopsies. ctDNA was detected at relapse in 19 of 35 (54%) patients with disease relapse and preceded clinical or radiological relapse detection in 17, with a median lead time of 10.3 months. In 13 (68%) patients, there was concordance with tumor mutations, and in seven patients, there was also concordance with metastasis. Among the relapse-free patients, seven were ctDNA-positive postsurgery, and only one of them had a match among the tumor variants. These findings suggest serial ctDNA analysis may enable earlier detection of systemic relapse in patients with operable breast cancer.
Immunotherapy has revolutionized cancer treatment; yet, a subset of patients with microsatellite instability-high (MSI-H) tumors fails to respond to treatment despite their elevated tumor mutational burden and immunogeni...Immunotherapy has revolutionized cancer treatment; yet, a subset of patients with microsatellite instability-high (MSI-H) tumors fails to respond to treatment despite their elevated tumor mutational burden and immunogenic potential. In a recent study, Xu et al. uncover a key mechanism of immune evasion in MSI-H tumors mediated by the exonuclease TREX1, which degrades cytosolic DNA and suppresses activation of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING)-type I interferon pathway. Loss of TREX1 restores cytosolic DNA sensing, promotes CD8 T and NK cell infiltration, and enhances antitumor immunity. These findings highlight TREX1 as a potential therapeutic target to overcome resistance to immune checkpoint blockade.
Penile carcinoma is a rare malignancy in developed countries but is more common in South America and East Africa. The small number of cases means there are limited resources to investigate disease pathogenesis. This repo...Penile carcinoma is a rare malignancy in developed countries but is more common in South America and East Africa. The small number of cases means there are limited resources to investigate disease pathogenesis. This report describes a method of generating primary cell lines from freshly isolated human penile tissue using a clonal expansion approach on mitotically inactivated fibroblasts. Matched normal and penile cancer cell lines from two patients were generated and characterised. Molecular karyotyping and targeted sequencing were performed to compare their genomic landscape. Gains in 8q13.3, 10q23.2, 10q25.1, 10q26.13,12p13.33, 20q13.33, Xq21.1 or losses in Yq11.23 were consistent in both tumour cell lines. Gains in 8q13.3 and Xq21.1 cytobands positively correlated with changes in the expression of nearby genes. The top 20 differentially expressed genes are involved in immune responses like interferon alpha/beta signalling. Additionally, there was an increase in integrin β1, transglutaminase 1, keratins 5, 10, 14 and 16, and a decrease in involucrin protein expression. The cell lines described in this study can provide an invaluable platform for new insights and testing of therapies for penile carcinoma.
Liquid biopsies containing circulating tumor DNA (ctDNA) are important biomarkers across several forms of cancer. The detection of mutations in cell-free DNA (cfDNA) indicates the presence of ctDNA. However, unsatisfacto...Liquid biopsies containing circulating tumor DNA (ctDNA) are important biomarkers across several forms of cancer. The detection of mutations in cell-free DNA (cfDNA) indicates the presence of ctDNA. However, unsatisfactory ctDNA mutation sensitivities, issues with sequencing errors, and clonal hematopoiesis variants have limited the clinical utility of mutation-based ctDNA assays. Recently, a new avenue of cfDNA assays has been developed, focusing on cfDNA epigenetics. Here, we outline the recent advancements in cfDNA epigenetics, focusing on cfDNA methylation, fragmentomics, and post-translational modifications (PTMs) of circulating nucleosomes. We present various methylation strategies concerning ctDNA detection and tissue of origin (TOO) analyses. cfDNA fragmentomics focuses on cfDNA fragment lengths, fragment end motifs, and nucleosome positioning to infer gene expression and estimate the ctDNA fraction. Lastly, we discuss the development of cell-free chromatin immunoprecipitation of circulating nucleosomes with PTMs. This method has been implemented to detect tumor gene expression, TOO, and treatment resistance. Combining the epigenetic features of cfDNA will expand the utility of liquid biopsies to give a more comprehensive insight into tumor biology, treatment response, and resistance.