Search

// Tareq A. Juratli 1, 2 , Christian Thiede 3 , Mara V.A. Koerner 1 , Shilpa S. Tummala 1 , Dirk Daubner 4 , Ganesh M. Shankar 1 , Erik A. Williams 5 , Maria Martinez-Lage 5 , Silke Soucek 2 , Katja Robel 2 , Tristan Penson 1 , Mechthild Krause 6, 7, 8 , Steffen Appold 6, 7, 8 , Matthias Meinhardt 9 , Thomas Pinzer 2 , Julie J. Miller 10 , Dietmar Krex 2, 7 , Heather A. Ely 11 , Ian M. Silverman 11 , Jason Christiansen 11 , Gabriele Schackert 2, 7 , Hiroaki Wakimoto 1 , Matthias Kirsch 2, 7, * , Priscilla K. Brastianos 12, * and Daniel P. Cahill 1, * 1 Translational Neuro-Oncology Laboratory, Department of Neurosurgery, Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, Massachusetts, USA 2 Department of Neurosurgery, Medizinische Fakultat Carl Gustav Carus, Technische Universitat Dresden, Dresden, Germany 3 Department of Medicine I, Medizinische Fakultat Carl Gustav Carus, Technische Universitat Dresden, Dresden, Germany 4 Institute of Neuroradiology, Medizinische Fakultat Carl Gustav Carus, Technische Universitat Dresden, Dresden, Germany 5 Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA 6 Institute of Radiooncology, Helmholtz-Zentrum Dresden - Rossendorf, Dresden, Germany 7 German Cancer Consortium (DKTK) Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany 8 Department of Radiation Oncology and OncoRay, Medizinische Fakultat Carl Gustav Carus, Technische Universitat Dresden, Dresden, Germany 9 Institute of Pathology, Medizinische Fakultat Carl Gustav Carus, Technische Universitat Dresden, Dresden, Germany 10 Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA 11 Ignyta, Inc., San Diego, California, USA 12 Department of Medicine, Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA * These authors have contributed equally to this work Correspondence to: Daniel P. Cahill, email: cahill@mgh.harvard.edu Priscilla K. Brastianos, email: PBRASTIANOS@mgh.harvard.edu Matthias Kirsch, email: Matthias.kirsch@uniklinikum-dresden.de Keywords: meningioma; telomere; heterogeneity; rearrangements; fusion Received: August 18, 2017 Accepted: October 30, 2017 Published: November 24, 2017 ABSTRACT Background: Recent studies have reported mutations in the telomerase reverse transcriptase promoter ( TERT p) in meningiomas. We sought to determine the frequency, clonality and clinical significance of telomere gene alterations in a cohort of patients with progressive/higher-grade meningiomas. Methods: We characterized 64 temporally- and regionally-distinct specimens from 26 WHO grade III meningioma patients. On initial diagnoses, the meningiomas spanned all WHO grades (3 grade I, 13 grade II and 10 grade III). The tumor samples were screened for TERT p and ATRX/DAXX mutations, and TERT rearrangements. Additionally, TERT p was sequenced in a separate cohort of 19 patients with radiation-associated meningiomas. We examined the impact of mutational status on patients’ progression and overall survival. Results: Somatic TERT p mutations were detected in six patients (6/26 = 23%). Regional intratumoral heterogeneity in TERT p mutation status was noted. In 4 patients, TERT p mutations were detected in recurrent specimens but not in the available specimens of the first surgery. Additionally, a TERT gene fusion ( LPCAT1-TERT ) was found in one sample. In contrary, none of the investigated samples harbored an ATRX or DAXX mutation. In the cohort of radiation-induced meningiomas, TERT p mutation was detected in two patients (10.5%). Importantly, we found that patients with emergence of TERT p mutations had a substantially shorter OS than their TERT p wild-type counterparts (2.7 years, 95% CI 0.9 – 4.5 years versus 10.8 years, 95% CI 7.8 -12.8 years, p=0.003). Conclusions: In progressive/higher-grade meningiomas, TERT p mutations are associated with poor survival, supporting a model in which selection of this alteration is a harbinger of aggressive tumor development. In addition, we observe spatial intratumoral heterogeneity of TERT p mutation status, consistent with this model of late emergence in tumor evolution. Thus, early detection of TERT p mutations may define patients with more aggressive meningiomas. Stratification for TERT alterations should be adopted in future clinical trials of progressive/higher-grade meningiomas.

The purpose of this study was to demonstrate the efficacy of an enrichment protocol for the detection of circulating carcinoma cells in the bloodstream of patients with various urologic cancers. Using 16-ml peripheral blood samples (BS) the mononuclear cells were isolated by density gradient centrifugation. The CD45 leukocyte depletion method based on a previous study was slightly modified and semi-automated by an immunomagnetic cell separation unit (autoMACS). Enriched tumor cells were analyzed on a single slide by cytokeratin (CK) immunocytochemistry. The number of recovered DU-145 prostate cancer cells in various spiking experiments was 70-88%. By the optimized tumor cell enrichment protocol 186 BS originated from 128 patients with different urologic cancers (60 prostate carcinoma, 34 bladder cancers, 24 renal cell carcinoma and 10 other tumors) were investigated before, during and after tumor surgery. In 59 BS from 52 patients on average 5 tumor cells were detected in each BS containing tumor cells. The median number of identified tumor cells was 8 cells per BS and patient. Tumor cells were found for the 3 tumor types with representative BS numbers in 29-39% of the investigated BS and in 38-53% of the affected patients. The detection rates increased in the order prostate carcinoma < renal cell carcinoma < bladder cancer. Surprisingly, in four bladder tumor cases with identified disseminated tumor cells in BS, the histopathological examination of the transurethral resection of bladder tumor specimens showed no evidence for tumor cells in situ but the affected patients had clinically known and histologically defined tumor residue or a bladder tumor recurrence during the follow-up. The semi-automated CD45 autoMACS depletion protocol for the enrichment and the detection of disseminated tumor cells in the peripheral bloodstream allows to study up to 20 BS per working day prospectively by one technician. The improved sensitivity and specificity might be of importance when applying the protocol to BS in future clinical studies.