bluuebunny/crossref_metadata_2025 huggingface dataset:
Real-time informatics to collect data, toxicities, image response assessments, and patient-reported outcomes (PROs) in a phase II clinical trial.
Authors: Pietanza Maria Catherine , Basch Ethan M. , Lash Alex , Schwartz Lawrence H. , Ginsberg Michelle S. , Zhao Binsheng , Shouery Marwan , Shaw Mary , Rogak Lauren J. , Wilson Manda , Gabow Aaron , Latif Marcia , Wu Quinfei , Kass Samantha Lindsay , Miller Claire , Tyson Leslie B. , Sumner Dyana K. , Berkowitz Alison , Sima Camelia S. , Kris Mark G.
Published: March 2020

e19647

Background: In clinical trials, delayed and disorganized data reporting due to traditional toxicity symptom monitoring methods, paper documentation and out-dated collection systems lead to inaccuracies of critical study information and inefficiencies in the process. Electronic systems offer an opportunity to collect this information in real-time from various sources. The feasibility of such an approach is unknown. Methods: We created a computer software system to collect PROs of symptomatic toxicities, automated RECIST-based tumor response, and lab data, and made this information available to investigators in real-time at the point of care during a phase II lung cancer trial. We assessed data completeness within 48hrs of each visit. Clinician satisfaction was measured. Results: We enrolled 44 patients from 1/23/09 to 09/20/11. There were a total of 725 visits, with mean and median being 13 (range, 1-64) and 10, respectively. At each visit, patients completed self-reports using a dedicated wireless laptop 99.6% of the time; only 3 reports were not completed due to lack of a laptop or technical issues with the institutional Intranet. All PROs were completely distributed in batch throughout the system within 24hrs of the visit, and similarly lab data were available for review with a mean of 26hrs from the time the laboratory received the specimen. Manual attribution to lab toxicities in the system took a median of 1 day. Automated RECIST tumor measurements were available to clinicians online with a median of 2 days from the time of imaging; only 8% obtained >7 days after scans were performed. Throughout the trial, there was improvement in data acquisition times (i.e., a “learning curve”). 89% (16/18) of clinicians and research study assistants felt there was greater accuracy in collection of PROs, radiographic responses and trial data. Conclusions: Existing data management systems can be harnessed to enable real-time collection and review of clinical information during trials. This approach facilitates reporting of information closer to the time of events and may improve accuracy and efficiency, as well as ability to make earlier clinical decisions.
Trabectedin in pretreated pediatric patients with relapsed or progressive advanced sarcomas: Toxicity and efficacy.
Authors: Palomo Colli Miguel Angel , Medina Sanson Aurora , Lezama -Valle Pablo , Sadowinski Pine Stanislaw , Pérez Villanueva Heynar
Published: March 2020

e20005

Background: Patients with advanced or metastatic soft tissue and bone sarcomas whose disease progresses after standard chemotherapy (CT) have few options and limited life expectancy. Trabectedin, an alkaloid derived from the sea squirt Ecteinascidia turbinate, has proven activity against some soft tissue (STS) and bone sarcomas. In pediatric patients there is limited information about its efficacy and toxicity profile. Purpose: To describe the toxicity and efficacy of trabectedin in a group of relapsed/refractory pediatric sarcomas. Methods: From January 2009 to December 2011, four relapsed/refractory pediatric sarcoma patients were treated with trabectedin in the Hospital Infantil de Mexico Federico Gomez. Trabectedin was administered as a 24-hour IV infusion every 21 days. Dose levels were 1.3-1.5 mg/m2. All received dexamethasone premedication. Grade 3-4 toxicity was evaluated according to the NCI criteria. Results: Diagnoses included a metastatic Ewing sarcoma, a triton tumor (TT), a metastatic alveolar soft tissue sarcoma and a phylodes sarcoma. All had previously failed to 2 or 3 CT regimens. In 3 patients radiotherapy had also been used and multiple surgical procedures were performed to treat recurrences, before trabectedin. A total of 13 courses of trabectedin were administered. Nausea/vomiting and fatigue were seen in 61.5% of the courses. Laboratory abnormalities included anemia (30.6%), neutropenia (15.3%), ALT (15.3%), AST (15.3%) and GGT (7.7%) elevations and lipasemia/amilasemia (7.7%). One patient presented significant transaminasemia, pancreatitis (probably steroid-related) and died after the first course due to IDC. The TT case, who had a massive unresectable lesion, exhibited a significant response, although her tumor progressed after 4 cycles and eventually died; none of the other patients responded to trabectedin. Conclusions: Trabectedin was generally well tolerated; however we faced a fatal event that might be not trabectedin related. The response observed in the TT case is an interesting finding considering the poor CT response of this aggressive tumor.
Long-term complications after staging laparotomy for Hodgkin lymphoma.
Authors: Rymeski Beth A , Burns Karen Cristly , Nagarajan Rajaram , Correll Judy , Kent Debra A. , O'Brien Maureen Megan , Perentesis John Peter , Dasgupta Roshni
Published: March 2020

e20007

Background: There are over 164,000 long-term survivors of Hodgkin lymphoma in the US. Awareness of the potential for preventable high risk late complications is critical for avoiding early mortality and insuring quality of life. We sought to identify key late complications in patients who had undergone staging laparotomy Methods: Retrospective review of hospital records and database of cancer survivors at our institution. Results: 95 patients with HL were identified; of these 24 patients with HL underwent staging laparotomy from 1971-1994. 18 patients had complete data available with median age at diagnosis of 13.7 years and median follow-up of 27.9 years. 16 patients underwent splenectomy, 12 patients received chemotherapy and 17 received XRT. Six patients (33%) had findings of intra-abdominal disease on laparotomy. Three patients (17%) required four repeat laparotomies for bowel obstruction more than 10 years after staging laparotomy. Of the 3 patients requiring repeat laparotomy for bowel obstruction, one had radiation to the inguinal region, one had mantle radiation (patient with 2 bowel obstructions requiring laparotomy and lysis of adhesions), and one had unknown radiation therapy history. In contrast, there were no cases of bowel obstruction in the other 71 patients who did not receive initial staging laparotomy. There were no documented cases of post-splenectomy sepsis and all post splenectomy patients received some prophylactic antibiotics. Conclusions: This is the first study to examine long term complications after staging laparotomy in a longitudinal cohort of survivors of HL. In long term follow-up, patients who underwent staging laparotomy for HL appear to have an increased incidence of repeat laparotomy, 3-4 fold higher than the expected rate of post laparotomy bowel obstruction incidence in the literature. Otherwise, the long term outcome of splenectomy in these patients appears to be very good with no episodes of post splenectomy sepsis, These findings underscore the importance of a high index of suspicion for bowel obstruction and related complications in HL survivors who had previous abdominal operative procedures. This work also underscores the importance of long term follow-up and screening in this population.
A cholesterol derivative (oxysterol; 24-ethyl-cholestane-3β, 5α, 6α-triol) with high antitumor activity against a variety of sarcomas.
Authors: Habib Nabil , Daaboul Hamid Elia , Hage Georges , Jabbour Abdo , Khalife Riad , Kassem Nader
Published: March 2020

e20500

Background: Oxygenated derivatives of cholesterol, oxysterols, have different physicochemical properties acting on cell membranes. Agents belonging to this class of compounds have been found to induce apoptosis and to harbor antitumor activity demonstrated in vitro and in vivo. 24-ethyl-cholestane- 3β, 5α, and 6α-triol is a new oxysterol developed in our lab. Unlike other derivatives, it is, to our knowledge, the first tested in the clinic. Methods: We have previously reported encouraging and rapid results observed in patients suffering from a variety of solid tumors with an improvement of their quality-of- life and without side- effects. We have treated eight patients suffering from different types of sarcomas on a compassionate basis because we did not have any ongoing trial in sarcomas. Furthermore, most of these patients would not have been eligible for a clinical trial because of their bad performance-status. Three patients were suffering from carcinosarcomas, one from angiosarcoma, one from osteosarcoma, one from chondrosarcoma, one from undifferentiated sarcoma and one from Ewing sarcoma. Most of them were pretreated with chemotherapy and radiotherapy and most of them were in bad clinical conditions. Seven patients were females and one male with ages ranging from 21 to 82 (median age 55y). Results: None of these 8 patients experienced any side-effect despite the fact that one of them was taking a mild chemotherapy in association with oxysterol. This patient was excluded from the evaluation of the response to therapy. Among the 7 patients evaluable for response, we observed 4 complete responses (one of them confirmed by PET scan), two stable diseases and one progressive disease. The complete responses were observed in one osteosarcoma, one Ewing sarcoma, one angiosarcoma and one carcinosarcoma. As with our previous experience with this drug, no clinical or biological side-effect was observed and symptom control was achieved rapidly in all 6 symptomatic patients. Conclusions: We believe that this new compound deserves to be tested in phase II trials in patients suffering from sarcomas.
Marginal resection for osteosarcoma: Long-term outcomes.
Authors: Yamamoto Norio , Takeuchi Akihiko , Shirai Toshiharu , Hayashi Katsuhiro , Kimura Hiroaki , Igarashi Kentaro , Tsuchiya Hiroyuki
Published: March 2020

e20505

Background: When a favorable response to chemotherapy is clinically obtained, we usually perform intentional marginal resection, which preserves the surrounding areas that are normally resected, such as ligament, nerves, blood vessels, and epiphysis region. We report on the long-term outcomes of osteosarcoma patients who underwent the combination therapy and subsequently underwent intentional marginal resection. Methods: The subjects were 43 patients with osteosarcoma. The mean age was 15 years and the mean follow-up period was 101 months. Tumors were Enneking’s surgical stage IIB in 36 patients and IIIB in 7 patients. The patients received preoperatively 3-5 courses of intraarterial fusion of caffeine combined with CDDP and ADM. The effectiveness of chemotherapy was evaluated using plain x-ray, angiography, MRI, and thallium scintigraphy. If the results of two or more imaging examinations showed effectiveness of the treatment, the patient was considered to have a complete response or partial response. Results: The treatment was effective in 14 patients (Grade III) and 29 patients (Grade IV) by histological evaluation (Rosen & Huvos) that correlated with the results of imaging evaluation. Limb function was evaluated according to ISOLS scoring system, excellent is 32 patients (74.4%), good in 7 patients (16.3%), fair in 3 patients (7.0%), and poor in 1 patient (2.3%). Local recurrence was observed in only 2 patients (4.7%). For stage IIB patients, both 5- and 10-year cumulative survival rates were 93.8%. The outcome (IIB + IIIB) was CDF in 26 patients, NED in 8 patients, AWD in 1 patient, and DOD in 7 patients. Conclusions: It is important to minimize surgical margin and to preserve healthy tissue and important structures for improving the function of the affected limb and providing optimal quality of life for the osteosarcoma patients. In the present study, intentional marginal resection was performed in osteosarcoma patients who were clinically obtained favorable responses to a combination of chemotherapy and caffeine administration. They had a good clinical course and there was no negative effect on the survival rate or local recurrence rate.
Risk of disease recurrence and mortality in gastrointestinal stromal tumor (GIST) patients receiving short-term versus long-term imatinib adjuvant therapy: A chart review analysis.
Authors: Conley Anthony Paul , Guerin Annie , Sasane Medha , Gauthier Genevieve , Schwiep Frances , Keir Christopher Hunt , Wu Eric Q.
Published: March 2020

e20511

Background: The benefits of long-term (36 months) use of adjuvant imatinib (IM) in high risk GIST patients (pts) have been demonstrated in a recent multicenter, prospective clinical trial that compared efficacy and safety of 3 years vs 1 year IM treatment. However, in clinical practice, there is no consensus on the optimal IM treatment duration after surgery. The objective of this retrospective observational study was to compare the risk of recurrence and survival among primary resectable Kit positive GIST pts treated with adjuvant IM for a short vs an extended period of time in a real-world setting. Methods: Information on adult pts with primary resectable Kit positive GIST initiating imatinib ≤84 days after surgery was collected from 248 U.S. oncologists using an online data collection form. Detailed pt information following first GIST diagnosis, including demographic, GIST-related characteristics (e.g., risk profile), comorbidity profile, IM treatment characteristics, disease recurrence and mortality was collected for pts with short-term (6-12 months) and long-term IM use (≥24 months). Disease recurrence and mortality rates were estimated from the 1st surgery date to the 1st evidence of recurrence, mortality, or end of observation period. Multivariate Cox proportional hazard models were used to compare recurrence and mortality rates between short vs long term IM use pts. Results: Among the 246 short-term and 395 long-term IM pts, the median follow up was 884 and 963 days, respectively. The average age was similar [59.0 (10.4) vs 58.1 (9.5); p=.23] but short-term pts had less males [57.7% vs 69.6% (p<.01)] and a lower Miettinen risk score [0.3 vs 0.4, p< .01)] than long-term pts. Disease recurrence [7.3 vs 1.8%, (p< .01)] and mortality rates [6.9% vs 2.3%, (p < .01)] were also higher in short- vs long-term pts. The adjusted risk of recurrence was 4.77 times [95% CI: 1.98 – 11.48, (p<.01)] higher and mortality risk was 3.44 times [CI: 1.53 – 7.75, (p<.01)] higher in short- vs long-term pts. Conclusions: Use of IM over an extended period of time is associated with a reduction in long-term risk of disease recurrence and mortality.
Prognostic relevance of induced and spontaneous apoptosis of disseminated tumor cells in primary breast cancer patients.
Authors: Hartkopf Andreas D. , Banys Malgorzata , Gebauer Gerhard , Krawczyk Natalia , Hahn Markus , Neubauer Hans , Wallwiener Markus , Taran Florin-Andrei , Wallwiener Diethelm , Fehm Tanja N.
Published: March 2020

e21003

Background: An imbalance between cell proliferation and programmed cell death leads to tumor growth. Although most cytostatic agents used in systemic therapy of breast cancer induce an apoptosis in tumor cells leading to their destruction, a high ratio of apoptotic cells in primary breast cancer is correlated with poor prognosis. The purpose of this study was to investigate the incidence and prognostic significance of apoptotic disseminated tumor cells (DTC) in bone marrow (BM) of primary breast cancer (PBC) patients. Methods: PBC patients who underwent primary surgery (PS) or were treated with neoadjuvant chemotherapy (NACT) between 01/2003 and 12/2007 were included into this prospective study. BM aspirates were analyzed by immunocytochemistry (M30 antibody) using ACIS system (Chromavision) according to the ISHAGE evaluation criteria. Patients without DTC in the BM were excluded from the study. Survival data were evaluated after a median follow up of 44 months. Results: BM aspirates of 383 PBC patients were analyzed. DTC were M30 positive in 82 of 298 (27%) patients who underwent PS and 41 of 85 (48%) patients treated with NACT. After NACT, the incidence of apoptotic cells in patients with complete/partial remission as compared to patients with stable/progressive disease was 63%/53% vs. 31%/0% (p=0.034). In the NACT group, M30-positive patients suffered less likely of a relapse than those without apoptotic DTC (7% vs. 23% of the events, p = 0.049). Inversely, overall survival in the PS group was significantly shorter in M30-positive as compared to M30-negative patients (75 months, 95% CI: 68-81 months vs. 84 months, 95% CI: 81-86 months; p = 0.008). Conclusions: Apoptotic DTC can be detected in breast cancer patients before and after systemic treatment. The presence of apoptotic DTC in BM of breast cancer patients can reflect the effects of neoadjuvant treatment. Spontaneous and chemotherapy-induced apoptosis may have different prognostic implications.
Activation by zoledoronic acid and IL-18 of γδ T cells from early-stage breast cancer patients in the context of helper NK cells.
Authors: Sugie Tomoharu , Murata-Hirai Kaoru , Iwasaki Masashi , Morita Craig T , Li Wen , Okamura Haruki , Minato Nagahiro , Toi Masakazu , Tanaka Yoshimasa
Published: March 2020

e21004

Background: Human γδ T cells display potent cytotoxicity against various tumor cells pretreated with zoledronic acid (Zol). Zol has shown benefits when added to adjuvant endocrine therapy for patients with early-stage breast cancer or to standard chemotherapy for patients with multiple myeloma. Although γδ T cells may contribute to this additive effect, the responsiveness of γδ T cells from early-stage breast cancer patients has not been fully investigated. In this study, we determined the number, frequency, and responsiveness of Vγ2Vδ2 T cells from early- and late-stage breast cancer patients and examined the effect of IL-18 on their ex vivo expansion. Methods: Breast cancer patients (n=80) were enrolled after institutional review board approval and with written informed consent. Peripheral blood mononuclear cells (PBMC) were purified and stimulated with Zol/IL-2 or Zol/IL-2/IL-18 for 2 to 10 days. The expanded cells were assessed on flow cytometry and the production of IFN-γ and TNF-α measured through ELISA. Results: The responsiveness of Vγ2Vδ2 T cells from patients with low frequencies of Vγ2Vδ2 T cells was significantly diminished. IL-18, however, enhanced ex vivo proliferative responses of Vγ2Vδ2T cells and helper NK cells (CD3-CD56brightCD11c+CD14-CD16+NKGD2+NKp44low) from patients with either low or high frequencies of Vγ2Vδ2 T cells. Cell-to-cell contact between γδ T and helper NK cells appeared to promote expansion of γδ T cells. Exogenous IL-18 markedly enhanced IFN-γ and TNF-α production from PBMC stimulated by Zol/IL-2, whereas the addition of an anti-IL-18Rα mAb reduced cytokine production. Conclusions: These results demonstrate that Zol elicits immunological responses by γδ T cells from early-stage breast cancer patients and IL-18 enhances proliferative responses and effector functions of γδ T cells in the context of helper NK cells.
A gene expression signature that distinguishes malignant from benign thyroid nodules.
Authors: Tian Song , DiCarlo John , Yu Jiaye , Quellhorst George J , Blanchard Raymond K , Fahey Thomas J , Crowley Michael J , Slepushkina Nadejda , Devgan Vikram , Zeng Xiao
Published: March 2020

e21011

Background: Thyroid nodules can be detected in as high as 67% of the population. Distinguishing thyroid cancers from benign lesions is crucial for determining an appropriate treatment plan. For years a gene expression signature for discriminating malignant from benign thyroid nodules has been sought by clinicians. In this study, multivariate bioinformatics tools were used to generate a qPCR based gene expression signature for determining malignancy in thyroid nodules. Methods: Multiple mathematical models, such as Random Forest, Support Vector Machine (SVM), and Nearest Shrunken Centroid (NSC), were used to analyze published microarray data sets and select 366 putative classifier (biomarker) mRNA targets. The selected 366 genes were further evaluated for their expression pattern by real-time PCR using a panel of 49 pathology assessed thyroid nodule samples (fresh frozen, 23 malignant and 26 benign). Results: Using the qPCR data set, Random Forest was compared with SVM and NSC classifier methods and was found to be more successful in finding genes with better discriminative powers. A Random Forest method identified a panel of 7 genes together with 5 reference genes as a gene expression signature for thyroid malignancy, which led to the development of a companion classifying algorithm to provide a probability score to assess malignancy of thyroid nodules. In our limited sample set, this signature was shown to distinguish malignant and benign thyroid nodules with 92% accuracy and 100% specificity. Conclusions: Our results suggest that a combination of multiple bioinformatics analysis tools is the proper approach for biomarker candidate selection from high-throughput gene expression data. As demonstrated here, panel of 12 genes and a companion classification algorithm has the potential to successfully discriminate malignant thyroid nodule with high accuracy and specificity. This panel of twelve genes is for molecular biology applications only.
Gene expression panel (TheraPrint) analyzed as predictors of response to neoadjuvant chemotherapy (NCT) in patients (pts) with stage II-III and inflammatory breast cancer (BC).
Authors: De Snoo Femke , Peeters Justine , Robinson Kim , Stork-Sloots Lisette , Simon Iris , Somlo George
Published: March 2020

e21013

Background: TheraPrint is a microarray-based gene expression panel of 125 genes identified as potential targets for prognosis and therapeutic response. These genes may hold the key to a greater level of personalized prognosis and therapy for BC pts. The aim of the current study was to assess the clinical relevance of the TheraPrint genes for either predictive and/or prognostic value in 2 patient cohorts treated with NCT. Methods: The 1st patient cohort are 68 Stage II-III BC pts treated with NCT. Expression data from Agilent full genome arrays, containing the MammaPrint, BluePrint and TheraPrint diagnostic profiles/probes (Somlo et al, 2009). Median FU 2.3 years. The 2nd patient cohort are 230 Stage I-III BC pts treated with NCT. Expression data from Affymetrix probe sets was publically available (Iwamoto et al, 2011). Median FU 5.2 years. To identify genes that are differentially expressed between responders (pCR/RCBI) and non-responders, a supervised analysis was performed. The analysis was performed across all pts and also within groups of HER2+ and HER2-. Univariate t-tests were performed, with results filtered by permutation p-value (p<0.05) and fold change of >1.5. Global test was also reported. In addition, survival data analysis was performed across all pts. Results: Overlapping genes between the 2 datasets that were significantly differentially expressed between responders and non-responders include: BCL2 (down-regulated) and CDH3, GRB7, KRT6B, KRT17 (up-regulated). When analysing the HER2- subgroup, 3 genes turned out to be differentially expressed between responders and non-responders in the 2 datasets: FLT1, PIK3R1 (down-regulated) and KRT6B (up-regulated). For the HER2+ subgroup, only one gene overlapped for the 2 datasets: IL2RA (up-regulated). The top canonical pathways for the significant genes have been analyzed, and in addition correlation of the TheraPrint gene expression with survival for these pt groups. Conclusions: This study has identified several genes from a panel of 125 TheraPrint genes with statistically significant correlation between expression and response to NCT.
Risk of venous thromboembolism in cancer patients treated with cisplatin: A systematic review and meta-analysis.
Authors: Seng Sonia Maria , Liu Ziyue , Chiu Sophia , Proverbs-Singh Tracey , Sonpavde Guru , Choueiri Toni K. , Tsao Che-Kai , Yu Menggang , Hahn Noah M. , Oh William K. , Galsky Matt D.
Published: March 2020

e21016

Background: Several reports suggest that cisplatin is associated with an increased risk of thromboembolism (TE). However, patients with solid tumors have multiple risk factors for TE and the excess risk of venous thromboembolic events (VTEs) with cisplatin-based chemotherapy as compared with non-cisplatin-based chemotherapy has not been well described. We performed a systemic review and meta-analysis of randomized controlled trials (RCTs) evaluating the incidence and risk of VTE associated with cisplatin-based chemotherapy. Methods: PubMed was searched for articles published from January 1, 1990 until December 31, 2010.The primary aim was to evaluate the association between treatment with cisplatin and VTEs in patients with cancer. Clinical trials that met the following criteria were included in the meta-analysis: (1) prospective randomized phase 2 and 3 trials of patients with cancer; (2) randomization to treatment with cisplatin versus a non-cisplatin containing chemotherapy regimen (3) available data on venous thromboembolic events. Data on all grade venous thromboembolic events was extracted. Study quality was calculated utilizing Jadad scores. Incidence rates, relative risks, and 95% confidence intervals (CIs) were calculated using a random-effects model. Subgroup analyses included the impact of publication year, tumor type, and cisplatin dose. Results: A total of 8216 patients with a variety of advanced solid tumors from 38 RCTs were included for analysis. Among patients treated with cisplatin-based chemotherapy, the summary incidence of VTE was 1.64% (95% CI, 1.06–2.25). Patients treated with cisplatin-based chemotherapy had a significantly increased risk of VTE with a relative risk of 1.65 (95% CI, 1.25–2.18; P = .01) compared with controls. Exploratory subgroup analysis revealed the highest relative risk of VTE in patients receiving a weekly equivalent cisplatin dose >30 mg/m2 (2.64; 95% CI, 1.18–5.77; P = .02) and in studies reported during 2000-2010 (1.70; 95% CI, 1.27–2.28; P = .01). Conclusions: Cisplatin chemotherapy is associated with a significant increase in the risk of VTE in patients with advanced solid tumors compared with non-cisplatin chemotherapy.
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.