Immune checkpoint inhibitors, such as pembrolizumab, are revolutionizing therapeutic strategies for different cancer types, including non‐small‐cell lung cancer (NSCLC). However, only a subset of ...patients benefits from this therapy, and new biomarkers are needed to select better candidates. In this study, we explored the value of liquid biopsy analyses, including circulating free DNA (cfDNA) and circulating tumour cells (CTCs), as a prognostic or predictive tool to guide pembrolizumab therapy. For this purpose, a total of 109 blood samples were collected from 50 patients with advanced NSCLC prior to treatment onset and at 6 and 12 weeks after the initiation of pembrolizumab. Plasma cfDNA was measured using hTERT quantitative PCR assay. The CTC levels at baseline were also analysed using two enrichment technologies (CellSearch® and Parsortix systems) to evaluate the efficacy of both approaches at detecting the presence of programmed cell death ligand 1 on CTCs. Notably, patients with high baseline hTERT cfDNA levels had significantly shorter progression‐free survival (PFS) and overall survival (OS) than those with low baseline levels. Moreover, patients with unfavourable changes in the hTERT cfDNA levels from baseline to 12 weeks showed a higher risk of disease progression. Additionally, patients in whom CTCs were detected using the CellSearch® system had significantly shorter PFS and OS than patients who had no CTCs. Finally, multivariate regression analyses confirmed the value of the combination of CTCs and cfDNA levels as an early independent predictor of disease progression, identifying a subgroup of patients who were negative for CTCs, who presented low levels of cfDNA and who particularly benefited from the treatment.
In the present work, we demonstrated that liquid biopsy, through the combinatory analyses of circulating free DNA (cfDNA) and circulating tumour cells and the monitoring of cfDNA during the treatment, could help to select advanced non‐small‐cell lung cancer patients who will benefit from pembrolizumab treatment as first line, providing a promising non‐invasive strategy for improving the clinical management of this tumour.
The lysyl oxidase-like protein LOXL2 has been suggested to contribute to tumor progression and metastasis, but
evidence has been lacking. Here we provide functional evidence that LOXL2 is a key ...driver of breast cancer metastasis in two conditional transgenic mouse models of PyMT-induced breast cancer. LOXL2 ablation in mammary tumor cells dramatically decreased lung metastasis, whereas LOXL2 overexpression promoted metastatic tumor growth. LOXL2 depletion or overexpression in tumor cells does not affect extracellular matrix stiffness or organization in primary and metastatic tumors, implying a function for LOXL2 independent of its conventional role in extracellular matrix remodeling. In support of this likelihood, cellular and molecular analyses revealed an association of LOXL2 action with elevated levels of the EMT regulatory transcription factor Snail1 and expression of several cytokines that promote premetastatic niche formation. Taken together, our findings established a pathophysiologic role and new function for LOXL2 in breast cancer metastasis.
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Background
Differences in cell‐free DNA (cfDNA) fragments have been described as a valuable tool to distinguish cancer patients from healthy individuals. We aim to investigate the concentration and ...integrity of cfDNA fragments in saliva from oral squamous cell carcinoma (OSCC) patients and healthy individuals in order to explore their value as diagnostic biomarkers.
Methods
Saliva samples were collected from a total of 34 subjects (19 OSCC patients and 15 healthy controls). The total concentration of salivary cfDNA (scfDNA) was determined using a fluorometry method and quantitative real‐time polymerase chain reaction (qPCR). To evaluate the scfDNA quantity and integrity, qPCR targeting Arthobacter luteus (ALU) sequences at three amplicons of different lengths (60, 115, and 247 bp, respectively) was carried out. ScfDNA integrity indexes (ALU115/ALU60 and ALU247/ALU60) were calculated as the ratio between the absolute concentration of the longer amplicons 115 bp and 247 bp and the total scfDNA amount (amplicon 60 bp).
Results
The total scfDNA concentration (ALU60) was higher in OSCC than in healthy donors, but this trend was not statistically significant. The medians of scfDNA integrity indexes, ALU115/ALU60 and ALU247/ALU60, were significantly higher in OSCC, showing area under the curve values of 0.8211 and 0.7018, respectively.
Conclusion
Our preliminary results suggest that scfDNA integrity indexes (ALU115/ALU60 and ALU247/ALU60) have potential as noninvasive diagnostic biomarkers for OSCC.
In the present study we investigated the prognostic value of Circulating Tumour Cells (CTC) and their utility for therapy monitoring in non-small cell lung cancer (NSCLC). A total of 43 patients ...newly diagnosed with NSCLC were prospectively enrolled. Blood samples were obtained before the 1st, 2nd and 5th cycles of chemotherapy and analyzed using CellSearch technology. Both CTC and CTC-related objects (not morphological standard or broken epithelial cells) were counted. At baseline 18 (41.9%) patients were positive for intact CTC count and 10 (23.2%) of them had ≥5 CTC, while CK positive events were found in 79.1% of patients. The group of patients with CTC ³5 at baseline presented worse PFS and OS than those with <5 CTC (p = 0.034 and p = 0.008, respectively). Additionally, high levels of total CK positive events were associated with poor prognosis in the group of patients with <5 CTC. Regarding therapy monitoring, patients presenting increased levels of CTC during the treatment demonstrated lower OS and PFS rates. All these data supported the value of CTC as a prognostic biomarker and as a surrogate indicator of chemotherapy effectiveness in advanced NSCLC patients, with the additional value of analyzing other "objects" such as apoptotic CTC or CK fragments to guide the clinical management of these patients.
Objectives
To provide a comprehensive characterization of DNA methylome of oral tongue squamous cell carcinoma (OTSCC) and identify novel tumor‐specific DNA methylation markers for early detection ...using saliva.
Material and Methods
Genome‐wide DNA methylation analysis including six OTSCC matched adjacent non‐tumoral tissue and saliva was performed using Infinium MethylationEPIC array. Differentially methylated levels of selected genes in our OTSCC cohort were further validated using OTSCC methylation data from The Cancer Genome Atlas database (TCGA). The methylation levels of a set of tumor‐specific hypermethylated genes associated with a downregulated expression were evaluated in saliva. Receiver operating characteristic (ROC) curves were performed to assess the diagnostic value of DNA methylation markers.
Results
A total of 25,890 CpGs (20,505 hypomethylated and 5385 hypermethylated) were differentially methylated (DMCpGs) between OTSCC and adjacent non‐tumoral tissue. Hypermethylation of 11 tumor‐specific genes was validated in OTSCC TCGA cohort. Of these 11 genes, A2BP1, ANK1, ALDH1A2, GFRA1, TTYH1, and PDE4B were also hypermethylated in saliva. These six salivary methylated genes showed high diagnostic accuracy (≥0.800) for discriminating patients from controls.
Conclusions
This is the first largest genome‐wide DNA methylation study on OTSCC that identifies a group of novel tumor‐specific DNA methylation markers with diagnostic potential in saliva.
Triple‐negative breast cancer (TNBC) is characterized by high rates of metastasis and no available molecular targets. CTCs derived xenografts (CDX) have demonstrated to be a promising tool for ...understanding cancer biology. In our study, a CDX from a TNBC patient was developed for the first time. After CDX characterization, WNT signaling was found as the main mechanism related with this tumor biology and potential CTCs markers were identified and subsequently validated in TNBC patients. In this cohort high levels of MELK expression were associated with poorer survival rates. Overall, our study demonstrates that CTCs from TNBC are tumorigenic and CDXs are a useful model to obtain valuable information about the tumor.
What's new?
Circulating tumor cells (CTCs) hold promise as a liquid biopsy and model to study tumor biology. However, technical limitations toward CTCs molecular characterisation and ex vivo expansion remain. This work describes for the first time the development of a CTCs‐derived xenograft (CDX) from a metastatic Triple Negative Breast Cancer (TNBC) patient, with WNT signalling emerging as the main tumor mechanism. Potential prognostic markers and therapeutic targets were further identified in CTCs from TNBC patients. This study demonstrates that CTCs from TNBC are tumorigenic and highlights CDXs as a unique tool to better understand TNBC biology and develop novel therapeutic approaches.
The analysis of mismatch repair proteins in solid tissue is the standard of care (SoC) for the microsatellite instability (MSI) characterization in endometrial cancer (EC). Uterine aspirates (UAs) or ...circulating‐DNA (cfDNA) samples capture the intratumor heterogeneity and provide a more comprehensive and dynamic molecular diagnosis. Thus, MSI analysis by droplet‐digital PCR (ddPCR) in UAs and cfDNA can provide a reliable tool to characterize and follow‐up the disease. The UAs, paraffin‐embedded tumor tissue (FFPE) and longitudinal plasma samples from a cohort of 90 EC patients were analyzed using ddPCR panel and compared to the SoC. A high concordance (96.67%) was obtained between the analysis of MSI markers in UAs and the SoC. Three discordant cases were validated as unstable by ddPCR on FFPE samples. Besides, a good overall concordance (70.27%) was obtained when comparing the performance of the ddPCR assay on UAs and cfDNA in high‐risk tumors. Importantly, our results also evidenced the value of MSI analysis to monitor the disease evolution. MSI evaluation in minimally invasive samples shows great accuracy and sensitivity and provides a valuable tool for the molecular characterization and follow‐up of endometrial tumors, opening new opportunities for personalized management of EC.
What's new?
The analysis of mismatch repair proteins in solid tumor tissue is the current standard of care for microsatellite instability (MSI) characterization in endometrial cancer. Our study demonstrates the accuracy and advantages of MSI analysis by droplet‐digital PCR in uterine aspirates and plasma samples from endometrial cancer patients. The new approach allows comprehensive tumor genotyping without the need for invasive procedures and improves the rate of MSI detection. The assay also has the capacity of monitoring tumor evolution in both localized and metastatic disease by assessing MSI markers. The approach could be easily implemented into the clinic as a follow‐up tool.
Circulating tumor cells (CTCs), proposed as major players in cancer dissemination, have demonstrated clinical prognostic significance in several cancer types. However, their predictive value remains ...unclear. Here we evaluated the clinical utility of six CTC markers (tissue specific and epithelial to mesenchymal transition transcripts) both as prognostic and predictive tools in metastatic colorectal cancer (mCRC) patients. CTCs were immunoisolated from blood in 50 mCRC patients at baseline and at 4 and 16 weeks after treatment onset. Expression levels of GAPDH, VIL1, CLU, TIMP1, LOXL3 and ZEB2 were determined by qualitative polymerase chain reaction and normalized to the unspecific cell isolation marker CD45. At baseline, median progression‐free survival (PFS) and overall survival (OS) for patients with high CTC markers were 6.3 and 12.7 months, respectively, versus 12.7 and 24.2 for patients with low CTC markers (PFS; p = 0.0003; OS; p = 0.044). Concerning response to therapy, PFS and OS for patients with increased CTC markers along treatment were, respectively, 6.6 and 13.1 months, compared with 12.7 and 24.3 for patients presenting CTC markers reduction (PFS; p = 0.004; OS; p = 0.007). Of note, CTC markers identified therapy‐refractory patients not detected by standard image techniques. Patients with increased CTC markers along treatment, but classified as responders by computed tomography, showed significantly shorter survival times (PFS: 7.8 vs. 13.2; OS: 14.4 vs. 24.4; months). In conclusion, we have generated a CTC marker panel for prognosis evaluation and the identification of patients benefiting or not from therapy in mCRC. Our methodology efficiently classified patients earlier than routine computed tomography and from a minimally invasive liquid biopsy.
What's new?
Circulating tumor cells (CTC) play a critical role in disseminating the primary tumor to distant metastases. In colorectal cancer, CTC quantification is used as prognostic marker for disease outcome. Here, the authors tested a panel of tissue‐specific and epithelial‐to‐mesenchymal transition transcripts (GAPDH, VIL1, CLU, TIMP1, LOXL3, ZEB2) in isolated CTCs to improve the prognostic value in patients with metastatic colorectal cancer. High CTC marker expression correlated inversely with patient survival, both before and during treatment, and was a better predictor of patient outcome than standard computed tomography. These data underscore the importance of CTCs and CTC‐derived biomarkers in the clinical management of patients with colon cancer.
Morbidity studies complement the understanding of hazards to raptors by identifying natural or anthropogenic factors. Descriptive epidemiological studies of wildlife have become an important source ...of information about hazards to wildlife populations. On the other hand, data referenced to the overall wild population could provide a more accurate assessment of the potential impact of the morbidity/mortality causes in populations of wild birds.
The present study described the morbidity causes of hospitalized wild raptors and their incidence in the wild populations, through a long term retrospective study conducted at a wildlife rehabilitation centre of Catalonia (1995-2007). Importantly, Seasonal Cumulative Incidences (SCI) were calculated considering estimations of the wild population in the region and trend analyses were applied among the different years. A total of 7021 birds were analysed: 7 species of Strigiformes (n = 3521) and 23 of Falconiformes (n = 3500). The main causes of morbidity were trauma (49.5%), mostly in the Falconiformes, and orphaned/young birds (32.2%) mainly in the Strigiformes. During wintering periods, the largest morbidity incidence was observed in Accipiter gentillis due to gunshot wounds and in Tyto alba due to vehicle trauma. Within the breeding season, Falco tinnunculus (orphaned/young category) and Bubo bubo (electrocution and metabolic disorders) represented the most affected species. Cases due to orphaned/young, infectious/parasitic diseases, electrocution and unknown trauma tended to increase among years. By contrast, cases by undetermined cause, vehicle trauma and captivity decreased throughout the study period. Interestingly, gunshot injuries remained constant during the study period.
Frequencies of morbidity causes calculated as the proportion of each cause referred to the total number of admitted cases, allowed a qualitative assessment of hazards for the studied populations. However, cumulative incidences based on estimated wild raptor population provided a more accurate approach to the potential ecological impact of the morbidity causes in the wild populations.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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