bluuebunny/crossref_metadata_2025 huggingface dataset:
Median progression free survival (PFS) for patients treated with everolimus (EVE) + exemestane (EXE) for HR+ mBC in routine clinical practice: Results of the 3rd interim analysis of the non-interventional trial BRAWO.
Authors: Jackisch Christian , Fasching Peter A. , Grischke Eva-Maria , Schuetz Florian , Decker Thomas , Uleer Christoph , Salat Christoph , Foerster Frank , Schmidt Marcus , Mundhenke Christoph , Tesch Hans , Wimberger Pauline , Aktas Bahriye , Zahn Mark-Oliver , Tome Oliver , Bloch Wilhelm , Hanson Sven , Schubert Joerg , Lueftner Diana
Published: September 2018

e12547

Background: BRAWO is a non-interventional study, which enrolled more than 2400 patients (pts) with advanced/metastatic, hormone-receptor-positive and HER2-negative breast cancer treated with EVE and EXE. Main objectives are a) the impact of physical activity on efficacy and quality of life, b) prophylaxis and management of stomatitis in clinical routine, and c) the sequence of therapy when EVE is used in daily clinical practice. We report updated data of the 3rd interim analysis, including PFS. Methods: This updated analysis (data cut-off 18 Oct 2016) covers data of the first 1345 documented pts with at least one follow up under therapy. Here we describe the baseline characteristics, safety and PFS as well as response rates. PFS was estimated using Kaplan-Meier estimator. Results: At the time point of this data cut-off 1289 pts (93.9%) had discontinued the documentation. The median time for pts since primary diagnosis was 7.1 yrs and 2.2 yrs, since first sign of relapse (local recurrence or metastases). At baseline, 54.1% presented with visceral metastases. 49.2% had an ECOG performance status of 0 and 74.8% of pts started with 10mg EVE, while 24.5% started with 5mg EVE. According to treatment lines we found 27% 1L (359 pts), 32% 2L (426 pts), 19% 3L (253 pts), 11% 4L (153 pts) and 11% 5L (154 pts) in our cohort Additional baseline and safety data will be presented. The Kaplan Meier estimate of the overall PFS is 6.9 months (95%CI 6.3-7.4). 2.2% (18 pts) of the pts experienced a complete and 17.8% (147 pts) a partial remission, while 57.4% (475 pts) remained stable as their best overall responses during the documentation period. 52.3% (718 pts) discontinued the treatment due to a progressive disease and 25.5% (350 pts) due to adverse events. 67.1% (902 pts) continued the antineoplastic treatment with a subsequent therapy. Conclusions: Here we report the PFS of pts treated with EVE + EXE in a real world scenario. The PFS of 6.9 months observed in our series matches somewhat perfect with the PFS of 7.8 months from the randomized Bolero-2 trial suggesting that these findings might be valid and useful for everyday routine. Clinical trial information: EUPAS9462.
Long-term disease control with capecitabine in advanced breast cancer.
Authors: Masci Giovanna , Ferraro Emanuela , Torrisi Rosalba , Giordano Laura , Zuradelli Monica , Losurdo Agnese , De Sanctis Rita , Santoro Armando
Published: September 2018

e12566

Background: Capecitabine is an active agent in the treatment of advanced breast cancer (ABC). It is commonly administered at the approved dosage of 1250 mg/m2 twice daily for 2 weeks followed by 1 week rest period. Methods: The study population included 162 pts retrospectively analyzed with ABC treated with capecitabine between 2006 and 2015 at our Institute. Capecitabine was given 14 day on,7 day off cycle at a daily dose of 1900 mg/m2, a modified schedule used to minimize side effects. The objective were overall response rate (ORR), median duration of treatment (MDT) progression-free survival (PFS), overall survival (OS). Results: The median age was 64 years (range 33-89). Pts with hormone receptor positive ABC were 133 (82%), 6 (4%) had HER-2 positive disease and 29 (18%) a triple negative profile. One hundred and thirty eight (85%) had already received chemotherapy in adjuvant and/or metastatic setting; anthracycline and taxanes were given in 64 pts (40%), anthracycline in 60 pts (37%) and taxane in 4 pts (2%), 9 pts (6%) received other regimens of chemotherapy. Twenty-five pts (15%) were chemo-naive. One hundred and thirty-three pts (82%) received endocrine therapy. Sixty-four pts (38%) had predominantly non-visceral metastases, 26 pts (16%) exclusive visceral involvement, 72 (44%) exhibited both characteristics. ORR was 58% (CR = 2%, PR = 16% SD = 40%) and MDT was 6.9 months. The median PFS and OS were 6.9 months and 21 months, respectively. We defined as “long responders” 19 pts (12%) with disease control interval > 24 months and, among them, as “very long responders”, 9 pts (6%) who exceeded 36 months. Efficacy was unrelated to biological profiles, sites of metastasis or previous therapies. No grade 3 and 4 adverse events occurred. Conclusions: Our results show that in pts with ABC a lower dose of capecitabine has a good toxicity profile and similar overall response rate and survival data in comparison to the approved dose. In addition, we identified a subset of long and very long responders but further studies are warranted to evaluate clinical/biological predictors of a long-term response.
Early detection of oral cancer in Portugal: The experience of UMEMD.
Authors: Estevinho Fernanda , Gouveia Margarida , Costa Marta , Moura Sara , Neves João Sérgio , Paulo-Subtil Domingos , Marinho Jorge , Leite-Moreira João
Published: September 2018

e13039

Background: Oral and pharyngeal cancer is the 11th most common malignancy worldwide. Although the oral cavity is a potentially accessible site for examination, up to 50% of oral cancers are detected at advanced stage. The Mobile Unit of Stomatology and Dental Medicine (UMEMD) of the Portuguese League Against Cancer (LPCC) and of the Portuguese Association of Hospital Dental Medicine (APMDH) has developed oral cancer early diagnosis consultations. This consultations were performed by a team of 50 doctors specialized in oral cancer diagnosis. Once a month the team of UMEMD went to a different region of the North of Portugal. Early oral cancer diagnosis consultations were also performed twice a week at the Portuguese League Against Cancer at Oporto.We present the experience of UMEMD in the early diagnosis consultation of oral cancer, since its foundation in August 2012 until the end of 2016. Methods: We performed a retrospective evaluation of medical records of patients observed in the early diagnosis consultation of oral cancer of UMEMD between August 2012 and December 2016. We assessed the demographic characteristics, risk factors, cancer history, likely diagnosis and the referral of patients. Results: 6,231 new patients were observed; median age of patients was 63 years-old [1;102], 93 patients were younger than 18 years-old. There was information about smoking habits of the majority of patients n = 4,521(72.6%): 597(9.6%) were smokers and 653(10.5%) former smokers. 228 (3.6%) patients had previous cancer. We detected: 963 lesions (15.5%); the majority of the lesions were at the tongue n = 263 (4.2%).The patients with suspicious lesions (n = 638; 10.2%) were referred to: Portuguese Institute of Oncology of Oporto: n = 146 (2.3%); Faculty of Dental Medicine of the University of Oporto: n = 353 (5.7%); Physicians at other hospitals: n = 139 (2.2%). Conclusions: Consultations for oral cancer early diagnosis performed by an experienced team has enabled the detection of suspicious lesions in 10.2% of the patients. Cooperation with other institutions, through protocols, allowed the referral and early treatment of these patients.
Low grade b-cell non-Hodgkin lymphomas (NHL) involving the central nervous system (CNS): An analysis from the National Cancer Database (NCDB).
Authors: Pophali Priyanka Avinash , Thanarajasingam Gita , Pulido Jose , Johnston Patrick B. , Go Ronald S.
Published: September 2018

e13083

Background: CNS involvement from low grade B-cell NHL is rare and has only been reported as case series. The distribution, demographics and outcomes of patients with low grade B-cell CNS NHLs have not been well characterized. Methods: The NCDB represents ~70% of cancer cases in the United States. Using the 2004-2013 NCDB extranodal NHL database, we identified all CNS B-cell NHLs based on ICD-O-3 site and histology codes. Primary or secondary CNS involvement could not be determined. Results: Out of 9435 CNS NHL cases, 475 [5.03%] had low grade histologies. In this group, the median age at diagnosis was 58 years [range 19-89]. Majority of the cases were female [56%], White, non-Hispanic [72%], privately insured [53%], with no comorbidities [74%] and treated in academic/research programs [38%]. Site of CNS disease was not specified in 22%. HIV status was known in 318 cases (6.3% positive). The brain [44%] was the most common site of involvement followed by spinal cord [19%] and meninges [15%]. Follicular lymphoma (FL) [48%] was the most common histology overall followed by marginal zone (MZL) [37%], small lymphocytic (SLL) [8%] and lymphoplasmacytic lymphomas (LPL) [7%]. MZL was the most common histology in the brain [44%] and meninges [61%] while FL was most common in the spinal cord [77%] and nervous system, NOS [69%]. Cranial nerves and eye (retina/optic nerve) involvement was very rare [2 and 1 case each- both MZL]. The overall survival (OS) of CNS B-cell NHL was significantly better if histology was low grade vs other [5-year OS 74% vs 32%, P < 0.0001]. Among CNS low grade B-cell NHLs, 5-year OS varied by histology [MZL 83%, FL 75%, LPL 56% and SLL 50%, P = 0.0003] and site of disease [spinal cord 89%, meninges 78% and brain 63%, P = 0.03] in addition to age at diagnosis and co-morbidities on both uni- and multivariate analysis. Survival was not influenced by sex, race, insurance, year of diagnosis, facility type or location. Conclusions: CNS involvement with low grade B-cell NHL is rare but has a relatively good outcome with most patients surviving beyond 5 years. FL and MZL are the more common low grade histologies. Both histology and disease site are important factors affecting survival.
Strategy for sensitive and specific somatic SNV/InDel calling: Leveraging different callers with in-house generated gold standard variant sets.
Authors: Huang Xiu , Sun Ruobai , Cingolani Pablo , Bjonnes Andrew , Lyle Stephen , Protopopov Alexei , Vuzman Dana
Published: September 2018

e13104

Background: Somatic mutation calling is critical for cancer genotyping. Although rapid development of mutation detection is witnessed with the maturity of NGS, the need for high sensitivity often results in compromised specificity and manual inspection. Here, we propose a methodology that leverages different variant callers to account for specificity without compromising sensitivity. Methods: We designed a cohort of training samples (n = 22), each with known set of SNVs/InDels that were discovered by KEW CANCERPLEX platform. We assessed the performance of four prevailing mutation callers that utilize different statistical approaches and therefore have different calls, using the training samples. We optimized the parameters of the four variant callers to detect all expected variants. We then examined the intersections of every combination of the four callers and identified the best one that eliminated the highest rate of false calls. A customized tool was developed for the intersection of component SNVs and InDels and the report differences among different callers. We also collected another set of validation samples (n = 28), each with true mutations that were both curated and orthogonally validated. We used this set to further test the efficacy of the refined calling strategy. Results: From the training samples, we chose the combination that provides highest sensitivity and specificity. This refined mutation calling strategy removed ~20% false SNV calls and ~50% false InDel calls in average for each sample. This result was further confirmed by the validation samples. Conclusions: The curated set of mutations from the genetic test platforms can provide valuable gold standard to test and tune mutation callers. The conclusions drawn from the curated variants are in line with the experimentally validated variants and showcase the validity of this practice, which we applied to demonstrate that the strategy of intersecting variants from optimized variant callers will generate mutation calls of higher specificity without compromising sensitivity. This methodology reduces the number of variants for curation and improves curation procedures and turn-around time.
Large germline next generation (NGS) sequencing panel of uterine serous carcinoma.
Authors: Romero Ignacio , Garcia-Casado Zaida , Lopez-Reig Raquel , Fernandez-Serra Antonio , Illueca Carmen , Poveda Andres , Lopez-Guerrero Jose Antonio
Published: September 2018

e13106

Background: Genetic predisposition to uterine serous carcinoma (USC) remains unclear and has been related to Lynch Syndrome (LS) and BRCA1/2 mutations. Wide NGS panels for this condition are starting to be implemented and data remain scarce. Methods: Consecutive USC were identified from a prospective endometrial cancer database (ECD) in a single institution. Clinical information including stage, follow-up, a three-generation pedigree, Microsatellite Instability (MSI) with 8 markers and the Mismatch Repair Protein (MMRPro) staining status, was retrieved from the ECD. Germline genetic testing was performed using a commercially available NGS panel including 80 familial cancer genes (Onco-GeneSGKit) on a MiSeq sequencer. Pathogenic or likely pathogenic variants were confirmed by Sanger sequencing. Results: Twenty-one women out of a total of 150 (14%) cases contained in the ECD were diagnosed with a USC. Median age was 65 (40-84 yo), median Body Mass Index of 26 (19-40), and histologically 18 were defined as pure and 3 as mixed USC. FIGO 2009 stages were I/II and III/IV in 42% and 58% of cases respectively. Adjuvant Chemotherapy (CT) was administered to 76%. MSI-High and MMRPro loss was described in 5%, 19% fulfilled Bethesda criteria, 23% had previous breast cancer and 5% personal Lynch related cancer history, 28% presented with pathological features of Lynch Syndrome such as dedifferentiation, Tumor Infiltrating Lymphocytes (14%), peritumoral lymphocytes or lower uterine location (14%). A median of 5 [range: 2-8] genetic alterations (pathogenic, variants of uncertain significance (VUS) and not described) per patient were observed, including 3 (14.3%) pathogenic mutations, 18 VUS and 43 not described. Among the three pathogenic mutations one was in MSH6 (p.Glu255Ter), another one in DKC1 (p.S280A) and the last one in the recessive gene MUTYH (p.Glu466del). VUS were detected in 57% of cases among 12 different genes: ATM (n = 3), AXIN2 (n = 2), BARD1 (N = 2), RAD50 (n = 2), RET (n = 2) and one case each of BMPR1A, BRIP1, CHEK2, MLH1, MSH6, SLX4 and TSC2. Conclusions: In our series 14% of USC harbored mutations in cancer predisposition genes.VUS are as high as 57% and represent a clinical challenge.
Breast cancer phenotype in patients with hereditary gene mutations other than BRCA1 and BRCA2.
Authors: Arun Banu , Elsayegh Nisreen , Lin Heather Y. , Clifton Katherine , Gutierrez-Barrera Angelica M. , Litton Jennifer Keating , Albarracin Constance T.
Published: September 2018

e13121

Background: Use of multi-gene panel (MGP) testing for hereditary breast cancer (BC) has increased significantly. Unlike in case of TNBC diagnosed before age 60 and ordering BRCA testing; for MGP testing, healthcare providers depend on personal and/or family history of cancer as tumor phenotype and on patient characteristics for non-BRCA hereditary BC genes which are not well described. Therefore, we planned to evaluate tumor phenotype of BCs associated with hereditary gene mutations other than BRCA 1 and BRCA2. Methods: Consecutive patients with invasive BC who are in a prospective cohort study and who underwent MGP testing based on published criteria as clinically indicated were included in the study. All patients’ breast pathology slides were reviewed by breast pathologists at our institution. For statistical analysis: Patients’ demographic and clinical characteristics were summarized using descriptive statistics such as frequency distribution, mean (± s.d.) and median (range). SAS version 9.4 and S-Plus version 8.04 were used to carry out the computations for all analyses. Institutional IRB approval was obtained. Results: Between 2013 and 2017, 1607 patients underwent MGP testing and 347 were found to be positive for a pathogenic variant. For the study purposes we included only patients with invasive breast cancer (N = 146). Median age was 45.1 yrs (Range: 25-78). Genetics testing results were as follows: BRCA1: 38 (26%), BRCA2: 37 (25%), CHEK2 15 (10%), PALB2: 12 (8%) , ATM: 12 (8%), TP53: 9 (6%), CDH1 : 5 (3%), PTEN: 3 (2%), BRIP1: 3 (2%), and 12 patients (8%) positive for other genes (CDKN2A, MUTYH, PMS2, APC, BARD1, MLH1, NBN, RAD51C, and SDHD). Tumor phenotype by gene mutation in shown in table 1. Conclusions: Several hereditary BC genes are associated with ER positive BC which could have implications for chemoprevention, while others associated with ER neg/TNBC that could have therapeutic implications. These findings and implications need to be further studied in larger cohorts. [Table: see text]
The MetAction project: Biomarker-directed molecularly matched therapy for end-stage cancer implemented in clinical practice.
Authors: Ree Anne Hansen , Flatmark Kjersti , Nygaard Vigdis , Heinrich Daniel , Boye Kjetil , Dueland Svein , Nygaard Vegard , Hovig Eivind , Beiske Klaus , Lund-Iversen Marius , Flørenes Vivi A , Johansen Christin , Bergheim Inger Riise , Sathermugathevan Menaka , Nakken Sigve , Geitvik Gry A , Lingjærde Ole Christian , Børresen-Dale Anne-Lise , Russnes Hege G , Mælandsmo Gunhild M ,   The MetAction Clinical Trial Team
Published: September 2018

e14033

Background: The MetAction project consists of two clinical trial phases. The completed first phase established the required diagnostic infrastructure, implemented security-approved systems for handling of sensitive information, educated the entire project staff within the context of tumor boards, and estimated costs of the initiative within public health services. The endeavor enabled expedite and safe mutation profiling of metastatic tumors in order to offer biomarker-based treatment with molecularly matched medication to patients with end-stage cancer, as reported in Ree et al., ESMO Open 2017. The ongoing second trial phase investigates the feasibility of the established MetAction pipeline in clinical practice. Methods: An eligible patient has end-stage metastatic disease from a solid cancer. Metastatic tissue is analyzed by DNA sequencing (Ion Oncomine™ Comprehensive Panel), where called variants are filtered prior to assessment and prioritization, supplemented with fluorescence in situ hybridization to cover relevant biomarkers. The Molecular Tumor Board interprets the findings within the likelihood of signaling pathway activity, and the sequential Clinical Tumor Board (CTB) may conclude on treatment with any systemic tumor-directed medication. Results: At the time of writing, 19 patients enrolled onto the second trial phase have accomplished the diagnostic procedures from sampling of metastatic tissue to CTB conclusion. Biopsy procedures were undertaken at lung or pleural sites (five cases), liver or superficial or deep peritoneal sites (13 cases), and an inguinal lymph node (one case) and did not cause adverse events. Histologic entities were 12 adenocarcinomas and one case each of squamous cell and undifferentiated carcinoma, cholangiocarcinoma, and four different sarcoma entities. Twelve patients have been found eligible for off-label use of molecularly matched therapy (inhibitor of ALK-, BRAF-, EGFR-, FGFR-, mTOR-, PARP-, or PD-1-mediated signaling). Conclusions: We will report on patient outcome (progression-free survival, overall response rate, and tolerance) to this biomarker-directed treatment in end-stage cancer. Clinical trial information: NCT02142036.
Lipid-insertion to enable targeting functionalization of paclitaxel loaded erythrocyte membrane nanoparticle by tumor-penetrating bispecific recombinant protein.
Authors: Sha Huizi , Chen Hong , Liu Baorui
Published: September 2018

e14047

Background: There is a great interest in targeting and penetrating of cancer cells for research or therapeutic purposes. Red blood cells (RBCs) are readily available and fully biocompatible long-circulating intravascular carriers that are amenable to chemical modifications, drug loading and reinjection. The purpose of this study was to design a tumor-targeting biocompatible drug delivery system for delivery of antitumor drugs. Methods: DSPE-PEG-MAL, phospholipid derivatives was used to insert into erythrocyte membrane nanoparticles. To make nanoparticles active targeting to the tumor site, a tumor-penetrating bispecific recombinant protein named anti-EGFR-iRGD was used. The characterization, bio-distribution, tumor targeting ability and antitumor activity of paclitaxel loaded anti-EGFR-iRGD modified erythrocyte membrane nanoparticle were evaluated. Results: In this study, anti-EGFR-iRGD-RBC-PTX nanoparticles was successfully constructed with a size of around 100 nm. A lipid-insertion method is employed to functionalize these nanoparticles without the need for direct chemical conjugation. It showed signifcantly targeted skill and increased cytotoxic effect toward both nontargeted RBC-PTX and combination of anti-EGFR-iRGD and RBC-PTX. The tissue distribution and antitumor assays in mice bearing gastric cancer xenograft confrmed the superior penetration tumor effcacy and antitumor activity of anti-EGFR-iRGD-RBC-PTX. Conclusions: We designed and successfully prepared a novel anti-EGFR-iRGD decorated, erythrocyte membrane sourced nanoparticle for targeted drug delivery, with enhanced tumor targeting and anti-tumor effect. Anti-EGFR-iRGD-RBC-PTX represents a potential effective nanomedicine against gastric cancer.
Combined treatment using the anti-p53 drug, APR-246 and eribulin: Synergistic growth inhibition in p53-mutated breast cancer cells.
Authors: Synnott Naoise C , Murray Alyson M. , O'Donovan Norma , Duffy Michael J. , Crown John
Published: September 2018

e14098

Background: TP53 is the most frequently mutated gene in triple-negative breast cancer, being present in approximately 80% of cases. APR-246 is a novel anticancer drug that acts by reactivating the mutant p53 protein, thereby converting it to a form with wild-type properties. Previously, we showed that APR-246 had antiproliferative, anti-migratory and pro-apoptotic activities in a panel of 23 breast cancer cell lines, including triple-negative (TN) cell lines. The aim of this study was to investigate if combined treatment with APR-246 and different cytotoxic agents resulted in enhanced growth inhibition. Methods: Cell viability was determined using the MTT assay. Combination index (CI) values were calculated using Calcusyn software, based on the Chou-Talalay method. Apoptosis was detected using Annexin V-FITC Apoptosis Detection Kit followed by FACs analysis. Results: Highly synergistic cell growth inhibition was found when APR-246 was combined with eribulin (Eisai Ltd.) in 6 different p53-mutated cell lines (mean CI values range from 0.38 to 0.77). In contrast, enhanced growth inhibition was not found using this combination in the 3 p53-WT cell lines investigated (mean CI values ranged from 1.13 to 2.9). Overall, p53 mutated cell lines had a significantly lower CI values than p53 wild-type cells (p = 0.008). In all the 4 p53-mutated cell lines investigated, a significant increase in apoptosis was also seen when APR-246 was combined with eribulin. This enhanced apoptosis appeared to result from increased mRNA expression of the pro-apoptotic factors PUMA and NOXA by the drug combination compared to either compound alone. In contrast to our findings with eribulin, combined treatment with APR-246 plus docetaxel, doxorubicin, cisplatin or carboplatin was cell line-dependent. Thus, docetaxel plus APR-246 was synergistic in 1/6 cell lines, while doxorubicin, cisplatin or carboplatin plus APR-246 was synergistic in 3/6 cell lines. Conclusions: Clinical trials investigating the combination of APR-246 and eribulin should be considered in patients with a p53 mutation such as triple-negative breast cancer.
Targeting mutant p53 with PK11007: A new approach for the treatment of patients with triple-negative breast cancer?
Authors: Synnott Naoise C , Bauer Matthias R , Madden Stephen F. , Murray Alyson M. , Klinger Rut , O'Donovan Norma , O'Connor Darran , Gallagher William M. , Crown John , Fersht Alan R , Duffy Michael J.
Published: September 2018

e14099

Background:The identification of a targeted therapy for patients with triple-negative breast cancer (TNBC) is one of the most urgent needs in breast cancer therapeutics. Since the p53 gene is mutated in approximately 80% of TNBC patients, it is a potential therapeutic target for this form of breast cancer. PK11007 is a 2-sulfonypyrimidine that stabilizes and reactivates mutant p53 (Bauer et al, PNAS 2016). The compound recently was reported to preferentially decrease viability in p53-compromised cancer cells. The aim of this investigation was to evaluate PK11007 as a potential new treatment for TNBC. Methods: Cell viability was determined using the MTT assay. Apoptosis was detected using Annexin V Apoptosis Detection Kit. Migration was determined by Transwell migration assay. Knockdowns of p53 protein were carried out using predesigned Flexitube sequences (Qiagen). Results: IC50 values for inhibition of proliferation by PK11007 in the panel of 17 breast cell lines ranged from 2.3 to 42.2 μM. There were significantly lower IC50values for TNBC than for non-TNBC cell lines (p = 0.03) and for p53-mutated cell lines compared with p53 WT cells (p = 0.003). Response to PK11007 however, was independent of ER or HER2 status of the cells. In addition, PK11007 induced apoptosis and inhibited migration in p53 mutant cell lines. Using RNAseq and gene ontogeny analysis, we found that PK11007 altered the expression of genes enriched in pathways involved in regulated cell death, regulation of apoptosis, signal transduction, protein refolding and locomotion. To establish if PK11007 acts by targeting mutant p53, we used siRNA to knockdown p53 in 3 p53-mutated TNBC cell lines. Reduction in p53 protein levels resulted in a significant decrease in the growth inhibitory effects of PK11007, in all 3 cell lines investigated, suggesting that PK11007 mediates growth inhibition via p53. The observations that PK11007 inhibited cell growth, induced apoptosis, blocked cell migration and altered genes involved in cell death, are all consistent with the ability of PK11007 to activate mutant p53. Conclusions: Based on our data, we conclude that targeting mutant p53 with PK11007 is a potential approach for treating p53-mutated TNBC.
ASN003, a highly selective BRAF and PI3K inhibitor: Preclinical and phase 1 clinical data in patients with advanced solid tumors.
Authors: Rasco Drew W. , Sullivan Ryan J. , Lakhani Nehal J. , Reddy Sanjeeva , Rao Niranjan Sathyanarayana , Denis Louis J. , Tolcher Anthony W. , Flaherty Keith
Published: September 2018

e14102

Background: RAS-RAF-MEK and PI3K-AKT-mTOR are two major signaling pathways involved in tumorigenesis. Components of these two pathways are frequently mutated in a broad range of tumors. ASN003 is a novel and highly selective small-molecule inhibitor of the RAS-RAF-MEK and PI3K pathways. Methods: The activity of ASN003 was determined using PI3K and BRAF enzymes, and efficacy was studied in human tumor xenograft models in mice. ASN003 is currently being investigated in patients with solid tumors in a Phase 1 trial using an accelerated dose titration design. In Part A, safety and tolerability of ASN003 is being studied in patients with advanced solid tumors. In Part B, safety, tolerability and preliminary efficacy of ASN003 will be evaluated in melanoma, CRC and NSCLC patients with a BRAF, PIK3CA or PTEN mutation. Pharmacokinetic (PK) profile and the pharmacodynamic (PD) effects of ASN003 on biomarkers such as pERK and pS6 are investigated in both parts of the study. Results: ASN003 showed potent and highly selective inhibition of BRAF and PI3K-α and -δ, and low affinity for PI3K-ß. ASN003 showed strong antiproliferative activity in cell lines and caused significant tumor growth inhibition in xenograft models harboring BRAF and PIK3CA or PTEN mutations. ASN003 showed antiproliferative activity in B-RAF and MEK inhibitor resistant cell lines. ASN003 had a strong antitumor activity in a BRAFV600mutant melanoma PDX model resistant to BRAF inhibitors, vemurafenib and dabrafenib. In humans, to date, ASN003 was well tolerated at 10 and 20 mg QD. Adverse events were mild and peak plasma level of 120 nM at 10 mg QD was achieved with a half-life of > 12 h. Dose escalation is ongoing. Conclusions: ASN003 is a unique small molecule, with highly selective and potent inhibition of BRAF, PI3-α and -δ kinases. ASN003 has strong antitumor activity in various xenograft tumor models harboring both BRAF and PIK3CA/PTEN mutations, and in a BRAF inhibitor resistant melanoma PDX model. To date, ASN003 was well tolerated and achieved good systemic exposure. Updated and detailed clinical, PK and PD results will be presented. Clinical trial information: NCT02961283.
Evolution of radiomic features on serial CT scans as an imaging based biomarker for evaluating response in patients with non-small cell lung cancer treated with nivolumab.
Authors: Xie Yuanqi , Khunger Monica , Thawani Rajat , Velcheti Vamsidhar , Madabhushi Anant
Published: September 2018

e14534

Background: Nivolumab is a PD-1 inhibitor that is FDA approved for treatment of chemotherapy refractory advanced NSCLC. The current standard clinical approach to evaluating tumor response is sub-optimal in evaluating clinical benefit from immunotherapy drugs. Our study aims to explore whether changes in radiomic features of the tumor between baseline and 2-week post-treatment CT scans can predict treatment response. Methods: 41 NSCLC patients treated with nivolumab were included in this study. 22 patients with pre- and post-nivolumab CT scans were used as a learning set and the remaining 19 for independent testing. Patients who did not receive nivolumab after 2 cycles due to lack of response or progression as per RECIST were classified as ‘non-responders’, and patients who had radiological response as per RECIST, or stable disease as per RECIST and clinical improvement were classified as ‘responders’. Lung nodules on pre-treatment CT scans were annotated with 3D SLICER software by a radiologist. 312 texture features of lung nodules were extracted and investigated in the study. The percent difference of each extracted feature was calculated based on the baseline and 2 week post-therapy CT scan. In the learning set, the six features that most significantly changed between baseline and post-treatment scans and also maximally differentially expressed between responders and non-responders were identified. Unsupervised clustering was applied on the set of 6 features for the 19 patients in the test set to predict which patients did and did not respond. Results: The top 6 features predictive of response corresponded to the Haralick, Gabor and Laws texture families. Unsupervised clustering yielded an accuracy of 78.95%. Conclusions: Our results suggest that changes in certain radiomic texture features between baseline and post-treatment CT scans following nivolumab could identify early clinical response to treatment. Additional validation of these novel quantitative imaging based approaches is warranted to accurately define clinical benefit from immunotherapy.
Tumor-penetrating peptide iRGD conjugation effectively potentiates intratumor T cell infiltration.
Authors: Ding Naiqing , Su Shu , Meng Fanyan , Sha Huizi , Chen Fangjun , Wei Jia , Chen Hong , Du Shiyao , Liu Baorui
Published: September 2018

e14543

Background: Poor infiltration of activated lymphocytes into solid tumors can be a fundamental factor limiting their efficacy. The purpose of this study was to determine whether a cycled tumor-penetrating peptide, iRGD, with a well-defined role in delivering drugs into extravascular tumor tissues, could facilitate lymphocytes infiltration into tumors in both the iRGD combination regimen and conjugated pattern. Methods: We used polyethylene glycol-conjugated phospholipid (PEG-lipid) derivatives, a time-efficient and versatile platform, to immobilize iRGD on T cell membrane, and examined the ability of iRGD to improve lymphocytes infiltration into tumors in multicellular spheroids and two xenograft mouse models. Furthermore, the synergy effect of iRGD modification and PD-1 gene knock out in adoptive T cell transfer immunotherapy was evaluated in a xenograft model of EBV associated gastric cancer. Results: In this study, we first showed that systematic injection of iRGD could double the number of CD3+ T cells in 4T1 tumor in BALB/C mouse and the synthetic iRGD-PEG-lipid, which could spontaneously transfer from solution to cell surfaces, had no effect to cell vitality, proliferation, phenotype and function. In adoptive transfer studies, the infiltration of T cells could be promoted by the co-administration with iRGD while T cells modified with iRGD could spread more extensively throughout both the multicellular spheroids and tumor mass. Furthermore, We demonstrated that iRGD modified T cells had superior antitumor efficiency to iRGD combination regimen due to increased T cell infiltration, and exhibited robust synergy effect with PD-1 gene knock out. Conclusions: Our study offers a simple method to target immune cells into tumors and may have the potential to enhance the clinical efficacy of adoptive T cell immunotherapy.
Target cell killing effects of CD20 targeting chimeric antigen receptor T cells derived from the type II anti-CD20 antibody.
Authors: Yao Yihong , Yao Xin , Zhu Shigui , Zhu Wei , Li Zhiyuan , Wang Qingxia , Zhu Lin , Ma Anyun , Li Yanfeng , Wei Yutian , Dai Chengxiang , Zhang Li , Huang Jiaqi , Liu Bizuo
Published: September 2018

e14548

Background: Chimeric Antigen Receptor T cells (CAR-Ts) targeting CD19 have shown very promising clinical outcomes in treatment of B-cell linage hematological malignancies. However, many patients with relapsed diseases were found to have down-regulated/loss of CD19 surface expression after CD19 CAR-T therapy. To solve this issue of CD19 single-targeting escape, we explored the application of another B-cell antigen, CD20, for targeted CAR-T therapy. Methods: We constructed four CD20 targeting CARs (all with 4-1BB co-stimulatory signaling) base on single-chain variable fragments (scFV) derived from four well-studied CD20 specific antibodies: Leu16, Rituximab, Obinutuzumab, and Ofatumumab. Leu16, Rituximab, and Obinutuzumab belong to the type I anti-CD20 antibody family and appear to bind to different epitopes located on the large loop of CD20, whereas Ofatumumab is the type II anti-CD20 antibody which has been shown to interact with the hydrophobic residues on the small loop surrounding a deep binding cleft. Results: All four CAR-T cells can specifically recognized CD20 positive target cells in our pre-clinical studies. They all showed up-regulated antigen-specific cell activation and high level of IFN-g release upon CD20 stimulation, and CAR-T20-Ofatumumab cells appeared to have significantly higher cell activation and more than 2-fold increase in IFN-g release compared to the other three CAR-T20 cells with their scFVs deriving from type I anti-CD20 antibodies. CAR-T20-Ofatumumab cells also showed higher degranulation upon stimulation, and it displayed ~50% of increase in ability to kill CD20 positive cells in cytotoxicity assays. Conclusions: Our data suggested that CAR-T20-Ofatumumab has better in vitro function and appears to be a CAR superior to those derived from other three antibodies. A possible explanation for this observation is that Ofatumumab interacts with the hydrophobic residues on the small loop, which is very close to cell membrane and confers more extensive binding to the small loop with striking slow off-rate. Our results suggest that CAR-Ts targeting CD20 with the scFVs from the type II anti-CD20 antibody may have superior cell killing effects.
HVEM: A novel cosignaling molecule of major interest in melanoma.
Authors: Malissen Nausicaa , Macagno Nicolas , Granjeaud Samuel , Granier Clemence , Moutardier Vincent , Gaudy Marqueste Caroline , Habel Nadia , Mandavit Marion , Guillot Bernard , Pasero Christine , Tartour Eric , Ballotti Robert , Grob Jean Jacques , Olive Daniel
Published: September 2018

e14591

Background: Herpes Virus Entry Mediator (HVEM) is a member of the TNF superfamily, its ligation of the immunoglobulin family member B-lymphocyte and T-lymphocyte attenuator (BTLA) activates inhibitory signaling in T-cells and could play a role in evading host anti-tumor immunity. Ectopic overexpression of HVEM is associated with poor survival in some solid tumors. Our objectives were: -to evaluate HVEM expression in melanoma (MM) -to explore potential link of its overexpression with clinical outcomes -to understand the mechanisms by which HVEM was ectopically expressed in MM, and how HVEM-BTLA may play a role in MM. Methods: HVEM expression was analyzed by IHC in formalin-fixed samples of mm metastases and was correlated with OS in 130 patients. To better understand how HVEM could interfere with prognosis, relation of HVEM with its ligand BTLA was studied in the tumor and tumor microenvironment on 15 fresh metastases by flow cytometry after tumor dissociation. Bioinformatic studies based on TCGA and CCLE data combined to targeting of candidate genes by siRNA were used to investigate HVEM regulation on MM. Results: Patients with high HVEM expression on mm cells in their metastases have a significantly (p = 0.0142) poorer overall survival than those with a low expression. TCGA transcriptomic data support these results. From the mechanistic point of view, we could show that 1-HVEM expressed at the tumor cell surface interacts with BTLA expressed by tumor-infiltrated lymphocytes. 2-HVEM expression is neither linked to the mm mutational status nor inducible by IFN 3-Genes co-regulated with HVEM are associated with an aggressive gene signature 4- HVEM strongly correlate with MITF expression, MITF binds HVEM promoter, and its downregulation by siRNA results in a decrease in HVEM expression. Conclusions: A high expression of HVEM by mm metastases seems to be a pejorative prognostic marker. HVEM-BTLA interaction is a co-signaling system similar to the PD1-PDL1 one, but seem to be constitutive rather than inducible. HVEM and its co-regulated genes may constitute a signature of aggressiveness associated to MITF. High HVEM expression on mm cells may dampen anti-tumor immune responses, making HVEM and BTLA potential new targets for “checkpoint blockade” therapy.
Measurement of PD-1, TIM-3 and LAG-3 protein in non-small cell lung carcinomas (NSCLCs) with acquired resistance to PD-1 axis blockers.
Authors: Datar Ila , Mani Nikita , Henick Brian S. , Wurtz Anna , Kaftan Edward , Herbst Roy S. , Rimm David L. , Gettinger Scott N. , Politi Katerina A. , Schalper Kurt Alex
Published: September 2018

e14611

Background: PD-1 axis blockade induces lasting clinical responses in ~20% of patients with advanced NSCLC. However, most patients eventually develop resistance. Acquired resistance is poorly understood, but may be mediated by alternative immune suppressive pathways. Methods: Using multiplex immunofluorescence we simultaneously measured levels of DAPI, CD3 (D7AE6), PD-1 (EH33), TIM-3 (D5D5R) and LAG-3 (17B4) in 11 whole tissue sections obtained from patients with NSCLC before PD-1 axis blockade and after acquired resistance (8 cases with progression on-treatment and 3 with progression off-therapy). Markers were measured in the whole tumor area or in CD3+ T-cells using fluorescence co-localization. The association between markers and changes upon acquired resistance were studied. Results: Expression of PD-1, TIM-3 and LAG-3 was seen in all cases with membranous staining pattern and signal predominantly located in CD3+ T-cells. Levels of TIM-3 and LAG-3 in T-cells were significantly correlated (Spearman’s R = 0.65, P = 0.001), but were not associated with PD-1 (R = -0.03, P = 0.86 for TIM-3 and PD-1; and R = 0.24, P = 0.28 for LAG-3 and PD-1). Compared to pre-treatment samples, 6 cases (55%) showed significantly higher levels of PD-1 or LAG-3 on acquired resistance and 5 cases (45%) had higher TIM-3. Of these, 4 cases had higher levels of the 3 markers and were on-therapy at progression. Lower levels of PD-1, TIM-3, and LAG-3 were found on acquired resistance in 5 (45%), 6 (55%), and 4 (36%) cases, respectively. Four of these cases showed lower levels of all inhibitory receptors, 3 of which were off-therapy at progression. Only one case had no change in LAG-3 levels. Conclusions: PD-1, TIM-3 and LAG-3 were expressed in the majority of NSCLCs with signal predominantly located in T-lymphocytes. Among acquired resistance cases, higher levels of PD-1, TIM-3 and LAG-3 were associated with progression on-treatment. Lower levels of the markers were associated with progression off-therapy. Although multiple mechanisms may exist, up-regulation of alternative immune inhibitory receptors such as TIM-3 and LAG-3 could mediate acquired resistance to PD-1 axis blockers in a proportion of NSCLCs.
Cancer peptide vaccine to suppress postoperative recurrence in esophageal SCC patients with induction of antigen-specific CD8+ T cell.
Authors: Yasuda Takushi , Nishiki Kohei , Yoshida Koji , Shiraishi Osamu , Iwama Mitsuru , Kato Hiroaki , Yasuda Atsushi , Shinkai Masayuki , Chiba Yasutaka , Okuno Kiyotaka , Nakamura Yusuke
Published: September 2018

e14635

Background: This is a following report of the exploratory phase II study of adjuvant peptide vaccine for thoracic esophageal squamous cell carcinoma (TESCC) (abstract #3087 in ASCO2016), focusing on an association of immunological responses with clinical outcomes. Three HLA-A-24-restricted epitope peptides derived from cancer-testis antigens, up-regulated lung cancer 10 (URLC10), cell division cycle associated 1 (CDCA1) and KH domain-containing protein overexpressed in cancer 1 (KOC1), were used in this study. Methods: TESCC patients who underwent neoadjuvant therapy followed by curative resection (Dec./2009 to Sep./2014) and were found LN metastasis were enrolled. One mg each of three peptides mixed with Montanide was injected to patients with HLA-A*2402 20 times in a 6-months period. No other adjuvant therapy was given until recurrence occurred. Primary endpoint is relapse-free survival (RFS). Immunological responses were examined by ELISPOT assay and immunohistochemistry (IHC). Results: 33 patients were enrolled in the vaccine group (VG) and 30 with non-HLA-A*2402 were used as controls (CG). There was no significant difference in clinical backgrounds between the two groups. 5-year RFS in the VG was 44.6% while that in the CG group was 31.6% (p = 0.207); patients who showed CD8+ CTL induction to multiple peptides tend to show better RFS rate [3/2/1 peptides = 69.0(n = 20)/46.8(n = 11)/0%(n = 2), respectively]. Five-year esophageal cancer-specific survival (ECSS) were 58.4% in the VG compared with 35.4% in the CG (p = 0.156); 90.0% (n = 11) vs. 50.0% (n = 15) and 43.8% (n = 22) vs. 24.0%(n = 15) in pN1 and pN2-3 groups, respectively. Regarding the CD8+ positivity, ECSS was improved only in the negative group (56.6% in VG vs. 26.7% in CG). Classification by T cell positivity and PD-L1 expression [type I(+/+), II(-/-), III(-/+) and IV(+/-)] showed 5-year ECSS to be 100%(n = 3), 66.4%(n = 19), 20.0%(n = 5) and 44.4%(n = 6). Conclusions: Our cancer peptide vaccines induced the antigen-specific CD8+ T cell highly and efficiently, and suppressed recurrence with the strong immune responses. Clinical trial information: UMIN000003557.
Tumor bulk as an independent marker of anti-EGFR therapeutic benefit in metastatic colorectal cancer.
Authors: Kratz Jeremy D. , LoConte Noelle K. , Lubner Sam Joseph , Mulkerin Daniel , Matkowskyj Kristina , Uboha Nataliya Volodymyrivna , Lubner Meghan G. , Kim David H. , Pickhardt Perry J. , Deming Dustin A.
Published: September 2018

e15054

Background: Molecular testing and location of the primary tumor (right-sided (R) versus (vs) left-sided (L)) are useful in predicting the clinical benefit of the anti-epidermal growth factor receptor (EGFR) antibodies, cetuximab (cet) and panitumumab (pan), in metastatic colorectal cancer (mCRC). We hypothesized that tumor bulk might also be predictive of treatment benefit given the potential for increased intra-tumor heterogeneity and reduced penetration of antibodies into bulkier lesions. Methods: A single institution retrospective cohort of 69 patients (pts) with KRAS wild-type mCRC were identified who received either cet or pan in the late-line setting +/- chemotherapy. Metastatic sites were cataloged including independent review of CT imaging prior to initiation of anti-EGFR therapy. Disease bulk was defined categorically as single metastatic lesion with diameter measuring > 3.0 cm. Results: This cohort represents a diverse group having received varying prior lines of therapy and having assorted disease sites including mediastinal, pulmonary, hepatic, omental, and osseous lesions. When treated with anti-EGFR therapies, pts with pre-treatment metastases ≤3.0 cm in diameter had significant improvement in median progression free survival (mPFS) (6.2 months (mos)) vs pts with metastases > 3.0 cm (3.9 mos, p < 0.01). A trend towards improvement in overall survival was observed for pts with non-bulky (15.8 mos) vs bulky disease (8.6 mos, p = 0.08). A trend towards increased mPFS existed in L (5.1 mos) vs R cancers (2.5 mos, p = 0.11). In R cancers, no significant difference in mPFS was noted between bulky (2.5 mos) and non-bulky disease (2.9 mos, p = 0.53). Non-bulky L cancers had a significantly improved mPFS of 7.5 mos compared to 3.9 mos with L bulky disease (p < 0.01). Non-bulky L cancers trended towards improved overall survival of 17.0 mos compared to 9.6 mos with L bulky disease (p = 0.08). Conclusions: Despite limited small sample size, these data indicate that tumor bulk is a potential predictor of the therapeutic benefit of anti-EGFR agents in left-sided mCRCs across a heterogeneous pt population. Disease bulk deserves further evaluation in larger datasets of mCRC across targeted therapeutics.