Lukasz Gasiorowski

A novel method for the in vivo isolation of circulating tumor cells from peripheral blood of cancer patients using a functionalized and structured medical wire

The isolation of circulating tumor cells (CTCs) from the blood of patients afflicted with solid malignant tumors becomes increasingly important as it may serve as a ‘liquid biopsy’ with the potential of monitoring the course of the cancer disease and its response to cancer therapy, with subsequent molecular characterization. For this purpose, we functionalized a structured medical Seldinger guidewire (FSMW), normally used to obtain safe access to blood vessels and other organ cavities, with a chimeric monoclonal antibody directed to the cell surface expressed epithelial cell surface adhesion molecule (EpCAM). This medical device was optimized in vitro and its biocompatibility was tested according to the regulations for medical devices and found to be safe with no noteworthy side effects. Suitability, specificity and sensitivity of the FSMW to catch and enrich CTCs in vivo from circulating peripheral blood were tested in 24 breast cancer or non-small cell lung cancer (NSCLC) patients and in 29 healthy volunteers. For this, the FSMW was inserted through a standard venous cannula into the cubital veins of healthy volunteers or cancer patients for the duration of 30 min. After removal, CTCs were identified by immunocytochemical staining of EpCAM and/or cytokeratins and staining of their nuclei and counted. The FSMW successfully enriched EpCAM-positive CTCs from 22 of the 24 patients, with a median of 5.5 (0–50) CTCs in breast cancer (n=12) and 16 (2–515) CTCs in NSCLC (n=12). CTCs could be isolated across all tumor stages, including early stage cancer, in which distant metastases were not yet diagnosed, while no CTCs could be detected in healthy volunteers. In this observatory study, no adverse effects were noted. Evidently, the FSMW has the potential to become an important device to enrich CTCs in vivo for monitoring the course of the cancer disease and the efficacy of anticancer treatment.

The early and late results of combined off-pump coronary artery bypass grafting and pulmonary resection in patients with concomitant lung cancer and unstable coronary heart disease

OBJECTIVE The aim of this study was to analyze the early and long-term results of simultaneous surgical treatment of both coronary heart disease (CHD) and lung cancer. METHODS Twenty-five patients with the diagnoses of both non-small cell lung cancer (NSCLC) and unstable angina were operated on between 2001 and 2006 in the Department of Cardiothoracic Surgery at the University of Medical Sciences in Poznań, Poland. Myocardial revascularization was performed simultaneously with the lung resection. The mean patient age was 63 years. The majority (18 patients) were male and the stage of lung cancer was predominantly AJCC II. Most of the patients were classified as stage II or III CCS and the predominant pathology was a two-vessel disease. Fifteen lobectomies, six pneumonectomies and four wedge resections were performed together with the aortocoronary graft implantation (mean: 1.9 graft per patient). RESULTS There were no postoperative deaths or perioperative myocardial infarctions (MI). The most frequent complications were as follows: atrial fibrillation (24%), atelectasis (12%) and residual pneumothorax (12%). All the patients were followed up for 8-60 months. Within this period, eight patients (32%) died, mostly due to the cancer relapse. The local recurrence of lung cancer and distant metastases were the only factors statistically influencing the late survival. No patient in the entire follow-up period had a MI. In three patients, the symptoms of recurrent angina occurred and one of them underwent a coronary stent implantation. CONCLUSIONS Simultaneous off-pump myocardial revascularization and lung resection is a safe and effective treatment when unstable CHD and lung cancer coexist. In selected patients, this combined procedure may be an alternative to the two-stage approach, surgical or non-surgical (cardiologic) interventions preceding the pulmonary resection. The only statistically significant factor having an impact on long-term survival is the recurrence of the cancer.

The prognostic value of DNA content analysis in patients with squamous cell lung cancer treated surgically.

The aims of the study were to assess the degree of ploidy and determine whether it had any influence on the remission time and survival of surgically treated patients with squamous cell lung cancer. The results were then related to the clinical staging, grading, size and location of the tumor. Tissue samples of squamous cell lung carcinoma (n=80) resected between 1995 and 1996 in the Department of Thoracic Surgery at University of Medical Sciences in Poznan were prepared using the modified Hedleys method. The measurements were made by means of a Cytoron Absolute flow cytometer. Abnormal (aneuploid) DNA was found in 45% of the tumors. In the 2-year observation period significantly more patients with aneuploid tumors died (75%) than those with diploid tumors (43.2%), P<0.05. No significant correlation was found between the ploidy and frequency of metastasis to regional lymph nodes, tumor size, location or grading. Estimation of the DNA content in cancer cells appears to be a significant prognostic factor. Furthermore measurement of the DNA content can be useful after surgery to estimate the risk of recurrence.

Can the condition of the cell microenvironment of mediastinal lymph nodes help predict the risk of metastases in non-small cell lung cancer?

BACKGROUND The aim of this study was to analyze the properties of the immune cell microenvironment of regional lymph nodes (LNs) positive for lung cancer. METHODS Twenty-four patients operated on for stages T1 and T2 of the NSCLC, were enrolled in the study. Peripheral blood and LN tissue were obtained from different lymph node sites and levels. As a control, LN tissue was taken from patients diagnosed with emphysema or pneumothorax. The cells from randomly chosen LN were tested by multi-color flow cytometry. Separate portions of LN were snap-frozen and examined for the presence of cytokeratin positive cells (CK). Propensity for apoptosis, level of TCR zeta chain expression of T cells and the number and maturation status of dendritic cells were confronted with the presence of CK-positive cells. RESULTS The presence of metastases correlated with the downregulation of TCR zeta, especially CD8(+) T cells. The most striking feature was the reduction in the number of myeloid CD11c(+) dendritic cells in the LN of patients with LN metastases. This could be a reflection of the immunodeficient state observed in lung cancer patients. Even in the absence of metastases in the regional LN, the same type of changes in the LN microenvironment were observed in those LN located nearer the primary tumor. CONCLUSIONS The preliminary results of this study suggest that this approach may be helpful as an independent tumor staging factor. It is also worth noting that part of the staging process could also be based on features describing the immune cells in the peripheral blood.

High-fidelity simulation — the first DCD-ECMO procedure in Poland

Mateusz Puslecki, Marcin Ligowski, Marek Dabrowski, Maciej Sip, Sebastian Stefaniak, Tomasz Klosiewicz, Lukasz Gasiorowski, Marek Karczewski, Tomasz Malkiewicz, Malgorzata Ladzinska, Marcin Zielinski, Aleksander Pawlak, Agata Dabrowska, Piotr Ziemak, Bartlomiej Perek, Marcin Misterski, Slawomir Katarzynski, Piotr Buczkowski, Wojciech Telec, Ilona Kiel-Puslecka, Michal Kiel, Michael Czekajlo, Marek Jemielity Poznan University of Medical Sciences, Department of Cardiac Surgery and Transplantology, Clinical Hospital SKPP, Poznan, Poland Poznan University of Medical Sciences, Department of Rescue and Disaster Medicine, Poznan, Poland Polish Society of Medical Simulation, Poland Poznan University of Medical Sciences, Center for Medical Simulation, Poznan, Poland Poznan University of Medical Sciences, Department of Intensive Care and Pain Treatment, Poznan, Poland Poznan University of Medical Sciences, Department of Transplantology, General, Vascular and Plastic Surgery, Poznan, Poland Poznan University of Medical Sciences, Department of Anesthesiology and Intensive Care, Clinical Hospital H. Święcickiego, Poznan, Poland Voivodeship Emergency Medical Services, Poznan, Poland Poznan University of Medical Sciences, Department of Palliative Medicine, Poznan, Poland ZF RTW, Częstochowa, Poland Hunter Holmes McGuire VA Medical Center, Department of Surgery, Richmond, United States of America Lublin Medical University, Lublin, Poland

Prognostic significance of DNA ploidy in squamous cell lung carcinoma: is it really worth it?

BACKGROUND Many previous studies attempted to associate DNA quantification with prognosis for lung cancer, but there is a divergence of opinion about its value. The purpose of the study was to evaluate the frequency of abnormal DNA content in squamous cell cancer (SCC) and analyze its correlation with late survival rates after surgical treatment. METHODS A group of 110 patients surgically treated because of SCC was analyzed. Paraffin-embedded pathologic material underwent cytometric analysis. Postoperative follow-up was performed with standard follow-up visits. The statistical analysis was carried out using Mann-Whitneys U and chi2 tests to compare various variables in both groups. The survival curves were drawn using the Kaplan-Meier method. Clinical staging, regional metastasis, and ploidy, were analyzed with multivariate analysis for having a great impact on survival rates. RESULTS Fifty (45%) tumors were DNA aneuploid. The survival rate in patients with aneuploid cancers was worse than in those with diploid tumors and the most frequent cause of death was local recurrence (p < 0.05). CONCLUSIONS DNA content abnormalities were found to be an important prognostic factor in patients with SCC. The DNA quantification can select a group of high risk of recurrence even after a radical procedure and set new guidelines for adjuvant therapy.

Abstract 1583: Circulating tumor cells isolated from non-small cell lung cancer patients using the in vivo CellCollector Technology

Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA Access to tumor tissue is often limiting and archival samples may not be reflective of the current disease. Alternative approaches using liquid biopsies are being explored to support personalized treatment of cancer patients. Circulating tumor cells (CTCs) detached from primary tumors or metastases into the blood may reflect the current molecular tumor profile providing the opportunity to monitor molecular changes in the tumor. As CTCs are rare cells in the bloodstream and most in vitro approaches for capturing CTCs are based on small samples volumes (5 - 10 ml), accurate and reliable quantification of CTCs is difficult to achieve. We report on a comprehensive clinical study using a novel CellCollectorTM for in vivo isolation of CTCs. Eighty four stage I - IIIB non-small cell lung cancer (NSCLC) patients and 7 healthy individuals were screened for CTCs using a CellCollectorTM (Gilupi, Germany) device. Molecular analysis of a patient with known mutation in the KRAS gene (G12D) was performed to confirm the predictive value of CTCs as “liquid biopsy”. CTCs were detected in 65 out of 84 NSCLC patients (77.4%) (median: 13 CTCs; range from 0 - 300). For comparison, using the U.S. Food and Drug Administration-cleared CellSearch® system only fifteen samples (17.8%) in this cohort were positive (median: 0 CTCs; range from 0-300). In all paired samples, the CellCollectorTM detected the same or higher numbers of CTCs. Healthy individuals were devoid of any cells classified as CTC. Investigating the KRAS status on CTCs captured by the CellCollectorTM, we were able to substantiate the G12D mutation that has earlier on been identified in the tissue of the primary lung tumor.Conclusions: The CellCollectorTM is a promising novel device for the in vivo isolation of CTCs and the device effectively isolated CTCs from NSCLC patients of all stages including the non-metastatic setting. The implementation of the CellColletorTM into clinical practice has the potential for early disease detection. Furthermore, this approach allows the molecular characterization of CTCs for possible therapeutic targets and early response to treatment which might be used in the setting of personalized cancer therapies and clinical development. Citation Format: Thomas Krahn, Lukasz Gasiorowski, Wojciech Dyszkiewicz, Grzegorz Dworacki, Maciej Zabel, Dave S. Hoon, Stefanie Herold, Bjoern Nowack, Johannes Tucholsky, Christian Schumann, Klaus Lucke. Circulating tumor cells isolated from non-small cell lung cancer patients using the in vivo CellCollector Technology. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1583. doi:10.1158/1538-7445.AM2015-1583