bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of the multikinase inhibitor regorafenib in neuroblastoma.
Authors: Zage Peter E. , Subramonian Divya , Mo Qianxing , Huang Shixia
Published: September 2018

10553

Background: Neuroblastoma (NB) is the most common extracranial solid pediatric tumor, and children with high-risk NB have poor survival rates and need novel treatment strategies. Regorafenib, a multi-receptor tyrosine kinase (RTK) inhibitor approved for treating adult solid tumors such as advanced metastatic colorectal cancer and gastrointestinal stromal tumors, inhibits many RTKs, including PDGFR-β, VEGFR1-3, RET, c-Kit and FGFR family members. Based on the potential roles for these targets in neuroblastoma pathogenesis, we explored the therapeutic potential of Regorafenib alone and in combination with 13-cis-retinoic acid against neuroblastoma cells. Methods: We treated NB cell lines with increasing concentrations of Regorafenib and measured cell viability using MTT assays. We further measured the occupied percent confluence over time using continuous live cell imaging. We performed Western blots for caspase cleavage to measure apoptosis and flow cytometry to determine cell cycle expression. We performed Reverse Phase Protein Array (RPPA) analysis of neuroblastoma cells before and after treatment with regorafenib combined with 13- cis-retinoic acid. Results: IC50values for the tested cell lines ranged between 2.5mcM and 12.5mcM after 72 hours of exposure to Regorafenib, and decreased viability was due to a combination of apoptosis and cell cycle arrest. RPPA analysis identified alterations in multiple proteins and pathways after Regorafenib with retinoic acid treatment, including the PI3K/Akt/mTOR and Jak/Stat pathways. Phosphorylation of Erk1/2, S6, Akt, and c-Jun were decreased, while protein expression of GATA3 was increased in a dose-dependent manner. Conclusions: Regorafenib treatment results in reduced neuroblastoma cell viability and increased apoptosis via effects on several signaling pathways. Effects on intracellular signaling pathways associated with responses to the combination of regorafenib plus retinoic acid represent opportunities to develop novel combination therapies, representing potential new therapeutic strategies for children with neuroblastoma.
Risk factors associated with metastatic site failure in patients with high-risk neuroblastoma.
Authors: Lucas John Thomas , Wakefield Daniel Victor , Doubrovin Michael , Li Yimei , Cooper David , Manole Bogdan-Alexandru , Santiago Teresa , Federico Sara Michele , Merchant Thomas E. , Davidoff Andrew M. , Krasin Matthew J. , Shulkin Barry L. , Santana Victor M. , Furman Wayne Lee
Published: September 2018

10557

Background: This retrospective study sought to identify predictors of metastatic site failure (MSF) in patients with high-risk (HR) neuroblastoma (NB). Methods: Seventy-six patients with HR NB treated on prospective trials at from 1997 to 2014 were eligible for inclusion. All patients were treated with induction chemotherapy (chemo) with surgery followed by myeloablative chemo & stem cell rescue. Primary & metastatic site (MS) RT were applied according to institutional protocol. CT & I-123 MIBG scans were used to assess Curie scores at diagnosis, post-induction, post-transplant & failure. Overall (OS), & progression-free survival (PFS) were described using the Kaplan-Meier estimator. Cox proportional hazards frailty (cphfR) & CPH regression (CPHr) were used to identify covariates predictive of MSF & new site MSF. Results: Forty-two (55%) patients had documented MSFs. Consolidative MS RT was applied to 30 MSs in 10 patients. Original site MSF occurred in 146 of 383 (38%) & 18 of 30 (60%) non-irradiated & radiated MSs respectively. Original site MSF occurred in post-induction MIBG avid lesions in 68 of 81 (84%) & 12 of 14 (85%) non-irradiated & radiated MSs respectively. The median OS & PFS were 61 mo (95% CI 42.6-NR) & 24.1 mo (95% CI 16.5-38.7). Univariate cphfR identified an increased hazard for original MSF when MIBG avid following induction chemo (HR 4.9, 95%CI 1.1-20.9, p = 0.03) & transplant (HR 7.3 95%CI 1.8-30.2, p = 0.006) relative to lesions that cleared after induction. Notably, MS RT nor site location did not modify the hazard for MSF. Multivariate CPHr identified inability to undergo transplant (HR 32.4 95%CI 9.3-96.8, p < 0.001) &/or maintenance chemo (HR 5.2, 95%CI 1.7-16.2, p = 0.005) & the presence of lung metastases (HR 4.4 95%CI 1.7-11.1, p = 0.002) at diagnosis as predictors of new site MSF. The new MSF free survival at 3 years was 25% vs. 87% in patients with high-risk factors relative to those without the risk factors suggesting limited benefit of consolidative MS RT in this population. Conclusions: Metastatic lesions that remained MIBG avid following induction chemo & post-transplant had an increased hazard for MSF. Consolidative site RT likely has limited benefit in patients with HR features.
Is incomplete estradiol suppression during aromatase inhibitor treatment in post-menopausal patients with breast cancer due to insufficient systemic drug concentrations?
Authors: Hertz Daniel Louis , Kidwell Kelley M. , Speth Kelly A , Gersch Christina L , Desta Zeruesenay , Storniolo Anna Maria , Stearns Vered , Skaar Todd C , Hayes Daniel F. , Henry Norah Lynn , Rae James M.
Published: September 2018

1063

Background: Aromatase inhibitors (AI) suppress estrogen biosynthesis and are effective treatments for estrogen receptor (ER)-positive breast cancer. In a prospectively enrolled cohort we observed a subset of post-menopausal women who exhibit high plasma estradiol (E2) concentrations during AI treatment, which could potentially contribute to treatment failure. We tested the hypothesis that incomplete E2 suppression is due to insufficient systemic AI concentrations. Methods: Five hundred post-menopausal women with ER-positive breast cancer were randomized to daily exemestane (Exe) 25 mg or letrozole (Let) 2.5 mg. Plasma E2 was measured using GC/MS/MS (lower limit of quantification (LLOQ) = 1.25 pg/mL) at baseline and after 3 months. Let and Exe plasma concentrations measured after 1 or 3 months were compared with the magnitude of E2 depletion using four complementary statistical procedures to assess associations of drug concentrations with: 1) a binary outcome of E2 suppression below LLOQ (logistic regression), 2) 3-month E2 concentrations (linear regression), 3) absolute change from baseline in E2 concentrations (Spearman correlation), and 4) an ordinal outcome defined by E2: decreased to below LLOQ, decreased but not to LLOQ, stayed the same, or increased from baseline (cumulative logistic regression). Results: 397 patients with E2 and AI concentration measurements were evaluable (Exe n = 199, Let n = 198). Thirty (7.6%) patients (Exe n = 13, Let n = 17) had E2 concentrations above the LLOQ at 3 months (range: 1.42-63.8 pg/mL). Exe and Let concentrations were not associated with achievement of unmeasurable E2 concentrations, on-treatment E2 concentrations, E2 change from baseline, or ordinal groupings of E2 change (all p > 0.05). In a parallel analysis there was no association of estrone-sulfate and drug concentrations (data not shown). Conclusions: Our results suggest that circulating drug concentrations do not explain incomplete E2 suppression in women receiving AI therapy. Additional studies are underway to determine whether age, body mass and genetic variation in the aromatase enzyme influence AI treatment response.
Primary surgery versus no surgery in synchronous metastastic breast cancer: Patient-reported outcomes of the ABCSG 28 Posytive trial.
Authors: Bjelic-Radisic Vesna , Fitzal Florian , Steger Guenther G. , Marth Christian , Balic Marija , Knauer Michael , Singer Christian F. , Hubalek Michael , Schrenk Peter , Haid Anton , Wette Viktor , Swoboda Ursula , Fuegger Reinhold , Greil Richard , Soelkner Lidija , Gnant Michael ,   ABCSG
Published: September 2018

1074

Background: The ABCSG 28 Posytive trial compared primary surgery versus primary systemic therapy without surgery in synchronous metastatic breast cancer. The primary aim of the study was to investigate whether immediate resection of the primary tumor followed by standard systemic therapy improves median survival compared with no surgical resection (NCT01015625). This report describes quality-of-life (QoL) results. Methods: Patients were randomized between 2011 and 2015. Patients completed the EORTC QLQ-C30 and EORTC QLQ-BR 23 before treatment and every 6 months during follow-up. Results: 90 patients (45 with surgery, 45 with primary systemic therapy without surgery) from 15 centers were included in the QoL analysis. At 6 months patients after surgery reported more insomnia, breast symptoms and arm symptoms than patients without surgery, but these differences were no longer present at later follow-up visits. In the univariate and multivariate analysis the global health status and physical functioning scales of EORTC QLQ C30 were statistically significant predictors for OS and TTP (p < 0.05). Over a 24-month follow-up, patients > 60 years showed more QoL impairments than those < 60 years, independent of treatment. Patients < 60 years had better physical functioning and less fatigue, appetite loss, constipation and breast symptoms than older patients. There were no differences in QoL between patients with bone metastases vs those with visceral ± bone metastases. Conclusions: In patients with primary metastatic breast cancer primary surgery does not appear to improve QoL. Global health status and physical functioning scales of EORTC QLQ-C30 appears to be a predictive factor for OS and TTP. Clinical trial information: NCT01015625.
A phase I trial of mifepristone (M), carboplatin (C), and gemcitabine (G) in advanced breast and ovarian cancer.
Authors: Stringer Erica Michelle , Saha Poornima , Swoboda April , Kocherginsky Masha , Baker Gabrielle , Olberkyte Simona , Owusu Nana , Dyer Brittany , Olalde Lauren , Olszewski Elizabeth , Hahn Olwen Mary , Hoffman Philip C. , Phillips Brooke Elizabeth , Suh Grace K. , Conzen Suzanne D. , Fleming Gini F. , Nanda Rita
Published: September 2018

1083

Background: Glucocorticoid receptor (GR) activity inhibits chemotherapy-induced apoptosis, and GR antagonism with m enhances chemotherapy sensitivity in GR+ breast (B) and ovarian cancer (OC) cells. C+G is a commonly used regimen for B and OC. We report the results of a phase I trial of the GR antagonist m plus C+G in patients with advanced B and OC. Methods: A standard “3+3” dose escalation phase I study was performed. Objectives were to assess the safety and tolerability of the regimen, and to determine the recommended phase 2 (RP2D) dose of M+C+G. C+G was administered on days 1 and 8 of a 21 day cycle, and m was administered the day prior to and the day of chemotherapy. The starting dose level (DL) was 1, with additional DLs as follows in the table. Results: 31 patients (pts) with a median age of 54 years (range 32-76) were enrolled. 18 pts had BC (3 ER+, 15 triple-negative), and 13 had high grade serous OC (11 platinum-sensitive, 2 platinum-resistant). The median number of prior therapies for advanced BC was 1 (range 0-5) and for OC was 2 (range 1-3). Dose de-escalation was necessary due to the DLT of neutropenia. After DL -3, prophylactic G-CSF (PGF) was instituted. The RP2D was C AUC 2, G 600 mg/m2, m 300 mg with PGF administered on day 9. Of the BC pts, 2 had a complete response (CR), 2 had a partial response (PR), 8 had stable disease (SD), 4 had progressive disease (PD). Of the OC pts, there was 1 CR (CR2 lasted > 27 mos; CR1 lasted only 8 mos), 1 PR, 6 SD, and 3 PD. 4 pts were inevaluable for response. Conclusions: These data suggest that M+C+G is safe and tolerable, and the most common DLT is neutropenia. This was easily managed with the institution of PGF. Studies correlating tumor GR expression with response are ongoing, and may help identify patients who are most likely to benefit from this combination. Clinical trial information: NCT02046421. [Table: see text]
Weekly paclitaxel (WP) +/- bevacizumab (B) in angiosarcoma (AS) patients (pts): Analysis of prognostic/predictive factors from a randomized phase 2 trial.
Authors: Lebellec Loic , Bertucci Francois , Tresch-Bruneel Emmanuelle , Ray-Coquard Isabelle Laure , Le Cesne Axel , Bompas Emmanuelle , Piperno-Neumann Sophie , Italiano Antoine , Chevreau Christine , Cupissol Didier , Bay Jacques-Olivier , Collard Olivier , Saada-Bouzid Esma , Isambert Nicolas , Delcambre Corinne , Blay Jean-Yves , Goncalves Anthony , Penel Nicolas
Published: September 2018

11024

Background: WP is an active regimen for treatment of AS pts (Ray-Coquard JCO 2015). We report here the correlative analysis conducted during a phase 2 trial assessing WP +/- B. Methods: Circulating pro/anti-angiogenic factors (FGF, PlGF, SCF, Selectin, thrombospondin, VEGF, VEGF-C) were collected at D1 and D8. Prognostic value for PFS was assessed using Cox model (biomarkers as continuous variables). We attempt to identify subgroups of pts benefiting from adding B using interaction tests (predictive factors). Results: Among the 51 pts enrolled in this trial, 45 were analyzable: 20 in Arm A (WP without B) and 25 in Arm B (with B). Median PFS was 5.5 and 6.1 months, respectively (p = 0.84). Samples were collected in 45 pts at D1 and 42 pts at D1 and 8. Baseline biomarkers were similar in both arms (excluding Selectin, significantly lower in arm A: median of 25 vs. 35 ng/mL, p = 0.03). In arm A, there was no significant difference between values at D1 and D8. In arm B, there were a significant decrease in VEGF (from a median of 0.49 to 0.08 ng/mL; p < 0.01) and selectin (from a median of 35.3 to 31.7 ng/mL; p < 0.01), and a significant increase in PlGF (from a median of 16.1 to 30.0 pg/mL; p < 0.01). In univariate analysis, factors associated with PFS were: de novo vs. radiation-induced AS (HR = 2.39 (p < 0.01), visceral vs. superficial AS (HR = 2.04; p < 0.03), VEGF-C at D1 (HR = 0.77; p < 0.03), FGF at D8 (HR = 1.17; p < 0.01), difference in FGF D8-D1 (HR = 1.24; p < 0.01), and PlGF value at D1 (HR = 1.02; p < 0.05). In multivariate analysis, factors associated with PFS were: de novo AS (HR = 2.39; p = 0.03), VEGF-C at D1 (HR = 0.73; p < 0.02) and FGF difference between D8 and D1 (HR = 1.16; p < 0.02). None of these factors were associated with benefit of adding B. Conclusions: Baseline VEGF-C levels and change in FGF were independent prognostic factors in pts with or without B. Addition of B significantly decreased the level of circulating VEGF and selectin and increased the level of circulating PlGF in AS patients. We did not identify subgroup of pts benefiting from adding of B to WP. Clinical trial information: NCT01303497.
The genomic and evolutionary landscape of osteosarcoma progression and lung metastasis.
Authors: Wang Jin , Bao Hua , Xu Huaiyuan , Shek Tony WH , Wu Xue , Tang Qinglian , Zhang Huizhong , Han Anjia , Yin Junqiang , Zou Changye , Wang Yongqian , Huang Gang , Batalha Sellma , Xie Xianbiao , Lu Jinchang , Song Guohui , Kang Yao , Shao Yang , Lam YingLee , Shen Jingnan ,   Bioinformaticians from Nanjing Geneseeq Technology Inc. and Pathologists from Queen Mary Hospital, Hong Kong
Published: September 2018

11029

Background: Osteosarcoma (OS) is a primary malignant bone tumor that has a high potential to metastasize to lungs. Recent studies have characterized somatic mutations of primary OS tumors. Nevertheless, lung metastases of OS are poorly studied, and whether they harbor distinct genetic alterations beyond those observed in primary tumors is largely unknown. Methods: We performed whole-exome sequencing (WES) of matched primary tumors and lung metastases in a cohort of 15 OS patients. Somatic single nucleotide variations (SNV) and copy number alterations (CNA) were analyzed to characterize the genomic and evolutionary landscape of metastatic OS. Results: Compared to matched primary tumors, lung metastases exhibited higher transversion rate for base substitution, and poor overlap ( < 10%) of genetic alterations was observed between primary and metastasis tumors. Multiple novel significantly mutated genes were identified, including ZNF717 in lung metastases, SPDYE1 in primary tumors, and CRIPAK in both. Copy number analysis indicated recurrent CNAs, including NEURL1B deletion and FLG amplifications in lung metastases, GSTT1 deletion in primary tumors, and CEACAM gene family deletion in both. Furthermore, phylogenetic analyses revealed that paired primary tumors and metastases underwent parallel evolution with few ubiquitous clonal mutations, suggesting that OS metastases are likely to be derived from primary tumors at a very early stage of their evolution. Conclusions: This study for the first time provides important evidence that OS metastases harbor distinct genetic alterations compared with primary tumors. Our findings strongly support a parallel evolution model of primary and metastatic tumors. Moreover, several novel CNAs and significantly mutated genes that are specifically associated with lung metastases may provide future therapeutic insight for OS.
Analysis of tumor-infiltrating immune cells in gastrointestinal stromal tumors (GIST) after tyrosine kinase inhibitor therapy.
Authors: Hohenberger Peter , Zhao Wenyi , Deligianni Maria , Simon-Keller Katia , Cao Hui , Marx Alexander
Published: September 2018

11042

Background: There is initial evidence that immune-infiltrates of tumor may correlate with the clinical course of patients with GIST beyond the response to tyrosine kinase inhibitors (TKI) according to the mutational status. We were interested to analyze the composition of tumor-infiltrating immune cells in GIST tumor tissues after different TKI regimens vs untreated controls. Methods: From 40 GIST patients who were treated with imatinib alone (neoadjuvant group) and other TKIs (M1 HEP group, progressing on more than ≥2 TKI inhibitors) prior to tumor removal, surgical specimens were available at ten 10 cases each,. They were compared with 20 untreated primary tumors graded for malignant behaviour (low vs high risk ) acc. to Miettinen&Lasota 2006. 2-μm sections of formalin-fixed, paraffin-embedded tissue samples were used for IHC-staining anti-CD68 (Dako); anti-CD11c, anti-CD11b, anti-FOXP3 (all Abcam); anti-CD163, anti-CD4 , anti-CD8 (all Leica). Evaluation was performed with Dako REAL EnVision Detection System (K5007). The phenotypes of immune cells were compared in the defined GIST groups. In the neoadjuvant group we used recurrence-free survival (RFS) for correlation, and overall survival (OS) after TKI failure in the M1 HEP group. Results: Foxp3+ Tregs, CD163+ M2 macrophages and CD11b+ myeloid cells are significantly higher in TKI resistance or neoadjuvant (viable cells > 30%) tissues compared with neoadjuvant (viable cells < 10%) cases. The rate of CD11c+ M1 macrophages is higher in low-risk GISTs compared to high risk GISTs, but CD11b+ myeloid cells are exactly the opposite. Kaplan-Meier curves show high CD163+ M2 macrophages or CD11b+ myeloid cells to correlate significantly with worse RFS in after neoadjuvant IM (p 0.01). In the TKI resistant condition, patients with high CD11c+ M1 macrophages show better median OS (p = 0.07, ns). Conclusions: Our study reveals dynamic changes of tumor-infiltrating immune cell protagonists in GISTs after different TKI therapeutic regimen and after neoadjuvant treatment. We could not detect a different immune infiltrate in untreated primary tumors of different risk categories.
Prognostic factors of recurrence and survival of gastrointestinal stromal tumors: First multicentric study of Mexico.
Authors: Medrano Guzman Rafael
Published: September 2018

11045

Background: Gastrointestinal stromal tumors are mesenchymal lesions arising from the interstitial cells of Cajal. In GIST the location, size, number of mitosis and risk group are accepted as prognostic factors; some factors in which there are still controversies about their value as prognostic factors include male gender, tumor cellularity, the margins of resection, the presence or absence of p16 or intraperitoneal tumor breakdown. Methods: Observational, retrospective and longitudinal study. Patients admitted to the Oncology Hospital, CMN Siglo XXI of January 1, 1991 to April 30, 2012. Results: We identified 384 patients, the mean follow-up time was 55.86 months, the mean age was 58 years, 80.3% had symptoms and only 4.54% the finding was incidental. The most common site was the stomach (66.6%), followed by small intestine (28.7%) and colon (1.54%). The 7.57% had metastases at diagnosis, 4.54% in liver and 3.03% in the peritoneum. Were expressed by abdominal pain (39.39%) and gastrointestinal bleeding (30.30%), intestinal obstruction in 1.5%. The most common site was the stomach (66.6%), followed by small intestine (28.7%) and colon (1.54%). The 7.57% had metastases at diagnosis. The average tumor size was 10.84 cm (2.2 to 38 cm). Immunohistochemical markers were studied: 94.28% positive for CD117, CD34 74.28%, 11.42% S100 protein, desmin 5.71% and 51.72% for smooth muscle actin (AML). Overall rate at 5 years and 82% rate of recurrence-free survival at 5 years and 61% survival. The location of the tumor (p = 0.0002), size (p = 0.03145), the number of mitosis (p = 0.008), risk group (p = 0.020) and adjuvant treatment with Imatinib showed a statistically significant difference for Survival Recurrence-free. For overall survival, lesion location was the only factor that showed statistical significance with p = 0.0054. Conclusions: The tumor location, size, number of mitosis, the risk group and adjuvant treatment with Imatinib were statistically significant prognostic factors for disease recurrence. The location of the lesion was the only factor that showed statistical significance as a predictor of overall survival.
A pilot trial of irinotecan, temozolomide and bevacizumab (ITB) for treatment of newly diagnosed patients with desmoplastic small round cell tumor (DSRCT).
Authors: Magnan Heather D. , Price Anita , Chou Alexander J. , Riedel Elyn , Wexler Leonard H. , Ambati Srikanth R. , Slotkin Emily Kanaya , Ulaner Gary , Modak Shakeel , La Quaglia Michael P. , Meyers Paul A.
Published: September 2018

11050

Background: DSRCT is a rare tumor with a dismal prognosis in the setting of current treatment options. Preclinical data suggested that VEGF-dependent angiogenesis is important for DSRCT tumor biology and that targeting angiogenesis with bevacizumab in combination with irinotecan was more effective than treatment with irinotecan alone. This pilot study was designed to explore the safety and feasibility of adding ITB to the existing “P6- like” regimen used to treat DSRCT. Methods: Fifteen patients with newly diagnosed DSRCT were enrolled onto this single-institution study. They began treatment with 2 cycles of irinotecan (20 mg/m2/dose x 10 days) and temozolomide (100 mg/m2/dose x 5 days). Bevacizumab 10 mg/kg q2 weeks was added after sufficient time had passed from initial biopsy or surgery. Patients were then treated with cycles of alkylator based chemotherapy (3 cycles of cyclophosphamide, doxorubicin, vincristine and 3 cycles of ifosfamide, etoposide). An initial surgical resection was performed after cycle 5 and a second resection or second look surgery after cycle 8. Toxicity was graded according to CTCAE v.4.0. Secondary efficacy objectives were assessed using RECIST 1.1 criteria and the Kaplan Meir method. Results: 14 of 15 patients completed planned protocol therapy. One patient was taken off study due to complications associated with surgery after cycle 5 of chemotherapy. Stopping rules for unacceptable toxicity were not met. No patients experienced toxicity attributed to bevacizumab, and surgical morbidity was no greater than expected. Grade 3 diarrhea associated with irinotecan was experienced by 2 patients. Expected toxicities with “P6-like” cycles included grade 3/4 hematologic toxicity and admissions for febrile neutropenia in all patients. Response rate to the ITB cycles was 27% (95% CI 8-55%) and to the 5 pre-resection cycles was 73% (95% CI 45-92%). Median time to progression was 18.1 months. Overall survival at 1 year was 100% and 3 years 61% (95% CI 25-84%). Conclusions: The combination of ITB is active in patients with DSRCT, and it is feasible to combine these agents with standard chemotherapy without greater than expected toxicity. Clinical trial information: NCT01189643.
Genetic variations within the vitamin C transporter genes to predict outcome in metastatic colorectal cancer patients treated with first-line FOLFIRI and bevacizumab: Data from FIRE-3 trial.
Authors: Berger Martin D. , Stintzing Sebastian , Yang Dongyun , Cao Shu , Miyamoto Yuji , Suenaga Mitsukuni , Soni Shivani , Gopez Roel , Melendez Elizabeth , Zhang Wu , Heinemann Volker , Lenz Heinz-Josef
Published: September 2018

11507

Background: Vitamin C is involved in many critical metabolic processes. Beside its major role as an antioxidant and free radical scavenger vitamin C exerts a regulatory influence on angiogenesis. Additionally, epidemiologic studies show an association between vitamin C levels and incidence of cancer.We therefore hypothesize that variations in genes encoding for vitamin C transporter proteins may predict outcome in patients (pts) with metastatic colorectal cancer (mCRC) treated with FOLFIRI and bevacizumab (bev). Methods: Theimpact of 3 functional SNPs within the SVCT1, SVCT2 and Glut1 genes on outcome was evaluated in 292 pts with mCRC treated with first-line FOLFIRI/bev in the randomized phase III FIRE-3 trial. 294 pts receiving FOLFIRI and cetuximab (cet) (FIRE-3) served as a negative control. Genomic DNA was extracted from formalin fixed paraffin embedded tissue and SNPs were analyzed by PCR-based direct sequencing. Results: Baseline characteristics in the FOLFIRI/bev arm were as follows: female/male 99/193; median age = 65y andmedian PFS/OS = 10.1/24.2 months (mts). The SVCT1 rs11242462 SNP showed significant association with PFS. T allele carriers had a longer median PFS compared to those with a C/C genotype (10.7 vs 9.7 mts) in both univariate (HR 0.77, p = 0.046) and multivariate analysis (HR 0.73, p = 0.028). The effect on outcome was most significant among KRAS mutant pts. Here, T allele carriers showed a markedly prolonged PFS and OS compared to pts with a C/C genotype (12.5 vs 7.0 mts, HR 0.50, p = 0.018 and 32.8 vs 14.7 mts, HR 0.45, p = 0.009). These associations remained significant in multivariate analyses (p = 0.009 and p = 0.021, respectively). However, the favorable impact on outcome was not observed among T allele carriers treated with FOLFIRI/cet. Conclusions: Our results provide the first evidence that the SVCT1 polymorphism rs11242462 might serve as a predictive marker in pts with mCRC treated with FOLFIRI/bev in the first-line setting. Targeting vitamin C transporter proteins might be a promising approach to further improve treatment options against mCRC and to overcome resistance to anti-angiogenic therapy.
Evaluation of clinical outcomes by analysis of mutations in tumor tissue and circulating plasma DNA using next-generation sequencing (NGS) from STEAM, a prospective, randomized, multicenter study in metastatic colorectal cancer (mCRC).
Authors: Lee John J. , Palma John F. , Yao Lijing , Lovejoy Alexander F. , Yaung Stephanie , Zhang David , Wingate-Pearse Nasiema , Yau May , Williams Carolyn , Pimentel Monica , Munoz Abraham , Mayol Katrina , Mancao Christoph , Nicholas Alan , Sommer Nicolas , Bendell Johanna C. , Hurwitz Herbert , Rohr Ulrich Peter
Published: September 2018

11510

Background: STEAM (NCT01765582) assessed the efficacy and safety of concurrent (c) and sequential (s) FOLFOXIRI-bevacizumab (BEV) vs FOLFOX-BEV for first-line treatment of mCRC. Methods: The AVENIO ctDNA Expanded Kit (Research Use Only) was used to assess somatic mutations in 77 cancer-related genes by NGS in tissue, and both pre- and post-induction plasma samples (n = 182, 150 and 118 respectively) from STEAM. Four mutation classes including single-nucleotide variants (SNVs), indels, copy number amplifications (CNAs) and fusions were identified. SNVs and indels were called in tissue and plasma at allele frequencies of 5% and 0.25% respectively. Results: Overall concordance of mutations in pre-induction plasma with tissue was 83%. Concordance for the seven most mutated genes ranged from 91.5%-100%. In pts with matched samples (n = 118), RAS WT pts showed significantly longer progression-free survival (PFS) in both cFOLFOXIRI-BEV (A) and sFOLFOXIRI-BEV (B) arms versus FOLFOX-BEV (C), using genotyping of either tissue or plasma. This was not seen in RAS MUT pts. In contrast, TP53 WT showed no significant treatment differences while TP53 MUT showed longer PFS for cFOLFOXIRI-BEV versus FOLFOX-BEV. A list of mutation frequencies for all samples, as well as hierarchical clustering analysis of tissue mutations will be presented. Conclusions: The AVENIO ctDNA Expanded Kit identified mutations in 77 cancer-related genes, in both plasma and tissue, with high overall concordance. Compared to FOLFOX-BEV, longer PFS was observed for c- or s- FOLFOXIRI-BEV in RAS WT pts and for cFOLFOXIRI-BEV in TP53 MUT pts, irrespective of sample type. These results are hypothesis generating and require further clinical validation. Clinical trial information: NCT01765582. [Table: see text]
Inter-tumor validation, through advanced MRI and circulating biomarkers, of plasma Tie2 as the vascular response biomarker for bevacizumab.
Authors: Jayson Gordon C. , Zhou Cong , Horsley Laura Hope , Marti Kalena , Shaw Danielle , Mescallado Nerissa , Clamp Andrew R. , Saunders Mark P. , Valle Juan W. , Backen Alison Catherine , Simpson Kathryn , Little Ross , Watson Yvonne , Cheung Susan , Roberts Caleb , Manoharan Prakash , Jackson Alan , O'Connor James , Parker Geoff J M , Dive Caroline
Published: September 2018

11521

Background: VEGF inhibitor (VEGFi) use is compromised by lack of predictive/ response biomarkers. Previously, we identified plasma Tie2 (pTie2) as a vascular response biomarker (VRB) for bevacizumab (bev) in ovarian cancer (OC). Here, we applied dynamic contrast-enhanced MRI (DCE-MRI) and circulating biomarkers in colorectal cancer (CRC), to validate pTie2 as the first tumor VRB. Methods: Seventy patients were recruited, with untreated, mCRC and ≥1 lesion of 3-10cm diameter for DCE-MRI. Patients received bev 10mg/kg for 2 weeks to elicit a biomarker response and then FOLFOX6/bev until progressive disease (PD) Thirteen circulating and 6 imaging biomarkers were measured before and during treatment and at PD. Unsupervised correlation analysis identified bev-induced biomarker correlations. Biomarkers were evaluated by clustered parameter-time course studies to determine their epithelial or vascular origin. Clinical significance was determined by relating the biomarker data to tumor 3D volumetric change assessed by MRI and PFS. The emergent vascular biomarker signal was modelled with epithelial biomarkers to assess the independent contribution of the vascular compartment to PD. Results: Bev induced significant correlations between pTie2, Ang2 and Ktrans. Cluster analysis of Tie2 concentration-time course curves showed that pTie2 reflected tumor Ktransbut not CK18, an epithelial antigen, i.e. changes in pTie2 reflected tumor vascular biology Patients who had the greatest area under the pTie2-time curve had tumors with high Ktransand/or low pVEGFR2, pre-treatment. They also had the greatest reduction in tumor volume and longest PFS. Fusion of pTie2 and CK18 data significantly improved modelling of PD. Conclusions: Bev impacts tumor vasculature causing proportional changes in pTie2. Information from pTie2 adds clinical value to that derived from the epithelial compartment. Thus (i) pTie2 is the first vascular response biomarker for bev and probably all VEGFi and (ii) demonstration of separate vascular and epithelial compartments in ovarian and CRC validates the vascular compartment as a target. This work identifies the first assay that could optimise use of VEGFi. Clinical trial information: 2009-011377-33.
Early detection of competing resistance mutations using plasma next-generation sequencing (NGS) in patients (pts) with EGFR-mutant NSCLC treated with osimertinib.
Authors: Guibert Nicolas Marie , Paweletz Cloud , Hu Yuebi , Feeney Nora B. , Plagnol Vincent , Poole Valerie , Jones Greg , Oxnard Geoffrey R.
Published: September 2018

11529

Background: In pts with EGFR+ NSCLC, genotyping of plasma cell-free DNA (cfDNA) has become a routine option for non-invasive detection of EGFRT790M. We hypothesized that serial NGS of cfDNA would allow early detection of co-existent resistance mutations during osimertinib treatment. Methods: Serial plasma samples were collected from pts with advanced EGFR+ NSCLC and T790M+ acquired resistance treated with osimertinib. Up to 4 specimens were analyzed, blinded to tumor genotype: baseline, initial 2 follow-ups, and progression. Plasma NGS was performed using enhanced tagged amplicon sequencing of hotspots and coding regions from 36 genes. Diagnostic accuracy was compared to tumor genotype (including NGS when available) and droplet digital PCR (ddPCR) of cfDNA. Results: 94 specimens from 26 pts underwent plasma NGS. Studying 26 baseline specimens, plasma NGS was more sensitive than ddPCR for known EGFR driver mutations (100% vs 88.5%). In 8 pts with pretreatment tumor NGS, 5 of 6 TP53 mutations were detected; one plasma positive/tissue negative result was seen across 36 genes, a PIK3CA mutation (0.6% AF) which was confirmed using ddPCR (99.6% specificity). Quantitative concordance of AF compared to ddPCR was high (R = 0.94). 21 pts had detectable EGFR driver mutations at resistance. Among 6 pts with maintained T790M, 4 acquired C797S and 2 of these additionally acquired low level KRAS mutations (G13D, Q61K). Among 15 pts with loss of T790M, 7 had competing non- EGFR alterations identified: MET amp, PIK3CA E545K, BRAF V600E (n = 2), HER2 amp, KRAS G12S, and FGFR1 amp. Resistance mutations detected at AF > 0.3% were confirmed with ddPCR. In 3 pts, a competing resistance mutation (1 KRAS, 2 BRAF) could be detected in plasma NGS pretreatment and reemerged as putative drivers at time of resistance. Conclusions: In this retrospective blinded validation, tagged amplicon-based plasma NGS was more sensitive than ddPCR with high specificity and quantitative concordance. In a subset of cases, serial plasma NGS can detect emergence of competing resistance mutations, creating an opportunity for the study of osimertinib-based targeted therapy combinations.
CCL5 expression and tumor infiltrating immune cells in triple negative breast cancer.
Authors: Araujo Jhajaira , Gomez Andrea C. , Salgado Roberto , Balko Justin M. , Aguilar Alfredo , Doimi Franco , Morante Zaida , Gomez Henry , Pinto Joseph A.
Published: September 2018

11553

Background: CCL5 is a chemo-attractant of regulatory T cells, promoting tumor immune avoidance and related to a poor outcome in several malignancies; however, in triple negative breast cancer (TNBC), it is related to a better outcome. Our aim was to evaluate the correlation between CCL5 and tumor infiltrating immune cells and their prognosis value. Methods: We evaluated 72 TNBC patients with residual disease after neoadjuvant chemotherapy with matched data of tumor infiltrating lymphocytes (TILs) count and CCL5 expression (profiled with NanoString). CCL5 expression levels were log2 transformed and median centered. Correlation between TILs (log2 transformed) and CCL5 was evaluated with the Spearman’s rank test. Cox PH model was used to investigate the effect of CCL5 (median as cutoff) and TILs ( < 20% and ≥20%) in distant-recurrence free survival. We used the CIBERSORT platform to evaluate the immune cells composition according to the expression of CCL5 (higher vs. lower or equal than median) in 3 independent TNBC datasets (GSE25066, GSE58812 and GSE76124). Results: There was a significant correlation between TILs and residual tumor size (P = 0.017) and CCL5 (ρ = 0.347, P = 0.003). In univariate analysis, TILs (HR = 0.276, 95%IC: 0.128-0.593; P = 0.001) and CCL5 (HR = 0.401; 95%CI: 0.206-0.781; P = 0.007) were both associated with outcome. In a multivariate analysis with CCL5 expression and TILs count, TILs was the only significant marker with a P = 0.008 (HR = 0.336; 95%CI: 0.150-0.753), in contrast to CCL5 (HR = 0.573; 95%CI: 0.285-1.154; P = 0.124). CIBERSORT analysis suggested that high CCL5 expression is associated with recruitment of CD8 cells (13% v 6%, P < 0.001; 6% v 1%, P < 0.001 and 12% v 8%, P = 0.003), activated CD4 memory T cells (4% vs. 2%, P < 0.001; 5% vs. 0%, P < 0.001; 3% vs. 0%, P < 0.001) and Macrophages M1 (9% vs. 7%, P = 0.022; 13% vs. 8%, P = 0.005; 11% vs. 5%, P < 0.001) in GSE25066, GSE58812 and GSE76124 datasets, respectively. Conclusions: TILs was the stronger and more significant prognostic immunological marker, even than CCL5 expression. High CCL5 expression was associated with enrichment of CD8 cells, activated CD4 memory T cells and Macrophages M1. Role of these cells in TNBC should be explored more deeply.
Effect of chemoradiation for cervical cancer on transient decrease and variable expansion in T-cell infiltrate and diversity of T-cell repertoire.
Authors: Colbert Lauren Elizabeth , Dorta-Estremera Stephanie , Previs Rebecca A. , Eifel Patricia J. , Jhingran Anuja , Ramondetta Lois M. , Hahn Stephen M. , Welsh James William , Jazaeri Amir A. , Frumovitz Michael M. , Sastry Jagannadha , Klopp Ann
Published: September 2018

11557

Background: The effect of fractionated radiation on intratumoral immune infiltrate is unclear. The purpose of this study was to characterize local immune changes and treatment response during chemoradiation (CRT). Methods: Cervical cancer patients underwent cisplatin based CRT over 5 weeks with brachytherapy. Cervical DNA swabs and cytology brushings were collected at baseline, one week, three weeks and five weeks. Deep T cell receptor β sequencing (TCR; Adaptive, Seattle WA) and multiparametric flow cytometry (MPFC) were performed for each time point. T cell density (TCD) and productive clonality (PC) were analyzed. Cells separated from the tumor brushings were stained and fixed with antibodies to T cell subsets with activation and suppressor markers including CD3, CD4, CD8, Ki67, PD-1, CTLA-4, and ICOS. Changes in T cell subsets were evaluated as percentage of live lymphocytes. Results: Eight patients were evaluated using MPFC. CD4 and CD8 percentages were lowest at one week and subsequently expanded. The percentage of proliferating CD4 (CD4+ Ki67+) was highest at week 5 (1.19%). There was no change in percentage of CD8+ cells expressing PD1, CTLA4 or ICOS over the course of treatment. TCR diversity was assessed for 9 patients. At baseline, week 1, week 3 and week 5, median TCD for all patients were 0.046 (IQR 0.008 to 0.097), 0.021 (IQR 0.005 to 0.043), 0.035 (IQR 0.015 to 0.083), and 0.033 (IQR 0.017 to 0.110). Median productive clonality at each point was 0.05 (IQR 0.06 to 0.02), 0.03 (IQR 0.06 to 0.02), 0.04 (IQR 0.05 to 0.02) and 0.03 (IQR 0.01 to 0.05). PC fold count increased (1.69, SD 1.3) for complete response (CR) patients and decreased 0.3 (SD 0.008) for patients with recurrent (REC) disease (p = 0.1). One TCR sequence was common in 4/9 patients at the end of treatment and six sequences were common in 3/9 patients. Conclusions: Chemoradiation induces a transient decline in tumor infiltrating CD8+ and CD4+ cells, followed by a variable expansion in T-cells at the end of treatment with an increase in proliferation phenotypes. TCR sequencing revealed increase in productive clonality during radiation for patients with a complete response to treatment.
The use of 18F-fluoroestradiol (FES) and 18F-fluorodeoxyglucose (FDG) PET in the evaluation of breast cancer heterogeneity.
Authors: Peterson Lanell M , Kurland Brenda F , Novakova Alena , Lee Jean H , Specht Jennifer M. , Muzi Mark , Romine Perrin , Wu Vicky , Mankoff David A. , Kinahan Paul , Linden Hannah M.
Published: September 2018

11572

Background: 18F-Fluoroestradiol (FES) is an estrogen analogue that has been shown to be a promising biomarker in ER imaging of breast cancer. FES uptake correlates to ER expression, provides qualitative and quantitative assessment of multiple tumor sites simultaneously, and can predict response to endocrine therapies. Tumor heterogeneity is a known feature of metastatic breast cancer. Our work and others has shown that patients that have tumors with high FDG-PET SUV and low FES-PET SUV uptake have a poorer prognosis. Biopsies of metastatic disease may be done initially for diagnosis of metastatic disease, but are generally only performed in the setting of target identification for clinical trials. A change in ER or HER2 expression, however, can result in a change in therapy. FES-PET imaging offers a virtual biopsy and can reveal heterogeneity of the entire tumor burden. Methods: We reviewed our prior evaluations of tumor heterogeneity with FES-PET from 3 different studies. 46 breast cancer patients with metastatic disease (de novo or recurrent) who had biopsy proven ER+ primary breast cancer underwent FES-PET and FDG-PET imaging and a biopsy of a metastatic lesion prior to therapy initiation. ER and HER-2 expression was reviewed. Results: Of the 46 patients, 5 (11%) had ER- metastatic biopsies. One (2%) biopsy changed from ER+/HER-2 neg to ER-/HER-2+, and one (2%) biopsy changed from ER+/HER-2+ to ER+/HER-2-. All 5 patients (11%) with changes in ER/HER2 expression underwent a change in therapy due to the unexpected findings by metastatic biopsy. FDG findings helped to guide selection of biopsy sites. FES quantitative measures correlated with biopsy findings. Conclusions: Biopsy resulted in a change in therapy for > 10 % of patients enrolled in trials of FES imaging. Imaging can help identify heterogeneous tumor locations to assist identification of evolving tumor targets in breast cancer. In addition, FES imaging may reveal a change in tumor phenotype that can ultimately affect choice of therapy. Research Support: P01CA42045, R01CA72064
Persistence of AKT1 low quiescent cancer cells after neoadjuvant chemotherapy in triple negative breast cancer patients.
Authors: Kabraji Sheheryar Kairas , Sole Xavier , Huang Ying , Bango Clyde , Bowden Michaela , Bardia Aditya , Sgroi Dennis , Loda Massimo , Ramaswamy Sridhar
Published: September 2018

11579

Background: The mechanisms that allow triple negative breast cancer (TNBC) tumors to survive neoadjuvant chemotherapy (NACT) are incompletely understood. Evidence suggests that proliferative heterogeneity may contribute to primary chemotherapy resistance in patients with localized triple negative breast cancer. However, the detailed characterization of a drug-resistant cancer cell state in residual TNBC tissue after NACT has remained elusive. AKT1lowquiescent cancer cells (QCCs) are a quiescent, epigenetically plastic, and chemotherapy resistant subpopulation initially identified in experimental cancer models. Here, we asked whether AKT1low QCCs actually exist in primary tumors from patients with TNBC and persist after treatment with NACT. Methods: We identified QCCs in primary and metastatic human breast tumors using automated, quantitative, immunofluorescence microscopy coupled with computational and statistical analysis. We obtained pre-treatment biopsy, post-treatment mastectomy, and metastatic specimens from a retrospective cohort of TNBC patients treated with neoadjuvant chemotherapy at Massachusetts General Hospital (n = 25). Using automated quantitative immunofluorescence microscopy, QCCs were identified as AKTlow / H3K9me2low / HES1high cancer cells using prespecified immunofluorescence intensity thresholds. QCCs were represented as 2D and 3D digital tumor maps and QCC percentage (QCC-P) and QCC cluster index (QCC-CI) were determined for each sample. Results: We found that QCCs exist as non-random and heterogeneously distributed clusters within primary tumors. In addition, these QCC clusters are enriched after treatment with multi-agent, multi-cycle, neoadjuvant chemotherapy in both residual primary tumors as well as nodal and distant metastases in patients with triple negative breast cancer. Conclusions: Together, these data qualify QCCs as a non-genetic mechanism of chemotherapy resistance in triple negative breast cancer patients that warrants further study.
Rates of PD-L1 expression testing in U.S. community-based oncology practices (USCPs) for patients with metastatic non-small cell lung cancer (mNSCLC) receiving nivolumab (N) or pembrolizumab (P).
Authors: Khozin Sean , Abernethy Amy Pickar , Nussbaum Nathan C. , Zhi Jizu , Curtis Melissa , Tucker Melisa , Lee Shannon E , Light Dave , Blumenthal Gideon Michael , Pazdur Richard
Published: September 2018

11596

Background: FDA approved N for previously treated squamous mNSCLC in 3/2015 and non-squamous mNSCLC in 10/2015 regardless of PD-L1 expression. P was approved for previously treated mNSCLC in 10/2015 for patients whose tumors express PD-L1 as determined by an FDA-approved companion diagnostic (CoDx). Post-FDA approval treatment patterns of N and P, including PD-L1 testing patterns, remain unclear. Methods: We conducted a retrospective analysis using de-identified patient-level electronic health record (EHR) data aggregated from USCPs by Flatiron Health in accordance to an institutional review board approved protocol. Data was abstracted using both structured and unstructured EHR content with 10% duplicate abstraction to confirm data quality. All patients in the database with clinically-confirmed mNSCLC diagnosed from 1/1/11 to 3/31/16 and documented order or administration of N or P were included in the analyses. PD-L1 expression testing results were abstracted from unstructured data in EHR documents. Results: Within the first year of FDA approval, 1,362 mNSCLC patients received N or P (96.2% N; 3.3% P, 0.5% N+P). The median duration of treatment was 113 days. Overall, 11.3% of patients were tested for PD-L1 expression, 42.2% of whom tested positive. PD-L1 expression testing increased quarterly (Q) from 3.0% in Q2 2015 to 12.8% in Q1 2016. An FDA-approved CoDx was used 30.5% of the time. Among patients who received N, 90.5% were not tested for PD-L1 expression, 3.2% tested positive, and 4.7% tested negative for PD-L1. Among patients who received P, 32.7% were not tested for PD-L1 expression, 50.0% tested positive, and 1.9% tested negative. Conclusions: We observed low rates of PD-L1 expression testing in USCPs for patients with mNSCLC receiving N or P. Testing rates increased quarterly following FDA-approval. The majority of patients were not tested using FDA-approved CoDx. Most patients who were not tested for PD-L1 expression or tested negative received N. Clinician education will be important as data on optimal PD-L1 testing strategies guiding immunotherapy treatment decisions accumulates.