bluuebunny/crossref_metadata_2025 huggingface dataset:
FDG PET/MR for therapy response assessment in sarcoma.
Authors: Richter Stephan , Beuthien-Baumann Bettina , Kotzerke Joerg , Laniado Michael , van den Hoff Joerg , Ehninger Gerhard , Schuler Markus Kajo , Platzek Ivan
Published: March 2020

e21518

Background: Magnetic resonance imaging is the preferred modality for local staging of sarcoma. Positron emission tomography (PET) allows for the evaluation of tissue metabolism. Recently, PET and MR can be performed as a combined examination on a single scanner. The aim of this pilot study was to evaluate the agreement of PET and MR regarding therapy response in patients who received PET/MR for sarcoma staging and assess the diagnostic value of PET/MR in contrast to standalone MR. Methods: Thirteen patients with soft tissue sarcoma (7 m, 6 f, mean age 50 y, age range 18-80 y) received FDG PET MR on to occasions to assess therapy response. Average time between the two scans was 47 d (21-98 d). Average time between tracer injection and the beginning of the PET scan was 70 min (58-106 min). 4.5 MBq 18F-FDG /kg body weight were applied before the examination (252-331 MBq 18F-FDG /patient, 289 MBq on average). Image evaluation was performed separately by a radiologist and a nuclear medicine physician, respectively, who were blinded for other imaging procedures or clinical data. On MR images therapy response was determined according to the RECIST criteria. PET data evaluation included the calculation of mean an maximum SUV (=standardized uptake value) and metabolic volume. In this case, an increase or decrease of 30% or more of the base value was considered a progressive disease or positive response, respectively. Agreement of PET and MR regarding response was evaluated using Cohen's kappa. Results: The primary tumor showed increased FDG uptake in all patients. Response assessment based on RECIST and SUVmean differed in six of 13 cases (k=0,25). Assessment based on RECIST and metabolic volume showed slightly better agreement (k=0,35). Assessment based on RECIST and SUVmax differed in x cases (k=0,34). Conclusions: Sarcoma therapy response based on morphologic MR imaging and PET parameters yield differing results in many cases. FDG positive sarcoma may show a decrease of metabolic parameters on PET scans, while they appear largely unchanged on MR scans. Due to additional information about tumor metabolism provided by PET, PET/MR is a promising modality for sarcoma imaging. Further studies are necessary to evaluate the prognostic significance of PET findings in sarcoma patients.
Comparison of traditional Chinese medicine (TCM) herbal mixture LQ and paclitaxel on the phase of cancer cell cycle blockage and cancer cell invasion.
Authors: Hoffman Robert M. , Wu Chengyu , Yano Shuya , Zhang Lei
Published: March 2020

e22014

Background: Although Traditional Chinese Medicine (TCM) has been used to treat cancer for thousands of years, the mechanisms of action are either poorly understood or unknown. In the present report, we use state-of-the-art technology to investigate the anti-cancer mechanism of the TCM herbal mixture LQ. Methods: Fluorescence ubiquitination-based cell cycle indicator (FUCCI) was used to monitor cell cycle arrest HeLa cells after LQ treatment. FUCCI-HeLa cells were cultured in two dimensional (2D) monolayer, Matrigel and 3D Gelfoam. Changes of cell cycle status were observed using the Olympus FV1000 confocal imaging system whereby cycling cells fluoresce green and quiescent cells red. Paclitaxel (Taxol) was used as the positive control. Results: Paclitaxel induced a G2/M cell cycle block. In contrast, LQ blocked FUCCI-HeLa cells in the G0/G1 phase of the cell cycle in all 3 culture models. In monolayer culture, the paclitaxel positive control had approximately 45% of the cells in G2/M phase. In contrast, the LQ-treated cells were mostly in the G0/G1 phase (>90%). In Matrigel culture, HeLa cells formed spheres. The spheres in the paclitaxel control group had 40% of the cells in G2/M phase, but only 15% in LQ-treated cultures. In 3D Gelfoam culture, cells grew along the structures of the Gelform. The paclitaxel positive control culture had approximately 45% of cells in G2/M phase. In contrast, the cells in LQ-treated culture were mostly in G0/G1 phase (>80%). The cells in pacilitaxel control group invaded to 250~300 µm deep in the Gelfoam, but only 150~200 µm deep in LQ-treated culture. Conclusions: The anti-cancer mechanism of TCM herbal mixture LQ involves cancer cell cycle arrest at G0/G1 and inhibition of cancer cell invasion.
Circulating tumor cells (CTCs) and epithelial mesenchymal transition (EMT) in primary breast cancer.
Authors: Karaba Marian , Mego Michal , Minarik Gabriel , Benca Juraj , Sedlackova Tatiana , Manasova Denisa , Sieberova Gabriela , Hlavata Zuzana , Jurik Andrej , Minarik Tomas , Macuch Jan , Gronesova Paulina , Oravcova Eva , Svabova Veronika , Sufliarsky Jozef , Pechan Juraj , Reuben James M. , Mardiak Jozef
Published: March 2020

e22023

Background: CTCs are prognostic in breast cancer (BC) patients and represent a heterogeneous population of cells with different phenotypes and biological value. EMT gives rise to cells with stem cell-like properties with increased chemotherapy resistance and may be responsible for under detection of CTCs by currently approved assays. The aim of this study was to characterize CTCs based on the expression of epithelial and mesenchymal markers in non-metastatic BC patients. Methods: This prospective ongoing study included 124 patients (pts) with non-metastatic BC enrolled from March 2012 to December 2012. CTCs were detected before surgery (124 pts), before 1st cycle (28 pts) and before 2nd cycle (23 pts) of adjuvant therapy. Isolated peripheral blood mononuclear cells (PBMC) were depleted of hematopoietic cells using RossetteSep kit negative selection with anti-CD45 antibody. RNA extracted from CD45-depleted (CD45-) PBMC was interrogated for expression of EMT-inducing transcription factors (TWIST1, SNAIL1, SLUG, ZEB1) and epithelial (CK19) gene transcripts by qRT-PCR. Expressions of gene transcripts in CD45- PBMC of patients were compared to those of CD45- PBMC of 60 healthy donors. Results: CTCs were detected in 26.6% pts before surgery, in 21.4% pts before 1st cycle and in 43.5% of pts before 2nd cycle of adjuvant therapy. Before surgery, CTCs exhibiting only epithelial makers were present in 12.9% pts, whereas CTCs with only EMT markers were observed in 12.1% of pts and CTCs coepressing both markers were detected in 1.6% pts. There was a trend for higher CTC positivity in high grade, triple negative tumors and in the presence of lymphovascular invasion. We detected CTCs with only an EMT phenotype more often in pts with N+ than in N0 disease (20.4% vs. 10.2%; p = 0.03) and in pts after 1stcycle of adjuvant chemotherapy compared to before surgery (30.4% vs. 12.1%; p = 0.03). There was no association between epithelial CTCs and analyzed clinico-pathological variables. Conclusions: Our data suggest that CTCs with EMT phenotype are involved in tumor dissemination and treatment resistance, and support the role of EMT in cancer pathogenesis. Moreover, these results underlay the concept of CTCs heterogeneity.
Prognostic role of microvascular density (DMV), VEGF, EGFR, and HIF-1 α expression and their relationship with epitelial mesenquimal transition (EMT) in patients with locally advanced cervical cancer (LACC) treated with chemoradiotherapy.
Authors: Rojas Puentes Luis Leonardo , Cardona Zorrilla Andres Felipe , Cetina Lucely , Otero Jorge Miguel , Carranza Hernan , Vargas-Baez Carlos A.
Published: March 2020

e22028

Background: In low and middle income countries cervical carcinoma is the second leading cause of death among women. Methods: To describe the prognostic value of MVD and protein expression of several genes related to survival and proliferation and their relationship with EMT of LACC. Overall response rates (ORR), clinical benefit (CB), progression-free survival (PFS), and overall survival (OS) were estimated for women suffering from LACC treated with chemoradiotherapy followed by high dose rate brachytherapy (HDRB). Results: Sixty-one patients were included (mean age 52±10); all of them had LACC (2.3% 2A/47.5% 2B/4.9% 3A/37.7% 3B/3.3% 4A/3.3% not defined), a tumour mean size of 6.4cm (SD±1.8cm) and HPV infection in 46% of the cases. Fifty-eight patients (95%) had a squamous pattern, two were adenocarcinomas and >50% presented moderated or poorly differentiated neoplasms. All of them were treated with chemotherapy (transitory interruption of external radiotherapy was documented in 19% due to toxicity and in 21.4% of cases by other causes/mean cycles of platinum administered during radiotherapy 4.8 ± 1.0) and brachytherapy (77% completed all planed intracavitary treatment). The median PFS and OS was 6.6 months (4.0-9.1) and 30 months (11-48) respectively. None of the variables had effect on PFS, whilst multivariate analysis revealed that VEGF (p=0.026) and EGFR expression levels (p=0.030) and <6 cm tumour volume (p=0.02) positively influenced the OS. TWIST expression and E-cadherine lost expression (E-) also influence OS (p=0.001) and (p=0.02) respectably and are related with VEGF and EGFR expression. When EGFR or VEGF are coexpressed with TWIST or related with E- expression the patients shows the worst OS (median 14 m CI 95% 3-24 m, p=0.02 for EGFR (+) and E- ; VEGF (+) and E - median 14 m CI 95% 3-25 m p=0.0001; EGFR (+) and TWIST (+) 19 m CI 95% 15-80 m, p=0.006; VEGF (+) and TWIST (+) 18 m CI 95% 15-80 m, p=0.0001. Conclusions: Classifying LACC patients treated with cisplatin-based chemoradiotherapy by protein expression and EMT markers had a positive influence on prognosis.
In vivo isolation of circulating tumor cells in prostate cancer patients by a medical device.
Authors: Theil Gerit , Hoda M.Raschid , Fischer Kersten , Schumann Andre , Haubold Kathrin , Boehm Catrin , Krahn Thomas , Luecke Klaus , Morgenthaler Nils G. , Fornara Paolo
Published: March 2020

e22055

Background: CTCs are discussed as a prognostic and stratification biomarker, and may help to assess the treatment efficacy. The aim was to demonstrate proof of concept of in vivo CTC isolation in Prostate cancer patients (PCa). Methods: A medical device (CellCollector) was inserted in a cubital vein for 30 minutes. The interaction of target CTCs with the CellCollector was mediated by antibodies to the epithelial cell adhesion molecule (EpCAM). To confirm binding of CTCs to the wire, immunohistochemical staining against Cytokeratin and CD45 was performed. There were 226 applications of the device in 16 metastasized PCa (PCa-m) with up to 8 wire applications per patient, 24 localized PCa (PCa-l) with up to 3 wire applications per patient as well as 19 men with benign prostate hypertrophy (BPH) and 21 women as control group with only one wire application. CTC counts from 22 PCa patients with 58 applications were directly compared to the CellSearch technology. Results: We obtained in vivo isolation of CTCs in 73 of 98 applications to PCa patients (74.5 %). The sensitivity for metastasized PCa was (86.2%) and for local PCa (57.5%), respectively. Follow up data of one patient with PCa-m: 2009 diagnosed PCa with multiple metastases. 2 years later PSA of 11.2ng/mL and 25 CTCs. After transurethral resection and removal of a brain metastasis, CTCs decreased to 20 cells with no effect on PSA levels. 4 months later we observed 187 CTCs and a PSA level of 76ng/mL, which was correlated with the patient condition. In PCa-l patients we observed prior radical prostatectomy (n=18) and 6 (n=15) and 12 (n=7) months after surgery CTC numbers on average of 18.2, 2.8 and 1.4, respectively. In the direct comparison of the CellCollector and CellSearch we showed detection rates of 65% (13/20) and 20% (4/20). Conclusions: We demonstrate the use of CTCs besides PSA during therapy adjustment in PCa patients In summary the CTC detection rate of the CellCollector in PCa-m and PCa-l in comparison to the CellSearch method increased by 26% and 45%. Further molecular genetic CTC characterization as well as multiple antibody coating at the CellCollector will assume crucial information for therapy decisions. Clinical trial information: ISRCTN10403616.
The sensitivity and specifity of DR-70 as a tumor marker for gastrointestinal cancers.
Authors: Erkol Burcak , Oven Basak Bala , Bayoglu Vedat , Paker Nurcan , Kochan Koray , Ozkara Selvinaz , Tilki Metin , Surmelioglu Ali , Yesil Atakan , Gonen Can
Published: March 2020

e22061

Background: There is a great need to detect gastrointestinal cancers at an early stage, reduce morbidity and mortality. CEA and CA 19-9 are widely used serological markers for gastrointestinal cancers during follow-up. Their utility are limited because of their low sensitivity. In this study, a novel tumor marker DR-70 is evaluated for the sensitivity and specificity, relationship between clinical parameters and compared with CEA and CA 19-9. Methods: Blood sera of 101 histologically proven gastrointestinal cancer patients (38 female, 63 male; 42 colon, 25 rectum, 18 stomach, 9 pancreas, 4 esophagus and 3 cholangiocellular carcinomas) and 105 healthy blood donors were included. The TNM stage of the disease and histologic grade was shown. All patients and controls were also tested for CEA and CA 19-9 levels. In DR-70 immunoassay, the fibrin degradation products were quantitatively measured using ELISA DR-70 kits. Results: The median DR-70, CEA and CA19-9 levels were 1.6 µg/mL, 3.05 ng/mL and 22.2 IU/L in cancer patients, they were lower in healthy control 0.5 µg/mL, 0.9 ng/mL, 1.9 IU/L; p < 0.001. While the stage and grade increased, the levels of DR-70, CEA and CA-19-9 (p < 0.05) were increased significantly. DR-70 and CA19-9 values were highest in patients with pancreas, and lowest in rectum cancer, there was no statistically significant difference between tumor localization and CEA levels.The sensitivity, specificity and the cut-off value of DR-70, in patients with malignancy by using ROC analysis were found as 97%, 95% and < 0.75 µg/mL, respectively. The positive and negative predictive values were 95%, and 97% and the efficacy was 96%. Sensitivities of CEA (cut off value 3ng/mL) and CA 19-9 (cut off value 37 U/ml) were 52% and 39% respectively, the specificities were 94% and 99% for CEA and CA 19-9, respectively; sensitivity increased by combined use of both of them. Conclusions: Due to its high sensitivity and specificity, high positive and negative predictive values DR-70 can be used more frequently as a tumor marker for detection of gastrointestinal cancer. It can also be used in tumor progression and treatment follow-up because of positive correlation between DR-70 levels and the tumor stage.
Prediction of therapy failure after VEGF-based first-line treatment of kidney cancer patients by DNA hypermethylation biomarkers.
Authors: Peters Inga , Dubrowskaja Natalia , Abbas Mahmoud , Seidel Christoph , Kogosov Michael , Scherer Ralph , Gebauer Kai , Merseburger Axel S. , Kuczyk Markus A. , Grünwald Viktor , Serth Juergen
Published: March 2020

e22076

Background: Kidney Cancer (RCC) is one of the top ten causes of cancer dependent death. Anti-VEGF targeted therapy has demonstrated increased progression-free (PFS) and overall survival in RCC patients, though the clinical outcome is still poor overall. Stratification of patient treatment currently depends on different clinical scoring systems such as the Memorial Sloan Kettering Cancer Center score. However, identification of molecular phenotypes of RCC patients could improve stratification of targeted therapies and prediction of clinical course. We investigated whether DNA hypermethylation-based markers can predict the PFS following first-line therapy. Methods: We examined primary tumor tissues obtained from 18 patients receiving anti-VEGF targeted therapy. Quantitative methylation-specific PCRs were carried out for CpG island (CGI) methylation analysis of 5 candidate genes CST6, GATA5, LAD1, hsa-miR124-3 and hsa-miR9.1 identified in advance as potential prognosticators for RCC. For Kaplan Meier survival analysis relative methylation values were uniformly dichotomized and logrank statistics calculated. The analyses of sensitivity and specificity were carried out using a cutoff of 6 months for PFS to categorize for therapy failure and response. Results: Hypermethylation of CST6 and LAD1 shows a highly significant association with a shortened PFS following first-line therapy (p=0.009, p=0.004, logrank test). The median survival observed for the high and low methylation group was 2.0 and 11.4 months for CST6 and LAD1. Moreover, LAD1 methylation showed a specificity of 1.0 (0.65-1.0, 95%CI) and a sensitivity of 0.73 (0.43-0.90, 95%CI) for detection of therapy failure. For CST6a specificity of 0.86 (0.49-0.97, 95%CI) and a sensitivity of 0.82 (0.52-0.95, 95%CI) was detected. Conclusions: The analysis of our discovery cohort revealed that CGI methylation of CST6 and LAD1 predicts a shortened PFS of patients after anti-VEGF therapy. Provided, these results can be confirmed in an independent larger evaluation cohort, detection of hypermethylated loci in primary RCC would allow, to our knowledge for the first time, a molecular-based clinical management of patients.
Effect of exogenous triiodothyronine [T3] on free thyroxine depletion and tumor response: A compassionate care study.
Authors: Hercbergs Aleck
Published: March 2020

e22077

Background: A thyroid hormone receptor on plasma membrane integrin av β3 initiates the actions of the hormone thyroxine [T4] on cancer cell proliferation and angiogenesis, initiated nongenomically, independently of the classical thyroid nuclear receptor. In studies of recurrent glioma, induced hypothyroidism significantly prolonged survival but associated fatigue has restricted acceptance of this approach. Triiodothyronine[T3] is the metabolically dominant hormone but in physiological extracellular concentrations is 10-100 fold less mitogenic than T4 in solid tumor models (R Meng et al., PLoS 2011; 6[11]:e27547). Exogenous T3 also maintains eumetabolism but inhibits TSH release to induce free T4 depletion [FT4D] defined as[< 0.93 ng/dL.] Maximal therapeutic free T4 depletion [FT4D] by T3 may therefore be feasible if combined with methimasole[MET]. Aims of study: Retrospective analysis of 1.) exogenous T4 withdrawal and 2.) daily exogenous T3 [ET3] and MET on tumor response and survival Methods: 21 Patients at geographically dispersed sites with recurrent or stage IV solid tumors: GBM 8 patients, brain stem glioma 1, pancreas 1, salivary gland 2, Lacrimal duct 1, breast 1, mesothelioma 1, ovarian 4, lung 2. 1.) In 7 patients exogenous T4 was terminated and replaced by exogenous T3 [ET3] 15 to 30μg daily in three divided doses. 2.) 15 euthyroid patients received 15-30μg ET3 and 20-40 mg. methimasole [MET] daily to induce [FT4D] usually obtained within 6-10 weeks. Results: There was no treatment related toxicity. 3/7 patients had PR to withdrawal of T4 only. 20/21 patients experienced PR[16] or CR[4]. to concurrent or sequential chemo,hormonal or radiation therapy for an overall response rate of 90% CHT elicited repeated responses in some patients. The 6 month PFS for the GBM cohort [n=8] was 70%. Two survived 36 and 53 months. For other solid tumor types survival was above /in the 90th percentile of reported survival studies. Conclusions: A high response rate was observed in advanced compassionate care cancer patients undergoing exogenous T3 modulated FTD. Tumor response to exogenous T4 withdrawal is a novel observation with potentially important therapeutic implications.
Development of a high-performance, blood-based screening diagnostic to detect early-stage breast cancer.
Authors: Wagner Joseph , Chapman Karen , Prendes-Garcia Maria , Lacher Markus , Kidd Jennifer , Sheibani Nadia , Tam Orlena , West Michael
Published: March 2020

e22083

Background: Limitations of current screening mammography, particularly in younger women, demonstrate the need for an alternative breast cancer screening strategy. A non-invasive, easily interpreted and low cost test should address this need. Methods: Gene expression microarray analysis was carried out on 128 individual tumor samples representing over 20 tumor types, 86 samples representing 31 diverse normal tissue types, 68 tumor cell lines and 97 diverse normal primary cell cultures. Genes were ranked for elevated expression in either: i) a large number and variety of tumors relative to normal tissues, or ii) in breast tumors. Elevated expression was verified for a subset of genes using qPCR in a set of independent RNA samples. Proteins coded by genes elevated in breast cancer samples were analyzed in a retrospective training set of breast cancer patient sera samples with cancer-free patient and benign pathology controls using ELISA or bead-based detection assay. Results: Based on availability of suitable reagents, 25 candidate biomarkers were assessed in patient sera samples (31-227 patient samples per biomarker) using ELISA or bead-based assays. Individually, the performance of individual markers varied (ROC AUC, 0.51 - 0.88); however, when expression levels of the best performing markers were combined, the multiplex test demonstrated high-sensitivity (>80%) and specificity (>90%) in identifying early-stage breast cancer patients. Conclusions: A multiplex, proteomic-based approach may provide for a high-performance, blood-based screening diagnostic for breast cancer.
Effect of a small-molecule MDM2/MDMX inhibitor on cell cycle arrest and apoptosis in breast cancer cells.
Authors: Geng Qian Qian , Li En Xiao , Dong Dan Feng , Wu Yin Ying , Wang Jie , Li Yi
Published: March 2020

e22096

Background: The MDM2 inhibitor which disrupted the MDM2-p53 interaction made little effort on the activation of p53 for breast cancer treatments, due to MDMX over expression. Previously a small molecule inhibitor targeting at MDM2 and MDMX had been successfully synthesized. We tested anti-tumor activity of the small molecule MDM2/MDMX inhibitor, compared with nutlin-3α, a well characterized MDM2 inhibitor, in the wild-type (wt) and mutant (mt) p53 breast cancer cell lines. Methods: Human breast cancer cell lines MCF-7 (wt-p53), ZR-7530 (wt-p53), BT-474 (mt-p53) and MDA-MB-231 (mt-p53) were cultured and treated with the small molecule MDM2/MDMX inhibitor, nutlin-3α or phosphate buffer solution (PBS) for 48 hrs separately. MTT for cell viability, FCM for cell cycle arrest and Annexin V FITC/PI for cell apoptosis were performed. The mechanism of antitumor activity of the small molecule MDM2/MDMX inhibitor was determined by Western-Blot analysis. Results: The inhibitor of MDM2 and MDMX inhibited cell growth and induced cell cycle arrest and apoptosis in mt-p53 breast cancer cells while nutlin-3α cannot. In the breast cancer cells with wt-p53, both of them inhibited cell growth, induced cell cycle arrest and apoptosis, but MDM2/MDMX inhibitor was proven to be more effective. Western Blot revealed that there was higher level of p53 expression in breast cancer cells treated with MDM2/MDMX inhibitor than nutlin-3α. The marked increases in p21, Bax and PUMA expressions were abserved in both wt-p53 and mt-p53 breast cancer cells which indicated that the small molecule MDM2/MDMX inhibitor induced cell apoptosis through p21, Bax and PUMA over expressions. Conclusions: The small molecule MDM2/MDMX inhibitor would be able to suppress cell proliferation, induce cell cycle arrest and apoptosis, activate p53 more effective than nutlin-3α in breast cancer cells, no matter with p53 status. P21, Bax and PUMA were involved in the mechanism of apoptosis induction. The inhibitor of targeting at both MDM2 and MDMX will be a novel treatment for breast cancer p53-independent status in the future.
Using low sensitivity of direct sequencing to detect K-ras mutations? A new method for improving diagnostic accuracy.
Authors: Chen Chi-Kuan
Published: March 2020

e22100

Background: Identifying K-ras mutation has became a standard procedure in cancer treatment. Colorectal cancer patients with K-ras mutation are likely to poorly respond to cetuximab. Therefore, detecting K-ras gene mutations should be suggested before the selection of personalized treatment in colorectal cancer. To date, general molecular biology techniques contain HRM, PCR-RFLP, TaqMan PCR and CE-IVD-validated Cobas 4800 KRAS (Roche Diagnostics) are used for K-ras mutation detection in molecular diagnosis, but the sensitivity limitation of these method is approximately 1%. Methods: Therefore, we used a new approach, a universal genetic detecting method (FemtoPath), which improves sensitivity of K-ras mutation detection and the limitation of sensitivity is closed to 0.1% of mutation type. Results: We compared the sensitivity between FemtoPath/direct sequencing test and Cobas KRAS real-time PCR. Cobas real-time PCR identifies mutations in 21 (40.38%) of the 52 tumors. Surprisingly, the FemtoPath/direct sequencing test identified mutations in 40 (76.92%) of the 52 tumors. Our data showed that the FemtoPath/direct sequencing test can identify 19 additional mutation samples. In addition, the FemtoPath/direct sequencing test can identify more unknown K-ras mutations adjacent to codon 12 and 13. Conclusions: Ensure the most timely and appropriate therapy for cancer patients is the first priority of clinical application. FemtoPath/direct sequencing test is more sensitive, accurate and inexpensive and needs fewer sample amount than Cobas real-time PCR.
Evaluation of proliferation and apoptosis markers in circulating tumor cells (CTCs) of women with early breast cancer who are candidates for tumor dormancy.
Authors: Spiliotaki Maria , Markomanolaki Haris , Kallergi Galatea , Papadaki Maria A. , Georgoulias Vassilis , Mavroudis Dimitrios , Agelaki Sofia
Published: March 2020

e22101

Background: Clinical dormancy is frequently observed in breast cancer (BC). In the present study, we aimed to characterize CTCs in dormancy candidates (DC) with BC in terms of proliferation and apoptosis. Methods: Cytospins of peripheral blood mononuclear cells (PBMCs) were obtained from DC (n=122) disease-free for ≥5 yrs and from metastatic pts (n=37) on relapse that occurred ≥5 yrs after surgery. Sequential samples (n=27) of 8 DC with late relapse and from 8 relapse-free (n=38), were also analyzed. 106 PBMCs were stained with pancytokeratin antibody along with ki-67 and M30 as proliferation and apoptosis markers, respectively. Results: CTCs were identified in 40 (32%) of 122 DC. In 25 (61.5%), all CTCs were ki-67(-)/M30(-), 7 (17.5%) had ki-67(+), 4 (10%) M30(+) CTCs and 4 had both phenotypes. Among 243 CTCs detected, 201 (82.5%) were ki-67(-)/M30(-), 14 (5.7%) ki-67(+) and 29 (11.9%) M30(+). Of 13 (35%) CTC(+) metastatic pts, 6 (46%) had ki-67(+) CTCs (p=0.037) whereas 54 (40%) of total CTCs were ki-67(+) (p<0.001). No M30(+) CTCs were detected. When sequential samples of the 8 DC who relapsed were analyzed, 2 (25%), had only ki-67(-)/M30(-) CTCs, 6 (75%) had ki-67(+) and 4 (50%) M30(+) CTCs. The respective numbers in the non-relapsed group were 4 (50%), 3 (37.5%) and 3 (37.5%). Moreover, ki-67(+) CTCs were more frequently observed in samples of the relapsed pts (29.6% vs 13.1%). Among 382 CTCs detected in all sequential samples from the relapsed group, 337 (88.14%) were ki-67(-)/M30(-), 26 (6.7%) ki-67(+) and 20 (5.1%) M30(+), whereas the respective percentages among 78 CTCs in non-relapsed pts were 77.6%, 6.5% and 15.8% (p=0.001). Conclusions: The great majority of CTCs detected in DC with BC express neither proliferation, nor apoptosis markers. However, the apoptotic index in CTCs is increased during clinical dormancy, whereas the proliferation index is increased on relapse. In addition, apoptotic CTCs are more frequently encountered during follow-up in DC free of relapse. The above observations suggest that monitoring proliferation and apoptosis in CTCs during clinical dormancy could be used to predict subsequent late relapse.
Impact of a multigene prognostic assay on postoperative management of early-stage non-small cell lung cancer.
Authors: Dormady Shane , Jahan Thierry , Mann Michael J. , Jablons David , Broder Michael S. , DeCristofaro Alison H. , Anene Ayanna
Published: March 2020

e22124

Background: Many stage II and most stage I non-small cell lung cancer (NSCLC) patients forgo adjuvant chemotherapy, despite the likely presence of occult metastasis in the 30-60% of these patients who die within 5 years of surgery, and despite demonstration that patients with undetected, i.e. very early, metastasis can achieve long-term survival with this intervention. A multi-gene assay using quantitative RT-PCR and run on formalin-fixed, paraffin-embedded samples (Pervenio Lung RS) has been independently validated in two large international studies to better differentiate high- and low-risk non-squamous NSCLC patients than either traditional risk assessment criteria or conventional staging. A survey of physicians who used this multi-gene assay probed whether better risk discrimination changed post-operative management. Methods: Physicians were queried via a web-based survey on practice patterns, patient characteristics and treatment recommendations. The survey was developed through cognitive interviews with oncologists and thoracic surgeons. Physicians were allowed to respond regarding up to 4 patients (stage I or II) for whom the multi-gene test was ordered. Results: Forty-one physicians (12 surgeons, 29 oncologists/internists) of 61 contacted (67%) provided responses on 77 patients. Community practices accounted for 83%, and the average years in practice was 15. Seventy-one percent provided responses on either 1 or 2 patients. Mean patient age was 66±10 years, with stage I and stage II comprising 73% and 27%, respectively. Prior to obtaining assay results, chemotherapy was recommended in 30% of patients, including 5 stage I patients. The assay characterized 5-year mortality risk as high, intermediate and low in 52%, 23% and 25% of patients, respectively. Treatment recommendations were changed post-assay in 36% of patients, including 73% of high-risk patients for whom observation was recommended initially. Change in treatment intensity correlated with the degree of assay-assessed risk. Conclusions: Refinement of risk assessment achieved via a multi-gene prognostic assay resulted in changes in treatment recommendations for a substantial number of early stage NSCLC patients.
Towards qualification of FDG PET as biomarker of response to neo-adjuvant therapy: A meta-analysis.
Authors: Vincent Andrew D. , Rizivi Saiyada N. F. , van Tinteren Harm , Riphagen Ingrid , Hoekstra Otto S.
Published: March 2020

e22128

Background: For FDG PET to qualify as a biomarker of response to cancer therapy beyond proof of principle, the evidence should be aggregated. Methods: A systematic literature search (Jan 1995-May 2011) for studies in solid extracerebral tumors with distinct cohorts of n≥10 patients, PET before and after cytotoxic neoadjuvant systemic therapy (with or without radiotherapy), dichotomized histopathological (PA) response. For model development we selected studies reporting individual or dichotomous aggregated level data, and performed bivariate SROC meta-regression. Results: 52 studies were identified, comprising data from 1,710 patients: rectal (32%), esophageal (21%), sarcoma (20%), breast (13%) and other cancers (14%). Most PET measures were SUVmax (65%) and other SUV measures (32%). Treatment type ChT (38%) and ChRT (62%); 46% were PA responders. The meta-regression indicated a weak increase in specificity for chemotherapy studies over chemoradiotherapy studies (p=0.08), however this disappeared after adjusting for publication bias. High baseline uptake rates improved diagnostic sensitivity (p=0.01). The SROC-AUC after adjusting for publication bias was 0.77. Maximal sensitivity and specificity was achieved with a PET reduction of 60%. Conclusions: There is a moderate association between FDG-PET change and histopathological response. This relation appears to be a function of baseline uptake, which may (at least in part) relate to repeatability issues.
DNA repair gene promoter methylation in non-small cell lung cancer (NSCLC) patients treated with DNA damaging agents.
Authors: Buckingham Lela , Pelkey Gregory , Fidler Mary J. , Bonomi Philip D.
Published: March 2020

e22130

Background: Approximately 75% of NSCLC patients are advanced at time of diagnosis. Treatments include regimens of DNA-damaging and alkylating agents. Previous in vitro studies have shown that loss of BRCA1 and BRCA2 function through promoter methylation increases sensitivity to sapacitabine, suggesting that tumor cells may be protected by repair systems including BRCA-RAD50 and MGMT pathways induced by DNA damage. The purpose of this study is to investigate the effect of DNA repair gene promoter methylation on outcome of NSCLC patients treated with DNA damaging agents. DNA repair gene promoter hypermethylation may sensitize tumor cells to DNA damaging agents. Methods: The patient group of 240 patients included 135 early stage and 105 late stage (III/IV) cases. Chemotherapy regimens of late stage patients included carboplatin, cisplatin and gemcitabine, as well as other non-DNA damaging agents. Mean overall survival (OS) was 43.7 months. DNA was extracted from micro-dissected primary tumor, bisulfite converted and assessed by pyrosequencing to quantify methylation of cytosine nucleotides along the MGMT, BRCA1 and BRCA2 promoters. Results: Overall average methylation values were 8.4%, 7.3% and 8.3% for MGMT, BRCA1, BRCA2 respectively (compared to 3.2%, 3.6% and 7.3%, respectively, in nonmalignant lung tissue). Percent promoter methylation levels were not significantly correlated with age, gender, smoking status nor histology, with the exception of higher MGMT promoter methylation in smokers. Hypermethylation (greater than 10%) of BRCA1 and BRCA2 promoter was not significantly associated with survival in late stage patients (6.6 mos vs 6.8 mos; p=0.533) in this patient group, however, superior survival was observed with BRCA2 promoter hypermethylation when early stage patients were included (29.3 mos vs median not reached; p=0.045). Promoter hypermethylation of MGMTwas significantly associated with lower five-year survival rate in early stage patients (p=0.022). Conclusions: These data suggest epigenetic control of DNA repair gene expression can affect response to therapy. Further studies on specific treatment regimens will be required to definitively assess these effects.
The key role of correlation between MR spectroscopy choline peak and calcium sensing receptor for breast cancer: Diagnosis by a 3T MR scanner.
Authors: Baio Gabriella , Valdora Francesca , Pace Daniele , Salvi Sandra , Villosio Nicolo , Truini Mauro , Calabrese Massimo , Neumaier Carlo Emanuele
Published: March 2020

e22132

Background: The Ca2+-sensing receptor (CaSR) regulates the Ca2+ signaling and its expression is present in both normal and malignant breast tissues. Recently, we demonstrated in a pre-clinical setting the CaSR expression in breast cancer mouse model by manganese enhanced MRI (Baio G et al. Eur Rad 2011). Alteration in choline phospholipid metabolism as detected by MRI, is a common feature of breast and many other cancer cells or tumors. Since CaSR-ChoK signaling plays an important role in breast cancer, we compared the MRS (MR spectroscopy) choline peak by a 3T MR scanner between calcium sensing receptor positive and negative breast cancers. Methods: Breast MRI of 16 consecutive patients with breast lesions were performed by a 3T MR and MRS findings were defined as positive by the visual inspection for the presence or the absence of choline peak. MRS imaging were compared with histologic findings and the pattern of CaSR by immunohistochemistry. The immunohistochemical results were qualitatively classified according to intensity and pattern of the staining, using a 6-point scale to score the intensity of the CaSR expression (0, absent; 5, intense, widespread expression). Results: A choline peak was present in 10 of 15 malignant lesions and the CaSR expression was between 3 to 5 score at immunohistochemestry analysis (p<.0006). In 4 of 15 malignant lesions the choline peak was absent or not consistent at the visual analysis with a CaSR patterns of 1-2. In one benign lesion (papillary hyperplasia), the choline peak was consistent and the CaSR pattern was with score 3. Conclusions: In this study we evaluated if MRS presence or absence of choline peak well correlated with CaSR positive and negative breast cancers. We observed a choline peak with an high level expression of CaSR (score 3-5) in 67% of the breast lesions, while in 27% of the lesions the choline peak was absent with a low CaSR expression (score 1-2). These preliminary results support the hypothesis that CaSR represents an important role in the production of choline in breast cancer, determining an increase of his production when CaSR is expressed at high level and implicating and absence of choline peak when CaSR is not expressed.
A single-center experience with CELLSEARCH system circulating tumor cell test on patients with metastatic breast cancer.
Authors: Montero Alberto J. , Eapen Sara , Andrews Ryan M. , Gorin Brian
Published: March 2020

e22135

Background: Clinical studies have shown that circulating tumor cell (CTC) detection via the CELLSEARCH assay is a validated prognostic marker that is a predictor of overall survival (OS) in metastatic breast cancer (mBC) patients. By analyzing real-world observational data in one center, we evaluated the impact of CTC testing on predicting OS. Methods: Electronic medical records (EMR) were used to identify patients who were: (i) women > 18 years diagnosed with mBC on or after their date of registration, (ii) registered to the EMR system on or after 4/1/2010, and (iii) followed for at least 2 months from the time of metastases. Patients who had CTC testing were required, on average, to have at least 1 valid CTC test result every 3 months. OS from the date of initial diagnosis of metastases among mBC patients with CTC testing was compared to a cohort of patients without CTC testing (non-CTC) using a Cox model controlling for covariates such as age, race, hormone receptor status, and scanning frequency. Results: Mean (SD) ages of CTC (N=79) and non-CTC (N=1,146) patients were 65 (13) and 64 (14) years, respectively. ER or PR+ patients comprised 67% of the CTC population, and 64% of the non-CTC population. The majority of patients in both cohorts were Caucasian (89% - CTC, 57% - non-CTC). The Cox model showed that the group where CTC testing occurred had a statistically significant 40% reduction in risk of death (HR=0.60, p-value=0.03); this reduction was even greater among ER or PR+ patients who received CTC testing compared to those who did not (51%, HR=0.49, p-value=0.03). Conclusions: This retrospective analysis shows that the CTC population had a better prognosis than the non-CTC population. Thus, the assessment of CTCs could be associated with improved monitoring and prediction of cancer progression, and improved evaluation of response to therapy in mBC patients, possibly guiding more informed care decisions. The reliance on a single center and the relatively small number of CTC patients limits the ability to rule out other potential confounders in the analysis. Further studies using larger CTC samples and data from other centers with CTC testing are needed to verify and better understand this result.
Correlation between biopsy type and insufficient tissue availability for biomarker testing in five solid cancer types.
Authors: De Richter Pieter , Ilacqua Jackie
Published: March 2020

e22136

Background: Limited tissue availability is a known barrier to conducting biomarker testing on cancer tissue specimens. It has been assumed that the frequency of insufficient tissue being available is correlated with type of tissue biopsy performed, but this has not previously been quantified across multiple cancer types. Methods: A panel of pathologists in the US (N=90) were asked to report on recent cancer tissues they prepared/handled for biomarker testing. The physicians were asked to report on the cancer type, biopsy method, molecular diagnostics tests conducted, and whether sufficient tissue was available for all requested tests. If insufficient tissue was available, they were asked to report on which tests they were unable to perform as a result. All data was collected through an online survey between August and November 2012. Results: In total, 99 breast cancer samples, 85 NSCLC samples, 60 CRC samples, 30 gastric cancer samples and 40 melanoma samples were reported on. Insufficient tissue was available in 5 (6%) of all NSCLC samples, 2 of which were obtained through fine needle aspirations (14% of all FNAs), a further 2 through core biopsies (5%), and a further 1 through a bronchoscopy (8%). Insufficient tissue was available in 1 (1%) of all breast cancer samples (core biopsy, 1% of all core biopsies), 1 (3%) of all melanoma samples (core biopsy, 13% of all core biopsies) and 1 (2%) of all CRC samples (core biopsy, 11% of all core biopsies). Conclusions: Insufficient cancer tissue for biomarker testing occurred across 4 out of 5 cancer types reported on, though was rare (<5%) in cancers other than NSCLC. The majority of tissue samples in which insufficient tissue was present were acquired through core biopsies (67% of all cases) or FNAs (22% of all cases).
Effect of tumor-treating fields on DNA repair in cancer cell lines.
Authors: Kirson Eilon David , Giladi Moshe , Schneiderman Rosa S. , Porat Yaara , Munster Mijal , Weinberg Uri , Palti Yoram
Published: March 2020

e22138

Background: TTFields Therapy is an antimitotic treatment modality approved by FDA for the treatment of patients with recurrent glioblastoma (GBM). TTFields act by disruption of spindle microtubule arrangement during metaphase and interference with cytokinesis during anaphase and telophase. These effects are the result of rotation of charged and/or polar macromolecules in the direction of the applied antimitotic fields. We hypothesized that the negatively charged, double stranded DNA fragments induced by ionizing radiation (RT) or ultraviolet light (UV) undergo similar rotation. Double strand break repair relies on proper alignment of the strands during the process of homologous recombination. Thus application of TTFields during the DNA repair process holds the potential of interfering with normal DNA repair. Methods: Cancer cells (786-O renal cell carcinoma) were exposed to TTFields for various durations up to 4 hours after UV or RT treatment. The extent of DNA repair over time was evaluated using the alkaline comet assay (Trevigen, USA). This assay allows the computation of the relative proportion of intact DNA strands and DNA fragments in individual cells after inducing DNA breaks. Results: Exposure of UV treated cells to TTFields for 4 hours led to a significant increase in the content of DNA fragments per cell compared to the content without TTFields exposure (45 + 14%; p=0.012). This increase was accompanied by a significant decrease in the intact DNA content per cell compared to the content of cells not exposed to TTFields (47 + 14%; p=0.005). Inhibition of DNA repair was apparent 1-4 hours after irradiation depending on the extent of DNA damage. Similar results were seen after exposure of the cultures to 2Gy RT (17% increase in DNA fragment content and 14% decrease in intact DNA content at 4 hours). Interestingly, while TTFields did not lead to any DNA damage when applied alone, the application of TTFields for one hour after UV exposure led to a 27 + 29% (p=0.02) increase in the extent of the DNA damage (fragment content). Conclusions: This is the first demonstration that, in addition to its known anti-mitotic effect, TTFields Therapy also inhibits DNA repair and may thus lead to an increase in the cytotoxic effects of therapeutic irradiation.
Demonstration of the efficacy of compound CFAK-C4 targeting FAK-VEGFR3 protein-protein interaction in gastric cancer.
Authors: Kurenova Elena V. , Sanghera Sartaj Singh , Liao Jianqun , Yemma Michael , Cance William G.
Published: March 2020

e22143

Background: While the emerging data strongly suggest that FAK is an excellent target for developmental therapeutics of cancer, kinase inhibitors of FAK have shown crossreactivity with other protein kinases and toxicity in preclinical and clinical studies. It is known that FAK acts pleiotropically, as a kinase and as a scaffolding protein, and our goal is to explore targeting the scaffolding function of FAK to inhibit protein-protein interactions important for tumor progression. Previously, we have shown that FAK physically interacts with VEGFR3 and we identified small molecule inhibitor CFAK-C4 that targets this site of interaction. Both of these kinases are overexpressed in gastric cancers and were found to be independent poor prognostic factors. The prognosis of patients with gastric cancer remains unfavorable and molecular based treatments are necessary for a potential breakthrough in the therapy of this disease. We hypothesize that FAK-VEGFR3 interaction provides essential survival signals for gastric tumor growth and that simultaneous inhibition of these signals will inhibit tumor progression. Methods: Effects of CFAK-C4 on gastric cancer cell lines AGS and NCI-N87 were examined by MTT assay (viability), colony formation assay and Western blotting (phosphorylation, apoptosis). Subcutaneous mouse model was used to demonstrate effect of CFAK-C4 in vivo. Results: CFAK-C4 specifically blocked phosphorylation of VEGFR3 and FAK, directly inhibited cell viability (p<0.05), increased cell detachment and inhibited colony formation in a dose-dependent manner (range 1-100µM). CFAK-C4 (50mg/kg, IP) effectively caused tumor regression in vivo, when administered alone and its effects were synergistic (p<0.05) with chemotherapy. In vivo effects of C4 were confirmed by a decrease in tumor FAK and VEGFR3 phosphorylation, and disruption of their complexes. Conclusions: In this study we have shown that CFAK-C4 inhibits FAK-VEGFR3 signaling in gastric cancer cells and affects tumor growth. This result demonstrates that targeting the scaffolding function of FAK is a unique approach of highly-specific molecular-targeted therapy and can be used to develop oral-based cancer therapeutics.