bluuebunny/crossref_metadata_2025 huggingface dataset:
Outcomes of multivisceral resection of gastric gastrointestinal stromal tumors.
Authors: Ganai Sabha , Posner Mitchell , Prachand Vivek N. , Alverdy John C. , Choi Eugene A. , Waxman Irving , Patti Marco G. , Roggin Kevin K.
Published: March 2020

10090

Background: Despite the recent introduction of imatinib and laparoendoscopic techniques to the management of gastric gastrointestinal stromal tumors (GISTs), outcomes remain uncertain in the setting of multivisceral involvement. Methods: We conducted a retrospective review of 73 consecutive patients who underwent resection of gastric GISTs from October 2002 through December 2011. Median follow-up was 22 months (interquartile range [IQR] 6-37). Results: Patients were 51% female, with a mean age of 65 ± 12 years and BMI of 30 ± 8 kg/m2. Patients undergoing multivisceral resection (n=14) had a longer interval from diagnosis to surgery (7.3 [IQR 1.9 – 15.0] vs. 1.3 [IQR 0.7-4.2] months, p<0.01), greater use of neoadjuvant imatinib (64% vs. 3%, p<0.0001), and greater preoperative tumor size (12 ± 8 vs. 4 ± 3 cm, p<0.0001) in comparison to gastric-only resections (n=59). Patients were less likely to be managed laparoscopically (7% vs. 71%, p<0.0001), had a longer operative time (310 ± 117 vs. 145 ± 62 min, p<0.0001), and were less likely to be R0 (71% vs. 98%, p<0.001). While patients undergoing multivisceral resection were more likely to have a pathological complete response to therapy (29% vs. 0, p<0.001), they were also more likely to have metastatic disease present (29% vs. 0, p<0.001). Hospital length of stay was greater (median 8 [IQR 6-9] vs. 3 [IQR 2-6] days, p<0.0001). There were no significant differences in grade or mitotic index between groups. There was greater use of adjuvant imatinib (64% vs. 25%, p<0.05). Overall survival was less in patients undergoing multivisceral resection (64% vs. 87% at 3 years, p<0.05), as was disease-free survival (52% vs. 71% at 3 years, p<0.05). Median overall and disease-free survival were 43 and 22 months after multivisceral resection for gastric GISTs. Controlling for tumor size, grade, resection status, and the use of neoadjuvant imatinib, multivisceral resection and use of adjuvant imatinib were both independent predictors of disease-free survival (p<0.05). Conclusions: Multivisceral involvement is associated with tumors of greater size and, despite an increased use of neoadjuvant imatinib, it is associated with poor outcome for patients with gastric GISTs.
Analysis of the quality of reporting surgical procedures in patients undergoing resection for primary gastrointestinal stromal tumors: A reporting tool derived from the EORTC–STBSG 62024 trial.
Authors: Hohenberger Peter , Rutkowski Piotr , Bonvalot Sylvie , van Coevorden Frits , Stoeckle Eberhard , Olungu Christine , Ouali Monia , Casali Paolo Giovanni , Gronchi Alessandro
Published: March 2020

10096

Background: Features of surgery for the primary tumor (rupture, extent and completeness of resection) contribute significantly on the indication for adjuvant treatment in patients with GIST. No reporting tool for surgery for a primary GIST has been established. EORTC 62024 is a phase III trial evaluating the use of adjuvant imatinib in GIST and was used for an analysis of the data of surgical procedures. Methods: 908 pts. from 11 countries were recruited. At study entry, a copy of the surgical record and pathology report had to be submitted in addition to the case record forms (CRFs) which included a surgical questionnaire on 19 items on the circumstances of the resection (elective/emergency, GIST diagnosis established preop., tumor rupture, R-status, concordance of preop. and intraop. staging, surgical technique, complications). We then reviewed the original documents and compared them with the documentation on the CRFs. The full set of data was available from 793/906 patients (87.3%). Results: The diagnosis of GIST was known preop. in 58% of the cases, and 12% of the pts. had been treated as an emergency. The R0 resection rate was 87%. The incidence of tumor rupture was 9% and 24% of the pts. had undergone lymphadenectomy. The consistency between pre- and intraoperative findings was 82%. The concordance between CRFs and original documents was 77%. Concordance of the original documents with the surgical questionnaire (CRFs) was critical in 4/19 items. Discrepancies and misinterpretation rates were highest when data managers from medical oncologists had completed the CRFs based on letters of patient discharge. Based on the evaluation, the surgical reporting tool can be refined and shortened to 13 items for future applications. Conclusions: Findings at surgery for the primary tumor not properly reported might significantly influence the decision of whether or not to treat patients with adjuvant imatinib. The refined surgical reporting tool could be helpful to gather all relevant information about the intraoperative elements of surgery for the primary GIST. Surgeons are to be asked to report those data themselves.
ABT-888 (veliparib) in combination with carboplatin in patients with stage IV BRCA-associated breast cancer. A California Cancer Consortium Trial.
Authors: Somlo George , Sparano Joseph A. , Cigler Tessa , Fleming Gini F. , Luu Thehang H. , Hurria Arti , Mortimer Joanne E. , Frankel Paul Henry , Chew Helen K. , Nanda Rita , Ma Cynthia X. , Chen Alice P. , Garcia Agustin , Vahdat Linda T. , Gandara David R. , Weitzel Jeffrey N.
Published: March 2020

1010

Background: Platinum and PARP inhibitors have both shown activity in BRCA-associated breast cancer (BC) patients (pts). We have conducted a phase I trial of carboplatin (Carb) and velapirib [V], a PARP inhibitor, to define dose limiting toxicities [(DLT) during cycle (C) 1] and the maximum tolerated dose (MTD). Methods: BRCA 1 or 2 carriers with stage IV BC were eligible. Carb starting at an AUC of 6 was given IV every 21 days (length of planned C) and V was administered orally, BID at dose levels (L) L1 through L5 (highest L planned). Results: 22 pts (21 eligible/evaluable, 20 with measurable BC) carrying BRCA1 (10) or BRCA2 (11), or both (1) mutations were accrued. Median age: 45 years, (32-65); 68% of BCs were ER+, and 10% were HER2+. In the table below are the schema, incidence of DLTs, and # of Cs on study. Toxicities: At L1, grade ¾ DLTs with C 1 were seen in 2/6 evaluable pts (1 pt w/grade 3 hyponatremia, pleural effusion, and dehydration, and 1pt w/grade 4 thrombocytopenia [PLT]), leading to deescalation of carb (AUC 5) for pts treated at Ls 2-5. At L2, 1 of 6 pts had grade 4 PLT. There were no DLTs at Ls 3 and L4. L5 is currently being expanded to 6 pts (3 currently enrolled, 1 pt with grade 4 granulocytopenia (Gr) and grade PLT reached DLT). Non-DLT dose delays mostly due ≥ grade 2 Gr or PLT were needed at 60%, 53%, 53%, and 43% of Cs in pts treated on Ls 1-4. Response: In 12 eligible pts treated at Ls 1 and 2, 2 complete and 6 partial responders (67%) and a clinical benefit (CB) of 75% were seen. All pts at Ls 3-5 are still being treated, and in pts treated at Ls 3 and 4, 2 unconfirmed PRs, and 4 cases of stable disease were seen, with L5 too early to assess. Conclusions: The combination of Carb at an AUC of 5 and daily V at doses 150 to 200 mg BID is feasible and the MTD is being defined. In preliminary analysis, response and CB rates are better than expected with the individual agents alone, providing justification to proceed with a planned phase II randomized single agent versus combination trial. [Table: see text]
SOLTI NeoPARP: A phase II, randomized study of two schedules of iniparib plus paclitaxel and paclitaxel alone as neoadjuvant therapy in patients with triple-negative breast cancer (TNBC).
Authors: Llombart Antonio , Lluch Ana , Villanueva Cristian , Delaloge Suzette , Morales Serafin , Balmaña Judith , Amillano Kepa , Bonnefoi Herve R. , Casas Ana Maria , Manso Luis , Roche Henri Hubert , Gonzalez-Santiago Santiago , Gavila Joaquin , Sánchez-Rovira Pedro , Di Cosimo Serena , Charpentier Eric , Garcia-Ribas Ignacio , Penault-Llorca Frederique Madeleine , Aura Claudia , Baselga José ,   SOLTI Group
Published: March 2020

1011

Background: Iniparib is an anticancer agent with a mechanism of action still under investigation. A phase 2 randomized neoadjuvant study in patients (pts) with TNBC was designed to explore the activity and tolerability of two schedules of iniparib with weekly paclitaxel (PTX). Here we report the efficacy and safety results from a planned interim analysis (IA). Methods: The trial accrued a total of 141 pts in October 2011, of whom, 74 are included in this IA. All were chemo-naive, histologicallyconfirmed Stage II-IIIA TNBC (IIA 47%; IIB 35%; IIIA 16%) with a median age of 50 yr. Triple negative status was centrally confirmed [ER/PR <10%, HER2 IHC (0+, 1+) or FISH negative]. Pts were randomized (1:1:1) to receive weekly PTX (80 mg/m2, IV, d 1; N=25) alone or in combination with iniparib, either on a once weekly (QW) (11.2 mg/kg, IV, d 1; N=25) or twice weekly (BIW) (5.6 mg/kg, IV, d 1, 4; N=24) schedule. The total planned treatment duration was 12 wks. The IA endpoint is pathological complete response in the breast (pCR) as assessed by independent pathologists. Results: Two/2/3 pts in the PTX/QW/BIW arms, respectively, discontinued due to progressive disease per RECIST. Another 3/2/2 pts, respectively, discontinued due to investigator decision or an adverse event (AE). Thirteen pts presented with Grade 3/4 Treatment Emergent AE: 3 pts in PTX arm (1 neutropenia, 1 presyncope, 1 ALT elevation), 3 in QW arm (1 lymphopenia, 1 hyperkalemia, 1 pulmonary embolism), and 8 in the BIW arm (1 febrile neutropenia, 3 neutropenia, 1 aphonia, 1 syncope, 1 radius fracture and 1 vertigo). Laboratory Grade 3/4 neutropenia occurred in 4% of pts in PTX, 0% in QW and 21% of BIW arms, with 1/2/3 pts, respectively, requiring G-CSF usage. There were 4/7/6 pts in the PTX/QW/BIW arms with PTX dose modifications. Four pts (16%) in PTX arm, 4 pts (16%) in the QW arm and 6 pts (25%) in the BIW arm had confirmed pCR in the breast. Conclusions: In this IA population, the addition of iniparib regardless of the schedule to weekly PTX did not seem to add clinically significant toxicity. pCR rate in the breast is similar across treatment arms at this IA. NCT01204125.
Tesetaxel: Activity of an oral taxane as first-line treatment in metastatic breast cancer.
Authors: Seidman Andrew David , Schwartzberg Lee Steven , O'Shaughnessy Joyce , D'Andrea Gabriella , Rubin Peter , Katz Seth , Danesi Hassan , Itri Loretta , Hudis Clifford
Published: March 2020

1016

Background: Tesetaxel, unlike standard taxanes (docetaxel, paclitaxel), is not a substrate for Pgp, a major cause of taxane resistance in tumor models. In a DU4475 breast cancer xenograft that overexpresses Pgp, tesetaxel induced a 94% reduction in tumor size, markedly exceeding the activity of docetaxel (46%) and paclitaxel (26%). Tesetaxel is associated with substantially less neuropathy preclinically than equi-myelotoxic doses of docetaxel. In clinical studies to date, tesetaxel is not associated with hypersensitivity reactions (0% incidence in > 450 patients [pts]), thus eliminating the need for premedication and extended observation. In a prior study, tesetaxel (27-35 mg/m2 Q3 wks) achieved a 38% partial response (PR) rate as 2nd-line therapy in pts with metastatic breast cancer (MBC) who had progressed after multidrug anthracycline-containing regimens. To extend these data, we initiated a phase 2 study of tesetaxel as 1st-line therapy in women with MBC. Methods: Eligibility included MBC; HER2-; ECOG PS 0-1; and adequate organ function. Adjuvant chemotherapy (including taxanes) was allowed. Tesetaxel was administered orally without anti-allergic premedication at a starting dose of 27 mg/m2 once every 3 wks. Overall response rate (ORR; RECIST) was the primary endpoint. Results: All 45 pts have been accrued. Median age is 58 y (range 36-80); median time from diagnosis, 4.0 y (range 0-21); triple negative status, 8 pts at diagnosis, 14 at time of metastasis. Metastatic sites are lung (22 pts), lymph nodes (22), liver (24), and bone (21). Prior treatment includes anti-estrogen therapy (32 pts), adjuvant chemotherapy (31), prior taxane (25), and radiotherapy (28). ORR in 24 pts evaluable for response is 50% (1 CR [4%], 11 PR [46%]); 5 responding pts had prior taxane therapy. Neutropenia is the most common ≥ Grade 3 adverse event with 50% of cases observed after dose escalation to 35 mg/m2; febrile neutropenia and Grade 3 peripheral neuropathy occurred in 2 pts each. There were no hypersensitivity reactions. Conclusions: Tesetaxel is highly active in 1st-line MBC and overcomes multiple limitations of standard taxanes. Updated ORR and PFS for all pts will be presented. Weekly dosing will be evaluated in a new cohort of pts.
Everolimus with paclitaxel plus bevacizumab as first-line therapy for HER2-negative metastatic breast cancer (MBC): A randomized, double-blind, placebo-controlled phase II trial of the Sarah Cannon Research Institute (SCRI).
Authors: Yardley Denise Aysel , Bosserman Linda D. , Peacock Nancy Walker , Favret Anne , Morgan Susan Kay , Priego Victor M. , Bass J. David , Griner Paula L. , Burris Howard A. , Hainsworth John D. , O'Shaughnessy Joyce
Published: March 2020

1018

Background: Constitutive activation of mTOR and amplified PI3K/Akt/mTOR signaling are common in MBC, and increase as treatment-resistance is acquired. Everolimus (E), an mTOR inhibitor, has single agent activity, combines well with paclitaxel (P) and bevacizumab (B), and prolonged PFS when added to AI therapy in BOLERO-2. In this randomized phase II trial, E was added to P/B as first-line treatment of HER2-negative MBC. Methods: Women with untreated HER2-negative MBC were randomized (1:1) to P 90mg/m2 IV (days 1, 8, and 15) and B10mg/kg IV (days 1 and 15) q28 days with E 10mg PO (Arm 1) or placebo PO (Arm 2) daily. Response assessment was performed q8 weeks until progression or intolerable toxicity. Primary endpoint was PFS. Secondary endpoints: safety, overall response rate, response duration, overall survival. 110 pts allowed detection of improvement in median PFS from 11 to 16 months with 70% power. Results: Between 8/2009 and 6/2011, 112 pts were randomized (Arm 1=55; Arm 2=57). Median age: 58 years (range: 25-79). 88% were ER+ or PR+. Pts received a median 5 treatment cycles (range: <1- 26+); 18 (16%) pts remain on treatment (Arm1, 9; Arm 2, 9). Median PFS were 8.8 months (Arm 1) and 7.1 months (Arm 2) (95% CIs 7.4-9.6; 5.5-9.1 months, p=0.79, HR 0.94). Complete responses were observed in 5% [Arm1, 4 (7%); Arm 2, 2 (4%)] with partial responses in 49% [Arm1, 29 (53%); Arm 2, 26 (46%)]. Responses rates in taxane pretreated pts Arm 1 22%, Arm 2 12%. Hematologic toxicity was similar in both arms; grade 3 mucositis occurred in 13% of E pts. Dose reductions (47% vs 25%) and interruptions (42% vs 26%) were more frequent with E due to mucositis and rash. Treatment discontinuation rates were similar. Conclusions: The addition of E did not result in a significant improvement in the efficacy of weekly paclitaxel/bevacizumab in the first-line treatment of HER2-negative MBC although response rates and median PFS were better with E. Possible explanation for these results may include lower dose intensity in the E arm, treatment of less resistant pts, or intrinsic differences in E activity when added to antiestrogen vs chemotherapy.
NSABP FB-6: Phase II trial of weekly paclitaxel (WP) and pazopanib following doxorubicin and cyclophosphamide (AC) as neoadjuvant therapy for HER2-negative locally advanced breast cancer (LABC).
Authors: Tan Antoinette R. , Buyse Marc E. , Rastogi Priya , Jacobs Samuel A. , Robidoux Andre , Flynn Patrick J. , Thirlwell Michael P. , Fehrenbacher Louis , Stella Philip J. , Goel Rakesh , Julian Thomas B. , Provencher Louise , Bury Martin Joseph , Paik Soonmyung , Geyer Charles E. , Swain Sandra M. , Mamounas Eleftherios P , Wolmark Norman
Published: March 2020

1025

Background: Pazopanib is an oral, small molecule inhibitor of VEGFR-1, -2, and-3, PDGFR-α, and -β, and c-kit tyrosine kinases. The purpose of this trial was to determine the activity and safety profile of pazopanib when added to neoadjuvant WP following AC in LABC. The primary endpoint was pathologic complete response in the breast and nodes (pCR-BN). Methods: Women with HER2-negative stage IIIA-IIIC breast cancer were treated with AC (60 mg/m2/600 mg/m2) for 4 cycles every 3 weeks followed by WP 80 mg/m2 on days 1, 8, and 15 every 28 days for 4 cycles concurrently with pazopanib 800 mg orally daily prior to surgery. Postoperatively, pazopanib was given for 6 months. The regimen would be considered active if ≥14 responses (16% pCR rate in breast and nodes) were observed in 87 evaluable patients. Patients were considered evaluable if they received at least 1 dose of pazopanib. Results: Between July 2009 and March 2011, 101 pts (median age 51 yrs, range 30-71) were enrolled; 56% had stage IIIA, 34% stage IIIB, and 10% stage IIIC disease. 74 pts (73%) had ER-and/or PR-positive tumors and 27 pts (27%) were triple negative. 8 patients did not begin pazopanib. The pCR-BN rate in evaluable patients for whom surgical information was known was 18% (16/89). The pCR-BN rate in ER positive disease was 9% (6/65) and was 42% (10/24) in TNBC. Toxicities observed with WP and pazopanib included diarrhea (gr 2/3, 10%/5%), hand-foot syndrome (gr 2/3, 11%/1%), hypertension (gr 2/3, 12%/3%), neuropathy (gr 2/3, 14%/1%), and neutropenia (gr 3/4, 25%/1%). Liver toxicity during WP and pazopanib included ALT (gr 2/3/4, 13%/7%/1%), AST (gr 2/3, 7%/7%), and total bilirubin (gr 2, 2%). Conclusions: A regimen of WP and pazopanib following AC was active as neoadjuvant therapy in women with LABC and met the pre-specified criteria of interest. The activity in TNBC was notable. The toxicity profile of WP and pazopanib was consistent with previous experience. Support: GlaxoSmithKline.
Molecular comparison of human triple-negative inflammatory breast cancer (TN-IBC) and human triple-negative noninflammatory breast cancer (TN-non-IBC).
Authors: Masuda Hiroko , Baggerly Keith , Wang Ying , Iwamoto Takayuki , Brewer Takae , Pusztai Lajos , Kai Kazuharu , Woodward Wendy A. , Reuben James M. , Van Laere Steven J. , Bertucci François , Hortobagyi Gabriel N. , Ueno Naoto T.
Published: March 2020

1047

Background: IBC has a poor prognosis because of its high rate of recurrence. There is an urgent need to define the biology of IBC to develop molecular-based targeted therapies for this disease. TNBC has a similar poor prognosis. Recently, Lehmann et al. (JCI, 2011) identified 7 subtypes of TNBC: basal-like (BL) 1 and 2, immunomodulatory (IM), mesenchymal (M), mesenchymal stem-like (MSL), luminal androgen receptor (LAR), and “unknown.” In light of these findings, we hypothesized that the distribution of TNBC subtypes differs between TN-IBC and TN-non-IBC. Methods: We qualitatively reproduced the Lehmann et al. experiments using Affymetrix CEL files from the same datasets to ensure the reproducibility of their findings. We quantified all arrays using frozen robust multiarray analysis with a linear model adjustment to account for batches. Then validated the results in a TNBC cohort from the World IBC Consortium for which IBC status was known (41 cases of TN-IBC; 53 cases of TN-non-IBC). We used the Fisher exact test to determine associations between TNBC subtypes and IBC status. We used Kaplan-Meier curves and log-rank tests to compare clinical outcomes between TNBC subtypes. Results: We found the 7 subtypes for both TN-IBC and TN-non-IBC. While the correspondence between our findings and those of Lehmann et al. was not perfect, there was a very significant correlation (P<2.2x10-16). We found no association between TNBC subtype and IBC status (P=.5023). As expected, we found that patients with IBC had significantly worse recurrence-free survival (RFS) than a comparison cohort of patients with advanced non-IBC that included not only patients with TNBC but also patients with ER-positive and HER2-amplified tumors (P=.0054). However, TNBC subtype did not predict RFS. IBC status was not a significant predictor of RFS or overall survival in the TNBC cohort. Conclusions: Both TN-IBC and TN-non-IBC are heterogeneous. TNBC subtypes are unrelated to IBC status. TN-IBC and TN-non-IBC have the same subtypes and clinical outcome. [Table: see text]
Evaluation of a multiparametric system able to predict nonsentinel lymph node status in breast cancer patients with a micrometastatic sentinel node assessed by the one step nucleic acidamplification (OSNA) assay.
Authors: Buglioni Simonetta , Mottolese Marcella , Casini Beatrice , Gallo Enzo , Terrenato Irene , Pescarmona Edoardo , Di Filippo Simona , Marandino Ferdinando , Ferretti Gianluigi , Di Filippo Franco
Published: March 2020

1050

Background: Axillary lymph node dissection (ALND) may not be necessary in women with breast cancer (BC) who have micrometastasis in a sentinel lymph node (SLN), owing to the low risk of non-SLN (NSLN) involvement. In our Institute we validated and adopted the molecular diagnostic tool OSNA based on the quantitative measurement of Cytokeratin 19 (CK19) mRNA.The aims of our work in a subgroup of women with micrometastatic SLN, were: 1) to correlate the copy numbers of CK19 mRNA with the risk of additional positive NSLNs; 2) to assess the relationships between the molecular subtype classification and the probability of a positive ALND; 3) to verify whether a combination of the new above mentioned parameters is able to identify a subgroup of patients with a micrometastatic SLN and a negligible risk of positive NSLNs in whom ALND may be avoided. Methods: The SLN lysates from 709 patients were analyzed by OSNA assay. We considered only patients with a micrometastatic SLN (copy numbers between 250 and 5000/μL) and the probability of having a positive ALND was calculated by the logistic regression model. This series of BC patients were divided into four main subtypes taking in account the BC classification as defined by a combination of estrogen, progesteron receptors and HER2 status. Results: OSNA positivity for micrometastasis was reported in 91/709 cases (12,8%).The number of patients with positive ALND was 20 (22%). The statistical analyses showed that the metastatic involvement of NSLNs is associated with SLNs with a high copy numbers (>2000) of CK19 mRNA together with HER2 subtype. Otherwise none of the luminal A patients with a positive SLN but presenting a copy number <1000, had a positive NSLNs. Conclusions: We showed that biologically-driven analyses may be able to build new models with higher performance in terms of breast cancer axillary status prediction after positive SLN biopsy for micrometastasis. The copy numbers of CK19 mRNA and the molecular subtypes are more advantageous than traditional parameters because they are not pathologist-dependent and therefore they are more reliable and reproducible.
DEAR1, a novel tumor suppressor, negative regulator of epithelial-mesenchymal transition, and prognostic factor in non-small cell lung cancer.
Authors: Quintas-Cardama Alfonso , Post Sean M. , Solis Luisa , Xiong Shunbin , Wistuba Ignacio I. , Lozano Guillermina
Published: March 2020

10506

Background: DEAR1, also known as TRIM62, maps to 1p.35, a genomic interval within which loss of heterozygosity occurs at high frequency in carcinomas. DEAR1 has been proposed as a regulator of cell polarity in vitro. Methods: We genetically engineered mice lacking DEAR1 alleles at murine chromosome 4. To study the impact of DEAR1 loss in lung cancer, DEAR1 deficient mice were crossed to mice carrying the latent mutant K-rasG12D allele (K-rasLA1). Results: No survival difference was observed between DEAR1+/─ and DEAR1─/─ mice (729 vs 699 days, p=0.98) but they were markedly shorter than that of DEAR1 wild-type littermates (803 days, p=0.01). Over 90% of mice developed tumors, mainly adenocarcinomas, including invasive and/or metastatic lung cancer in 23% of cases. DEAR1-deficient K-rasLA1 mice had reduced life span compared with DEAR1+/+:K-ras+/LA1 littermates (184 vs 291 days, p<0.0001). The survival of DEAR1+/─:K-ras+/LA1 and DEAR1─/─:K-ras+/LA1 mice were similar (180 vs 153 days, p=0.38) and qPCR and IHC analyses suggested loss of the wild-type DEAR1 allele. The survival of DEAR1-deficient K-rasLA1 mice was also shorter than that of control p53+/─:K-ras+/LA1 mice (240 days; p=0.01). The metastasis incompetent K-rasLA1–positive LKR13 cell line after shRNA DEAR1 knock-down and cell lines derived from DEAR1-deficient K-rasLA1 tumors recapitulated an invasive phenotype in transwell migration and 3D culture assays and a metastatic phenotype on SC or IV injection into immunocompetent wild-type littermates. The latter was associated with epithelial-mesenchymal transition involving loss of E-cadherin and DEAR1 expression and upregulation of vimentin, Twist, and CD44. DEAR1 downregulation was very prevalent in a non-small cell lung cancer (NSCLC) tissue array with 214 samples. Patients with early stage NSCLC and loss of DEAR1 expression had a markedly shorter time to relapse compared to those retaining DEAR1 expression (5.1 vs 2.87 years, p=0.049) and worse 5-year relapse-free and overall survival rates. Conclusions: DEAR1 is a novel tumor suppressor that negatively regulates EMT and promotes lung cancer metastasis. DEAR1 loss is a poor prognostic factor in NSCLC.
Mutational analysis of circulating tumor cells in breast cancer patients by targeted clonal sequencing.
Authors: Xu Xunhai , Shen Yufeng , Dewitt William , Pandya Deep , Bischoff Farideh Z. , Crew Katherine D. , Hershman Dawn L. , Maurer Matthew A. , Parsons Ramon , Kalinsky Kevin
Published: March 2020

10516

Background: The molecular signature of circulating tumor cells (CTCs) may serve as a surrogate marker for accurate description of the metastatic tumor of interest, especially in the setting of treatment response and selection. We present a method for mutational analysis of CTCs in metastatic breast cancer (MBC) patients by using an emulsion-formulated, semiconductor-based, targeted clonal sequencing platform. Methods: CTCs in MBC patients were enriched by a microfluidic OncoCEE device using antibodies against both epithelial and mesenchymal markers. Genomic DNA was extracted from enriched CTC samples. Emulsion-based multiplex-PCR targeted for various cancer genes was performed, after which semiconductor-based deep sequencing was completed. The read error rates were analyzed based on quality score and context of sequence including homopolymers. Statistical significance for each mutational analysis was assessed using a method based on beta-binomial distribution. Results: Of the 17 patients samples obtained, we were able to enrich CTC samples in 9 of them (CTC range 1-1063, median=12). Multiplex targeted sequencing was performed on DNA from the enriched CTC patient samples (purity range 0.3-6%). Greater than 3000-fold coverage was accomplished. Missense mutations at E545D on PIK3CA (p= 2.0e-07), F354L on STK11 (p=2.0e-04), and Q61R on NRAS (p=2.0e-07) were detected. Novel mutations of L540F and Q1033K within the hot spot regions of PIK3CA were observed (p=1.3e-04). Genomic DNA from WBCs from a healthy female was analyzed concurrently as a negative control, in which none of the mutations were observed. Conclusions: Mutational analysis of CTCs in MBC patients can be accomplished by deep sequencing. We developed a de novo protocol for clonal mutation analysis on CTCs in MBC and detected various significant and novel mutations. We anticipate reporting sequencing results on CTCs and matched WBCs, as negative controls, from 40 MBC patients. This will provide the foundation for the future studies in which we will compare the mutational profile between CTCs and primary/metastatic tumors. We intend to validate clonal mutational analysis of CTCs as a predictive blood-based biomarker in subsequent trials.
Intraoperative detection of lymph node metastasis using one-step nucleic acid amplification (OSNA) in breast cancer patients: Effect on second surgery rate and delay for adjuvant therapy.
Authors: Klingler Sophie , Marchal Frederic , Rauch Philippe , Kenouchi Ouarda , Chrétien Anne-Sophie , Genin Pascal , Leroux Agnès , Merlin Jean-Louis
Published: March 2020

10517

Background: Sentinel lymph node (SLN) analysis is conventionally analysed using HES and CK19 immunohistochemistry. In case of SLN involvement, a second surgery is required for axillary lymph node (ALN) resection thus delaying the initiation of adjuvant therapies. Methods: 381 pts with invasive breast cancer were considered in this retrospective study. SLN were detected using combined radio-isotope and color detection. SLN involvement was analysed using OSNA for CK19 mRNA, in 100 pts (group 1) and compared to conventional histopathology in 281 pts (reference population, group 2). In all cases, control cytology was performed. Results: No significant difference was found between group 1 and 2 regarding patients characteristics, tumor localization, size, grade, steroid receptors and HER2 expression and the mean number of SLN analysed per pt, 2.4 (range 1-7) and 2.5 (range 1-8), respectively. Considering positive SLN as “++” (CK19 mRNA copy number>5000), “+” (250 < CK19 mRNA copy number < 5000) and positive by inhibition in group 1 and macro-, micrometastases and isolated tumor cells in group 2, no difference in lymph node involvement rate was found between the two groups with 29.0 and 29.9% of positive SLN, respectively. Only one discordance case was observed with negative OSNA analysis and positive cytology with isolated tumor cells. Using OSNA intraoperatively, the mean time to process SLN was 42 min (range 10-104) allowing immediate ALN resection. Significant (P<.01) reduction of re-intervention rate was observed when OSNA was used (9 vs 39%), including margins insufficiency (5 vs 13%), ALN resection (3 vs 15%) or both (1 vs 11%). Delay before adjuvant therapy was also significantly (P <.01) reduced when OSNA was used with 43 (range 20-82) vs 59 (range 14-212) days. Conclusions: Results achieved with OSNA are fully consistent with those achieved using conventional immunohistochemistry analysis. SLN analysis using OSNA avoids a second operation in most patients for ALN resection and shortens the delay for adjuvant therapy.
Reproducibility, performance, and clinical utility of a genetic risk prediction model for prostate cancer in Japanese patients.
Authors: Akamatsu Shusuke , Takata Ryo , Takahashi Atsushi , Inoue Takahiro , Kubo Michiaki , Kamatani Naoyuki , Inazawa Johji , Fujioka Tomoaki , Habuchi Tomonori , Ogawa Osamu , Nakamura Yusuke , Nakagawa Hidewaki
Published: March 2020

10520

Background: Prostate specific antigen (PSA) is widely used as a diagnostic biomarker for prostate cancer (PC). However, due to its low predictive performance, many patients without PC suffer from the harms of unnecessary prostate needle biopsies. The present study aims to evaluate the reproducibility and performance of a genetic risk prediction model and estimate its utility as a diagnostic biomarker in a clinical scenario. Methods: We created a logistic regression model incorporating 16 SNPs that were significantly associated with PC in a genome-wide association study of the Japanese. The model was validated by two independent sets of samples comprising 3,294 cases and 6,281 controls. Various cut offs were evaluated to be used in a clinical scenario. Results: The area under a curve (AUC) of the model was 0.679, 0.655, and 0.661 for the samples used to create the model and those used for validation respectively. The AUC of the model was not significantly altered in samples with PSA 1-10 ng/ml. 24.2% and 9.7% of the patients had odds ratio <0.5 (low risk) or >2 (high risk) in the model, and assuming the overall positive rate of prostate needle biopsies to be 20% in PSA gray zone (PSA 2-10 ng/ml), the positive biopsy rates were 10.7% and 42.4% respectively for the two genetic risk groups. Conclusions: The genetic risk prediction model was highly reproducible, and its predictive performance was not influenced by PSA. The model could have a potential to affect clinical decision when it is applied to patients with gray-zone PSA, which should be confirmed in future clinical studies.
Frequency and clinical characterization of NSCLC patients harboring PIK3CA mutations identified within a regional screening network.
Authors: Gardizi Masyar , Scheffler Matthias , Heukamp Lukas Carl , Bos Marc Christiaan Allardt , Albus Kerstin , Hayn Birgit , Ko Yon-Dschun , Schlesinger Andreas , Brockmann Michael , Serke Monika Heidi , Gerigk Ulrich , Hekmat Khosro , Stoelben Erich , Reiser Marcel , Schnell Roland , Schmitz Stephan H. , Frank Konrad , Buettner Reinhard , Wolf Jurgen , Zander Thomas
Published: March 2020

10526

Background: PIK3CA mutations are a rare oncogenic event of potential therapeutic relevance in NSCLC. Here we report frequency and characteristics of patients with PIK3CA mutated lung tumors. Methods: Patients with NSCLC and PIK3CA mutations were identified within our regional Network for Molecular Screening in Lung Cancer. We further analyzed the presence of BRAF, KRAS, EGFR mutations as well as ALK translocation, ERBB2 and FGFR1 amplifications in PIK3CA mutated samples. Clinical data on age, sex, TNM classification and tumor stage, histological type, grading, overall survival, smoking status, comorbidity, BMI and secondary malignancies were retrieved from clinical charts in accordance with the local ethics committee. Results: PIK3CA mutations were detected with a frequency of 3.7% (24% exon 20,76% exon 9) in 1000 patients. Histologically 32% were defined as squamous cell carcinoma, 48% as adenocarcinoma and 18% other histological subtypes or NSCLC-NOS. Exon 9 mutations were present in the acinar and lepidic subtype, whereas exon 20 mutations were seen in the papillary and solid subtype. Cooccuring genetic lesions were observed in 16% (mutations in KRAS=2, EGFR=1, BRAF=1; FGFR1 amplification=2). 14 were female, 23 male with a mean age of 69 years. 21 of these patients were further clinically annotated. 11 patients presented with stage IIIb/IV eligible for palliative treatment and 10 stage I – IIIa eligible for surgical therapy +/- adjuvant therapy. All but 1 patient were smokers with an average BMI of 26,2kg/m2 with a typical high load of comorbidity mainly of cardiovascular diseases, 8 of 21 patients showed prior malignancies in their medical history. The median overall survival within this population has not been reached yet. Conclusions: Screening for PIK3CA mutations is feasible. A high proportion (38%) of patients with PIK3CA mutated lung cancer have prior malignancies and show a high load of comorbidity. Furthermore PIK3CA mutations are not exclusive to KRAS, EGFR or BRAF mutations or FGFR1 amplifications. Successful identification of patients with oncogenic lesions in lung cancer in a screening network might allow future personalized treatment of these patients.
Circulating melanoma cells isolated from clinical blood samples and characterized by full-length mRNA sequencing at single-cell level.
Authors: Wang Yu-Chieh , Ramskold Daniel , Luo Shujun , Li Robin , Deng Qiaolin , Daniels Gregory A. , Khrebtukova Irina , Loring Jeanne F. , Laurent Louise C. , Schroth Gary P. , Sandberg Rickard
Published: March 2020

10539

Background: Melanoma is the most aggressive type of skin cancer. Late-stage melanoma is highly metastatic and currently lacks effective treatment. This discouraging clinical observation highlights the need for a better understanding of the molecular mechanisms underlying melanoma initiation and progression and for developing new therapeutic approaches based on novel targets. Although genome-wide transcriptome analyses have been frequently used to study molecular alterations in clinical samples, it has been technically challenging to obtain the transcriptomic profiles at single-cell level. Methods: Using antibody-mediated magnetic activated cell separation (MACS), we isolated and individualized putative circulating melanoma cells (CMCs) from the blood samples of the melanoma patients at advance stages. The transcriptomic analysis based on a novel and robust mRNA-Seq protocol (Smart-Seq) was established and applied to the putative CMCs for single-cell profiling. Results: We have discovered distinct gene expression patterns, including new putative markers for CMCs. Meanwhile, the gene expression profiles derived of the CMC candidates isolated from the patient’s blood samples are closely-related to the expression profiles of other cells originated from human melanocytes, including normal melanocytes in primary culture and melanoma cell lines. Compared with existing methods, Smart-Seq has improved read coverage across transcripts, which provides advantage for better analyzing transcript isoforms and SNPs. Conclusions: Our results suggest that the techniques developed in this research for cell isolation and transcriptomic analyses can potentially be used for addressing many biological and clinical questions requiring genomewide transcriptome profiling in rare cells.
HER2 expression on circulating tumor cells (CTC) in patients with early HER2-positive breast cancer: Results of the German SUCCESS B trial.
Authors: Jaeger Bernadette Anna Sophia , Rack Brigitte Kathrin , Neugebauer Julia Katharina , Andergassen Ulrich , Melcher Carola Anna , Mouarrawy Doraid , Ziemendorff Gabriele , Clemens Michael , von Abel Ekkehard , Heinrich Georg , Schueller Katharina , Schneeweiss Andreas , Lichtenegger Werner , Beckmann Matthias W. , Pantel Klaus , Sommer Harald Leo , Friese Klaus , Janni Wolfgang
Published: March 2020

10540

Background: Recent reports showed a discrepancy in the Her2-status of metastases or minimal residual disease in blood and bone marrow compared to the primary tumor in patients with breast cancer. The Her2-status of CTC was prospectively evaluated in the German multicenter SUCCESS B study. Methods: The SUCCESS B trial is a randomized Phase III study comparing FEC-Docetaxel (Doc) vs. FEC-Docetaxel-Gemcitabine (Doc-G) as well as Her2 targeted therapy with Trastuzumab as adjuvant treatment in patients with early, Her2 positive, node positive or high risk node negative primary breast cancer. As part of the translational research program, 23ml of peripheral blood were drawn before adjuvant chemotherapy. In 638 samples CTC and Her2-status were assessed using the CellSearch System (Veridex, USA). After immunomagnetic enrichment with an anti-Epcam-antibody, cells were labeled with anti-CK8/18/19, anti-CD45 antibodies as well as a fluorescein conjugate antibody for Her2 phenotyping. Cutoff for CTC-positivity was ≥1 CTC and for Her2 ≥1 CTC with strong Her2-staining (+++). Results: 40.2% of pts (n=257) were positive for CTC (mean 4.52; range 0-1689). The number of detected CTC was distributed as follows: 1 CTC (n=112; 43.6%), 2 CTC (n=65; 25.3%), 3 CTC (n=36; 14.0%), 4 CTC (n=12; 4.7%) and ≥5 CTC (n=31; 12.1%). Her2 status on CTC was negative or weak in 12.5% (n=32) and 8.9% (n=23) of CTC positive patients respectively and therefore categorized as Her2 negative. In 21.4% (n=55) we detected moderate and in 57.2% (n=147) strong Her2-staining of ≥1 CTC per sample. Therefore 57.2% of CTC-positive samples were diagnosed as Her2 positive and 21.4% as questionable. No association was found between the detection of CTC or the Her2-status on CTC with tumor size, histopathological grading, hormone receptor status or axillary lymph node involvement. Conclusions: The data of this trial confirms the frequent discordance of Her2 expression on CTC compared to the primary tumor. Survival data within the Success B trial will give further insight into the tumor biology of Her2 positive disease and the role of the Her2-status on CTC to predict treatment efficacy.
Combined targeting of LSD1 (KDM1A) and histone deacetylases exerts superior efficacy against human AML.
Authors: Fiskus Warren , Sharma Sunil , Rao Rekha , Balusu Ramesh , Venkannagari Sreedhar , Ganguly Siddhartha , Bearss David , Bhalla Kapil N.
Published: March 2020

10549

Background: LSD1 (KDM1A) is FAD-dependent histone H3K4Me2 demethylase. Inhibition of LSD1 increases H3K4Me3-a permissive mark for gene expression, and inhibits growth of pluripotent cancer cells. We have previously noted that treatment with the histone deacetylase (HDAC) inhibitor panobinostat (PS) depletes EZH2 (the catalytic subunit of the polycomb repressive complex 2, PRC2) and disrupts its interaction with the other PRC2 proteins, attenuates LSD1, and de-represses growth inhibitory and pro-apoptosis genes. Methods: In the present studies, we determined the chromatin effects and the pre-clinical in vitro and in vivo anti-AML activity of the novel, non-monoamine oxidase, reversible inhibitor of LSD1, HCI2509 (100 to 500 nM), alone and in combination with PS, utilizing cultured (OCI-AML3 and HL-60) and primary human AML blast progenitor cells. Results: Treatment with HCI2509 dose-dependently increased the levels of H3K4Me3, p16, p27, and CEBPα in cultured AML cells. This correlated with inhibition of cell growth and induction of morphologic differentiation in AML cells. Exposure to HCI2509 disrupted the binding of LSD1 with the co-repressor CoREST and HDAC1. Treatment with PS (10 to 50 nM) dose-dependently depleted not only LSD1 but also EZH2, SUZ12 and BMI1 in AML cells. Co-treatment with PS enhanced the chromatin modifying effects of HCI2509. The combination also synergistically induced loss of viability of cultured and primary AML cells (combination indices, CI <1.0). Following tail vein infusion and establishment of AML by OCI-AML3 cells in NOD-SCID mice, treatment with HCI2509 (25 mg/kg, b.i.w, IP, for 3 weeks) improved survival, as compared to the control mice (p <0.001). HCI2509 treatment (15 mg/kg IP) also dramatically improved survival of NSG mice with established human AML following tail-vein injection of primary AML blasts expressing FLT3-ITD. Survival was further improved upon co-treatment with HCI2509 and PS (5 mg/kg IP, MWF). Mice did not experience any toxicity or weight loss. Conclusions: Combined epigenetic therapy with HCI2509 and PS exerts superior pre-clinical activity, warranting further in vivo development and testing of HCI2509 against human AML.
Prognostic role of microRNA polymorphisms in advanced gastric cancer patients.
Authors: Stenholm Linnea , Stoehlmacher-Williams Jan , Al-Batran Salah-Eddin , Heussen Nicole , Akin Sema , Pauligk Claudia , Lehmann Sylvia , Senff Tina , Ehninger Gerhard , Brummendorf Tim H. , Goekkurt Eray
Published: March 2020

10552

Background: As little is known about the impact of microRNA (mir) polymorphisms on prognosis of advanced gastric cancer (AGC) we sought to determine their impact on overall survival (OS) in these patients (pts). Methods: Pts were treated with either 5-fluorouracil, leucovorin and oxaliplatin (FLO) or with additional docetaxel (FLOT) in frame of a phase III or 3 phase II trials of the Arbeitsgemeinschaft Internistische Onkologie. All pts consented genetic analyses. DNA was extracted from whole blood samples (n=515) and polymorphisms of mir26a1 (rs7372209), mir27a (rs895819), mir100 (rs1834306), mir125a (rs12975333), mir146a (rs2910164), mir196a2 (rs11614913), mir219-1 (rs107822) and mir423 (rs6505162) were genotyped using PCR-based methods. Hetero- and homozygous variant genotypes were grouped versus homozygous wild type genotypes. Selection of clinical and genetic parameters for the final multivariate model was based on univariate and multivariate Cox-regression analyses with a cut-off value of p < 0.25 and included 498 pts. Results: Median age was 67 (range 23 to 86) years. 42% were treated with FLO and 58% with FLOT with a median OS of 14 months. Multivariate analyses revealed following associations: Genetic factors significantly associated with OS were mir26a1 variant genotypes (HR 1.3 [95%CI (1.01;1.6), p=.025]), mir27a variant genotypes (HR 1.3 [95%CI (1.01;1.6), p=.036]) and mir196a2 variant genotypes (HR 0.8 [95%CI (0.6;0.99), p=.046]). Clinical factors with significant impact on OS were ECOG 2 performance status (HR 1.9 [95%CI (1.3;2.8), p=.002]), curative surgery of advanced disease (HR 0.3 [95%CI (0.1;0.5), p<.001]) and locally advanced disease (HR 0.5 [95%CI (0.3;0.7), p<.001]). Combined analyses of the identified adverse genotypes of mir26a1, mir27a and mir196a2 resulted in a median OS of 9, 13, 14 and 17 months for pts with 0 (n=72), 1 (n=193), 2 (n=171) and 3 (n=54) adverse genotypes (p=.029), respectively. Conclusions: In addition to established clinical factors, mir26a1, mir27a and mir196a2 polymorphisms were significantly associated with OS. Our data suggest a strong impact of these mir polymorphisms on prognosis in AGC and might be of help for a better guidance of treatment decisions.
Activation of and dependence on ataxia telangiectasia mutated kinase in PTEN-deficient tumor cells.
Authors: McCabe Nuala , Walker Steven M. , Goffard Nicolas , Wikstrom Katarina , Greenan Caroline , McLean Estelle , McDyer Fionnuala A , Keating Karen E , James Iain , Harrison Timothy , Mullan Paul B , Harkin D. Paul , Chen Clark , Kennedy Richard D.
Published: March 2020

10553

Background: Loss of PTEN function has been widely reported to cause up-regulation of the PI3K/AKT signalling pathway resulting in increased cell growth, proliferation and survival. More recently it has been reported that PTEN null cells demonstrate genomic instability and have increased production of reactive oxygen species (ROS) and oxidative stress induced DNA damage. Ataxia Telangiectasia Mutated (ATM) is the primary response kinase, which responds to stalled DNA replication and DNA double strand breaks due to oxidative DNA damage. Methods: A metagene representing ATM activation was generated from cell line data and used to perform hierarchical clustering analysis of public DNA microarray profiling datasets of breast cancer, ovarian cancer and glioblastoma with known PTEN IHC/mutation status. Furthermore, we ask if ATM activation may be therapeutically exploited in PTEN null tumours using ATM specific siRNA and compounds in 2 PTEN isogenic cell line model systems. Results: We show that PTEN null cells have elevated levels of ROS, DNA damage and have endogenous activation of ATM, an enzyme important in responding to DNA damage resulting from oxidative stress. We hypothesised that PTEN deficient tumours may rely on ATM enzyme for survival. To investigate this we generated a 189-gene list representing ATM activation and used this to perform hierarchical clustering analysis of a breast cancer DNA microarray dataset. This list was able to significantly cluster tumours with known loss of PTEN expression (p=0.004). Furthermore, this gene list was able to segregate PTEN null/mutant tumours from PTEN wild-type tumours in 2 independent datasets of glioblastoma and ovarian cancer (p=0.015 and p=0.012). In addition, we found that inhibition of ATM using the selective inhibitor KU-55933 caused DNA damage, cell cycle arrest and apoptosis specifically in PTEN deficient cells when compared to PTEN wild-type cells. Conclusions: These observations suggest that ATM may represent a therapeutic target in PTEN deficient tumours and furthermore ATM activation may be the basis of a biomarker of PTEN status in human cancers.