MicroRNA-126 selected with broad-spectrum analysis of microRNAs - a new predictive factor for the effectiveness of immunotherapy or chemoimmunotherapy in advanced NSCLC patients?

Introduction: Expression of PD-L1 on cancer cells is the only validated predictive factor for immunotherapy in NSCLC (Non-Small Cell Lung Cancer) patients. However, on this basis, it is difficult to predict the occurrence of resistance to immune checkpoint inhibitors (ICIs). MicroRNAs are widely studied as biomarkers of cancers. Our study was designed to determine whether microRNAs can be sensitive predictive factors in the qualification of NSCLC patients to first-line immunotherapy or chemoimmunotherapy. Material and methods: The two-stage research on validation group (n=20) and study group (n=35) of patients with advanced NSCLC was conducted. Analysis of microRNAs expression by qPCR in plasma collected prior to the start of immunotherapy (pembrolizumab) or chemoimmunotherapy (combination of pembrolizumab with chemotherapy) was made. Broad-spectrum analysis of microRNAs expression was used in the studied group. Three microRNAs selected in that group as important for the effectiveness of ICIs were then examined in the validation group. Results: In the studied group, significantly higher expression of miRNA-126-3p, miR-144-3p and miR-146-5p was observed in patients with long PFS compared to those with short PFS. In the validation group, low miRNA-126 expression indicated lower median progression-free survival and overall survival (2.3 vs. 5.0 months and 5.2 vs 11.2, respectively). These patients had a significantly higher risk of progression (HR= 2.92, 95% CI: 1.01 to 8.40, p=0.04) and death (HR=3.64, 95% CI: 1.22 to 10.84, p=0.02). Conclusion: Our study showed that the expression of miR-126 in blood plasma may be a predictive factor for the effectiveness of first-line immunotherapy or chemoimmunotherapy in advanced NSCLC patients.

Numerical and physical modeling of breast cancer based on image fusion and artificial intelligence.

PURPOSE: The key problem raised in the paper is the change in the position of the breast tumor due to magnetic resonance imaging examinations in the abdominal position relative to the supine position during the surgical procedure. Changing the position of the patient leads to significant deformation of the breast, which leads to the inability to indicate the location of the neoplastic lesion correctly. METHODS: This study outlines a methodological process for treating cancer patients. Pre-qualification assessments are conducted for magnetic resonance imaging (MRI), and 3D scans are taken in three positions: supine with arms raised, supine surgical position (SS), and standing. MRI and standard ultrasonography (USG) imaging are performed, and breast and cancer tissue are segmented from the MRI images. Finite element analysis is used to simulate tissue behavior in different positions, and an artificial neural network is trained to predict tumor dislocation. Based on the model, a 3D-printed breast with a highlighted tumor is manufactured. This computer-aided analysis is used to create a detailed surgical plan, and lumpectomy surgery is performed in the SS. In addition, the geometry of the tumor is presented to the medical staff as a 3D-printed element. RESULTS: By utilizing a comprehensive range of techniques, including pre-qualification assessment, 3D scanning, MRI and USG imaging, segmentation of breast and cancer tissue, model analysis, image fusion, finite element analysis, artificial neural network training, and additive manufacturing, a detailed surgical plan can be created for performing lumpectomy surgery in the supine surgical position. CONCLUSION: The new approach developed for the pre-operative assessment and surgical planning of breast cancer patients has demonstrated significant potential for improving the accuracy and efficacy of surgical procedures. This procedure may also help the pathomorphological justification. Moreover, transparent 3D-printed breast models can benefit breast cancer operation assistance. The physical and computational models can help surgeons visualize the breast and the tumor more accurately and detailedly, allowing them to plan the surgery with greater precision and accuracy.

Assessment of child safety on bicycles in baby carriers - The importance of evaluating both head and neck injuries.

INTRODUCTION: The paper addresses an important accident type that involves children in bicycle seats - the bicycle fall over. It is a significant and common accident type and many parents have been reported to experience this type of "close call." The fall over occurs at low velocities and even while a bicycle is standing still, and may result from a split-second lack of attention on behalf of the accompanying adult (e.g. while loading groceries, i.e. while not being exposed to traffic per se). Moreover, irrespective of the low velocities involved, the trauma that may result to the head of the child is considerable and may be life-threatening, as shown in the study. METHOD: The paper presents two methods to address this accident scenario in a quantitative way: in-situ accelerometer-based measurement and numerical modeling approaches. It is shown that the methods produce consistent results under the prerequisites of the study. They are therefore promising methods to be used in the study of this type of accident. RESULTS: The importanance of the protective role of a child helmet is without discussion in everyday traffic.However, this study draws attention to one particular effect observed in this accident type: that the geometry of the helmet may at times expose the child's head to considerably larger forces, by having contact with the ground. The study also highlights the importance of neck bending injuries during bicycle fall over, which are often neglected in the safety assessment - not only for children in bicycle seats. The study concludes that considering only head acceleration may lead to biased conclusions about using helmets as protective devices.

Stabilization of after-traumatic flail chest by minimally invasive modified NUSS procedure in patient with starting respiratory insufficiency.

A flail chest is one of the possible medical conditions suffered by individuals who were injured in traffic accidents, caused by multiple fractures of the ribs and sternum. Which often results in paradoxical chest movements. The consequence may be respiratory failure and need for long-term mechanical ventilation. Such treatment require Intensive Care Unit and may be associated with the possibility of numerous complications.Modified Nuss procedure was performed in 79-year-old man, a victim of a car crash to obtain stabilization of the flail chest. After compensation of paradoxical movements on the third day it was possible to end mechanical ventilation. A quick procedure dedicated to the congenital deformation of the chest made it possible to avoid long, expensive intensive therapy with possible respiratory complications.The NUSS procedure enables the effective and safe treatment of a flail chest in a selected group of patients.

Methylation of DROSHA and DICER as a Biomarker for the Detection of Lung Cancer.

Background: Lung cancer is the leading cause of cancer-related deaths. Early diagnosis may improve the prognosis. Methods: Using quantitative methylation-specific real-time PCR (qMSP-PCR), we assessed the methylation status of two genes (in two subsequent regions according to locations in their promoter sequences) related to carcinogenesis, DICER and DROSHA, in 101 plasma samples (obtained prior to the treatment) of lung cancer patients and 45 healthy volunteers. Results: The relative level of methylation of DROSHA was significantly lower (p = 0.012 for first and p < 0.00001 for the second region) and DICER significantly higher (p = 0.029 for the first region) in cancer patients. The relative level of methylation of DROSHA was significantly (p = 0.037) higher in patients with early-stage NSCLC (IA-IIIA) and could discriminate them from healthy people with a sensitivity of 71% and specificity of 76% (AUC = 0.696, 95% CI: 0.545-0.847, p = 0.011) for the first region and with a sensitivity of 60% and specificity of 85% (AUC = 0.795, 95% CI: 0.689-0.901, p < 0.0001) for the second region. Methylation analysis of the first region of the DICER enabled the distinction of NSCLC patients from healthy individuals with a sensitivity of 96% and specificity of 60% (AUC = 0.651, 95% CI: 0.517-0.785, p = 0.027). The limitations of the study include its small sample size, preliminary nature, being an observational type of study, and the lack of functional experiments allowing for the explanation of the biologic backgrounds of the observed associations. Conclusion: The obtained results indicate that the assessment of DICER and DROSHA methylation status can potentially be used as a biomarker for the early detection of lung cancer.

Quantitative Assessment of the State of Threat of Working on Construction Scaffolding.

Working at height, and especially on construction scaffolding, is one of the most accident-prone situations on a construction site. The article attempts to assess the state of threat of working on scaffolding on the basis of the proposed coefficients concerning the possibility of an occupational accident occurring. The article presents the analysis of 10 parameters, which were classified into three groups of factors that cause accidents: technical, organizational, and human factors. In order to assess the state of threat of working on scaffolding, partial hazard factors and a simplified and accurate factor of the state of threat of working were proposed. The coefficients were determined on the basis of the data collected from post-accidental control reports on occupational accidents occurring on scaffolding in the construction industry that took place in Poland in five voivodeships in the years 2008-2017, and also on the basis of the obtained results of research on 120 scaffoldings conducted in the years 2016-2018. Based on the determined factors, it was possible to determine the probability of an undesirable event, in other words, an occupational accident. In addition, the developed test method proposed numerical scales for assessing the state of threat of working on scaffolding. The form proposed in the article for assessing the state of threat of working on scaffolding, which was developed using a spreadsheet, can provide support for people managing work at workstations involving scaffolding, for example, construction directors, construction engineers, work managers, or construction managers.

Tissue MicroRNA Expression as a Predictor of Response to Immunotherapy in NSCLC Patients.

INTRODUCTION: Expression of PD-L1 protein on tumor cells, which is so far the only validated predictive factor for immunotherapy, is regulated by epigenetic and genetic factors. Among the most important ones that regulate gene expression are microRNAs. MATERIALS AND METHODS: The study included 60 patients with NSCLC who underwent first or second line immunotherapy with pembrolizumab or nivolumab. FFPE materials were collected before the start of immunotherapy. We examined relative expression of microRNAs (miR-141, miR-200a, miR-200b, miR-200c, miR-429, miR-508-3p, miR-1184, miR-1255a) and PD-L1 mRNA expression. Copy number variation (CNV) of PD-L1 gene by qPCR and FISH methods were assessed. Two single nucleotide polymorphisms (SNPs) in promoter region of PD-L1 gene (rs822335 and rs822336) were examined. Expression of PD-L1 protein on tumor cells was assessed by immunohistochemistry (IHC). The response rate to immunotherapy and progression free survival (PFS) measured in weeks and overall survival (OS) measured in months from the start of immunotherapy were evaluated. RESULTS: Response to immunotherapy was observed in nine patients (15%, including one complete response), disease stabilization in 22 patients (36.7%), and progression in 29 patients (48.3%). Significantly higher (p=0.015) expression of miR-200b and significantly lower (p=0.043) expression of miR-429 were observed in responders compared to patients who did not respond to immunotherapy. The median PFS in the whole group of patients was 16 weeks, and the median OS was 10.5 month. In univariate analysis, the median PFS was significantly higher in patients with high miR-200b expression (HR=0.4253, 95%CI: 0.1737-1.0417, p=0.05) and high miR-508 expression (HR=0.4401, 95%CI: 0.1903-1.0178, p=0.05) and with low expression of miR-429 (HR=0.1288, 95%CI: 0.01727-0.9606, p=0.0456) compared to patients with low and high expression of these molecules, respectively. The median OS was higher in patients with low expression of miR-429 (HR=0,6288, 95%CI: 0,3053-1,2949, p=0.06) compared with patients with high expression of this microRNA. In multivariate analysis, we found that patients with PD-L1 expression on ≥1% of tumor cells compared to patients without PD-L1 expression on cancer cells had a significantly lower risk of progression (HR=0.3857, 95%CI: 0.1612-0.9226, p=0.0323) and death (HR=0.377, 95%CI: 0.1636-0.8688, p=0.022). CONCLUSION: The miR-200b and miR-429 molecules in tumor cells seem to have greatest impact on the effectiveness of immunotherapy in NSCLC patients.

Elective lung resection increases spatial QRS-T angle and QTc interval.

BACKGROUND: Lung resection changes intra-thoracic anatomy, which may affect electrocardiographic results. While postoperative cardiac arrhythmias have been recognized after lung resection, no study has documented changes in vectorcardiographic variables in patients undergoing this surgery. The purpose of this study was to analyse changes in spatial QRS-T angle (spQRS-T) and corrected QT interval (QTc) after lung resection. METHODS: Adult patients undergoing elective lung resection under general anaesthesia were studied. The patients were allocated into four groups: those undergoing (1) left lobectomy (LL); (2) left pneumonectomy (LP); (3) right lobectomy (RL); and (4) right pneumonectomy (RP). The spQRS-T angle and QTc interval were measured one day before surgery (baseline) and 24, 48 and 72 h after surgery. RESULTS: Seventy-one adult patients (47 men and 24 women) aged 47-80 (65 ± 7) years were studied. In the study group as a whole, lung resection was associated with significant increases in spQRS-T (p < 0.001) and QTc (p < 0.05 at 24 and 48 h and p < 0.01 at 72 h). The greatest changes were noted in patients undergoing LP. Postoperative atrial fibrillation (AF) was noted in 6.4% of patients studied, in whom the widest spQRS-T angle and the most prolonged QTc intervals were also noted. CONCLUSIONS: Lung resection widens the spQRS-T angle and prolongs the QTc interval, especially in patients undergoing LP. While postoperative AF was a relatively rare complication after lung resection in this study, it was associated with the widest spQRS-T angles and most prolonged QTc intervals.

Promoter polymorphisms of TOP2A and ERCC1 genes as predictive factors for chemotherapy in non-small cell lung cancer patients.

BACKGROUND: Topoisomerase 2-alpha (TOP2A) is an enzyme that controls topologic changes in DNA during transcription and replication. ERCC1 is an enzyme that takes part in DNA repair processes. The purpose of this study was to assess the predictive role of particular single nucleotide polymorphisms (SNPs) in the promoter regions of TOP2A and ERCC1 genes in non-small cell lung cancer patients (NSCLC) treated with chemotherapy. MATERIALS AND METHODS: We enrolled 113 NSCLC patients treated in the first line with platinum-based chemotherapy. Effectiveness was available for 71 patients. DNA was isolated from whole blood using the Qiamp DNA Blood Mini kit (Qiagen). We examined five SNPs: rs11615 (ERCC1), rs3212986 (ERCC1), rs13695 (TOP2A), rs34300454 (TOP2A), rs11540720 (TOP2A). Quantitative PCR using the TaqMan probe (ThermoFisher) was performed on a Eco Illumina Real-Time PCR system device (Illumina Inc). RESULTS: Patients with the A/A genotype in rs11615 of the ERCC1 gene had significantly longer median progression free survival (PFS) (8.5 months; P = .0088). Patients with the C/C genotype in rs3212986 of the ERCC1 gene had longer median PFS (7 months; P = .05). Patients with the C/C genotype in rs34300454 of TOP2A gene had significantly higher median PFS (7.5 months; P = .0029). Carriers of the C/C genotype in rs34300454 of the TOP2A gene had significantly longer median OS (15.5 months; P = .0017). Patients with the A/A genotype in rs11615 of the ERCC1 gene had significantly higher risk of neutropenia (P = .0133). CONCLUSIONS: Polymorphisms of the TOP2A and ERCC1 genes may be a predictive factor of toxicities and survival for chemotherapy in NSCLC patients.

Air pollution: how many cigarettes does each Pole 'smoke' every year and how does it influence health, with special respect to lung cancer?

INTRODUCTION: Air pollution is one of the most important issues of our times. Air quality assessment is based on the measurement of the concentration of substances formed during the combustion process and micro-particles suspended in the air in the form of an aerosol. Microscopic atmospheric particulate matters (PM) 2.5 and 10 are mixtures of organic and inorganic pollutants smaller than 2.5 and 10 µm, respectively. They are the main cause of negative phenomena in the earth's atmosphere of Earth and human health, especially on the respiratory and cardiovascular systems. Particulates have the ability to cause permanent mutations of tissue, leading to neoplasms and even premature deaths. Nitrogen dioxide (NO2) is one of the main pollutants which arises mainly during the burning of fossil fuels. Based on numerous scientific researches, it has been proved that long-term exposure to NO2 could increase morbidity of cancer due to inflammatory processes increasing abnormal mutations. MATERIAL AND METHODS: Data available in the Polish National Cancer Registry, Chief Inspectorate for Environmental Protection and Map of Health Needs in the Field of Oncology for Poland, WHO Air Quality Guidelines 2005 were analyzed. Air pollution was also evaluated: PM2.5, PM10, NO2, and compared with lung cancer morbidity. RESULTS AND CONCLUSIONS: Based on the available data and literature, it can be concluded that in 2009-2017, on average, each Pole smoked ten cigarettes a day +/- 2. Therefore, it can be estimated that after 60 years everyone had 30 package-years of smoking, leading to a high risk of lung cancer and other smoking related diseases. Additionally air quality in Poland is not satisfactory, exceeding the standards presented in the WHO Guidelines 2005. It can be assumed that this may translate into an additional, independent continuous increase in morbidity and mortality dependent on smoking.

PD-L1 gene copy number and promoter polymorphisms regulate PD-L1 expression in tumor cells of non-small cell lung cancer patients.

Most drugs targeting PD-1 or PD-L1 are more effective when cancer cells of non-small cell lung cancer (NSCLC) patients express PD-L1 protein. The polymorphisms of PD-L1 gene and PD-L1 gene copy number could be responsible for PD-L1 mRNA and protein expression. We analyzed PD-L1 protein expression using two IHC assays, mRNA (PD-L1) expression by qRT-PCR, PD-L1 gene promoter region polymorphisms (rs822335 and rs822336) by qPCR and PD-L1 gene copy number by fluorescence in situ hybridization method. Patients with CC genotype in rs822335 had significantly (p = 0.043) higher percentage of tumor cells with PD-L1 expression (test with 22C3 antibody) than patients with CT or TT genotypes. PD-L1 gene copy number significantly positively correlated with percentage of tumor cells with PD-L1 expression detected in tests with 22C3 antibody (p = 0.005, R = +0.442) and with SP142 antibody (p = 0.021, R = +0.369). PD-L1 gene copy number did not correlate with PD-L1 mRNA expression. Patients with PD-L1 expression tested with 22C3 antibody had significantly higher expression of PD-L1 mRNA (p = 0.023), number of chromsosme 9 centromeres (p = 0.023) and PD-L1 gene copy number (p = 0.003) than patients without PD-L1 expression on tumor cells PD-L1 gene polymorphisms and PD-L1 gene copy number may be a predictor for PD-L1 protein expression on tumor cells.

MicroRNAs aid the assessment of programmed death ligand 1 expression in patients with non-small cell lung cancer.

The qualification of patients with non-small cell lung cancer (NSCLC) for anti-programmed cell death 1 (PD-1) or anti-programmed death ligand 1 (PD-L1) antibody therapy is based on an immunohistochemistry (IHC) assessment of PD-L1 expression. Immunological checkpoint inhibitors improve the overall survival of patients with expression of PD-L1; however certain PD-L1-negative patients may also benefit from immunotherapy. This indicates the requirement for novel predictive factors for the qualification of immunotherapy. It is also necessary to understand the mechanisms that effect the expression of PD-L1 in tumor cells. The expression of PD-L1 in 47 formalin-fixed, paraffin-embedded, NSCLC specimens was assessed using IHC and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The expression of 8 microRNAs (miRNAs, miRs) complementary to PD-L1-mRNA was also evaluated using RT-qPCR. A positive correlation was revealed between the expression level of PD-L1-mRNA and 2 miRs, miR-141 (R=0.533; P=0.0029) and miR-1184 (R=0.463; P=0.049). There was also a positive correlation between the percentage of PD-L1-positive tumor cells and the expression levels of miR-141 (R=0.441; P=0.0024), miR-200b (R=0.372; P=0.011) and miR-429 (R=0.430; P=0.0028), and between the percentage of the tumor area with immune cell infiltration and the expression levels of miR-141 (R=0.333; P=0.03) and miR-200b (R=0.312; P=0.046). Additionally, the percentage of tumor cells expressing PD-L1 positively correlated with miR-141 expression (R=0.407; P=0.0055). Correlations between the expression of the investigated miRs (particularly miR-141) and PD-L1 indicated that miRs may regulate PD-L1 expression at a post-transcriptional level.

Investigation of biomechanics of skull structures damages caused by dynamic loads.

PURPOSE: The aim of the study was to examine the influence of cranial sutures on the crack behaviour of a human skull after the impact. The authors focused on the assessment of skull breaking nature, based on a real-world vehicle-to-bicyclist accident. In the state of the art, there is still no consensus about sutures mechanical properties. Currently, most of the numerical head models do not have distinguished cranial sutures. METHODS: The authors compared different elastic properties for cranial sutures and their influence on the nature of the skull fracture. The mathematical and numerical modelling have been applied to mimic the nature of the skull fracture. The LS-DYNA explicit code with material models featuring the erosion of finite elements was used. The models of the skull with different cranial sutures properties were impacted against a validated front-end of a vehicle. RESULTS: Various fracture patterns were obtained for different material properties of the sutures and the results were compared to a model without the cranial sutures. Based on the results, a graph was plotted to indicate differences in sutures energy absorption capabilities. The numerical results were supported by the mathematical modelling. The developed diagram may enable better understanding of the complex mechanical phenomena on the suture interface. CONCLUSIONS: Biomechanical evidence was provided for the important role of the sutures in numerical models as well as their significant influence on the biomechanics of skull fractures caused by dynamic loads.

Screening for ALK abnormalities in central nervous system metastases of non-small-cell lung cancer.

Anaplastic lymphoma kinase (ALK) gene rearrangement was reported in 3%-7% of primary non-small-cell lung cancer (NSCLC) and its presence is commonly associated with adenocarcinoma (AD) type and non-smoking history. ALK tyrosine kinase inhibitors (TKIs) such as crizotinib, alectinib and ceritinib showed efficiency in patients with primary NSCLC harboring ALK gene rearrangement. Moreover, response to ALK TKIs was observed in central nervous system (CNS) metastatic lesions of NSCLC. However, there are no reports concerning the frequency of ALK rearrangement in CNS metastases. We assessed the frequency of ALK abnormalities in 145 formalin fixed paraffin embedded (FFPE) tissue samples from CNS metastases of NSCLC using immunohistochemical (IHC) automated staining (BenchMark GX, Ventana, USA) and fluorescence in situ hybridization (FISH) technique (Abbot Molecular, USA). The studied group was heterogeneous in terms of histopathology and smoking status. ALK abnormalities were detected in 4.8% (7/145) of CNS metastases. ALK abnormalities were observed in six AD (7.5%; 6/80) and in single patients with adenosuqamous lung carcinoma. Analysis of clinical and demographic factors indicated that expression of abnormal ALK was significantly more frequently observed (P = 0.0002; χ2 = 16.783) in former-smokers. Comparison of IHC and FISH results showed some discrepancies, which were caused by unspecific staining of macrophages and glial/nerve cells, which constitute the background of CNS tissues. Their results indicate high frequency of ALK gene rearrangement in CNS metastatic sites of NSCLC that are in line with prior studies concerning evaluation of the presence of ALK abnormalities in such patients. However, they showed that assessment of ALK by IHC and FISH methods in CNS tissues require additional standardizations.

Prevalence of NRAS, PTEN and AKT1 gene mutations in the central nervous system metastases of non-small cell lung cancer.

Somatic mutations in NRAS, PTEN and AKT1 genes are rarely (~1%) reported in primary NSCLC, but their role in carcinogenesis have been proven. Therefore, we assessed the frequency of them in 145 FFPE tissue samples from CNS metastases of NSCLC using the real-time PCR technique. We identified four (two NRAS and single AKT1 and PTEN) mutations in CNS metastases of NSCLC. All mutations were observed in current male smokers (4% out of the male group; 4/100 and 4.25% out of smokers; 4/94). Three mutations have been detected in patients with SqCC (10.3% out of SqCC patients; 3/29), and only one mutation in the NRAS gene-in a patient with adenocarcinoma (1.25% out of AC patients; 1/80). The examined genes were mutually exclusive in terms of molecular background in KRAS; EGFR; DDR2; PIK3CA; HER2 and MEK1 genes that were evaluated in our previous studies. The OS of the patients who harbored NRAS, AKT1 and PTEN mutations was 10.1, 12.1, 7.3 and 4 months, respectively (vs 13.5 months of the studied group). Our results suggest that the presence of NRAS, PTEN and AKT1 gene mutations may have an influence on the occurrence of CNS metastases in patients with SqCC.

PIK3CA Mutations Detected in Patients with Central Nervous System Metastases of Non-small Cell Lung Cancer.

BACKGROUND: In non-small cell lung cancer (NSCLC) the phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) gene mutations have been reported in fewer than 5% of primary tumors. MATERIALS AND METHODS: We assessed PIK3CA gene mutations in 145 tissue samples from central nervous system (CNS) metastases of NSCLC using three polymerase chain reaction (PCR) techniques: high resolution melting-PCR (HRM-PCR), allele-specific-quantitative PCR (ASP-qPCR) and TaqMan PCR. RESULTS: HRM analysis allowed us to select three PIK3CA-positive specimens (2.1% of the studied group) and ASP-qPCR techniques identified them as one E542K and two H1047R substitutions, which were confirmed by TaqMan probes. The PIK3CA mutations were indicated only in males (3% of all males). One of the patients was reported to be a non-smoker with adenocarcinoma (AC; 2.5% of the AC group), however, the other two patients were smokers with squamous cell carcinoma (SCC; 3.4% of SCC group). CONCLUSION: This is the first report of the presence of PIK3CA gene mutation in CNS-metastatic lesions of NSCLC worldwide that could broaden therapeutic choices in such patients.

Analysis of KRAS and BRAF genes mutation in the central nervous system metastases of non-small cell lung cancer.

KRAS mutations are associated with tumor resistance to EGFR TKIs (erlotinib, gefitinib) and to monoclonal antibody against EGFR (cetuximab). Targeted treatment of mutated RAS patients is still considered as a challenge. Inhibitors of c-Met (onartuzumab or tiwantinib) and MEK (selumetinib-a dual inhibitor of MEK1 and MEK2) signaling pathways showed activity in patients with mutations in KRAS that can became an effective approach in carriers of such disorders. BRAF mutation is very rare in patients with NSCLC, and its presence is associated with sensitivity of tumor cells to BRAF inhibitors (vemurafenib, dabrafenib). In the present study, the frequency and type of KRAS and BRAF mutation were assessed in 145 FFPE tissue samples from CNS metastases of NSCLC. In 30 patients, material from the primary tumor was simultaneously available. Real-time PCR technique with allele-specific molecular probe (KRAS/BRAF Mutation Analysis Kit, Entrogen, USA) was used for molecular tests. KRAS mutations were detected in 21.4 % of CNS metastatic lesions and in 23.3 % of corresponding primary tumors. Five mutations were identified both in primary and in metastatic lesions, while one mutation only in primary tumor and one mutation only in the metastatic tumor. Most of mutations were observed in codon 12 of KRAS; however, an individual patient had diagnosed a rare G13D and Q61R substitutions. KRAS mutations were significantly more frequent in adenocarcinoma patients and smokers. Additional analysis indicated one patient with rare coexistence of KRAS and DDR2 mutations. BRAF mutation was not detected in the examined materials. KRAS frequency appears to be similar in primary and CNS.

Sensitive methods for detection of the S768R substitution in exon 18 of the DDR2 gene in patients with central nervous system metastases of non-small cell lung cancer.

Discoidin death receptor 2 (DDR2) receptor belongs to a DDR family that shows a tyrosine kinase activity. The somatic mutations in DDR2 gene, reported in non-small cell lung cancer (NSCLC), are involved in up-regulation of cells' migration, proliferation and survival. A S768R substitution in DDR2 gene was commonly reported in squamous cell lung carcinoma. Clinical data of patients carrying the DDR2 gene mutation suggest that its presence can be independent of gender and age. The effectiveness of an oral dual-specific (Src and Abl) multikinase inhibitors-dasatinib-was observed in different cell lines and in some NSCLC patients with identified DDR2 mutation. In the present study, we have used three molecular methods (ASP-real-time PCR, ASP-DNA-FLA PCR and direct sequencing) to detect the DDR2 gene mutation in 143 patients with NSCLC metastases to the central nervous system (CNS). The prevalence of the DDR2 gene mutation was correlated with the occurrence of mutations in the EGFR, KRAS, HER2 and BRAF genes. We identified three patients (2.1% of studied group) with DDR2 mutation. The mutation was observed in two patients with low differentiated squamous cell lung cancer and in one patient with adeno-squamous cell carcinoma (ADSCC). In ADSCC patients, DDR2 mutation coexisted with G12C substitution in KRAS gene. According to the current knowledge, examination of the presence of the DDR2 gene mutation in metastatic lesion is the first such report worldwide. The information, that these driver mutations are present in CNS metastases of NSCLC, could broaden therapeutic choices in such group of patients.

Rare co-existence of mutation in KRAS and ALK gene re-arrangement in an adenocarcinoma patient--a case report.

Anaplastic lymphoma kinase (ALK) gene re-arrangements are present in approximately 4% of patients with non-small cell lung cancer (NSCLC), mostly in non-smokers with adenocarcinoma. V-KI-RAS2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations are more common in smokers. These molecular lesions were usually described as are mutually exclusive. We herein describe a rare case of co-existence of ALK and KRAS abnormalities in adenocarcinoma tumor with massive local growth (disproportionality of clinical symptoms) and rapid central nervous system (CNS) metastases spread. T3N1M0 stage tumor (size: 10×12×13 cm) in upper lobe of the right lung was diagnosed in a 56-year-old Caucasian male smoker. Adenocarcinoma of solid predominant was surgically resected with chest wall reconstruction. One month after surgery, CNS metastases were diagnosed and subsequently treated with radiotherapy. We noted an 8-month overall survival from tumor resection. In the case of comorbidity of disorders in the ALK (uncertain prognostic significance) and KRAS gene (described as unfavorable prognostic factor), these abnormalities may ultimately decide the course of the disease in the form of brain metastases.