The clinical implementation of circulating tumor cells (CTCs) as a predictive tool for therapy efficacy in the growing field of precision oncology likely requires advanced CTC phenotyping. While their predictive value fo...The clinical implementation of circulating tumor cells (CTCs) as a predictive tool for therapy efficacy in the growing field of precision oncology likely requires advanced CTC phenotyping. While their predictive value for disease progression and therapy response has extensively been shown, the low number of CTCs that are obtained from whole blood tubes limits their clinical application. The magnetic enrichment of CTCs using the FDA-cleared CellSearch system is limited by the expression of the epithelial cell adhesion molecule (EpCAM) on CTC, and an increase in CTC yield may be obtained through enhanced capture of low EpCAM expressing cells. In this study, we compared the Flow Enabled Target Capture Halbach (FETCH) magnetic enrichment technology with the FDA-cleared CellSearch system for CTC capture using paired blood samples from 34 patients with non-small-cell lung, breast, and prostate cancer. Results show a statistically significant median 1.5-fold increase in CTCs captured with the FETCH system compared with CellSearch, accompanied by a median twofold reduction in non-specifically captured leukocytes. This increase in CTC capture can aid the comprehensive analysis of CTCs in a larger number of patients, further supporting clinical implementation of CTC-based diagnostics.
Schreuder A, de Lint K, Mỹ Phúc Võ H
… +8 more, Góis MM, Kampen RA, San Martin Alonso M, Nootenboom I, Garzero V, Wendel TJ, Wolthuis RMF, Noordermeer SM
With the recent rise in CRISPR/Cas9-mediated genome-wide synthetic lethality screens, many new synthetic lethal targets have been identified for diseases with underlying genetic causes such as tumours with BRCA1 mutation...With the recent rise in CRISPR/Cas9-mediated genome-wide synthetic lethality screens, many new synthetic lethal targets have been identified for diseases with underlying genetic causes such as tumours with BRCA1 mutations. Such screens often use full deficiency of a protein to identify novel vulnerabilities. However, patient-derived mutations not only result in full loss of the protein but often also concern missense mutations with hypomorphic phenotypes. Here, we study the genetic vulnerabilities of two previously described hypomorphic BRCA1 missense mutations and compare these to a BRCA1-depleted setting to study whether this affects screening for synthetic lethal interactions. Our research showed that BRCA1 mutated cells have very similar vulnerabilities to BRCA1 wild-type cells, confirming its low tumorigenic effect. In contrast, the BRCA1 mutation induced a more similar phenotype to BRCA1-deficient cells. For this mutation, we also unveiled a unique vulnerability to the loss of NDE1. Specifically in BRCA1 mutated cells, and not BRCA1-proficient or -deficient cells, NDE1 loss leads to increased genomic instability. Altogether, our findings highlight the importance to differentiate between patient-derived mutations when assessing novel treatment targets.
Genetic variants in the DNA polymerase enzymes POLE and POLD1 can affect protein function by altering stability, catalysis, DNA binding, and interactions with other biomolecules. Understanding the structural basis of the...Genetic variants in the DNA polymerase enzymes POLE and POLD1 can affect protein function by altering stability, catalysis, DNA binding, and interactions with other biomolecules. Understanding the structural basis of these variants is important for a comprehensive interpretation of variant impact. In this study, we used MAVISp, a modular structure-based framework, and molecular dynamics simulations to analyze over 60 000 missense variants of POLE and POLD1. By integrating results from changes in folding and binding free energies, local alterations in the proximity of the active or phosphorylation sites, we provided a detailed structural interpretation of variants reported across various databases, including ClinVar, COSMIC, and cBioPortal. Moreover, we predicted the functional consequences of variants not found yet in disease-related databases, thereby creating a comprehensive catalog for future studies. Of note, our approach enabled us to classify 364 Variants of Uncertain Significance (VUS) as PP3 evidence and 323 as BP4 evidence, in accordance with the American College of Medical Genetics and Genomics (ACMG) guidelines. Additionally, we identified a group of variants that could alter the native orientation of the residues within the catalytic site of the exonuclease domain, such as POLE variants P297S and P436R. Finally, we identified a group of variants predicted to affect DNA-binding affinity and rationalized their effects in terms of different energetic contributions and structural features. Collectively, our results not only advance our understanding of protein variant effects in POLE and POLD1 at the structural level but also support future studies aimed at variant classification, variant prioritization for experimental studies, and functional interpretation across diverse biological contexts.
Stergiopoulou D, Markou A, Nicolo E
… +10 more, Serafini MS, Zhang Q, Zhang Y, Gerratana L, Davis AA, Liu H, Gradishar WJ, Reduzzi C, Cristofanilli M, Lianidou E
Endocrine resistance is a complex phenomenon, including alterations of the ESR1 gene. The aim of this study was to simultaneously analyze ESR1 promoter methylation and ESR1 hotspot mutations in circulating tumor cells (C...Endocrine resistance is a complex phenomenon, including alterations of the ESR1 gene. The aim of this study was to simultaneously analyze ESR1 promoter methylation and ESR1 hotspot mutations in circulating tumor cells (CTCs) and paired plasma-circulating tumor DNA (ctDNA) from patients with estrogen receptor-positive (ER+) advanced breast cancer (BC). We retrospectively analyzed samples from 42 ER+ advanced BC patients characterized for CTCs and ctDNA at Northwestern University. CTCs were enumerated using the CellSearch® system, while ctDNA was analyzed with the Guardant360 NGS platform. Genomic DNA from CellSearch-enriched CTC fractions was amplified and analyzed using the ESR1-NAPA assay. ESR1 methylation analysis was performed in 34 samples. ESR1 mutations were detected in 59.5% CTC-derived samples, a significantly higher proportion than in paired plasma ctDNA (29.6%). ESR1 methylation was observed in 26.5% patients. Concurrent ESR1 mutations and methylation were identified in six cases, suggesting combined genetic and epigenetic mechanisms of endocrine resistance. Overall, CTC-derived genomic DNA showed higher sensitivity for detecting ESR1 mutations than plasma ctDNA, supporting the potential value of CTC analysis for characterizing endocrine resistance in advanced BC.
Cellular plasticity is central to non-small cell lung cancer (NSCLC) disease progression and linked to aberrant regulation of developmental programmes. Here, we investigated the contribution of two developmental programm...Cellular plasticity is central to non-small cell lung cancer (NSCLC) disease progression and linked to aberrant regulation of developmental programmes. Here, we investigated the contribution of two developmental programmes, alveogenesis (ALV) and branching morphogenesis (BM), to NSCLC disease progression. ALV and BM inversely correlated across multiple transcriptome datasets. In squamous carcinomas (LUSC), ALV suppression and BM activation were consistently observed relative to controls, but these features were not prognostic. In contrast, adenocarcinomas (LUAD) displayed heterogeneous BM activation, associated with poor overall survival in several observational cohorts (n = 5) and resistance to tyrosine kinase inhibitors or immune checkpoint blockade. Exome sequencing linked TP53 pathway mutations to BM activation in LUAD, which was validated in conditional Trp53 knock-out mouse models. Single-cell RNA-sequencing combined with multiplexed immunohistochemistry showed LUAD BM activation reflected increased morphological grade with tumour cells transdifferentiating to a basal-like cell state. Finally, 3D organotypic cultures identified type-I interferon signalling as a driver of BM activation in TP53-mutant LUAD. Collectively, our findings reveal a novel TP53-interferon axis that promotes transcriptomic plasticity in LUAD, with important implications for biomarker and therapeutic target discovery.
Aberrant signaling through insulin-like growth factor 1 receptor (IGF1R) and epidermal growth factor receptor (EGFR) drives glioblastoma (GBM) progression and therapy resistance. Herein, we describe the synthesis and bio...Aberrant signaling through insulin-like growth factor 1 receptor (IGF1R) and epidermal growth factor receptor (EGFR) drives glioblastoma (GBM) progression and therapy resistance. Herein, we describe the synthesis and biological evaluation of W1B, a novel styrylquinazolinone-based small-molecule inhibitor. In antiproliferative assays, W1B exhibited potent submicromolar activity against a panel of GBM cell lines. Kinase assays and binding studies confirmed strong inhibition and high binding affinity toward IGF1R. Molecular docking suggested possible interactions with both IGF1R and EGFR, with W1B adopting distinct binding poses in each kinase domain. In cellular studies, W1B reduced IGF1R and EGFR protein levels in LN229 cells and suppressed Akt phosphorylation. Under high-glucose conditions, however, W1B only retained inhibitory activity toward IGF1R, resulting in attenuated effects on the Akt/mTOR axis and underscoring the influence of glucose-dependent signaling rewiring on drug efficacy. Combination studies revealed that W1B acts synergistically with the EGFR inhibitor dacomitinib, effectively overcoming compensatory activation of parallel pathways. Biomimetic lipophilicity and in silico pharmacokinetic analyses indicated that styrylquinazolinone has the potential to cross the blood-brain barrier (BBB). The in vivo studies on Danio rerio have shown a good safety profile, as well as strong antitumor potential of the tested compound. Therefore, these findings establish W1B as a promising derivative for the development of next-generation dual IGF1R/EGFR inhibitors in GBM.
Xu J, Niklaus NJ, Schläfli AM
… +11 more, Tokarchuk I, Humbert M, La Manna F, Vu B, Maul D, Radpour R, Kruithof-de Julio M, Zlobec I, Dengjel J, Torbett BE, Tschan MP
The cyclin D binding Myb-like Transcription Factor 1 (DMTF1) is a haploinsufficient tumor suppressor in various tumors. Alternative splicing generates a dominant negative, truncated version of full-length DMTF1α, named D...The cyclin D binding Myb-like Transcription Factor 1 (DMTF1) is a haploinsufficient tumor suppressor in various tumors. Alternative splicing generates a dominant negative, truncated version of full-length DMTF1α, named DMTF1β. DMTF1β has so far been described as an oncogene in breast cancer development. However, a clear understanding of how DMTF1β contributes to carcinogenesis remains unknown. Analyzing DMTF1β protein expression in breast cancer cell lines, as well as a highly metastatic prostate cancer cell line, confirmed a positive correlation between DMTF1β expression and tumorigenic potential. Specifically, knocking down DMTF1β in aggressive MDA-MB-231 breast and PC3MPRO4 prostate cancer cells significantly reduced wound closure and tissue invasion. β-specific interactome and RNA-sequencing studies in DMTF1β overexpressing and knockdown cells, respectively, suggest that DMTF1β expression is associated with the autophagy recycling pathway. Depleting DMTF1β levels in cancer cells significantly decreased autophagic flux. Moreover, inhibiting autophagy led to decreased migration of DMTF1β expressing breast cancer cells. Mechanistically, DMTF1β protein interacts with and stabilizes the key autophagy protein ULK1. In conclusion, we identified a novel function for the alternatively spliced DMTF1 gene product in autophagy-dependent cancer cell motility.
Nishimura J, Kuribayashi T, Brägelmann J
… +17 more, Okawa S, Taoka M, Mori S, Nishimura T, Tanaka T, Makimoto G, Ninomiya K, Rai K, Ichihara E, Katayama R, Hotta K, Tabata M, Togashi Y, Maeda Y, Sos ML, Kiura K, Ohashi K
Epidermal growth factor receptor (EGFR)-mutant non-small-cell lung cancers (NSCLCs) lack effective immunotherapy due to a noninflamed tumor microenvironment (TME). We previously reported that EGFR tyrosine-kinase-inhibit...Epidermal growth factor receptor (EGFR)-mutant non-small-cell lung cancers (NSCLCs) lack effective immunotherapy due to a noninflamed tumor microenvironment (TME). We previously reported that EGFR tyrosine-kinase-inhibitor (TKI) induced CD8 T-cell immunity, which was insufficient for tumor eradication. We evaluated the potential of combining EGFR-TKI with stimulator of interferon genes (STING) agonists in activating a systemic antitumor response. Using a syngeneic mouse model of genetically engineered Egfr-mutant NSCLC, we evaluated the antitumor effects of STING agonist ADU-S100, alone and combined with osimertinib. Immunohistochemistry and flow cytometry were used to assess the TME. Osimertinib alone enhanced CD8 T-cell infiltration but not Natural Killer (NK) cell infiltration. ADU-S100 injection alone modestly suppressed tumor growth with increasing CD8/NK cell infiltration in the TME, but lacked an abscopal effect. Combining ADU-S100 with osimertinib significantly enhanced the antitumor effects and CD8/NK cell infiltration. Depletion of either CD8 or NK cells reduced the combination effect. Crucially, the combination induced an abscopal effect accompanied by PD-1/CD8 cell infiltration. Combining osimertinib with a STING agonist augmented innate and adaptive immunity, inducing systemic antitumor responses in EGFR-mutant NSCLC.
The 3'-5' exoribonuclease EXOSC10 degrades aberrant mRNAs and noncoding RNAs in cooperation with the nuclear RNA exosome. EXOSC10's localization and stability are regulated by sumoylation and proteasomal degradation in r...The 3'-5' exoribonuclease EXOSC10 degrades aberrant mRNAs and noncoding RNAs in cooperation with the nuclear RNA exosome. EXOSC10's localization and stability are regulated by sumoylation and proteasomal degradation in response to stress, and the protein is essential for cell growth and proliferation, fertility, hematopoiesis, and brain development. EXOSC10 is a cancer biomarker; its activity is inhibited by the widely used anticancer drug 5-fluorouracil (5-FU) and the protein's depletion sensitizes cells to 5-FU. We employed mass spectrometry to reveal EXOSC10's post-translational modifications, such as phosphorylation, acetylation and ubiquitination, and to explore its protein interaction network, which includes RNA exosome subunits and enzymes involved in protein degradation. Furthermore, we find that the EXOSC10 allele identified in colon cancer and located within a motif for targeted proteolysis is stable, nuclear but nonfunctional in vivo, since homozygous Exosc10 mice exhibit early embryonic lethality. We identified equivalent S402P/S402A variants and heterozygous loss-of-function (LoF) alleles in cancers and healthy individuals using public genomics data. Our findings suggest that recessive EXOSC10 LoF alleles may cause increased 5-FU sensitivity in tumors bearing de novo mutations and hypertoxicity in heterozygous carriers.
Schober AS, Moreno-Cugnon L, Martínez-Pérez L
… +14 more, Ercilla A, Zabala-Letona A, Vicente-Barrueco A, Fagoaga-Eugui M, Garcia-Longarte S, Calle-Ciborro B, Pérez-Andrés E, Carlevaris O, Pozo S, Mendizabal I, Mayor U, Carracedo A, Sée V, Berra E
Oxygen availability is frequently compromised in solid tumours, making intratumoural hypoxia a common feature of cancer. In prostate cancer (PCa), hypoxia is strongly associated with aggressive disease and poor prognosis...Oxygen availability is frequently compromised in solid tumours, making intratumoural hypoxia a common feature of cancer. In prostate cancer (PCa), hypoxia is strongly associated with aggressive disease and poor prognosis. Hypoxia-inducible factor (HIF) is the master transcriptional regulator mediating hypoxia adaptation and is mainly controlled through proteasomal degradation of its α-subunit by the ubiquitin-proteasome system (UPS). However, the contribution of deubiquitinases (DUBs) to HIF signalling in PCa remains largely unexplored. Using a computational strategy based on CA9 expression as a surrogate of HIF activity, we identified Ubiquitin-Specific Protease 29 (USP29) as a key regulator associated with hypoxia and tumour progression and severity in PCa. Mechanistically, USP29 functions as a noncanonical positive regulator of HIF-α stability in a catalytic-dependent manner. USP29 interacts with HIF-1α, reduces its poly-ubiquitination and protects it from proteasomal degradation across multiple cancer cell lines. Additionally, USP29 stabilizes HIF-2α acting on the C-terminal region of HIF-α. These findings uncover a novel regulatory layer of HIF signalling and highlight USP29 as a potential therapeutic target in hypoxia-driven PCa progression.
The insulin-like growth factor (IGF) axis regulates cancer cell proliferation, growth, invasion, and therapy resistance. Elevated expression of the type 1 IGF receptor (IGF-1R) is linked to radioresistance and biochemica...The insulin-like growth factor (IGF) axis regulates cancer cell proliferation, growth, invasion, and therapy resistance. Elevated expression of the type 1 IGF receptor (IGF-1R) is linked to radioresistance and biochemical recurrence in prostate cancer, yet the molecular mechanisms underlying IGF-1R-mediated DNA damage responses remain unclear. We investigated the role of IGF-1R in DNA double-strand break (DSB) repair by assessing chromatin recruitment of DNA repair proteins, repair pathway usage, and therapeutic sensitivity in cancer cell models with altered IGF-1R status. Loss of IGF-1R impaired DNA-dependent protein kinase catalytic subunit (DNA-PKcs) localisation to chromatin, resulting in defective non-homologous end-joining (NHEJ) and a compensatory reliance on alternative repair pathways, including microhomology-mediated end-joining (MMEJ). Modulating IGF-1R expression restored radiosensitivity in poly (ADP-ribose) polymerase (PARP) inhibitor-resistant breast cancer cells. IGF-1R inhibition compromises canonical DSB repair and re-sensitises resistant cancer cells to therapy, supporting its potential as a therapeutic strategy in homologous recombination-deficient tumours. Furthermore, IGF-1R mutant cancers may benefit from targeted inhibition of the MMEJ pathway.
Nykel A, Kubiak I, Rutkowska L
… +10 more, Kasprzyk Ż, Kępczyński Ł, Bednarek M, Sobieraj D, Wilkosz J, Kania P, Jędrzejczyk A, Kałużewski B, Gach A, Kałużewski T
Bladder cancer (BC) is the ninth most common cancer worldwide, with urothelial carcinoma accounting for approximately 90% of all cases and presenting predominantly as non-muscle-invasive disease. Due to its high recurren...Bladder cancer (BC) is the ninth most common cancer worldwide, with urothelial carcinoma accounting for approximately 90% of all cases and presenting predominantly as non-muscle-invasive disease. Due to its high recurrence rate and the need for long-term surveillance, BC is associated with the highest lifetime treatment costs per patient among all cancers, making its effective management a significant clinical and economic challenge. The most frequently identified variants in the TERT gene promoter are c.-124C>T (C228T) and c.-146C>T (C250T), located within a region characterized by high guanine-cytosine (GC) content, which makes amplification challenging. We aimed to validate the AbsoluteQ Digital PCR assay for the detection of urine-based TERT promoter variants for the diagnosis of urothelial bladder cancer and to assess its diagnostic performance in comparison with standard methods. Urine samples were collected from patients with histopathologically confirmed bladder cancer (n = 58) and compared with a control group (n = 55). The C228T and C250T variants were tested using the AbsoluteQ Digital PCR assay. Sensitivity, specificity, and predictive values were calculated to evaluate the performance of the assessed method. The AbsoluteQ Digital PCR demonstrated superior diagnostic performance compared to conventional Sanger sequencing for detecting TERT promoter variants, achieving a sensitivity of 89.65% (95% CI: 78.16-95.72) and a specificity of 100% (95% CI: 91.87-100), with no false positives observed. Given its robustness and clinical relevance, AbsoluteQ Digital PCR is emerging as a promising tool for non-invasive molecular diagnostics targeting TERT promoter variants.
Casas-Arozamena C, Lande KT, Diaz E
… +13 more, Vilar A, Cueva J, Arias E, Sampayo V, Abalo A, González N, Colás E, Gil-Moreno A, Abal M, Moreno-Bueno G, Sørlie T, Lindemann K, Muinelo-Romay L
Intratumor heterogeneity presents a major challenge in precision oncology for endometrial cancer (EC). Circulating tumor DNA (ctDNA) offers a minimally invasive method to monitor tumor evolution and therapeutic resistanc...Intratumor heterogeneity presents a major challenge in precision oncology for endometrial cancer (EC). Circulating tumor DNA (ctDNA) offers a minimally invasive method to monitor tumor evolution and therapeutic resistance. In this retrospective study, we evaluated a tumor-agnostic NGS panel to detect and track ctDNA in 18 EC patients and directly compared its performance with a tumor-informed ddPCR approach. ctDNA was detected by NGS in over 60% of plasma samples, while ddPCR showed higher positivity rates in paired samples (71.9% vs 62.5%), with overall concordance of 65.7% and fair agreement (Cohen's kappa = 0.23). The cfDNA-NGS panel identified a broad spectrum of alterations, including relapse-specific mutations indicative of clonal evolution, but showed lower sensitivity for low-frequency variants compared with ddPCR. Discordant cases, including false-negative results in both approaches, highlight the impact of assay sensitivity, target selection, and biological factors on ctDNA detection. ctDNA dynamics correlated with disease progression and treatment response, although detection was limited in cases with brain metastases. These findings support the utility of tumor-agnostic ctDNA monitoring in advanced EC and highlight the importance of assay quality and careful interpretation to address limitations such as clonal hematopoiesis and technical sensitivity.
Chen C, Cheng J, Hou R
… +12 more, Zheng X, Su L, Bjaanæs MM, Karlsson A, Planck M, Staaf J, Helland Å, Esteller M, Christiani DC, Chen F, Cao X, Zhang R
The MAPRE3 gene is aberrantly expressed in several cancers. We profiled DNA methylation in tumor tissues from early-stage non-small cell lung cancer (NSCLC) patients and assessed associations with overall survival (OS)....The MAPRE3 gene is aberrantly expressed in several cancers. We profiled DNA methylation in tumor tissues from early-stage non-small cell lung cancer (NSCLC) patients and assessed associations with overall survival (OS). Significant CpG probes were validated in The Cancer Genome Atlas (TCGA). The methylation level of cg12821679 showed significant associations with OS in lung squamous cell carcinoma (LUSC) (HR = 0.32, P = 6.55 × 10), but it was not observed in lung adenocarcinoma (LUAD). In LUSC, MAPRE3 expression was significantly correlated with cg12821679 (r = 0.17, P = 2.96 × 10) and potential trans-regulated genes were enriched in the Nicotine addiction pathway. Additionally, MAPRE3 expression showed significant associations with OS in both LUAD and LUSC (LUAD: HR = 2.28, P = 2.40 × 10; LUSC: HR = 1.61, P = 0.0244). The association between smoking cessation and overall survival was significantly modified by MAPRE3 expression (HR = 0.69, P = 0.0282). Smoking cessation improved OS only in patients with high MAPRE3 expression (HR = 0.56, P = 2.82 × 10). We conclude MAPRE3 may predict NSCLC prognosis and influence the prognostic benefit of smoking cessation.
The response to radiotherapy (RT) is influenced by the individual DNA repair capacity of both the tumor cells and the host. In this study, we assessed whether circulating tumor cell (CTC) enumeration, kinetics, and CTC v...The response to radiotherapy (RT) is influenced by the individual DNA repair capacity of both the tumor cells and the host. In this study, we assessed whether circulating tumor cell (CTC) enumeration, kinetics, and CTC viability (i.e., the apoptotic rate) could provide a more accurate tool for monitoring and stratifying the RT response. We analyzed CTCs and tumor-derived extracellular vesicles (tdEVs) during RT in 71 lung (n = 35) and breast (n = 36) cancer patients receiving RT treatment for brain (n = 54) and bone (n = 19) metastases. The DNA repair capacity of the host was assessed by ex vivo irradiation of peripheral blood mononuclear cells (PBMCs). RT treatment did not seem, in most cases, to cause a short-term release of CTCs. We found that both the number and apoptotic rate of CTCs before and after RT treatment is a powerful indicator of poor prognosis. Additionally, the fraction of apoptotic CTCs correlated with RT response and patient outcome. This study demonstrated that the RT response is associated with tumor-specific traits, which can be accessed via easily accessible liquid biopsy approaches.
Diffuse gliomas are primary brain tumors including glioblastomas (GB), astrocytomas, and oligodendrogliomas, the latter two harboring IDH1 mutations and exhibiting slower progression. Gliomas display cellular plasticity,...Diffuse gliomas are primary brain tumors including glioblastomas (GB), astrocytomas, and oligodendrogliomas, the latter two harboring IDH1 mutations and exhibiting slower progression. Gliomas display cellular plasticity, with transitions between astrocyte-like, oligodendrocyte-like, progenitor-like, and mesenchymal-like states driven by genetic alterations and microenvironmental signals. The proneural-to-mesenchymal transition (PMT), associated with increased malignancy, is tightly regulated by the tumor microenvironment, notably through cytokine signaling and non-tumor cell interactions. Endothelins (ET-1, ET-2, ET-3), vasoactive peptides mainly produced by vascular cells, signal through the G-protein-coupled receptors EDNRA and EDNRB and were previously suggested to promote glioma proliferation based on serum-based models. Here, we revisited endothelin signaling using eleven serum-free glioma lines and tumor samples. Multi-omics and electrophysiological analyses identified EDNRB as the predominant receptor, enriched in astrocyte-like cells, increased by BMPs or growth factor withdrawal, and repressed by interferons, IL-6 family cytokines, endothelins, and Hippo/YAP signaling. EDNRA was confined to a perivascular tumor subpopulation and induced by Notch signaling selectively in GB. Functionally, endothelins reduced proliferation while promoting migration and PMT via EDNRB-dependent Ca signaling, ERK/STAT3 activation, and apamin-sensitive SK2/SK3 potassium channel activity. Collectively these findings establish endothelin signaling as an important regulator of glioma cell plasticity and behavior.
Saleh A, Medina-Itzhaki N, Chekov M
… +10 more, Gal-Swisa E, Gabesh-Wahabi R, Savyon I, Naroditsky I, Kenny HA, Fares B, Korsensky L, Girsh E, Berger L, Perets R
Paired box protein Pax-8 (PAX8) is a critical lineage marker and master regulator of transcription in high-grade serous ovarian carcinoma (HGSC)-the most common subtype of epithelial ovarian cancer-driving cell prolifera...Paired box protein Pax-8 (PAX8) is a critical lineage marker and master regulator of transcription in high-grade serous ovarian carcinoma (HGSC)-the most common subtype of epithelial ovarian cancer-driving cell proliferation and migration and resisting apoptosis. This study aimed to elucidate the mechanism of action of PAX8 in this disease. By performing an unbiased analysis of PAX8-regulated genes, we discovered two PAX8-regulated genes-coiled-coil domain-containing protein 80 (CCDC80) and cluster of differentiation 276 (CD276) antigen (also known as B7-H3)-that mediate PAX8 activity and play key cancer cell autonomous roles in this disease. Our findings indicate that PAX8 negatively regulates CCDC80, a novel cell autonomous tumor suppressor in HGSC. We demonstrate that CCDC80, localized to the nucleus, significantly reduces HGSC tumor growth and metastasis in vivo in a mouse model. Notably, CCDC80 exerts its function by suppressing the expression of the immune checkpoint protein B7-H3. However, in HGSC, B7-H3 is predominantly cytoplasmic and promotes HGSC proliferation and migration independent of its immune role. Additionally, PAX8 positively regulates B7-H3 expression in a CCDC80-independent manner, underscoring the multifaceted oncogenic role of PAX8. This study highlights the complex regulatory network involving PAX8, CCDC80, and B7-H3 in HGSC progression. Targeting this signaling pathway may provide a novel therapeutic strategy to improve treatment outcomes for patients with epithelial ovarian cancer. B7-H3, which is currently targeted in clinical trials, shows promise as HGSC target for therapy.
CD47 is best known for its role in tumor immune evasion; however, studies in diverse cell models indicate that it also has cell autonomous, tumor-promoting functions which are cell type- and context-specific. Motivated b...CD47 is best known for its role in tumor immune evasion; however, studies in diverse cell models indicate that it also has cell autonomous, tumor-promoting functions which are cell type- and context-specific. Motivated by the prognostic and therapeutic significance of CD47 and the limited knowledge regarding its roles beyond immune evasion in non-small cell lung cancer (NSCLC), we sought to define the cellular and molecular processes driven by intrinsic CD47 signaling in NSCLC. Transcriptome profiling of CD47 wild-type and knockout NSCLC cells implicated its regulation of genes enriched for signatures of mitogen-activated protein kinase (MAPK) signaling and epithelial-to-mesenchymal transition (EMT). A significant positive association between CD47 and MAPK/EMT expression signatures was also evident in large cohorts of NSCLC cell lines and tumor tissues. Functional studies indicated that CD47 does not regulate cell proliferation in NSCLC cells like it does in other cancer types. Instead, CD47 regulates cell adhesion and migration through an ERK and EMT axis, validating our transcriptomic findings. Moreover, CD47 loss-of-function significantly diminished the ability of NSCLC cells to metastasize in vivo, demonstrating the physiological relevance of cell-intrinsic CD47 signaling in lung cancer cells. Our data reveal a novel role for CD47 in relaying signals through ERK to promote EMT expression programs and prometastatic phenotypes in NSCLC. Although additional mechanistic studies are needed to further decipher the CD47-ERK-EMT signaling pathway, our findings reinforce the therapeutic potential of CD47, rationalizing further research to develop CD47 blockade as a multimodal therapy for NSCLC.