The limited efficacy of immunotherapy in clinical trials in high-grade serous ovarian cancer (HGSOC) may improve by implementing experimental models that are more reflective of human biology into preclinical studies. To...The limited efficacy of immunotherapy in clinical trials in high-grade serous ovarian cancer (HGSOC) may improve by implementing experimental models that are more reflective of human biology into preclinical studies. To address this, we developed and validated a humanized patient-derived xenograft mouse model of HGSOC. Human hematopoietic stem cells and patient-derived HGSOC cells were engrafted into immunodeficient mice. The mice were administered durvalumab (anti-PD-L1) and/or oleclumab (anti-CD73) immunotherapy intraperitoneally twice a week for 5 weeks. The treatment showed good tolerability with no observed side effects, though it failed to elicit a measurable antitumor response. Leukocytes in primary tumors were analyzed immunohistochemically, and circulating T cells were characterized using spectral flow cytometry. All tumors exhibited an immune-excluded immunophenotype. No significant inter-group differences in disease burden, intratumoral leukocyte density, or circulating T cells were observed. In the durvalumab-only group, tumor burden significantly positively correlated with intratumoral cytotoxic and regulatory T-cell densities. This model reflects the immunotherapy resistance of human disease in line with clinical findings, providing a robust platform for studying tumor-immune interactions and immunosuppressive mechanisms in HGSOC. Impact statement Our results address the critical need for representative preclinical models for testing combination immunotherapy in HGSOC by providing a robust preclinical platform that can enhance the reliability of preclinical data and contribute to the improvement of the design and outcomes of future clinical trials.
Gene expression is tightly controlled by DNA elements called enhancers by associating with lineage-specific transcription factors. These enhancers transcribe noncoding RNAs (called enhancer RNAs or eRNAs). eRNA expressio...Gene expression is tightly controlled by DNA elements called enhancers by associating with lineage-specific transcription factors. These enhancers transcribe noncoding RNAs (called enhancer RNAs or eRNAs). eRNA expression is an early indicator of transcription factor activity and is associated with treatment response and survival in cancer patients. By analysing ~ 300 000 eRNA loci profiled using RNA-sequencing data sets from 975 breast cancer patients using machine learning approaches, we categorised eRNAs specific to breast cancer molecular subtypes and survival. We associated these eRNAs with subtype-specific mRNAs to define proximal co-expressed regulatory eRNAs (ProxCReAm), which are enriched in pathways characteristic of their respective subtypes. Interestingly, cistrome and transcription factor motif analyses on these eRNAs highlighted involvement of diverse nuclear receptors (GR/AHR for luminal and GR/RAR for basal) and FOX factors in luminal regions. Moreover, luminal eRNAs were associated with better outcomes and Her2 eRNAs with worse outcomes in patients. Overall, we demonstrate that machine learning approaches performed on RNA-seq data sets can classify functionally relevant subtype-specific and prognostic eRNAs, which can identify critical gene pathways and transcription factor networks in breast cancer.
Transcriptional heterogeneity in pancreatic ductal adenocarcinoma (PDAC) arises not only from changes in gene expression but also from dynamic rewiring of gene-gene coordination. Using a divergent-edge framework applied...Transcriptional heterogeneity in pancreatic ductal adenocarcinoma (PDAC) arises not only from changes in gene expression but also from dynamic rewiring of gene-gene coordination. Using a divergent-edge framework applied to 77 155 malignant cells from 42 tumors, we identified four reproducible adaptive modules-integrated growth-energy (IGE), stress-adaptive transcription (SAT), IL-2-linked immune evasion (IL2), and multi-pathway collective invasion (MPC)-that cut across canonical PDAC states and reflect distinct regulatory programs. Integrating these modules with CRISPR-Cas9 dependency profiles and PRISM drug-response data revealed that adaptive behaviors collapse into two higher-order axes: a biosynthetic-metabolic IGE axis enriched for translational and DNA-repair dependencies, and a broader SAT-IL2-MPC stress-immune-invasion axis characterized by proteostasis, cytokine-linked, and cytoskeletal vulnerabilities. This architecture emerges only when divergent-edge modules are mapped into functional genomics space. Module activity also carried clinical relevance in PDAC. SAT-high tumors showed poorer survival, while MPC-high tumors exhibited a similar adverse trend; together, these modules defined a stress-immune-invasion poor-prognosis axis. In contrast, IGE activity showed no overall risk association, although an optimal-cut point-defined IGE-high subgroup displayed modestly improved survival.
Medulloblastoma (MB) is a brain tumor for which current treatments cause serious side effects and are not curative for all patients, highlighting the need for more effective and brain-protecting therapies. Recently, we c...Medulloblastoma (MB) is a brain tumor for which current treatments cause serious side effects and are not curative for all patients, highlighting the need for more effective and brain-protecting therapies. Recently, we combined phosphoinositide 3-kinase (PI3K) inhibitor (BYL719), fibroblast growth factor receptor (FGFR) inhibitor (JNJ-42756493) and cyclin-dependent kinase (CDK)4/6 inhibitor (PD-0332991) in MB cell lines, and discovered synergistic effects. In the current study, we investigate the most efficient therapies in a normal/tumorigenic neural stem cell model. A sonic hedgehog (SHH)-MB model, including a Gorlin syndrome patient neuroepithelial stem cell line (NES) and its tumor derivative (tNES), was used to evaluate single and combined treatments of PI3K, AKT, FGFR, and CDK4/6 inhibitors (BYL719, AZD5363, JNJ-42756493, and PD-0332991, respectively). Effects on viability, cell confluence and apoptosis were tested on NES and tNES cells cultured as 2D monolayers and 3D spheroids. We found that 2D tNES cells were generally more sensitive to the inhibitory effects of both single and combination treatments compared to 2D NES cells. In the 3D setting, all single drugs were more effective against tNES than NES, except for JNJ-42756493, which showed the opposite trend. Drug combinations in 3D cultures generally resulted in synergistic or additive effects on cell viability in NES and tNES. This study illustrates that single and combined administrations of PI3K, FGFR, CDK4/6, and AKT inhibitors in a NES/tNES model have dose-dependent and additive/synergistic anti-MB activity impacting tumor growth. Their effects on tNES cells were generally more pronounced than on NES; however, the difference in proliferative capacity between the cells should be considered.
To explore the immune microenvironment of RAS-mutated (RASmt) microsatellite stable (MSS) colon cancer (CC), we retrospectively performed whole exome sequencing, RNA sequencing, and robust digital pathology analyses and...To explore the immune microenvironment of RAS-mutated (RASmt) microsatellite stable (MSS) colon cancer (CC), we retrospectively performed whole exome sequencing, RNA sequencing, and robust digital pathology analyses and studied immune markers in a cohort of 161 patients treated with standard-of-care therapies with early stage disease (both fresh frozen and formalin-fixed paraffin-embedded [FFPE] samples) or 121 patients with metastatic setting (primary tumor FFPE samples). Only a small proportion of cases exhibited a highly infiltrated immune microenvironment, with a strong association between Immunoscore (IS)-high (13% of the samples) and Tumor Lymphocytes Infiltrating Score (TuLIS)-high scores (25% of the samples). Immunoscore Immune-Checkpoint (ISIC)-high tumors (52% of the samples) shared a similar microenvironment composition to IS-high and TuLIS-like high tumors and displayed higher mutational burdens than ISIC-low tumors. In conclusion, a substantial proportion of MSS RASmt CCs exhibit high ISIC scores, meriting evaluation in prospective trials of immunotherapy-based combination regimens.
Liquid biopsies enable noninvasive tumor profiling and longitudinal disease monitoring. Their analytical performance is strongly influenced by pre-analytical factors, yet direct comparisons between cell-free DNA (cfDNA)...Liquid biopsies enable noninvasive tumor profiling and longitudinal disease monitoring. Their analytical performance is strongly influenced by pre-analytical factors, yet direct comparisons between cell-free DNA (cfDNA) and extracellular vesicle-derived DNA (evDNA) remain scarce. We prospectively evaluated four pre-analytical variables: processing delay, storage temperature, tube type, and plasma input volume, on cfDNA and evDNA from cancer patient plasma (n = 244) using ddPCR, Qubit, and TapeStation. Key findings were validated in archived plasma samples (n = 723). In the prospective cohort, cfDNA concentrations increased after 24 h and evDNA after 48 h at room temperature, while retrospective analysis revealed earlier changes (cfDNA: 6 h; evDNA: 24 h). Storage conditions influenced both analytes, as short-term refrigeration (4 °C) better preserved DNA quality than -80 °C freezing, while extracted DNA remained stable at -80 °C. Acid citrate dextrose (ACD) and KEDTA tubes performed similarly under prompt processing. Higher plasma volumes improved evDNA, but not cfDNA, for mutation detection. evDNA demonstrates greater resilience than cfDNA under suboptimal conditions. Standardized workflows and prompt processing are essential to ensure reliable mutation detection in clinical liquid biopsy applications.
Bielesch S, Vogel I, Nokodian S
… +11 more, Moeller J, Blechschmidt A, Hecht V, Kuentzel V, Thiedig K, Schwab M, Schilling O, Bronger H, Kiechle M, Magdolen V, Dreyer T
High-grade serous ovarian cancer (HGSOC) is the most prevalent and lethal subtype of epithelial ovarian cancer (EOC), characterised by extensive peritoneal metastasis. The intermediate filament keratin 19 (KRT19) has bee...High-grade serous ovarian cancer (HGSOC) is the most prevalent and lethal subtype of epithelial ovarian cancer (EOC), characterised by extensive peritoneal metastasis. The intermediate filament keratin 19 (KRT19) has been linked to tumour progression and chemoresistance in various cancers. However, its role varies across tumour types and remains unclear for HGSOC. We evaluated KRT19 protein expression in 199 HGSOC patients and correlated findings with clinical outcomes. In vitro, we assessed the effects of KRT19 on tumour-associated mechanisms, including proliferation, migration, adhesion, and spheroid formation. A xenograft mouse model was used to assess tumour burden in vivo. Publicly available datasets enabled in silico validation. KRT19 was significantly overexpressed in HGSOC, and high expression was associated with reduced overall survival. In vivo, KRT19-overexpression increased peritoneal tumour burden. In vitro and ex vivo, KRT19 induced a hybrid epithelial phenotype through enhanced epithelial-mesenchymal plasticity (EMP), promoting adhesion, migration, and spheroid integrity, thereby potentially supporting metastatic processes. Further, KRT19 could contribute to paclitaxel resistance. Altogether, KRT19 represents a potential independent prognostic marker and therapeutic target to inhibit metastatic dissemination.
The nucleoside analogue cytarabine (ara-C) is part of standard treatment against acute myeloid leukaemia (AML). The efficacy of this therapy is dependent upon accumulation of the active triphosphate metabolite ara-CTP, w...The nucleoside analogue cytarabine (ara-C) is part of standard treatment against acute myeloid leukaemia (AML). The efficacy of this therapy is dependent upon accumulation of the active triphosphate metabolite ara-CTP, which mis-incorporates into genomic DNA, triggering cell death. The deoxyribonucleoside triphosphate triphosphohydrolase (dNTPase) SAMHD1 can hydrolyse ara-CTP and thereby convert the active metabolite back to its inactive prodrug form. This constitutes a barrier to treatment efficacy and thus strategies to target SAMHD1 are warranted. SAMHD1 activity is allosterically regulated by nucleotides, which are synthesised in cells via distinct pathways. We screened a collection of drugs targeting nucleotide biosynthetic enzymes and identified that inhibition of inosine-5'-monophosphate dehydrogenase (IMPDH), responsible for catalysing the rate-limiting step in guanine nucleotide biosynthesis, sensitises AML cell lines to ara-C in a SAMHD1-dependent manner. We show that approved drugs inhibiting IMPDH-mycophenolic acid and ribavirin-imbalance deoxyribonucleoside triphosphate pools and increase ara-C efficacy in SAMHD1-proficient, but not deficient, leukaemic cells. Altogether, we provide insight into SAMHD1 regulation in leukaemic cells and show how this process can be exploited by approved drugs to improve ara-C therapy.
Myelodysplastic neoplasms (MDS) are heterogeneous malignancies originating in hematopoietic stem cells. In this explorative study, we carried out ultra-deep total RNA sequencing on FACS-sorted CD34+ bone marrow cells fro...Myelodysplastic neoplasms (MDS) are heterogeneous malignancies originating in hematopoietic stem cells. In this explorative study, we carried out ultra-deep total RNA sequencing on FACS-sorted CD34+ bone marrow cells from 71 patients and eight healthy age-matched controls. We investigated the expression of circular RNAs (circRNAs), a group of noncoding RNAs produced by back-splicing of nonadjacent splice sites. Key findings were further explored in an independent cohort of 118 patients with MDS and ring sideroblasts. circRNA abundance was higher in the disease groups than in controls, and different spliceosome mutations were associated with distinct circRNA expression patterns. Expression of the proliferation-related gene MKI67 was negatively correlated with circRNA abundance. High circRNA abundance was associated with a significantly increased risk of disease progression at 3 years. The majority of the 38 circRNAs that were significantly upregulated in MDS demonstrated highly correlated expression, and many were associated with risk of leukemic progression. Furthermore, we confirmed the specificity of circZEB1 expression to cases with SF3B1 mutations. We conclude that aberrant circRNA expression is found in MDS and displays associations with disease characteristics and patient outcomes.
Pancreatic ductal adenocarcinoma (PDAC) has poor prognosis as early-stage asymptomaticity leads to late-stage diagnoses. Strategies to detect PDAC earlier or identify high-risk individuals are therefore paramount. Here,...Pancreatic ductal adenocarcinoma (PDAC) has poor prognosis as early-stage asymptomaticity leads to late-stage diagnoses. Strategies to detect PDAC earlier or identify high-risk individuals are therefore paramount. Here, we report results from genetically engineered mice and PDAC patients that identify serum proteins associated with pancreatic intraepithelial neoplasms (PanINs), the most common PDAC precursor, and early-stage PDAC. Initially, we screened previously described PanIN-abundant mice, harbouring pancreatic and duodenal homeobox 1 (Pdx1)-Cre, Lox-STOP-Lox-Kras and floxed alleles of essential autophagy genes autophagy-related 7 (Atg7) or autophagy-related 5 (Atg5). Sera from these mice were assessed by proteomics and hits were compared to those in Lox-STOP-Lox-Kras Lox-STOP-Lox-Trp53 Pdx1-Cre (KPC) mice, which closely recapitulate human disease, and early-stage (I-II) PDAC patients. Levels of inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) were significantly elevated in all three screens, with complement C5, complement factors B and H (CFB/CFH), and monocyte differentiation antigen CD14 increased in KPC mice and PDAC patients; and all were significantly increased co-ordinately in PDAC according to disease stage. Serum levels of C5, CFH and CD14 together constitute a novel panel for identifying PanINs and early-stage PDAC with confidence, and when combined with additional screening, could help increase survival from this dismal disease.
Epithelial-to-mesenchymal transition (EMT), driven by cues from the tumor microenvironment, is a critical initiator of metastatic progression. In breast cancer patients, elevated expression of cartilage oligomeric matrix...Epithelial-to-mesenchymal transition (EMT), driven by cues from the tumor microenvironment, is a critical initiator of metastatic progression. In breast cancer patients, elevated expression of cartilage oligomeric matrix protein (COMP) is associated with shorter survival and increased metastatic risk. Here, we investigated the role of COMP in regulating EMT in breast cancer. Breast cancer cells treated with recombinant COMP or engineered to overexpress COMP exhibited a marked decrease in the epithelial marker CDH1 and an increase in mesenchymal markers such as VIM and VCAN. Consistent with these in vitro findings, COMP-expressing xenograft tumor tissues showed enhanced EMT characteristics. Functionally, COMP promoted increased migration and invasion of breast cancer cells in both autocrine and paracrine manners, dependent on its thrombospondin (TSP) and C-terminal domains. We further identified protein TMEPAI (encoded by gene PMEPA1) as a functional COMP-binding partner that mediates COMP-induced EMT, primarily through interaction with the TSP domain of COMP. Mechanistically, COMP shifted transforming growth factor beta (TGFβ) signaling from canonical phosphorylated mothers against decapentaplegic homolog 2/3 (pSMAD2/3) activation toward pSMAD1/5, likely through its interaction with PMEPA1. This study suggests the COMP-PMEPA1 axis as a new driver of EMT in breast cancer models.
Recurrent ovarian cancer (OC) remains a major cause of mortality due to chemoresistance and metastasis. Epigenetic aberrations, particularly dysregulated microRNA (miRNA) expression, contribute to disease progression and...Recurrent ovarian cancer (OC) remains a major cause of mortality due to chemoresistance and metastasis. Epigenetic aberrations, particularly dysregulated microRNA (miRNA) expression, contribute to disease progression and represent a promising therapeutic target. Here, we identify the miR-214-3p/miR-199a-5p cluster as a stage-associated, tumor-suppressive network that is lost in recurrent and chemoresistant OC but can be elevated using engineered small extracellular vesicles enriched with this miRNA cluster (m214-sEVs). Using a clinically relevant mouse model that recapitulates spontaneous OC relapse following platinum-based chemotherapy, we show that m214-sEVs are internalized by OC cells and niche fibroblasts, leading to increased intracellular levels of this cluster and suppression of key chemoresistance-associated pathways, including through downregulation of Toll-like receptor 4 (TLR4), β-catenin, and the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein YKT6. m214-sEV treatment reprograms secondary tumor-derived sEVs toward a less prometastatic cargo profile and decreases carboplatin resistance and cell migration. Enforced YKT6 overexpression abrogates these effects, establishing YKT6 as a key downstream effector. Collectively, these findings support engineered sEVs as a translatable strategy to overcome chemoresistance and disrupt pro-tumorigenic EV signaling in recurrent OC.
Receptor-interacting protein kinase 4 (RIPK4) has been implicated in the progression of numerous tumours. In nonmelanoma skin cancer, RIPK4 plays a suppressor role, whereas in melanoma, it functions as an oncogene that m...Receptor-interacting protein kinase 4 (RIPK4) has been implicated in the progression of numerous tumours. In nonmelanoma skin cancer, RIPK4 plays a suppressor role, whereas in melanoma, it functions as an oncogene that modulates key signalling pathways involved in melanoma cell survival and expansion. Increased RIPK4 levels in metastatic melanoma biopsies prompted us to investigate the consequences of RIPK4 loss for the invasive and metastatic phenotype of melanoma cells. Using an integrated approach involving clinical samples, in vivo xenograft models, transcriptomic analysis and 3D functional assays, we show that RIPK4 deletion significantly reduces pulmonary metastasis formation. This reflects its role in late-stage metastatic events, such as extravasation and colonization, particularly since this phenotype correlates with extensive transcriptional reprogramming of adhesion- and motility-related genes in melanoma cells, as evidenced by next-generation sequencing and functional validation in spheroid and collagen-based models. Despite exhibiting features of a partial shift towards an amoeboid phenotype such as membrane blebbing and increased MLC2 phosphorylation, RIPK4 knockout cells display impaired motility and invasion. Re-expression of RIPK4 restores mesenchymal morphology and migratory capacity. Together, our results establish RIPK4 as a critical regulator of melanoma invasion and metastasis. Nonetheless, they also demonstrate that the loss of RIPK4 function activates compensatory phenotypic shifts in melanoma cells that fail to fully rescue their invasive potential.
Targeted drug therapy is very important for the treatment of triple-negative breast cancer (TNBC), and the development of carrier systems to deliver apoptosis-inducing proteins such as TRAIL to cells is important in canc...Targeted drug therapy is very important for the treatment of triple-negative breast cancer (TNBC), and the development of carrier systems to deliver apoptosis-inducing proteins such as TRAIL to cells is important in cancer therapy. In this study, a nanosystem formulation (TRAIL-PEG-Apt-PLGA) encapsulating TNBC-targeted aptamer-bound-TRAIL protein was performed and the efficacy of this system was evaluated in a mouse tumor model. The characterization of TRAIL-PEG-Apt-PLGA was confirmed by FTIR, NTA and SEM microscopy. The efficacy of TRAIL-PEG-Apt-PLGA was evaluated by in vitro release assays and interactions with TNBC cells (MDA-MB-231) and healthy breast cells (MCF-10A). TRAIL-PEG-Apt-PLGA was administered intravenously to NOD/SCID gamma mouse breast tumors and evaluated in vivo. Pharmacokinetics, bioavailability testing, histological staining (DR4/DR5, TUNEL, HE staining) and molecular alterations with PCR array were evaluated in tumor tissues. TRAIL-PEG-Apt-PLGA induced apoptosis in both in vivo and in vitro studies. It was found that it regulated cellular responses along with apoptotic mechanisms in cells without developing resistance in suppressing tumor growth by making changes on Atf2, Casp8, Bcl2 and Irf5 genes and proteins. As a result, the biotechnological drug potential of TRAIL was discovered in an aptamer-bound nanosystem for the treatment of triple-negative breast cancer and innovative applications for clinical use.
Poturnajova M, Kozovska Z, Pos O
… +23 more, Pavlov K, Gulati S, Makovicky P, Jakic K, Burikova M, Sedlackova E, Svitkova B, Tyciakova S, Bystry V, Blavet N, Tichy B, Hrnciar M, Budis J, Tomas M, Dubovan P, Kolnikova G, Repaska V, Mojzesova N, Zomborska E, Pindak D, Mego M, Szemes T, Matuskova M
Colorectal cancer ranks third in global incidence and second in cancer mortality. Patient-derived models are irreplaceable for studying tumor biology. We established a human epithelial cell line from a rectal adenocarcin...Colorectal cancer ranks third in global incidence and second in cancer mortality. Patient-derived models are irreplaceable for studying tumor biology. We established a human epithelial cell line from a rectal adenocarcinoma overexpressing cancer stem cell marker ALDH1A1, and we investigated the effect of ALDH1A1 knockout on tumor cell traits. The cell line and its CRISPR-Cas9 ALDH1A1 knockouts were characterized by genomic and cytogenetic methods (CNV, WES, RNAseq, karyotype), in vitro (proliferation, response to chemotherapy, migration, invasion, apoptosis), and in vivo methods. We identified the landscape of somatic mutations and copy number alterations in the original tumor and the derived cell line. Genetic attenuation of ALDH1A1 was characterized by an increase in migratory potential and extensive metastatic ability, accompanied by reduced growth of subcutaneous xenografts and alterations in gene expression associated with inhibited proliferation and promoted invasion and metastasis, ultimately resulting in dysregulation of the Wnt signaling pathway. Increased metastatic potential was also confirmed in HT-29 cells after ALDH1A1 genetic attenuation. CRISPR-Cas9-mediated editing led to functional, cellular, and molecular changes confirming the role of ALDH1A1 in colorectal cancer carcinogenesis.
Despite having excellent prognosis when detected early, colorectal cancer (CRC) remains a leading cause of cancer-related deaths globally. Screening remains an important contributor to CRC survival, but the cost and inva...Despite having excellent prognosis when detected early, colorectal cancer (CRC) remains a leading cause of cancer-related deaths globally. Screening remains an important contributor to CRC survival, but the cost and invasiveness of traditional imaging methodologies can hinder patient compliance. A blood-based approach would be more convenient and accessible, but reliable serum markers are lacking. In this study, the peptide enteroendocrine hormone Fibroblast Growth Factor 19 (FGF19) was identified as an attractive marker for CRC through a meta-transcriptomic analysis. While its pro-tumor effects are documented, FGF19's utility as a blood serum marker for CRC is not well defined. Studies presented here show that FGF19 is constitutively expressed and secreted in 3 of 5 CRC cell lines, and secreted levels increase with seeding density. A subcutaneous CRC cell line-derived xenograft model revealed that FGF19 is detectable in serum of mice injected with FGF19-positive, but not negative, CRC cells at levels corresponding to tumor volume. Enteroendocrine effects of tumor-derived FGF19, including suppression of bile acid synthesis, are evident in liver samples via RNA sequencing and validated by RT-PCR. Notably, the hepatic response to CRC-secreted FGF19 has not been explored prior to this study even though FGF19 is a key regulator of hepatic cholesterol metabolism and bile acid homeostasis. Collectively, these findings support the clinical utility of FGF19 as a putative serum marker for CRC and provide important evidence that CRC-derived FGF19 can modulate liver physiology consistent with the enteroendocrine function of FGF19.
Cervical cancer remains a leading cause of mortality among women, particularly in low- and middle-income countries. Despite distinct prognoses and clinical outcomes between its main histological subtypes, squamous cell c...Cervical cancer remains a leading cause of mortality among women, particularly in low- and middle-income countries. Despite distinct prognoses and clinical outcomes between its main histological subtypes, squamous cell carcinoma (SCC) and adenocarcinoma (ADC), current treatment regimens remain largely similar, creating an urgent need for targeted therapeutic strategies based on molecular distinctions. To address this gap, the long noncoding RNA IGFL2-AS1 was investigated as a potential prognostic biomarker and therapeutic target in cervical adenocarcinoma. A translational approach was employed that integrated patient transcriptome data, in silico analysis from public databases, and in vitro validation. Using CRISPR/dCas9 technology, IGFL2-AS1 expression was modulated in HeLa (ADC) and SiHa (SCC) cell lines to assess its impact on cellular characteristics associated with tumorigenesis. In silico analysis revealed that IGFL2-AS1 expression was significantly reduced in ADC compared to SCC, and its low expression was consistently linked to poorer ADC prognosis and decreased patient survival. Notably, overexpression of IGFL2-AS1 in HeLa cells significantly reduced cell proliferation, migration, clonogenic survival, and enhanced sensitivity to cisplatin and doxorubicin. Conversely, IGFL2-AS1 repression in SiHa cells yielded no significant phenotypic changes, suggesting a context-dependent mechanism. IGFL2-AS1 is identified as a histological subtype-specific prognostic biomarker and promising therapeutic target for cervical adenocarcinoma.
Peroxidasin (PXDN), an extracellular matrix (ECM)-associated peroxidase, has been implicated in cancer progression. However, its roles in melanoma biology and therapeutic sensitivity remain unclear. Here, we demonstrate...Peroxidasin (PXDN), an extracellular matrix (ECM)-associated peroxidase, has been implicated in cancer progression. However, its roles in melanoma biology and therapeutic sensitivity remain unclear. Here, we demonstrate that elevated PXDN expression is associated with poor prognosis and reduced survival in melanoma patients. Functional studies revealed that PXDN depletion impairs melanoma cell proliferation, disrupts the cell cycle, and reduces melanoma cell invasive capacities. Furthermore, we found that secreted PXDN modulates anti-melanoma immunity by enhancing melanoma resistance to natural killer (NK)-cell-mediated cytotoxicity. Structural modeling identified a trimeric organization of PXDN, stabilized by disulfide-linked peroxidase domains. Molecular dynamics simulations identified a previously unknown inhibitory interaction between the PXDN N-terminal leucine-rich repeat domain and the NK cell-activating receptor NKG2-D type II integral membrane protein (NKG2D). These findings uncover a redox-independent role for PXDN in promoting immune evasion and tumor progression. Overall, our study highlights PXDN as a critical regulator of melanoma cell biology and a potential therapeutic target for NK-cell-based immunotherapy in melanoma and other solid cancers.
According to the seed and soil hypothesis, the organ specificity of metastasis is not a random process and depends on multiple tumor-intrinsic and microenvironmental factors. In this study, we characterized the mutationa...According to the seed and soil hypothesis, the organ specificity of metastasis is not a random process and depends on multiple tumor-intrinsic and microenvironmental factors. In this study, we characterized the mutational landscape of a large cohort of human metastatic samples to investigate whether mutational trends determine metastatic dissemination. Genomic data from nine cancer types (bladder, breast, colorectal, endometrial, melanoma, non-small cell lung cancer, ovarian, pancreatic, and prostate) including 19827 patients were obtained from a pan-cancer study. When restricting the analysis to driver mutations, no robust, recurrent mutational patterns associated with metastatic locations were identified across cancer types. However, when cancer types were analyzed separately, mutational trends associated with specific metastatic locations emerged. Considering the total tumor mutational burden (TMB), central nervous system (CNS)/brain and lung metastases harbored a higher TMB than other metastatic locations. Since higher TMB in CNS/brain metastases was also associated with improved prognosis, these findings may be pivotal in refining immunotherapy strategies. Indeed, this observation was confirmed in an independent dataset including patients treated with immunotherapy. In conclusion, our findings suggest that TMB may have greater influence on metastatic organotropism than driver mutational background.
Cervical cancer is highly prevalent in India, with most cases being diagnosed at advanced stages. Despite the standard concurrent chemoradiotherapy (CCRT), 30-40% of patients' experience treatment failure, underscoring t...Cervical cancer is highly prevalent in India, with most cases being diagnosed at advanced stages. Despite the standard concurrent chemoradiotherapy (CCRT), 30-40% of patients' experience treatment failure, underscoring the need for improved therapeutic strategies. Understanding resistance mechanisms and identifying predictive biomarkers are crucial to improve treatment efficacy and enable personalized medicine. We conducted a comprehensive genomic and proteomic analysis to identify molecular signatures associated with CCRT. We identified recurrent mutations in phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform (PIK3CA) and histone-lysine N-methyltransferase 2D (KMT2D), with mutation signature analysis revealing a prevalent DNA dC- > dU-editing enzyme, APOBEC mutagenesis signature. Distinct genomic alterations, including epidermal growth factor receptor (EGFR) amplification and serine/threonine kinase 11 (STK11) deletion, were exclusively observed in the chemoradiation-resistant cohort. Proteomic analysis identified 73 significantly dysregulated proteins, with syntaxin-3 (STX3), SERPINB7, lipopolysaccharide-binding protein (LBP), EMILIN2, and ribosyldihydronicotinamide dehydrogenase (quinone) (NQO2) being the top five upregulated proteins. Integrative pathway analysis highlighted an active DNA repair pathway in the resistant cohort. This study presents the first proteogenomic profiling of cervical cancer in the Indian population, linking molecular alterations to CCRT response. STK11 and STX3 emerged as predictive biomarkers for poor response, whereas EGFR presents as a promising therapeutic target in the resistant group.