bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of leukapheresis on efficient CTC enrichment for comprehensive molecular characterization and clinical diagnostics.
Authors: Stoecklein Nikolas H. , Niederacher Dieter , Topp Stefan A. , Zacarias Föhrding Luisa , Vay Christian , Knoefel Wolfram Trudo , Kasprowicz Nikola S. , Hepp Philip Gm , Mohrmann Svjetlana , Janni Wolfgang , Stratmann Antje , Krahn Thomas , Honisch Ellen , Raba Katharina , Fischer Johannes C. 
Published: March 2020

e21020 Background: Circulating tumor cells (CTCs) are promising biomarkers for diagnosis and systemic therapy. However, their infrequent detection rates limit currently their clinical use. Here we tested whether leukapheresis could be a suitable method to increase CTC yields and detection rates by increasing dramatically the analyzed blood volume. Methods: We screened 3x106 PBMNCs of 48 historical leukapheresis products harvested from 24 breast cancer patients in the setting of high-dose chemotherapy and from a non-cancer control group (n=10), respectively, with a standard immuno-assay using an anti-cytokeratin antibody (A45/BB3) to detect CTCs. Detected CTCs were isolated, their genomic DNA amplified, and the whole genome screened for chromosomal aberrations using comparative genomic hybridization (CGH) (n=48). To validate the CTC detection frequencies in leukapheresis products, we initiated a prospective pilot-study and performed leukapheresis in 13 cancer patients (breast and pancreatic cancer). 1 mL of the leukapheresis product (total volume: up to 150 ml) was analyzed using the CellSearch® system as well as 7.5 ml peripheral blood taken from the same patients. Results: 44/48 (91.7%) of leukapheresis samples contained CTCs with a median count of 3.0 in 3x106 PBMNC (range: 1-35 CTCs). None of the 10 analyzed healthy controls displayed CTCs. We successfully performed single cell CGH of 48 CTCs from 19 patients revealing aberrant CGH profiles in 56% with gains and losses typical for breast cancer. Interestingly, CTCs with high aberration numbers were associated with early metastatic relapse. The validation study using CellSearch® in 13 cancer patients demonstrated in leukapheresis products significantly higher CTC detection frequencies (13/13 vs. 5/13) and CTC numbers (median: 13, range: 1-51 vs. median 0, range: 0-7; p<0.001) compared to the matched peripheral blood samples. Conclusions: Our results indicated that leukapheresis provides CTC detection rates and yields at unprecedented scale. Thus we believe, that leukapheresis will help to exploit the full clinical potential of CTCs as biomarkers for diagnosis and systemic therapy.
Senescence, a new concept in pathologic response evaluation of rectal carcinomas (RC) after neoadjuvant treatment.
Authors: Valverde Claudia María , Aizpurua Miren , Capdevila Jaume , Morales Rafael , Nuñez Isaac , Suarez Cristina , Landolfi Stefania , de Torres Ines , Carles Joan , Moline Teresa , Romagosa Cleofe 
Published: March 2020

e21021 Background: Available protocols of response evaluation after therapy are based on the presence of viable cells, fibrosis and necrosis. Senescence is an irreversible cell cycle arrest that can be activated by different kinds of stress like DNA damage and cytotoxic drugs. Senescent cells are histologically very similar to viable neoplasic cells and usually evaluated alike. Our hypothesis is that neoadjuvant treatment can induce senescence as part of the therapeutic response and its presence may impact the prognosis. Methods: Tumor samples of 45 consecutive patients (pts) with RC treated between sept/02 and feb/2007 with fluorouracil-based chemotherapy and neoadjuvant radiotherapy, and of 33 non-treated colon cancer pts (controls), were selected. p53 and P16-ki67 double immunostaining were retrospectively evaluated in endoscopic-pretreatment biopsies, post-therapy specimens, and controls. P16+/ki67- malignant glands were considered as senescent-like glands. SPSS v20.0 (Kruskal-Wallis, Cox regression and log-rank test) was employed for analysis. Results: After a median follow up of 60 months 13 pts had relapsed. Significant differences in the percentage of senescent-like glands were observed between treated carcinomas and their pre-treatment samples (p=0.0001) and also with the colonic-non-treated carcinomas (p=0.0001). A tendency toward a better disease free survival (DFS) (p=0.236) in those patients with more than 30% of senescent glands after treatment was observed. No differences in p53 were found between the 3 groups. However, low levels of p53 expression in pretreatment-rectal biopsies correlate with a better pathologic response (GR) (p=0.029), and better DFS (p=0.0958). Conclusions: The number of senescent-like glands increases after neoadjuvant therapy. There was a tendency for a better DFS in the pts with treatment-induced senescence. Moreover, an inverse correlation between p53 expression in pretreated endoscopic-samples and pathologic response and DFS was observed. Further and larger studies should be performed to confirm the prognostic implications of evaluating senescence markers in treated rectal carcinomas.
Outcome and evaluation of 1p and 19q deletion by chromogenic in situ hybridization (CISH) in patients with diagnosis of oligodendroglioma (OD) and oligoastrocitoma (OA).
Authors: Bella Santiago Rafael , Llugdar Jose Roberto , Lingua Alejo , Theaux Ricardo Alejandro , Papalini Francisco , Olocco Ricardo , Lerda Daniel 
Published: March 2020

e21025 Background: In OD and OA, the 1p and 19q deletion has prognostic value in survival. It is also a predictive factor for response to chemotherapy. Fluorescence in situ hybridization (FISH) is the standard method for its evaluation. CISH could be an alternative that has already been validated in other neoplasias. In OD and OA, this combined deletion is present in about 50% of patients when analyzed with FISH. Methods: Patients resected at Clinica Reina Fabiola from january 2006 to january 2010 and diagnosed of OD and OA were propectivelly included. Paraffin-embebed tumor tissue was analyzed for 1p19q deletions by CISH. The results were correlated to the histology (OD and OA) and grade (II and III) of the tumors. Results: The demographic features of the patients from the present study coincide with literature. The 1p and 19q deletion was found in 3 of the 24 patients analyzed (13%). The combined deletion was only found in those with grade II OD. No combined deletion was found in patients diagnosed of grade III OD and grade II and III OA. In the subgroup of patients with grade II OD, the combined deletion was observed in 3 of 11 patients (27%). The 3 patients in which the deletion in both chromosomes was observed, received treatment with chemotherapy and radiotherapy, all of them with complete response. 5 years DFS was 90%-median follow up 36,8 (CI: 30,5-42,98) Conclusions: The detection of the combined deletion with CISH technique was inferior (13%) than the literature. We cannot demonstrate that CISH is a reliable method for the detection of the 1p and 19q deletion. The possible reasons of this difference could be attributed to the number of patients of the study, to deviations in the procedures of the test or to the fact that the CISH method is not coincident with FISH. This prospectivelly monoinstitutional results are also different to our previous report, and may be due to different pathological evaluation.
Improved techniques to localize foci of carcinoma within the prostate using high-resolution transrectal gamma imaging (TRGI) of monoclonal antibody directed at prostate specific membrane antigen (PSMA).
Authors: Franc Benjamin L. , Cui Yonggang , Rosenthal Seth A , Johnston Tammie , Poonamallee Uday , Wiesner Jason B , Vandewalker Kristen MN , Seo Youngho , Lall Terrance , James Ralph 
Published: March 2020

e21026 Background: As many as 40% of prostate cancers may be missed on biopsy, necessitating novel methods for biopsy guidance. There is also interest in mapping primary prostate cancers for treatment. The presented work evaluated the ability of a transrectal gamma-imaging (TRGI) device to detect and localize a prostate-specific membrane antigen (PSMA) - specific radiopharmaceutical in the prostate. Methods: The IRB approved this exploratory proof-of-concept study. Male patients with untreated, biopsy-proven prostatic adenocarcinoma received 5±0.5 mCi In-111 capromab pendetide (Prostascint, a PSMA-specific agent) i.v. and were imaged 4 days later using single photon emission computed tomography (SPECT-CT). A commercially available cadmium zinc telluride (CZT)-based endorectal gamma camera (ProxiScanTM, Hybridyne Imaging Technologies, Inc.) determined the radiopharmaceutical’s planar distribution (1.5x4 cm FOV; 5 min acquisition) in four prostatic quadrants. Safety assessments occurred day 1 and 7. MRI was acquired within 14 days. Pathology from TRUS biopsy or following prostatectomy identified prostatic quadrants containing adenocarcinoma. Results: Of 8 patients recruited, 6 completed imaging, all without adverse event. Subsequent treatment included prostatectomy (n=3) and radiation (n=3). Twenty-three prostatic quadrants were evaluated. In detecting a cancer-containing quadrant, TRGI, SPECT, and MRI had sensitivities of 100%, 75%, and 33%, respectively, and similar accuracy (61-70%). Target-to-background signal ratio of TRGI was independent of serum PSA level and not significantly different than SPECT. TRGI identified a greater number of foci in each quadrant when compared with SPECT (p<0.001). Lesion sizes on TRGI were smaller (p=0.04) and correlated with cancer size on pathology in patients treated surgically (r=0.69), identifying tracer uptake associated with tumors ≥ 5 mm. Conclusions: TRGI provides improved localization of prostate carcinoma compared with conventional SPECT with potential implications for the diagnosis and treatment of prostate cancer.
Pancreatic circulating tumor cells: Where are they?
Authors: Khoja Leila , Mitchinson Christy , Brown Damien , Zhou Cong , Morris Karen , O'Reilly Derek , Deshpande Rahul , de Liguori Carino Nicola , Clack Glen , Hughes Andrew M. , Valle Juan W. , Dive Caroline 
Published: March 2020

e21037 Background: We previously demonstrated that circulating tumor cell (CTC) numbers in advanced pancreatic carcinoma were low in contrast to other epithelial carcinomas. We hypothesized that pancreatic CTCs may sequest in the liver and conceived a study to explore CTC number in the peripheral and portal circulations of patients undergoing Whipple’s surgery. Methods: Seventeen patients undergoing Whipple’s surgery were recruited prospectively after informed written consent. Blood was collected intra-operatively from the portal vein prior to tumor mobilization and from the peripheral circulation. Paired samples were analysed for CTC number (CellSearch system, Veridex, USA), and M30/M65 biomarkers of cell death. Results: Of 17 patients 6 had cholangiocarcinomas and 11 had pancreatic adenocarcinomas (5 inoperable). CTC number in the portal circulation was consistently higher than in the peripheral circulation. All patients with cholangiocarcinoma had no CTCs in the peripheral circulation but two patients had 139 and 21 CTCs in the portal circulation. Five patients with pancreatic carcinoma had no CTCs in the peripheral circulation but 1 CTC in 2 patients, 4 CTCs in 2 patients and 32 and 731 CTCs were seen in 2 other patients in the portal circulation. Five patients had no CTCs in both circulations and the remaining patient had 1 CTC in the peripheral circulation and 4 CTCs in the portal circulation. Where present, circulating tumour microemboli were detected in the portal circulation only. M65 levels were higher in the portal circulation indicating higher levels of cell death. M30 levels did not differ significantly between circulations but appeared to correlate with portal CTC number. Conclusions: This is the first study to enumerate CTCs in peripheral and portal circulations in pancreato-biliary carcinomas. Higher CTC numbers are present in the portal circulation suggesting CTC sequestration in the liver. Five patients with pancreatic cancer (2 of whom were inoperable) had no CTCs in either circulation, a finding consistent with the poor vascularity and dense stromal characteristics of this carcinoma. Nonetheless the portal circulation may be a potential source for CTC characterisation including genomic analysis to further define tumor biology.
Excision repair cross-complementary group 1 (ERCC1) as a prognostic biomarker in patients (pts) with solid tumors: A meta-analysis on detection methods.
Authors: Mak Milena Perez , Takahashi Tiago Kenji , Hoff Paulo Marcelo , Castro Gilberto , Snitcovsky Igor M. L. 
Published: March 2020

e21040 Background: ERCC1 overexpression is associated with better outcomes in some cancers. The best method to assess its expression, immunohistochemistry (IHC) or polymerase chain reaction (PCR), is not established. To clarify such question, this meta-analysis was conducted. Methods: PUBMED, EMBASE and Cochrane databases were searched with terms: “ERCC1” and “IHC” and/or “PCR” and/or “mRNA”. Inclusion criteria: full papers in English, solid tumor pts, ERCC1 evaluated by IHC and PCR on the same samples, reported correlation between ERCC1 expression and overall survival (OS) – the primary endpoint. As secondary endpoint, correlation between IHC and PCR expression was assessed. Exclusion criteria: review articles and no reported endpoint. Two authors reviewed and classified all papers. Pooled HR and variance were calculated by standard methods, using CMA v2.2.064 (Englewood, USA). Results: 25 articles were retrieved, with 4 included for the primary endpoint (224 pts) and 4 (191) for the secondary endpoint. All were retrospective, with diverse primary sites. Except for one, platinum chemotherapy was employed. All studies used 8F1 clone, and in 2, FL297 clone was also evaluated. ERCC1 positivity was determined by H-score in 2 studies and by AQUA in 2. mRNA ERCC1 positivity was variably defined. In general, ERCC1 overexpression was correlated with longer OS (HR 0.569, 95% CI 0.436 - 0.743, p <0.001). Similar results were found in terms of OS either by IHC or PCR (HR 0.626, 95% CI 0.46 – 0.853, p 0.003 and HR 0.434, 95% CI 0.257 – 0.730, p 0.002; respectively). No correlation between IHC and PCR overexpression was found (coefficient 0.098 p 0.444, random effect). Heterogeneity was detected only in IHC analysis, when expression was detected by both FL297 and 8F1 antibodies (p 0.001, fixed effect). Conclusions: ERCC1 overexpression is associated with better OS, regardless of employed methodology. However, protein and mRNA expression are not correlated. Major limitations to our analysis include the variety of employed ERCC1 detection methods and retrospective nature of data. Definitive conclusions on the prognostic role of ERCC1 and best methodology remain to be answered.
Histology differentiation in non-small cell lung cancer using matrix-assisted laser desorption/ionization mass spectrometry.
Authors: Park Jun Won , Lee Geon Kook , Lee Hee Seok , Zo Jae Ill , Kim Young Hwan , Kim Kwang Pyo , Kim Hark K. 
Published: March 2020

e21043 Background: New developments in the treatment of lung cancer have necessitated the correct histologic differentiation between two major histologic types of NSCLC. Methods: Histology-directed tissue matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) has been demonstrated to be useful for molecular profiling of common solid tumors. In this approach, mass spectra are obtained from discrete locations on the thin sections of frozen tissues sections. In this study, we evaluated various MALDI MS protocols for the histopathologic classification of NSCLC. Results: When adenocarcinoma and squamous cell carcinoma biopsy samples were compared for protein profile using standard sinapinic acid matrix, the median prediction accuracy of the best histology classifier in test sets was 83.3% in 100 random training-to-test partitions (feature selection P <0.05). When 2,5-dihydroxybenzoic acid/α-cyano-4-hydroxycinnamic acid matrices were used for lipid MALDI MS, the median prediction accuracy of the best histology classifier in test sets was 85.7% in 100 random partitions (feature selection P<0.01). We then validated the discriminatory lipid profile using independent set of 51 NSCLC surgical samples. Eleven discriminatory lipids correctly classified the histology of 80.4% of independent NSCLC surgical tissue samples (41 out of 51) in validation set. Phosphatidylcholine 32:0 (m/z 756.68) was overexpressed in adenocarcinomas. Conclusions: Due to the small amount of tissue required for MALDI analysis, rapid experimental procedure, and high predictive values documented in this study, histology-directed MALDI MS, especially lipid profiling, may be a promising approach to the histologic classification of NSCLC (This work was supported by Converging Research Center Program through the Ministry of Education, Science and Technology of Korea (2010K001121)).
Ex vivo expansion of CTCs in culture isolated by size from patients with melanoma.
Authors: Benali-Furet Naoual , Wechsler Janine , Ye Fei , Ezaoui Sarah , Avril Marie-Françoise , Boitier Francoise , Carlotti Agnes , Paraiso Isbou , Cayre Yvon E 
Published: March 2020

e21046 Background: To date there are no studies demonstrating the ability of circulating tumor cells (CTCs) isolated from patients to grow ex-vivo in culture. Most of our knowledge on the mechanisms of cancer metastasis has been obtained from CTCs of mouse models. In human, a main issue for studying circulating melanoma cells subpopulations and constructing tumor models is to be able to grow CTCs from patients. Here we defined optimized conditions to isolate CTCs alive and to culture them using the ScreenCell technology. Methods: CTCs were isolated alive by size using a ScreenCell device from 6 ml blood of 11 patients with melanoma at diagnosis and before surgical excision. Cells were cultured directly on a porous and high adhesion filter with optimized culture conditions. The filter was also used as a crossing barrier for cell invasion assay. After 15 days culture, cells were fixed and immunohistochemically stained for proliferative, hypoxia and invasive phenotype markers. Results: After 5 daysunder specific culture conditions, CTCs were adherent to the filter with several shapes and size in all patients, while no cells were found in blood of healthy donors. After proximately 10 days in culture, a population of small cells started to growth while a subpopulation of large cells migrated through the pores of the filter to form spheroids into the Matrigel® located under the filter. After 15 days in culture the non-invasive and invasive cells were positively immunostained for expression of the Melan-A proliferative phenotype marker and for the Glut-1 and Wnt5A hypoxia and invasive phenotype markers, respectively. In parallel, the corresponding primary melanoma lesions were immunostained and compared to immunostaining data of CTCs. Conclusions: TheScreenCell technology preserves the viability, proliferative and invasion potentials of CTCs. Furthermore, CTCs can be maintained in culture allowing ex vivo expansion and characterization of subpopulations of CTCs with proliferative and invasive characteristics. Using the ScreenCell devices we have defined optimal technical and microenvironment conditions for CTCs isolation and culture. We are currently exploring the ability of CTCs to xenoengraft and develop tumor in immunocompromized mice.
The role of GNAS mutation in diagnosis of mucinous pancreatic cysts.
Authors: Ellsworth Eric M. , Schmidt C. Max , Bleicher Jamie F. , Smith Dennis M. , Finkelstein Sydney D. 
Published: March 2020

e21048 Background: Pancreatic cysts pose a challenge in patient management primarily due to difficulties distinguishing non-mucinous cysts from potentially pre-malignant mucinous ones, and in determining the malignant potential of mucinous cysts. Mutations to the GNAS gene have been identified as a marker for intraductal papillary mucinous neoplasm (IPMN). [1,2] We evaluated the significance of GNAS mutation in diagnosis of pancreatic cysts. Methods: We retrieved archival DNA from 237 cyst fluids. 200 specimens were chosen with 100 KRAS mutant and 100 KRAS wild type in a range of CEA values. 37 specimens were chosen with a molecular diagnosis of aggressive biological behavior, to enrich for malignant IPMNs. We sequenced each fluid’s DNA from for codon 201 of GNAS. Molecular criteria for mucinous cysts included KRAS mutation, elevated DNA, or ≥2 high clonality LOH mutations. Results: Of the 237 specimens, 25 were not amplifiable due to degraded DNA, 52 (25%) had a GNAS mutation, and 160 had no GNAS mutation. Of the 52 GNAS mutated specimens, 5 were diagnosed as biologically aggressive based on significant associated mutations, 28 as statistically indolent based on associated molecular changes, and 19 as benign (no accompanying mutations). The proportion of GNAS mutations decreased with increasing molecular changes linked to malignancy. Data from [1] shows no statistical difference in frequency of GNAS mutation in side branch vs. main vs. mixed IPMNs (p= 0.46). Similarly, in our cohort, 6 cases had clear imaging features of a side branch IPMN, and of these 2 (33%) had a mutation in GNAS. Using molecular criteria for identification of mucinous cysts, 166 were mucinous and 71 non-mucinous. Incorporation of GNAS into the molecular determination of mucinous etiology identified 13 additional cases (7%). Conclusions: Use of GNAS increases sensitivity for detection of mucinous lesions. GNAS does not appear to correlate with molecular measures of biological aggressiveness. These findings confirm those in [1], that GNAS should be used in conjunction with a panel of molecular markers for evaluating the malignant potential of pancreatic cystic lesions. References 1. Sci Transl Med. 2011 Jul 20; 3:92ra66. 2. PNAS 2011 Dec 27; 108:21188-93.