bluuebunny/crossref_metadata_2025_split huggingface dataset:
A sensitive QPCR assay for EGFR mutation in plasma samples of NSCLC patients.
Authors: Feng Qin 
Published: September 2018

2595 Background: Tyrosine kinase inhibitors (TKI) have improved the overall outlook and quality of life for most of EGFR mutation positive NSCLC patients. However, tumor tissues are often absent or insufficient for testing EGFR mutations to guide EGFR TKIs treatment of patients with nonsmall cell lung cancer (NSCLC). An assay that detects EGFR mutations in the plasma would provide a noninvasive technique to assess suitability for TK inhibitor therapy. The ACCB new EGFR Mutation Kit is a ARMS-PCR test for the qualitative detection of 45 mutations in exons 18, 19, 20, and 21 of the EGFR gene in DNA derived from human plasma from NSCLC patients. Methods: 10 mL tubes of blood were collected from patients who never had been treated by EGFR TKI, and plasma circulating tumor DNA were extracted from plasma by Biomark Circulating DNA Kit. Qubit 2.0 Fluorometer was used to make plasma circulating DNA tumor quantitation. The concentration of final DNA sample is ≦2ng/μl. Total, 272 plasma DNA samples from 246 lung adenocarcinoma, 23 lung squamous carcinoma and 3 lung adenosquamous carcinoma patients were collected. 104 paired tissue EGFR mutations were detected by the same kit. Results: EGFR mutation was detected in 88 from 272 plasma DNA samples, 86 lung adenocarcinoma, 1 lung squamous carcinoma, 1 lung adenosquamous carcinoma, with EGFR mutation rate were 35.0% (86/246), 4.3% (1/23), 33.3% (1/3) respectively. In 61 Ⅰ~Ⅲ and 211 Ⅳ stage NSCLC patients, EGFR mutation rate were 21.3% (13/61) and 35.5% (75/211) respectively. Conclusions: The ACCB new EGFR Mutation Kit is a sensitive, accurate, rapid, and reproducible assay capable of testing DNA extracted from human plasma from NSCLC patients.
Effect of chimeric antigen receptor (CAR) T cells on clonal expansion of endogenous non-CAR T cells in patients (pts) with advanced solid cancer.
Authors: Kim Rebecca H , Plesa Gabriela , Gladney Whitney , Kulikovskaya Irina , Levine Bruce L , Lacey Simon F. , June Carl H. , Melenhorst Jan J. , Beatty Gregory Lawrence 
Published: September 2018

3011 Background: CAR T cells have produced remarkable responses in heme malignancies, but efficacy in solid cancers is limited. Poor in vivo persistence and heterogeneous expression of the CAR target on tumors are potential barriers to the success of CAR T cell therapy. However, even with transient persistence, CAR T cells may elicit a “vaccine” effect by inducing cancer cell death and subsequent release of tumor antigens that could stimulate tumor-specific T cell activity. Methods: 6 pts with pancreatic ductal adenocarcinoma (PDAC) received repeated 3x per week intravenous (iv) infusions of mRNA-transfected mesothelin-redirected CAR T cells (CARTmeso). Pts with PDAC (n = 5), ovarian carcinoma (n = 5), and mesothelioma (n = 5) received iv infusion of lentiviral-transduced (lenti) CARTmeso with or without cyclophosphamide (Cy) preconditioning. Peripheral blood samples were collected from pts at baseline and defined time points after treatment. Genomic DNA from these samples or from pre-infused CAR T cell product was used for deep sequencing of the TCRbeta chain using the ImmunoSEQ platform. A TCRbeta clone was considered to have expanded from baseline to defined time points after treatment if it showed a two-fold change from baseline and met statistical significance by Fisher’s exact test (p < 0.05). Results: mRNA CARTmeso cells persisted in vivo for < 24 hrs. Unexpectedly, therapy induced clonal T cell expansion detected in the blood by day 14 in all 6 pts. Expanded clones underwent contraction by day 28 in 3 pts. In one pt, peripherally expanded clones were also detected in a tumor biopsy, but without significant intratumoral clonal expansion. Lenti CARTmeso therapy also induced peripheral expansion of T cell clones both present and not present in the infused CAR T cell product. However, with Cy preconditioning, clonal expansion seen after lenti CARTmeso therapy was predominately restricted to clones detected in the CAR T cell product. Conclusions: In pts with advanced solid cancers, CARTmeso stimulates clonal expansion of endogenous T cells, which is lost with Cy conditioning. Findings suggest that CAR T cells may elicit a “vaccine” effect with potential therapeutic implications.
Gene expression analysis of tumor biopsies from a trial of durvalumab to identify subsets of NSCLC with shared immune pathways.
Authors: Streicher Katie , Morehouse Chris , Sebastian Yinong , Kuziora Michael , Higgs Brandon W. , Ranade Koustubh 
Published: September 2018

3041 Background: We previously reported that NSCLC pts with high pretreatment tumoral IFNG mRNA signature (sig) expression have improved outcomes (ORR, PFS, OS) to the anti-PD-L1 therapy durvalumab (D). To explore the relationship of other immunotherapy targets with the IFNG sig, we evaluated expression of CD137, PD1, PDL1, CTLA4, GITR, OX40, TLR7, TLR8, CD73, TIM3, NKG2A, IDO1, CD40, LAG3, A2AR, CXCR4, iNOS, ARG1, and STAT3 in D-treated NSCLC. Methods: CP1108/NCT01693562 was a nonrandomized phase 1/2 trial evaluating D (10 mg/kg Q2W) in advanced NSCLC. As of 24OCT16, 304 primarily previously-treated pts were enrolled. RNA sequencing was conducted on available tumor specimens from 98 pts. ORR was evaluated using RECIST v1.1. 19 genes were z-scored, scaled and clustered across pt tumors. Each gene was coded binary using 0 as a cut point. The proportion of tumors with concordant or discordant over-expression between a gene and the IFNG sig was calculated. Results: In 43% of evaluable NSCLC pts, mRNAs for PD-1, IDO1, PD-L1, CD40, CTLA4, LAG3, TIM3, TLR8, NKG2A, and CD137 were co-expressed with IFNG sig (Spearman’s rho > = 0.7). In 34% of evaluable pts, TLR7, OX40, GITR, A2AR, and CXCR4 mRNAs had moderate concordance with IFNG sig (0.5 < rho < 0.7) and in 23% of evaluable pts iNOS, CD73, ARG1, and STAT3 had low concordance (rho < 0.5). Within the cluster of pts including high IFNG sig, a small subset expressed high iNOS and CD73; however, pts with high STAT3 or ARG1 formed a distinct cluster within the low IFNG sig subset. The subset with high IFNG sig had an ORR of 24% compared to only 10% in all other subsets combined. Conclusions: Our findings enrich understanding of the immune microenvironment of NSCLC by identifying three broad categories of tumors: tumors with pre-existing immunity that have high IFNg sig and select other IO pathways with enhanced responses to D; tumors with moderate expression of IO genes in which the local microenvironment is crucial; finally a distinct “cold” subset of tumors with high expression of STAT3 or ARG1 and characterized by low or no expression of IFNG sig and other IO genes. These results may aid in identifying the right pts for anti-PD-L1 and in prioritizing immunotherapy combinations.
Paracrine wnt-β-catenin signaling inhibition as a strategy to enhance the efficacy of anti-PD-1 antibody (Ab) therapy in a transgenic model of melanoma.
Authors: DeVito Nicholas C. , Xiao Christine , Zhao Fei , Evans Kathy S. , Theivanthiran Bala , Lewicki John , Hoey Tim , Hurwitz Herbert , Strickler John H. , Hanks Brent Allen 
Published: September 2018

3053 Background: Activation of the Wnt-β-catenin signaling pathway is associated with poor T cell infiltration of tumors. We have previously demonstrated that paracrine Wnt5a-β-catenin signaling is a critical trigger of dendritic cell (DC) tolerization and regulatory T cell (Treg) differentiation in the melanoma microenvironment. In a transgenic BRAFV600EPTEN-/- model, the genetic silencing of melanoma-derived Wnt5a potently enhances infiltrating CD8+T cell effector function and promotes responses to anti-PD-1 Ab therapy. Ipafricept (IPA) is a recombinant Wnt decoy receptor and Vantictumab (VAN) is a Fzd receptor monoclonal Ab. Both molecules inhibit Wnt-β-catenin signaling and have been well-tolerated in ongoing phase I/Ib clinical trials. We explored the ability of IPA/VAN to reverse tumor-mediated immune tolerance and enhance the efficacy of anti-PD-1 Ab immunotherapy in a pre-clinical model that closely recapitulates human melanoma. Methods: Both IPA and VAN were utilized to investigate Wnt-β-catenin inhibition as a strategy for suppressing melanoma-induced DC indoleamine 2,3-dioxygenase (IDO) expression and Treg differentiation in vitro. These agents were further tested for their ability to enhance anti-tumor T cell responses and to augment the efficacy of anti-PD-1 Ab therapy in syngeneic and autochthonous models of BRAFV600EPTEN-/- melanoma. Results: IPA and VAN effectively inhibit Wnt5a and melanoma-induced DC IDO expression and Treg differentiation in vitro. Further studies demonstrate that IPA and VAN significantly augment anti-PD-1 Ab-mediated suppression of primary and metastatic tumor progression in both syngeneic and autochthonous BRAFV600EPTEN-/- melanoma models. These anti-tumor effects correlated with suppressed IDO enzymatic activity, enhanced tumor-infiltrating CD8+T cell/Treg ratios, and increased activation of TRP2 antigen-specific effector T cells. Conclusions: The pharmacological inhibition of paracrine Wnt-β-catenin signaling with IPA and VAN augment the anti-tumor efficacy of anti-PD-1 Ab therapy and represent a promising strategy for further phase I testing in melanoma and other solid tumors.
Safety profile of avelumab in patients with advanced solid tumors: A JAVELIN pooled analysis of phase 1 and 2 data.
Authors: Kelly Karen , Infante Jeffrey R. , Taylor Matthew H. , Patel Manish R. , Gordon Michael S. , Wong Deborah Jean Lee , Iannotti Nicholas , Mehnert Janice M. , Loos Anja , Koch Helga , Speit Isabell , Gulley James L. 
Published: September 2018

3059 Background: Avelumab is a fully human IgG1 anti–PD-L1 antibody with clinical activity in several tumor types. Pooled safety data from a large phase 1 trial in various tumors and a phase 2 trial in Merkel cell carcinoma (NCT01772004, NCT02155647) were analyzed to further characterize the safety profile of avelumab. Methods: Patients (pts) received avelumab 10 mg/kg 1-hour IV Q2W until progression, unacceptable toxicity, or withdrawal. Treatment-related adverse events (TRAEs) were graded by NCI CTCAE. In post hoc analyses, immune-related adverse events (irAEs) were identified via an expanded AE list and medical review, and infusion-related reaction (IRR) events were identified based on prespecified MedDRA terms, occurring within 1 day or related symptoms that resolved within 2 days of infusion. Results: In 1,738 pts analyzed (phase 1, n = 1,650; phase 2, n = 88) who received ≥1 dose of avelumab for a median of 12 weeks (range 2-138), the most common any grade TRAEs were fatigue (n = 307, 18%), IRR (n = 295, 17%), and nausea (n = 150, 9%). 177 pts (10%) had a grade ≥3 TRAE; most common were fatigue and elevated lipase (17 [1%] each). TRAEs led to discontinuation in 107 pts (6%). Four pts (0.2%) died due to a TRAE. Any grade irAEs occurred in 247 pts (14%), which were grade ≥3 in 39 pts (2%) and considered serious in 43 pts (2%). The most common any grade irAEs were thyroid disorder (n = 98, 6%) and rash (n = 90, 5%). Other irAEs (eg, colitis, hepatitis, pneumonitis, adrenal insufficiency, myositis) each occurred in < 2%. irAEs led to discontinuation in 34 pts (2%). IRR or related symptoms (eg, chills, pyrexia, hypersensitivity) occurred in 439 pts (25%), which were grade 3 in 9 pts (0.5%) and grade 4 in 3 pts (0.2%). An IRR occurred at first infusion in 79% and within first 4 doses in 99%; 63/439 pts (14%) had IRR recurrence in later cycles. IRR led to dose interruption in 152 (9%), infusion rate reduction in 124 (7%), and discontinuation in 35 pts (2%). Conclusions: This large pooled analysis confirms that avelumab has an acceptable safety profile. A minority of pts experienced a grade ≥3 TRAE or irAE and discontinuation due to TRAEs was uncommon. IRRs mostly occurred at first infusion and the rate of recurrence was low. Clinical trial information: NCT01772004, NCT02155647.
Sustained oligoclonal T cell expansion correlates with durable response to anti-PD1 therapy.
Authors: Olugbile Sope Omowale , Kiyotani Kazuma , Park Jae Hyun , Hoffman Philip C. , Szeto Livia , Patel Jyoti D. , Vokes Everett E. , Nakamura Yusuke 
Published: September 2018

3061 Background: Immune checkpoint blockers have demonstrated durable response in many tumor types including lung cancer. This clinical benefit is however restricted to a subset of patients in whom longitudinal assessments of their tumor microenvironment have revealed increase CD8+ T cell infiltration. However detailed characterization of this T cell dynamics at clonal levels in a large population of patients is yet to be reported. Such analyses will provide vital insight into the mechanism of tumor eradication in responders. Methods: We performed next-generation sequencing of T cell receptor β chain in longitudinal tumor and peripheral blood samples obtained from lung cancer patients treated with immunotherapies. We first extracted RNA from these samples and synthesized cDNA with 5’RACE adapter using SMART library construction kit (Clontech). We then amplified TCR β gene products to prepare sequence libraries which were analyzed with MiSeq system (Illumina). Results: Here we report on TCR receptor β chain sequencing data for 29 patients in whom we had 3 or more serial samples. Thirteen (45%) of the selected patients in this cohort had durable response and seven (24%) had radiological complete response. We confirmed that there is concordance between the expanded T cell clone at tumor site and the peripheral blood. In one responder, we found expansion of a dominant T cell clone (20%) at a metastatic site where he had pathological complete response on day 17 of treatment and the same clone remained persistent in his peripheral blood (24%) at week 48 of therapy. Similarly in other responders, there were T cell clonal expansions detected as early as week 2 after only one cycle of treatment and such clones remained at high frequencies several months afterwards. Such pattern of early and sustained clonal expansions were absent among non-responders even while they remained on therapy. Conclusions: We found durable response to immune checkpoint blockade to correlate with early and sustained expansion of one to two dominant T cell clones in this selected patient cohort.
Relationship between liver metastases and PD-1 blockade in melanoma.
Authors: Lee James Chi-Chiang , Tsai Katy K. , Algazi Alain Patrick , Rosenblum Michael , Bluestone Jeffrey , Daud Adil 
Published: September 2018

3072 Background: WhilePD-1 blockade is effective in melanoma, durable responses remain elusive. We have previously reported that liver metastasis is associated with reduced response rates and that the fraction of CTLA4 hi/PD-1 hi CD8+ cells (“activated-exhausted” or T-ex cells) within the TIL is predictive of response to PD-1 blockade. Here, we explore the biology behind liver metastasis in human melanoma and in animal models. Methods: Patients with metastatic melanoma with or without liver metastasis were biopsied pre- PD-1 treatment and immune infiltrates were analyzed by FACS. The CD8 fraction was gated on CTLA4 and PD-1. C57BL/6 mice were implanted with a “primary” subcutaneous tumor and a “metastatic” tumor in the liver or the lungs (control), and given systemic PD-1 blockade therapy. Results: Patients with melanoma and liver metastasis (n = 25) had 15.2% T-ex cells while those without liver metastasis (n = 76) had 26.5 % T-ex cells, p = 0.0092. A T-ex fraction < 20% was significantly associated with lack of PD-1 response, p < 0.005. In C57BL/6 mice implanted with a B16 tumor (subQ & liver) treated with PD-1 antibody, 0/35 mice achieved subQ tumor rejection while in the SubQ only mice 9/30 mice (30%) rejected their tumors. The mean tumor size of mice with Sub Q+liver metastasis was 139.2 mm2 vs subQ only mice 23.4 mm2 at d 14, p = 0.002. Mice with liver metastasis showed a T-ex fraction 31.9% vs 67.3%without liver met, p = 0.0003. In contrast, in mice made lung metastatic, the subQ tumor rejection rate was 7/20 (35%), with T-ex infiltrate at 57.9%. The implantation of liver metastases from an unrelated MC38 tumor does not protect the subQ tumor from immune rejection. Conclusions: The presence of liver metastases is associated with reduced response to PD-1 blockade and reduced T-ex infiltrate in patients with stage IV melanoma. Mechanistic studies using a mouse model of syngeneic organ site specific metastasis confirms that the liver metastasis results in reduced antigen specific T cell at distant sites, resulting in reduced response. Site of metastasis may determine immune responsiveness in both mouse models and in humans with melanoma.
Phase 1 open-label, multiple ascending dose trial of AGEN1884, an anti-CTLA-4 monoclonal antibody, in advanced solid malignancies.
Authors: Wilky Breelyn A. , Kumthekar Priya , Wesolowski Robert , Hwang Jimmy J. , Raizer Jeffrey J. , Gentry Carleen , Ross Ainsley , Drouin Elise , Wilson Nicholas , Shebanova Olga , Goldberg John M. , Buell Jennifer , Stein Robert Benjamin , Cuillerot Jean-Marie 
Published: September 2018

3075 Background: AGEN1884 is a fully human IgG1 monoclonal antibody targeting the co-inhibitory protein cytotoxic T lymphocyte-associated protein 4 (CTLA-4). CTLA-4 blockade has been shown to augment T cell activation and proliferation, resulting in immune infiltration of the tumor and subsequent regression. Objectives: Assess the safety, maximum tolerated dose (MTD), and pharmacokinetic (PK) and pharmacodynamic (PD) characteristics of AGEN1884 in patients (pts) with advanced and refractory malignancies using a “3+3” trial design. Methods: Eleven pts have been enrolled and treated to date. AGEN1884 was administered intravenously q3w for 4 doses and then q12w. Three (0.1, 0.3 and 1 mg/kg) of six (3, 6 and 10 mg/kg) planned dose levels have been completed. Results: Five pts were accrued at 0.1 mg/kg dose level (2 were not DLT evaluable) and three pts each at doses of 0.3 mg/kg and 1 mg/kg. Median age was 56 years (range 26–70), ECOG 0–2, with a median of 4 (range 1–8) prior therapies. No DLT events have been observed thus far. Data from 5 pts were available for PK evaluation. Half-life of AGEN1884 post first dose was 8.8 and 9.6 days for 0.3 mg/kg and 0.1 mg/kg dose levels, respectively, as measured by ELISA. As of Jan 31, 2017, pts across cohorts were followed for a median of 6 weeks (range 0-28). Six pts (54.5%) have come off study due to disease progression, while 5 (45.5 %) remain on study. One confirmed partial response (80% reduction) by RECIST criteria was seen at 0.1 mg/kg in a patient with angiosarcoma. Conclusions: AGEN1884 is safe at 0.1 and 0.3 mg/kg dose levels. Dose escalation is ongoing and updated safety and PK data will be presented. Clinical trial information: NCT02694822.
Characterization of the T-cell receptor repertoire and immune microenvironment in patients with locoregionally advanced squamous cell carcinoma of the head and neck.
Authors: Saloura Vassiliki , Fatima Aiman , Zewde Makda , Kiyotani Kazuma , Brisson Ryan J. , Park Jae Hyun , Ikeda Yuji , Vougiouklakis Theodore , Bao Riyue , Seiwert Tanguy Y. , Cipriani Nicole , Lingen Mark W. , Vokes Everett E. , Nakamura Yusuke 
Published: September 2018

3080 Background: Immunotherapy with checkpoint blockade was recently approved for patients with recurrent/metastatic SCCHN, however it has not been investigated in the curative-intent setting yet. In this study, we investigated the T-cell receptor repertoire and the immune microenvironment in tumor tissues of SCCHN patients with locoregionally advanced disease. Methods: T-cell receptor sequencing and polymerase chain reaction for immune-related genes of tumor tissues from 44 patients with locoregionally advanced SCCHN prior to treatment with definitive chemoradiotherapy were conducted. T-cell receptor clonality and the mRNA expression levels of immune-related genes were correlated with various clinicopathological parameters. Results: In patients with locoregionally advanced SCCHN, tumor infiltrating T-cells clonally expand and GRZB mRNA levels were associated significantly with longer progression-free survival (PFS) (p = 0.003) independent of HPV status, tumor and nodal stage. The TCR-β DI was significantly lower in HPV-negative compared to HPV-positive tumors (p = 0.002), signifying more clonal T-cell expansion in HPV-negative tumors. A higher percentage of HPV-negative tumors expressed HLA-A protein compared to HPV-positive tumors (p = 0.049), suggesting that the greater T-cell clonal expansion might be due to more robust antigen presentation by HPV-negative tumors. Conclusions: This study suggests the pre-existence of clonally expanded T-cells in patients with locoregionally advanced SCCHN prior to treatment, and provides rationale to introduce immunotherapy in the curative-intent setting. The association of high GRZB mRNA levels with favorable PFS independent of HPV-status, tumor and nodal stage supports that the pre-existence of an intrinsically inflamed microenvironment enhances chemoradiotherapy effects. Finally, in HPV-positive tumors, the T-cell infiltrate seemed to be more diverse which could be secondary to virally-induced defective expression of HLA class I molecules.
Association of autologous AdHER2 dendritic cell vaccination with antitumor activity and number of circulating tumor cells.
Authors: Wood Lauren Virginia , Roberson Brenda D. , Agarwal Piyush K. , Apolo Andrea B. , Stroncek David , Weiner Louis M. , Morris John Charles , Terabe Masaki , Lee Min-Jung , Trepel Jane B. , Berzofsky Jay A. 
Published: September 2018

3089 Background: We achieved cure of large established tumors in syngeneic BALB/c mice using an adenoviral vector vaccine expressing rodent HER2 extracellular (EC) and transmembrane (TM) domains mediated by the induction of anti-HER2 antibodies. We report clinical translation utilizing an autologous adenoviral- transduced dendritic cell (DC) vaccine expressing human HER2 EC and TM domains (AdHER2ECTM) in adults with advanced metastatic tumors with 1-3+ HER2 expression. Methods: In this two-part phase I study (NCT01730118) subjects with HER2+ metastatic solid tumors naïve to HER2-targeted therapies (Part 1) or HER2+ treatment-experienced breast cancer (Part 2) received 5 doses of vaccine at Weeks 0, 4, 8, 16 and 24. Part 1 dose escalation involved 3 cohorts of 6 patients utilizing 5x106, 10x106 and 20x106 viable DCs per dose, respectively. With no DLTs or evidence of cardiac toxicity, dose escalation to 40x106 viable DCs per dose was allowed in the Part 1 Expansion Cohort and Part 2 treatment was begun at a starting dose of 20x106 viable DCs per vaccine. Adjuvant muscle-invasive bladder cancer patients were allowed to enroll with safety documented out to 12 weeks in > 10 treated patients. Re-staging was assessed using immune-related response criteria (irRC). Remaining study accrual: Part 1 (N = 9), Part 2 (N = 24). Results: A total of 27 cancer patients (7 colon, 6 breast, 5 ovarian, 3 bladder, 6 other) have received > 2 vaccine doses (median 4): 19F, 8M, median age 60 yrs, median 3 prior treatment regimens (range 1-11); HER2 IHC 1+ N = 6, 2+ N = 9, 3+ N = 12. Of metastatic patients receiving 10x106viable DCs per dose or higher, 7 of 19 (37%) had evidence of response by irRC: 1 CR (ongoing at 100 weeks), 1 PR (-71% lasting 44 weeks) and 5 SD (median duration 24 weeks, range 8 to 48 weeks). Two adjuvant bladder cancer patients remain without disease at 100 weeks. Adverse events were limited to local injection site reactions < G2. Of patients with paired baseline samples, 40% (8/20), 83% (5/6) and 100% (2/2) exhibited decreases in circulating tumor cells at 12, 28 and 48 weeks, respectively. Conclusions: AdHER2 DC vaccination is safe and is associated with evidence of clinical benefit and anti-tumor activity. Clinical trial information: NCT01730118.