Marcela Tomíšková

TaqMan based real time PCR assay targeting EML4-ALK fusion transcripts in NSCLC.

OBJECTIVES Lung cancer with the ALK rearrangement constitutes only a small fraction of patients with non-small cell lung cancer (NSCLC). However, in the era of molecular-targeted therapy, efficient patient selection is crucial for successful treatment. In this context, an effective method for EML4-ALK detection is necessary. We developed a new highly sensitive variant specific TaqMan based real time PCR assay applicable to RNA from formalin-fixed paraffin-embedded tissue (FFPE). MATERIALS AND METHODS This assay was used to analyze the EML4-ALK gene in 96 non-selected NSCLC specimens and compared with two other methods (end-point PCR and break-apart FISH). RESULTS EML4-ALK was detected in 33/96 (34%) specimens using variant specific real time PCR, whereas in only 23/96 (24%) using end-point PCR. All real time PCR positive samples were confirmed with direct sequencing. A total of 46 specimens were subsequently analyzed by all three detection methods. Using variant specific real time PCR we identified EML4-ALK transcript in 17/46 (37%) specimens, using end-point PCR in 13/46 (28%) specimens and positive ALK rearrangement by FISH was detected in 8/46 (17.4%) specimens. Moreover, using variant specific real time PCR, 5 specimens showed more than one EML4-ALK variant simultaneously (in 2 cases the variants 1+3a+3b, in 2 specimens the variants 1+3a and in 1 specimen the variant 1+3b). In one case of 96 EML4-ALK fusion gene and EGFR mutation were detected. All simultaneous genetic variants were confirmed using end-point PCR and direct sequencing. CONCLUSION Our variant specific real time PCR assay is highly sensitive, fast, financially acceptable, applicable to FFPE and seems to be a valuable tool for the rapid prescreening of NSCLC patients in clinical practice, so, that most patients able to benefit from targeted therapy could be identified.

Nutritional Risk Screening Predicts Tumor Response in Lung Cancer Patients

Background: Malnutrition in cancer patients may be associated with poor tolerance of chemotherapy and lower response rate after oncological treatment. Methods: Nutritional Risk Screening 2002 (NRS) adapted for oncological patients was used to assess the risk of undernutrition in a group of 188 patients with lung cancer. The risk was evaluated on a 6-point scale according to common signs of nutritional status (weight loss, body mass index, and dietary intake), tumor, and its treatment risk factors. A score of 3 or more (called “nutritional risk”) means significant risk of malnutrition and poor outcome. Results: Acceptable NRS score was found in 50.6%, and in 45.3% a score of 3–5 suggested the risk of malnutrition (nutritional risk). Unexpectedly, the toxicity of anticancer treatment was not significantly different between the subgroups (acceptable score vs nutritional risk). The rate of treatment response evaluated by imaging techniques was significantly higher in patients with an acceptable score compared to nutritional risk. Overall survival rate was significantly higher in cytostatically treated patients with lung cancer with an acceptable score. Conclusion: Nutritional risk screening is a significant predictor of tumor response in patients with lung cancer. Early detection of malnutrition is important to determine the prognosis of cancer patients as well as to plan effective supportive care.

Thyroid transcription factor 1 expression is associated with outcome of patients with non-squamous non-small cell lung cancer treated with pemetrexed-based chemotherapy

Pemetrexed is an antifolate cytostatic agent targeting several folate-dependent enzymatic pathways, widely used in the treatment of locally advanced or metastatic stage non-small cell lung cancer. Aside from the non-squamous histology, there is still no available molecular biomarker predicting treatment efficacy of pemetrexed-based chemotherapy. The aim of our retrospective study was to evaluate the association of thyroid transcription factor 1 expression with outcome of a large cohort of patients with non-squamous non-small cell lung cancer treated with pemetrexed. We retrospectively analysed clinical data of 463 patients with advanced-stage non-small cell lung cancer (IIIB or IV) treated with pemetrexed-based chemotherapy. Thyroid transcription factor 1 expression was assessed using indirect immunohistochemical detection in formalin-fixed paraffin-embedded tumour tissue at the time of diagnosis. Thyroid transcription factor 1 expression was detected in the tumour tissue from 76.0% of patients, and tumours from 24.0% of patients were thyroid transcription factor 1 negative. The median progression-free survival and overall survival for patients with thyroid transcription factor 1 positive tumours were 4.8 and 11.8 months compared to 2.8 and 8.3 months for those with thyroid transcription factor 1 negative tumours (p = 0.001 and p < 0.001). The multivariable Cox proportional hazards model revealed that thyroid transcription factor 1 expression was significantly associated with progression-free survival (hazard ratio = 1.57, p < 0.001) and also with overall survival (hazard ratio = 1.73, p < 0.001). In conclusion, the results of the conducted retrospective study suggest that the thyroid transcription factor 1 expression was independently associated with progression-free survival and overall survival in patients with advanced-stage non-squamous non-small cell lung cancer treated with pemetrexed-based chemotherapy.

Identification of atypical ATRNL1 insertion to EML4-ALK fusion gene in NSCLC

We herein present a rare case of an EML4-ALK positive patient. A 61-year-old man was diagnosed with locoregional non-small cell lung cancer (NSCLC). No EGFR mutations were detected, and therefore the ALK rearrangement was evaluated using immunohistochemistry (IHC), fluorescence in situ hybridization (FISH) and the reverse transcription PCR (RT-PCR) method for EML4-ALK. All methods showed a positive result and, therefore, the patient was treated with crizotinib with a good therapeutic response. However, a detailed RT-PCR analysis and sequencing revealed an unexpected 138 bp insertion of attractin-like 1 (ATRNL1) gene into the EML4-ALK fusion gene. In our case, the positive therapeutic response suggests that ATRNL1 insertion does not affect EML4-ALKs sensitivity to crizotinib. This case shows great EML4-ALK heterogeneity and also that basic detection methods (IHC, FISH) cannot fully specify ALK rearrangement but in many cases a full specification seems to be important for an effective TKI indication, and sequencing ALK variants might contribute to optimized patient selection.

[Effect of Erlotinib in 2nd and 3rd Line Anticancer Treatment in Patients with Squamous Cell Lung Cancer - Case Series].

BACKGROUND Squamous cell carcinoma of the lung (SCC) represents cca 30-40% of new cases of non-small cell lung cancer (NSCLC) in the Czech Republic. The tyrosine kinase inhibitor erlotinib is indicated as a 1st line treatment for patients with locally advanced and metastatic disease and activating mutations in endothelial growth factor receptor (EGFR), or as a 2nd or 3rd line treatment in EGFR-negative NSCLC patients after chemotherapeutic failure. OBSERVATION We present three case reports of patients with SCC treated with erlotinib as a 2nd or 3rd line of treatment. All patients were verified by histological analysis of tumor samples. EGFR mutation status was negative in one patient, while the other samples were not suitable for genetic screening. RESULTS The therapeutic response to erlotinib lasted for 68, 40, and 13 months, resp. The patient with the longest therapeutic response (patient no. 1) is still continuing erlotinib treatment (as of December 2016). The overall survival of the two patients who died was 50 and 43 months, resp. One patient died of an unknown cause with no signs of progression of the disease on CT scans. The other patient died of terminal progression of the oncological disease. All three patients experienced major therapeutic benefit from erlotinib treatment as shown by the long periods of progression-free survival and prolonged overall survival. CONCLUSION The three case reports demonstrate that erlotinib may be effective as a 2nd or 3rd line treatment in patients with SCC, especially in patients with limited alternative anticancer treatment options.Key words: non-small cell lung cancer - squamous cell carcinoma - erlotinib - treatment - tyrosine kinase inhibitor The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 5. 8. 2016Accepted: 14. 12. 2016.

Full Oral Vinorelbine (NVBO) on D1 and D8 With Carboplatin (CBDCA) as First Line Treatment in Advanced Non-small Cell Lung Cancer (NSCLC) Patients: Preliminary Results of a Prospective Sudy in Nonrandomized Population

The purpose of this trial was to evaluate the activity and feasibility of CBDCA together with NVBO as first line treatment in patients with advanced NSCLC. METHODS: Patients with advanced NSCLC received NVBO 80 mg/m2 on D1 and D8 with CBDCA AUC5 on D1 every three weeks. In stage III, chemotherapy was followed by external radiotherapy. The outcomes include following: response, median overall survival (mOS) and median progression free survival (mPFS). The difference in response relative to baseline characteristics was determined using Pearson Chi-square test. Differences in OS and PFS relative to baseline characteristics were evaluated for significance using Log-rank test. RESULTS: 259 patients were treated: 209 men (80,7%) and 50 women (19,3%), median age 65 years. ECOG performance status at inclusion was PS 0 in 47 (18,2% patients, PS 1 in 185 (71,7%) and PS 2 in 26 (10,1%) patients. Most patients had stage IIIB 97 (37,5%) and stage IV NSCLC 130 (50,2%), only 32 (12,4%) were stage IIIA . Adenocarcinoma was confirmed in 52 patients (20,1%), squamous-cell carcinoma in 152 (58,7%), large-cell carcinoma in 8 (3,1%) and other in 47 (18,4%). Complete response was confirmed in 1 (0,4%) patient, partial response in 121 (46,7%), stable disease in 58 (22,4%), 79 (30,5%) patients progressed. The regimen was well tolerated. Median cycles was 4, the dosage of NVBO was without changes in 159 (61%) patients, the dosage of NVBO was reduced in 12 (4,7%) and escalated in 64 (24,8%). In 23 (8,9%) of patients was the dosage of NVBO reduced after escalation. Major toxicities (Grade 3-4) were neutropenia in 69 (26,9%), leucopenia in 51 (19,8%), anemia in 7 (2,7%), and thrombocytopenia in 6 (2,3%) patients. Febrile neutropenia was observed in 17 (6,6%) patients. Gastrointestinal toxicity grade 3-4 was observed in 47 (18,4%) patients. The estimated mOS was 13,8 moths. and the estimated mPFS was 9,4 months by median follow-up 8,5 months. The differences between groups of pts according to PS (0+1 vs. 2) were statistically significant (p < 0,001) better for patiens with PS 0+1. CONCLUSIONS: In this group of 259 non-selected patients with advanced NSCLC was the treatment with full NVBO and CBDCA in first line more convenient and well tolerated with evidence of high antitumour activity. This combination was active in all groups patients according histology.

Intravenous vinorelbine (NVBiv) on D1 and oral vinorelbine (NVBo) on D8 in combination with carboplatin (CBDCA) as first-line treatment in advanced non-small lung cancer (NSCLC) patients: Results of a prospective study in nonselected population.

The use of intravenous vinorelbine (NVBiv) on D1 and oral vinorelbine (NVBo) on D8 in combination with carboplatin (CBDCA) as first-line treatment in advanced non-small lung cancer (NSCLC) patients: results of a prospective study in nonselected population are reported.

59P MARKERS OF PROLIFERATION ACTIVITY IN NON-SMALL CELL LUNG CANCER

A variety of parameters acknowledged to be markers of proliferation activity of non-small cell lung cancer were followed up and discussed in this paper.

6 Chemotherapy with paclitaxel, carboplatin and concurrent radiotherapy for advanced non small cell lung cancer

The authors report the results of a paclitaxel-caboplatin study in patients with advanced non small cell lung cancer with concurrent radiotherapy.

P-921 The 5-years results of the surgical treatment in patients with non-small cell lung cancer (NSCLC): An analysis of 540 patients

The 5-years results of the surgical treatment in patients with non-small cell lung cancer (NSCLS): An analysis of 540 patients