bluuebunny/crossref_metadata_2025 huggingface dataset:
Monitoring PD-L1 expression on circulating tumor cells (CTCs) in patients with NSCLC treated with PD1 inhibitors.
Authors: Guibert Nicolas Marie , Delaunay Myriam , Lusque Amellie , Gouin Sandrine , Boubekeur Nadia , Rouquette Isabelle , Clermont Estelle , Fourtoul Aurélien , Favre Gilles , Pradines Anne , Mazieres Julien
Published: February 2018

147

Background: Inhibitors of the immune checkpoint PD-1/PD-L1 have become a standard of care in NSCLC. Patient selection, currently based on PD-L1 expression in tumor tissue, is limited by its temporal and spatial heterogeneity. We hypothesized that monitoring PD-L1 staining of circulating tumor cells (CTCs) could represent a valuable non-invasive biomarker. Methods: Up to 3 blood samples were prospectively collected from patients with advanced NSCLC: i) pretreatment (nivolumab), ii) first follow-up, iii) progression. CTCs were isolated from 10 mL of blood using cell size-based technology (ISET, Rarecells). PD-L1 expression was assessed by immunofluorescence on CTCs and immunohistochemistry on tissue. Results: 162 samples from 96 patients were collected. PD-L1 expression could be assessed pretreatment on tissue and CTCs in 72% and 93%, respectively; and was ≥1% in 37% and 83%; ≥5% in 35% and 79%, respectively. No correlation between tissue and CTCs PD-L1 expressions was observed (Spearman coefficient correlation = 0.04, p = 0.77). At baseline, each 10/7.5 ml increase in CTCs count was associated with increased risk of death and progression (HR[95%CI]: 1.06 [1.005;1.117] p = 0.03 for OS and HR[95%CI]: 1.05 [1.01;1.10] p = 0.02 for PFS). The presence of PD-L1(+)CTC (≥1%) had no prognostic impact (OS: p = 0.89 and PFS: p = 0.55), but pretreatment PD-L1(+)CTCs were more frequent in the “non-responders” group (PFS < 6 months) (p = 0.04). Median CTC count was 30 (n = 96), 68.3 (n = 44) and 50.3 (n = 22) pretreatment, at the first follow-up and at progression, respectively. The changes in CTC count at the first follow-up had no impact on PFS (p = 0.45) or OS (p = 0.68). 96% of patients had PD-L1(+)CTCs at progression. Further analyses are ongoing to assess the prognostic value of the persistence of CTCs and PD-L1(+)CTCs at the first follow up and at progression. Conclusions: Analysis of PD-L1 expression on CTCs is highly feasible. PD-L1 expressions on tissue and CTCs are discordant, CTCs being more likely positives, suggesting false negatives occurring in small biopsies. Further analyses of the kinetics of this biomarker throughout ICI treatment, along with other circulating biomarkers, are ongoing. Clinical trial information: NCT02827344.
Clinical impact of 22C3 PD-L1 expression in non-small cell lung cancer.
Authors: Okabe Naoyuki , Takagi Hironori , Suzuki Hiroyuki
Published: February 2018

149

Background: Immune checkpoint inhibitors has been widely applied in the field of non-small cell lung cancer (NSCLC). Immunostaining of PD-L1 using 22C3 was carried out as a companion diagnosis for Pembrolizumab. To date, only PD-L1 expression thought to be a predictive biomarker approved in USA and JAPAN. However, clinical impact or characteristic of PD-L1 expression in NSCLC is still controversial. In this report, we tried to analyze the relationship between PD-L1 expression and clinic-pathological parameter in NSCLC. Methods: A total 110 of NSCLC samples (Biopsy tissue and surgical resection specimen) were stained with PD-L1 IHC assays by using 22C3 antibody. The expression rate of PD-L1 was examined and relationship between the PD-L1 expression and clinicopathological factors was statistically analyzed. Results: The frequency of Tumor Proportion Score (TPS) less than 1% was 50%, TPS 1 to 49% was 32.7% and TPS 50% or more was 19%. Compared with the previous clinical trials such as KEYNOTE 001 and the KEYNOTE 010, our data of positivity tended to be low. In comparison between histological subtype, the expression in squamous cell carcinoma was higher than non-squamous histology. Furthermore, in relation to smoking history, higher smoking subgroup tended to be higher than the group with low smoking subset. In addition, the specimen preserved for 3 years or longer had a low positive rate, suggesting the possibility of false negatives due to long term formalin fixation of tissue specimens. Conclusions: We examined the relationship between the expression rate of PD-L1 and clinicopathological factors in 110 tissue samples subjected to PD-L1 staining by 22C3 at our hospital. Our results consistent with recent previous studies. In considering to analyze the archived tissue, we recommend that more fresh sample should be use for PD-L1 testing.
Survival stratification using a baseline inflammatory physiology based scoring system in advanced non-small cell lung cancer (NSCLC) treated with anti-programmed cell death-1 (anti-PD-1) therapy.
Authors: Naqash Abdul Rafeh , Stroud Chipman Robert Geoffrey , Muzaffar Mahvish , Yang Li V. , Walker Paul R.
Published: February 2018

152

Background: Immune checkpoint blockade (ICB) has shown promise in NSCLC with improved survival and durability in disease control. Despite these advances, the response to ICB remains variable. Thus identifying easily available biomarkers that can assist in the optimal selection of patients for ICB holds paramount importance. Methods: Retrospectively we identified 87 stage III/IV NSCLC patients initiated on anti-PD-1 therapy from April 2015 to March 2017 after progressing on a platinum doublet. These patients were part of an ongoing prospective biomarker-based study at our institution. Follow up cutoff for survival analysis was set at October 1, 2017. Enrolled participants had inflammatory markers (C-reactive protein, absolute neutrophil count, absolute lymphocyte count, serum albumin) measured on the day of first dose of anti-PD-1 administration as well as subsequent doses. Using multivariate Cox analysis, factors demonstrating an association with overall survival after immunotherapy (OSI) were used to develop a composite score to stratify patient survival. Results: The median age was 64 years with predominant histology being adenocarcinoma in 46.0 % followed by squamous cell carcinoma (43.7 %). Stage IV disease was present in 70.1%, with skeletal involvement (54.1 %) and liver (27.9 %) being the most common metastatic sites. In the multivariate Cox regression with backward elimination, factors independently associated with OSI were noted to be: CRP, neutrophil-lymphocyte ratio, and prognostic nutritional index. A composite inflammatory biomarker score was developed using the B-coefficients from the Cox multivariate regression. A score > 1 demonstrated inferior OSI compared to a score of ≤ 1 [1.7 vs. 9.3 months; P < 0.001, HR 4.00, 95% CI (2.21-7.25)]. Conclusions: This study provides preliminary evidence in favor of a composite inflammation based score that can aid in survival stratification of these patients. Validation of this score in prospective NSCLC trials to elucidate its potential utility as a predictive or prognostic tool in facilitating optimal patient selection for ICB is required.
The effect of chemotherapy on the tumor immune microenvironment in non-small cell lung cancer (NSCLC): A single-institution retrospective analysis.
Authors: Devitt Michael Edward , Gade Kristine E , Hall Richard Delmar , Horton Bethany J , Stelow Edward B , Bullock Timothy , Gentzler Ryan D.
Published: February 2018

155

Background: Platinum-doublet chemotherapy (chemo) is routinely used for treatment of early stage and metastatic NSCLC. Immunotherapy is a proven treatment in these patients as well. The presence of tumor infiltrating lymphocytes (TILs) has been shown to be prognostic of this disease; however, the effect of chemo on TILs and the tumor immune microenvironment in NSCLC has not been well studied. We tested the hypothesis that chemo results in a decrease in the presence of TILs in NSCLC. Methods: We retrospectively identified patients treated with neo-adjuvant platinum-doublet chemo who underwent surgical resection from 2007-2015 at the University of Virginia. The specimens were sectioned and IHC stained for CD8, FoxP3, and PD-L1. Two independent investigators assessed a central and a peripheral high power field of tumor for number of CD8 and FoxP3 positive TILs. An independent pathologist assessed tumor cell PD-L1 expression. The nonparametric Wilcoxon Mann Whitney test was used to compare measurements of TILs in chemo-exposed tissue to a control cohort from a database of NSCLC surgical specimens that had not received pre-operative therapy. Results: Of the 17 patients who met eligibility criteria, 11 had sufficient tissue for evaluation. Results are shown in Table 1. There was a significant decrease in CD8 and FoxP3 TILs in both the center and periphery in patients exposed to chemo. There was no difference in PD-L1 expression. Conclusions: These data suggest that platinum-doublet chemo significantly decreases the presence of cytotoxic and regulatory subsets of TILs without effecting PD-L1 expression. This study is limited by the lack of pre-treatment tumor samples to compare pre- and post-treatment TILs in individual patients. Further studies are warranted to evaluate the impact of decreased TILs in the setting of concurrent or sequential immunotherapy treatment with chemo in NSCLC. [Table: see text]
Simultaneous, single cell, flow cytometric quantification of PD-L1/TILs/cell cycle in non-small cell lung cancer tissue biopsies: Concordance of PD-L1 expression detection between flow cytometry and immunohistochemistry.
Authors: Young Stephen , Griego-Fullbright Christen , Wagner Aaron , Chargin Amanda , Patterson Bruce Kendrick
Published: February 2018

159

Background: Tumor cell expression of PD-L1 leads to the inhibition of immune responses specifically inactivation of cytotoxic T-cells. PD-L1 expression on tumor cells by immunohistochemistry does not provide the complete picture of therapeutic (PD-L1) and prognostic (TILs, aneuploidy) markers. Here, we report a clinical assay that quantifies tumor infiltrating lymphocytes (TILs), cell cycle/DNA content as well as PD-L1 expression on tumor and aneuploid tumor cell populations. Methods: Punch biopsies were taken from 19 fresh tissues specimens and were processed into single cell suspensions using the IVD/CE-IVD IncellPREP Single Cell Preparation Kit. Cells were fixed and permeabilized in 1 mL IncellMax. Cells were tested with the OncoTect iO Lung Assay which contains antibodies directed against PD-L1 (clone 28-8), CD45, CD3, and CD8, and a cell cycle dye. Concordance to IHC was tested by the Dako PD-L1 IHC 28-8 PharmDx Kit. Immune cell populations were quantified and aneuploidy was determined using a DNA index > 1.05. Results: The number of CD8+ CTL were significantly increased (P = 0.02) in tumor compared to normal lung tissue (37.4% to 28.2%). The number of CTL in aneuploid tumors was twice the number of CTL in diploid tumors. The percentage of antigen presenting cells (CD45+, high SSC) was decreased in the tumor cell samples relative to the normal lung tissue (P = 0.01). PD-L1 expression varies in cells depending on the cell cycle. Conclusions: In this study, the concordance between Oncotect iO and IHC was 95% (NPA-97%, PPA-89%). PD-L1 expression varies with DNA content, PD-L1 expression decreases with increasing DNA content. Though CTL are increased in aneuploid tumors, PD-L1 expression decreasing with DNA content may contribute to the association of aneuploidy with distant metastases. PD-L1 expression is a powerful determinant of treatment and aneuploidy is a powerful prognostic factor that combined yield important clinical information.
Unleashing natural killer cells upon immunization with a carbohydrate mimetic peptide vaccine.
Authors: Kieber-Emmons Thomas , Makhoul Issam , Pennisi Angela , Emanuel Peter D. , Hutchins Laura Fulper , Siegel Eric R , Monzavi-Karbassi Behjatolah
Published: February 2018

16

Background: Immunotherapies that activate Natural Killer (NK) cells can lend to their tumor infiltration or help shape the adaptive response toward a T helper type 1 (Th1) profile thought to favor anti-tumor responses. Methods: We have analyzed the functional NK response to immunization with a carbohydrate mimetic peptide (CMP) vaccine in preclinical tumor models and in humans. This CMP, referred to as P10s, is the N-terminal half of a peptide vaccine named P10s-PADRE, the C-terminal half of which (PADRE) is a Pan-T-cell epitope. Syngeneic mouse models associated with mammary 4T1, mouse lymphoma cell line EL4, and human breast cancer MDA-MB-231 in athymic mice were immunized with the vaccine. NK infiltration was identified by staining mouse tumor tissue before and after immunization. For NK cell depletion studies Athymic mice were injected intraperitoneally with anti-asialo GM1 rabbit serum prior to challenge with MDA-MB-231 tumor cells after the last immunization and then every 4-5 days for the period of a month. A Phase 1 trial was conducted with the vaccine and the NK phenotype assessed by flow cytometry. Serum Th1/Th2 cytokine profile was determined using multiplex MSD® kits (Meso Scale diagnostics). Results: Immunization of murine models with the P10s vaccine results in infiltration of NK cells into tumors and depletion of NK cells in these models abrogate anti-tumor responses. An early phase clinical trial in Stage IV metastatic Breast Cancer patients indicated that the activates the natural cytotoxicity receptor NKp46 phenotype, which has a mouse Ortholog, with a Th1 cytokine environment marked by significant elevation of IFN-γ levels. While P10s-PADRE immunization induced proapoptotic antibodies that are caspase-3 dependent, the induced antibodies also led to ADCC targeting of human breast cancer tumor cell lines in vitro further emphasizing a role for activated NK cells. Conclusions: Our results reveal that immunization with a CMP vaccine can mediate the activation of an anti-tumor NK response that can affect tumor growth in preclinical models that might translate to humans. Clinical trial information: NCT01390064.
Efficacy of nivolumab in patients treated by corticosteroid due to immune-related adverse events.
Authors: Higashiyama Ryoko Inaba , Horinouchi Hidehito , Sekine Katsutoshi , Matsumoto Yuji , Murakami Shuji , Goto Yasushi , Kanda Shintaro , Fujiwara Yutaka , Yamamoto Noboru , Ohe Yuichiro
Published: February 2018

163

Background: Nivolumab is a standard treatment for metastatic or refractory non-small cell lung cancer (NSCLC). Because immune-related adverse events (irAEs) are common and can be severe, a number of these patients require systemic corticosteroids. irAEs are reportedly associated with improved survival in melanoma patients treated with nivolumab. To date, little information is available regarding the effect of corticosteroid on the efficacy of nivolumab in terms of progression-free survival (PFS) and overall survival (OS). Methods: We reviewed consecutive patients who received nivolumab for metastatic or refractory NSCLC between December 2015 and August 2017. Nivolumab was administered at the standard dose of 3 mg/kg every two weeks. We recorded the patient demographics, efficacy and safety of nivolumab, previous and subsequent treatments, information about irAEs, corticosteroid usage, and survival. PFS and OS were calculated from the start of nivolumab treatment. A Cox proportional hazards regression model was applied using variables with the potential to influence the PFS: sex, age, performance status (PS), smoking history, driver gene alterations, histology, line of nivolumab treatment, and irAEs. Results: A total of 184 patients received nivolumab. Among them, 117 (64%) were male, the median age was 64 years (range, 34-83 years), 23 (13%) had a PS of 2 or more, 125 (68%) were ex-smokers, 37 (20%) had squamous NSCLC, and 37 (20%) had some kind of driver gene alteration, mainly EGFR mutation. Fifty patients (27%) experienced some grade of irAEs, and 37 (20%) patients required corticosteroid treatment. A multivariate analysis identified the occurrence of irAEs (hazard ratio [HR], 0.55; 95% confidence interval [CI] 0.33-0.91) and smoking (HR, 0.60;95% CI, 0.36-0.99) as independent predictors of a favorable PFS. There were no apparent differences in the proportions of patients who survived with or without the use of corticosteroid for irAEs: 73% vs. 71% at 6 months and 46% vs. 45% at 18 months. Conclusions: The occurrence of irAEs was positively associated with the PFS, and corticosteroid treatment did not have an adverse effect on OS in patients with NSCLC who were treated with nivolumab.
Prognostic impact of smoking status in non-small-cell lung cancer patients treated with immune checkpoint inhibitors.
Authors: Kim Hyo Jung , Kim Jung Han
Published: February 2018

164

Background: Immune checkpoint inhibitors (ICIs) have emerged as a new treatment option for patients with advanced non-small-cell lung cancer (NSCLC). Some studies with ICIs in NSCLC suggested that smoking history was associated with improved survival outcomes. We conducted this meta-analysis to investigate if the survival benefits of ICIs in patients with advanced NSCLC are different according to smoking status. Methods: Electronic databases were searched for eligible studies. We included randomized controlled trials with the data of survival outcomes and extracted progression-free survival (PFS) or overall survival (OS) stratified by smoking status. Results: From 6 studies, 2,389 ever-smokers and 413 never-smokers were included in the meta-analysis. In first-line treatment setting, ICIs tended to improve PFS in patients with smoking history (HR = 0.85 [95% CI, 0.71-1.10], P = 0.07). For never-smokers with advanced NSCLC, chemotherapy, not ICIs, was significantly associated with improvement of PFS (HR = 2.30 [95% CI, 1.23-4.28], P = 0.009). In more than second-line setting, ICIs significantly prolonged OS over that with chemotherapy in ever-smokers (HR = 0.70 [95% CI, 0.63-0.79], P < 0.00001). For never-smokers with NSCLC, however, ICIs failed to significantly improve OS (HR = 0.79 [95% CI, 0.59-1.06], P = 0.12). Conclusions: This meta-analysis indicates that ICIs can prolong survival over that with chemotherapy in ever-smokers with advanced NSCLC. However, ICIs failed to improve survival in never-smokers. These results suggest that smoking status may be a predictive marker for survival benefits to ICIs.
Avelumab in patients with previously treated mesothelioma: Updated phase 1b results from the JAVELIN Solid Tumor trial.
Authors: Hassan Raffit , Thomas Anish , Nemunaitis John J. , Patel Manish R. , Bennouna Jaafar , Chen Franklin , Delord Jean-Pierre , Dowlati Afshin , Taylor Matthew H. , Powderly John D. , Vaishampayan Ulka N. , Verschraegen Claire F. , Grote Hans Juergen , von Heydebreck Anja , Chin Kevin M. , Gulley James L.
Published: February 2018

166

Background: Avelumab, a human anti-PD-L1 IgG1 monoclonal antibody, is approved for treatment of metastatic Merkel cell carcinoma (US and EU) and advanced urothelial carcinoma progressed on platinum therapy (US). Here, we report updated phase 1b data for avelumab in patients (pts) with previously treated mesothelioma. Methods: Pts with unresectable pleural or peritoneal mesothelioma whose disease had progressed after platinum and pemetrexed therapy received avelumab 10 mg/kg IV Q2W until progression, unacceptable toxicity, or withdrawal. Tumors were assessed every 6 wks (RECIST 1.1). Endpoints included objective response rate (ORR), progression-free survival (PFS), overall survival (OS), and adverse events (AEs; NCI CTCAE v4.0). Results: As of Dec 31, 2016, 53 pts were treated and followed for a median of 24.8 mos (range 16.8–27.8). Median age was 67 y (range 32–84). Pts had received a median of 2 prior lines of therapy (range 1–8). Confirmed ORR was 9.4% (95% CI 3.1–20.7; complete response in 1.9%, partial response in 7.5%). In pts with 1 (n = 18), 2 (n = 15) or ≥3 (n = 20) prior lines of therapy, ORR was 5.6%, 13.3% and 10.0% respectively. Median duration of response was 15.2 mos (95% CI 11.1–not estimable). 26 pts (49.1%) had stable disease as best response and the disease control rate was 58.5%. Median PFS was 4.1 mos (95% CI 1.4–6.2) and the 6-mo PFS rate was 38.0% (95% CI 24.2–51.7). Median OS was 10.9 mos (95% CI 7.5–21.0) and the 12-mo OS rate was 45.9% (95% CI 31.9–58.8). In evaluable pts with PD-L1+ (n = 16) and PD-L1− (n = 27) tumors (≥5% tumor cell cutoff), ORR was 18.8% (95% CI 4.0–45.6) and 7.4% (95% CI 0.9–24.3), and the 6-mo PFS rate was 37.5% (95% CI 14.1–61.2) and 42.0% (95% CI 23.1–59.8). 43 pts (81.1%) had a treatment-related (TR)AE, most commonly ( > 10%) infusion-related reaction (35.8%; all grade 1/2), chills (15.1%), fatigue (15.1%) and pyrexia (11.3%). 5 pts (9.4%) had a grade ≥3 TRAE. 14 pts (26.4%) had an immune-related AE, which was grade ≥3 in 3 pts (5.7%; pneumonitis, colitis, and type 1 diabetes mellitus). No treatment-related deaths occurred. Conclusions: Avelumab showed clinical activity and acceptable safety in pts with previously treated mesothelioma. Clinical trial information: NCT01772004.
Comprehensive analyses of the clinical benefits of immune checkpoint inhibitors in advanced non-small cell lung cancer.
Authors: Ou Qiyun , Fu Tuping , Yu Yunfang , Lin Dagui , Gao Quanlong , Zhang Yuxin , Luo Baoming
Published: February 2018

168

Background: Treatment with immune checkpoint inhibitors is becoming the standard of care for patients and it was approved for the treatment of non-small-cell lung cancer (NSCLC) growing at a rapid pace. However, selecting patients who are appropriate for therapy and which therapeutic strategies to use can be challenging. Methods: We searched for randomized clinical trials (RCTs) investigating immune checkpoint inhibitors versus observation in patients with advanced NSCLC until September 2017. Overall survival (OS) and progression-free survival (PFS) were pooled by meta-analysis. The GRADE system was used to describe the quality of evidence. Results: The analysis included 11 trials with 5,538 unique patients. High- to moderate-quality evidence indicated that immune checkpoint inhibitors extended NSCLC survival and PFS, expressed as hazard ratios (HRs) (OS: 0.79, P = 0.000; PFS: 0.78, P = 0.000). High- to moderate-quality evidence revealed prolonged OS and PFS were similar across preplanned subgroups in patients with squamous (0.77, P = 0.000 and 0.74, P = 0.001) or nonsquamous disease (0.76, P = 0.003 and 0.73, P = 0.041), EGFR wild-type positive status (0.67, P = 0.000 and 0.59, P = 0.010), current or former smokers (OS: 0.83, P = 0.009), and male (0.79, P = 0.000 and 0.67, P = 0.023). Increasing improvement in OS was associated with increasing PD-L1 expression (TC3 or IC3 HR 0.54, P = 0.000; TC2/3 or IC2/3 HR 0.62, P = 0.007; TC1/2/3 or IC1/2/3 HR 0.64, P = 0.000; TC0 and IC0 HR 0.72, P = 0.017; high- to moderate-quality evidence). In exploratory subgroup analysis suggest that there was advantageous of immune checkpoint inhibitors in previous definitive chemotherapy compared with chemoradiotherapy, with concurrent administration of chemotherapy (OS as HR: P = 0.001, P = 0.006, respectively). Conclusions: This large and comprehensive analysis produced firm evidence that immune checkpoint inhibitors extended advanced NSCLC survival and PFS, while in some patients with EGFR wild-type positive status, current or former smokers, male and higher PD-L1 expression had substantial benefit. The benefits from therapy appear to be influenced by preceding definitive therapy and concurrent chemotherapy.
RORγ agonist LYC-55716, a novel small molecule immunotherapy: Rationale for clinical evaluation in non-small cell lung cancer based on translational and bioinformatic evaluation.
Authors: Carter Laura , Liu Xikui , Li Hongxiu , Zawidzka Elizabeth , Gao Yilin , Fox Brian , Weems Garry , Wilkins H. Jeffrey , Hu Xiao
Published: February 2018

171

Background: Retinoic acid receptor–related orphan receptor γ (RORγ) agonists modulate immune cell gene expression to enhance effector function and decrease regulatory T cell formation and expression of checkpoint pathways. RORγ agonists have demonstrated antitumor activity in syngeneic tumor models. Translational and bioinformatic assessments were conducted to support inclusion of patients with non–small-cell lung cancer (NSCLC) in a Phase 2a expansion of clinical trial LYC-55716-1001 (NCT02929862). Methods: The Cancer Genome Atlas (TCGA) NSCLC dataset was evaluated for (a) expression of RORγ and RORγ-inducing cytokines and correlation with survival; (b) genes related to RORγ biology, biomarkers of endogenous RORγ agonists; and (c) tumor microenvironment (TME) immune profiles. RORγ expression and the in vitro effects of a RORγ agonist on peripheral blood mononuclear cells (PBMCs) from lung adenocarcinoma (LA) and squamous cell carcinoma (SCC) patients were assessed. Results: In TCGA, 25% of NSCLC tumors expressed moderate/high levels of RORγ, suggesting infiltration of Type 17 cells. There was a statistically significant correlation between high RORγ expression and improved survival in LA patients. Genes that support Type 17 cell formation (IL1B, IL23A, IL6) were expressed in ~50% of tumors. RORγ expression was confirmed in PBMCs isolated from LA and SCC patients. Analysis of TCGA data and patient samples identified low expression of sterol efflux and uptake genes, suggesting low levels of endogenous RORγ agonist in TME. In TCGA, high mutational burden and high expression of immune-related genes were found in NSCLC tumors. RORγ agonist treatment of PBMCs from LA and SCC patients increased IL-17A and IL-26 (LA and SCC) and decreased PD1 (LA). In Phase 1 clinical testing, LYC-55716 has been well tolerated, demonstrating long-term disease stabilization in heavily pretreated patients. Conclusions: Bioinformatic analyses of RORγ expression/biology, correlation with improved survival, plus in vitro findings support inclusion of NSCLC in an ongoing Phase 2 clinical trial of LYC-55716.
Efficacy of immune-checkpoint inhibitors (ICI) in non-small cell lung cancer (NSCLC) patients harboring activating molecular alterations.
Authors: Mhanna Laurent , Milia Julie , Lusque Amellie , Couraud Sebastien , Mascaux Celine , Veillon Remi , Frueh Martin , Moro-Sibilot Denis , Lattuca-Truc Mickaël , Tomasini Pascale , Barlesi Fabrice , Drilon Alexander E. , Gautschi Oliver , Mazieres Julien
Published: February 2018

172

Background: Two revolutions recently occur in the treatment of advanced NSCLC: the development of targeted therapies and of ICI). Both strategies have been shown to outperform chemotherapy. Patients with molecular alterations are usually considered as poor candidate for immunotherapy. Here, we aimed to analyze the efficacy of immunotherapy in NSCLC patients with oncogenic addiction. Methods: We conducted a retrospective multicentric study, on patients treated with ICI and carrying an activating molecular abnormality: EGFR, ALK, KRAS, ROS1, HER2, BRAF, MET and RET. We have collected anonymized data, which were evaluated for clinical characteristics and outcome (progression free survival duration of ICI treatment and overall survival since initiation of ICI). Results: 209 patients were registered from 7 centers in France and Switzerland. 198 patients had adenocarcinoma (95.2%), 6 large cell carcinoma (2.9%), 161 were former or current smokers (81.3%), 99 were female (47.4%), median age at diagnosis was 59 yrs. (range 30-79). 33 patients had EGFR mutations (15.8%), 132 KRAS mutation (63.2%), 6 ALK rearrangement (2.9 %), 2 ROS1 rearrangement (1 %), 5 HER2 mutation (2.4 %), 12 BRAF mutations (5.7 %), 7 MET alteration (3.3 %), 3 RET rearrangements (1.4 %), and 9 had concomitant multiple molecular alterations (4.3%). After validated treatment 200 patients were treated with PD1 inhibitors (nivolumab, 97.5% and pembrolizumab, 2.5%), 4 with anti CTLA4 (tremelimumab), 5 with anti PDL1 (4 atezolizumab). The median PFS was 2.8 m. [95%CI 2.3;3.5] for the whole population 2.1 for EGFR [1.7;2.8], 3.5 for KRAS [2.5;4.9], 2.7 for BRAF [1.5;NR] and not estimable for other alterations. Median exposure to ICI was 1.89 m. [0;18.5]. The median overall survival for the whole population was 13.0 m. [9.4; 15.6], for EGFR 13.3 m. [4.7; NR], KRAS 11.3 m. [8.2; 16], BRAF 10.7 m. [1.5; NR] and not yet estimable for the other alterations. Conclusions: PFS (2.8 m.) and OS (13 m.) are very similar from the ones observed in pretreated unselected NSCLC patients. ICI is associated with better PFS in patients with KRAS mutation than in EGFR mutated patients. Analysis of outcome in other molecular subgroups is ongoing.
Prognostic value of the expression of immunomodulatory targets in surgically resected lung adenocarcinoma.
Authors: Takamochi Kazuya , Hosoya Masaki , Mogushi Kaoru , Hayashi Takuo , Kawaji Hideya , Hayashizaki Yoshihide , Suzuki Kenji
Published: February 2018

173

Background: Therapies targeting immune checkpoints have recently shown encouraging activity in patients with non-small cell lung cancer. T-cell activation is controlled by the balance of co-stimulatory molecules that act positively in concert with T-cell receptor (TCR) signaling induced by antigen stimulation and co-inhibitory molecules (immune checkpoint molecules) that negatively regulate TCR signaling. Methods: We performed a Cap Analysis of Gene Expression (CAGE) analysis to quantify the gene expression of PD-L1, PD-L2, co-inhibitory molecules (CTLA-4, PD-1, TIM-3, BTLA, VISTA and LAG-3) and co-stimulatory molecules (CD28, OX40, GITR, CD317, CD27 and HVEM) using RNA extracted from 71 frozen tumor tissue samples of surgically resected lung adenocarcinomas, including 35 EGFR-mutated, 11 KRAS-mutated and 25 wild-type tumors. The correlations between the expression of the above immunomodulatory targets and the overall survival were analyzed. None of patients received immunotherapy. Results: Lung adenocarcinomas in this study were divided into two groups: tumors showing high expression of almost all immunomodulatory targets and those with low expression. The expression of CTLA-4, PD-1 and PD-L1 was not prognostic. The prognosis of patients with tumors showing high expression of TIM-3 was significantly worse than that of those with low expression, regardless of the EGFR and KRAS mutation status. The prognosis of patients with tumors showing high expression of VISTA was significantly better than that of those with low expression, regardless of the EGFR and KRAS mutation status. The prognosis of patients with tumors showing high expression of LAG-3 was significantly worse than that of those with low expression among overall patients. However, high expression of LAG-3 was an unfavorable prognostic factor in patients with EGFR-mutated or wild-type tumors, and it was a favorable prognostic factor in patients with KRAS-mutated tumors. Conclusions: Lung adenocarcinomas may be divisible into immunogenic and nonimmunogenic tumors. The prognostic value differs according to the immunomodulatory target and driver oncogene status.
Identification of mechanisms of immune regulation and exhaustion generated by tumor-derived exosomes in patients with advanced non-small cell lung cancer using the trans-vivo DTH assay.
Authors: Lema Diego , Jankowska-Gan Ewa , Burlingham William , Leal Ticiana A.B.
Published: February 2018

175

Background: The success of PD-1/PD-L1 inhibition is clearly dependent on optimal patient selection. Developing validated biomarkers that identify patients with an increased probability of response to these antibodies remains a challenge. DTH responses are a standard method to evaluate cellular immune responses in animal and human immunotherapy trials. The trans-vivo delayed-type hypersensitivity (tvDTH) methodology, using the SCID mouse footpad swelling response to human PBMC (peripheral blood mononuclear cells) + antigen injection, and using antibodies to block the regulatory pathways, was developed in our laboratory and has been used to characterize regulation systems in tolerant transplant recipients and more recently in prostate cancer. Methods: In this pilot project we aim to investigate the mechanisms of regulatory immune responses in patients with advanced NSCLC receiving anti-cancer therapies using the tvDTH, and to correlate tvDTH results with responses to checkpoint inhibitor blockade therapy. So far we have enrolled 4 NSCLC patients. PBMCs are obtained by blood draw and lung cancer antigen (LCA) is prepared from malignant pleural effusion cellular pellet or fresh frozen tumor. 1 of 4 patients’ sample was negative for malignancy. Results: Our preliminary data suggests that all NSCLC patients with pleural effusions positive for malignancy had a PD-1-dependent exhausted immune response towards their LCA. Remarkably, 1 of these patients not only showed PD-1-dependent exhaustion, but also PD-1-dependent bystander suppression, whereby a positive control immune response was reduced by co-injection of LCA, suggesting the existence of immunosuppressive mechanisms. Unlike our previous results with prostate cancer, no patient showed CTLA-4-dependent exhaustion or suppression. Importantly, an antigen preparation from a NSCLC that showed no malignant cells by cytology did not induce a swelling response. Conclusions: We plan to expand our study patient population and follow up our patients in order to correlate tvDTH results with clinical outcomes.
Evaluation of the clinical benefits of immune checkpoint inhibitors in patients with advanced melanoma.
Authors: Ou Qiyun , Yu Yunfang , Lin Dagui , Fu Tuping , Gao Quanlong , Luo Baoming
Published: February 2018

194

Background: Immune checkpoint inhibitors selection in advanced melanoma is complicated further, with choices among anti-CTLA4 or anti-PD-1 therapeutic antibodies options. We aimed to evaluate the clinical benefits of immune checkpoint inhibitors for discussing evidence-based treatment strategies by a meta-analysis of randomized clinical trials (RCTs) data. Methods: We searched for RCTs investigating immune checkpoint inhibitors in patients with advanced melanoma until September 2017. Hazard ratios (HRs) was estimated for overall survival (OS) and progression-free survival (PFS). The quality of the evidence was evaluated with the GRADE framework. Results: Overall, 18 RCTs including a total of 8,917 patients were identified. Immune checkpoint inhibitors versus placebo or observation prolonged PFS (HR = 0.59, 95% confidence interval (CI) 0.44 to 0.78, P < 0.0001) and OS (HR = 0.77, 95% CI 0.72 to 0.84, P < 0.0001). The combination immunotherapy had significantly higher benefit than monotherapy for PFS (HR = 0.75, 95% CI 0.61 to 0.92, P = 0.005) and OS (HR = 0.70, 95% CI 0.61 to 0.79, P < 0.0001). Treatment with anti-PD-1 monoclonal antibody was associated with improved PFS (HR = 0.61, 95% CI 0.59 to 0.69, P < 0.0001) and OS (HR = 0.66, 95% CI 0.58 to 0.77, P < 0.0001) compared with anti-CTLA-4 monoclonal antibody. According to BRAF status analysis, there was a PFS benefit of immune checkpoint inhibitors versus placebo or observation (mutant, HR = 0.58, 95% CI 0.35 to 0.96, P = 0.035; wild–type, HR = 0.52, 95% CI 0.43 to 0.69, P = 0.001), anti-PD-1 outperformed anti-CTLA-4 checkpoint inhibitor (mutant, PFS HR = 0.64, 95% CI 0.51 to 0.79, P < 0.0001; wild–type, PFS HR = 0.58, 95% CI 0.47 to 0.71, P < 0.0001); and combination compared with single-agent immunotherapy (mutant, HR = 0.38, 95% CI 0.15 to 0.98, P = 0.046; wild–type, HR = 0.40, 95% CI 0.23 to 0.69, P = 0.001). Conclusions: This analysis provides an evidence that immune checkpoint inhibitors enhanced OS and PFS in patients with advanced melanoma, as well as combination immunotherapy and anti-PD-1 monoclonal antibody appear to be clinically beneficial option preference.
Time to first-line (1L) therapy discontinuation in metastatic melanoma (MM).
Authors: Luke Jason J. , Ghate Sameer , Ionescu-Ittu Raluca , Ndife Briana , Burne Rebecca , Laliberté François , Duh Mei Sheng
Published: February 2018

196

Background: Nivolumab or pembrolizumab monotherapy (N/P) and dabrafenib+trametinib combination therapy (D+T) are currently the most commonly used regimens for the treatment of MM in anti-PD1 and BRAF inhibitor classes, respectively. We describe time to discontinuation of 1L therapy with N/P or D+T in patients (pts) with MM in a real-world setting. Methods: Adults with MM initiated on N/P or D+T in 1L were identified in Truven MarketScan databases (Q1/2014 - Q2/2016; n = 443). Outcomes included (a) distribution of time to 1L discontinuation among pts who discontinued 1L therapy during the study follow-up (reported by year of 1L start) and (b) rates of pts still on 1L therapy at different time points after 1L therapy start ("on treatment" rates) estimated from Kaplan-Meier analyses. Results: Of 443 pts, 243 (55%) were initiated on N/P and 200 (45%) on D+T. Pts initiated on D+T appeared to be younger, with more brain metastases, and higher use of emergency care in the 6 mo prior to 1L therapy start as compared to those initiated on N/P (Table). Time to 1L discontinuation and "on treatment" rates were similar between the treatment groups, with a tendency towards longer time to 1L discontinuation for D+T (Table). Conclusions: This real-world data study showed numerically longer time to 1L discontinuation for pts treated with D+T as compared to N/P, despite higher comorbidity burden. [Table: see text]
Real world treatment patterns of first-line combination therapies among BRAF+ metastatic melanoma patients.
Authors: Ghate Sameer , Tang Jackson , Li Zhiyi , Nakasato Antonio Reis
Published: February 2018

197

Background: For patients (pts) with metastatic melanoma (MM) and BRAF V600 mutation (BRAF+), options for first-line (1L) systemic combination therapy include immunotherapy (IO) or targeted therapy (TT). This study describes real world treatment patterns among BRAF+ MM pts treated with 1L ipilimumab+nivolumab (I+N) or dabrafenib+trametinib (D+T). Methods: This retrospective observational analysis used Flatiron Health’s electronic health record-derived database from Oct 2015 - Jul 2016. Oct 2015 was chosen as the start date as both combo use of I+N and D+T had been approved by the FDA. Pts were aged ≥18 years with a MM diagnosis, tested BRAF+ prior to therapy, and treated with either I+N or D+T as 1L therapy. Baseline demographic and clinical characteristics, and treatment patterns were collected from structured data and unstructured data in physician notes, and were descriptively assessed. Kaplan-Meier (KM) analysis measured time to discontinuation. Statistical inferences were not planned in this study. Results: Among 76 BRAF+ pts, 62% (47) were treated with D+T as 1L, and 38% (29) were treated with I+N as 1L. Compared to D+T pts, lower proportion of I+N pts had a history of brain metastases (31% vs. 34%) and elevated ( > 333 IU/L) lactate dehydrogenase (10% vs. 19%), while a higher proportion of I+N pts reported ECOG performance status score of zero (38% vs. 23%). The two cohorts were similar in age, gender and baseline comorbidities. Discontinuation among D+T and I+N was 43% (20) and 48% (14) respectively. The primary reason for discontinuation was progression among D+T pts (10/20) vs. toxic effects of therapy among I+N pts (8/14). KM median time to discontinuation was 213 days for D+T pts vs. 196 days for I+N pts. 55% of I+N pts did not complete full induction of 4 doses of ipilimumab, citing toxic effects of therapy. Conclusions: 1L D+T patients had higher tumor burden, elevated LDH levels, and higher ECOG performance status score, but lower discontinuation rate and longer time to treatment discontinuation relative to BRAF+ 1L I+N pts.
Immune checkpoint inhibitor responses in KIT-mutated metastatic melanoma.
Authors: McKean Meredith Ann , Oba Junna , Ma Junsheng , Haydu Lauren Elaine , Bassett Roland L. , Woodman Scott Eric
Published: February 2018

199

Background: KIT-mutated MM is a rare melanoma entity. Response rates of KIT-mutated MM to anti CTLA-4 and anti PD-1 have not previously been reported. Methods: A single-institution retrospective review identified patients (pts) with KIT-mutated MM treated with at least two doses of anti CTLA-4 or anti PD-1 between 2008-2017 with response determined by immune-related response criteria (irRC). Overall survival (OS) was from ICI start to death or last follow-up date, and progression-free-survival (PFS) was from ICI start to date of progression or death if pts deceased without disease progression. Results: Thirty-five pts treated with ipilimumab were identified. Median follow-up was 63.7 months. Median age at MM diagnosis was 65.4 years (range 26-81) and 54.3% were male. Subtypes were 51.4% mucosal, 22.9% acral-lentiginous, 22.9% cutaneous, and 2.9% unknown primary. KIT mutations were 57.1% exon 11, 25.7% exon 17, 11.4% exon 13, and 5.7% exon 2. Anatomical M stage at treatment initiation was 11.4% M1a, 34.3% M1b, and 54.3% M1c. Median OS was 11.8 months (8.4-35.6) and PFS was 3.0 months (2.8-5.8). Responses to ipilimumab were 8.6% complete response (CR), 11.4% partial response (PR), 28.6% stable disease (SD), and 51.4% progressive disease (PD) for a 48.6% clinical response rate (CRR). Twenty pts treated with anti PD-1 were identified. Median follow-up was 7.9 months. Median age at MM diagnosis was 66.7 years (range 42.6-86.8) and 70% were male. Subtypes were 40% mucosal, 30% cutaneous, 20% acral-lentiginous, and 10% unknown primary. KIT mutations were 45% exon 11, 35% exon 17, 10% exon 2, 5% exon 13 and and 5% exon 10. Anatomical M stage at treatment initiation was 10% M1a, 15% M1b, and 75% M1c. Median OS was 22.5 months (7.4-NA ) and PFS was 3.2 months (2.7-NA). Responses to anti PD-1 were 10.0% CR, 25.0% PR, 20.0% SD, and 45.0% PD for a 55.0% CRR. For both anti CTLA-4 and anti PD-1 treated cohorts, univariate analysis demonstrated no statistical significance between tumor response to ICI and clinical pt characteristics or KIT mutation exon. Conclusions: KIT-mutated MM demonstrated CRR of 48.6-55.0% to ICI. There was no correlation between pt characteristics or KIT exon mutation and response to therapy; however, analysis was limited by sample size.
Safety and clinical activity of pembrolizumab immunotherapy and multi-organ site ablative stereotactic body radiotherapy (iMOSART) in patients with advanced solid tumors.
Authors: Lemons Jeffrey , Luke Jason J. , Karrison Theodore , Pitroda Sean , Melotek James , Zha Yuanyuan , Al-Hallaq Hania A , Arina Ainhoa , Janisch Linda A. , Chang Paul J , Patel Jyoti D. , Fleming Gini F. , Moroney John William , Sharma Manish , White Julia R. , Ratain Mark J. , Gajewski Thomas , Weichselbaum Ralph R. , Chmura Steven J.
Published: February 2018

20

Background: Clinical responses to PD1 blockade have been linked to IFN-γ associated gene expression in the tumor microenvironment. Stereotactic body radiotherapy (SBRT) can stimulate innate and adaptive immunity to potentially augment immunotherapy. Anti-PD1 treatment outcomes are improved with lower disease burden and multi-site radiation is an emerging paradigm for eradicating metastatic disease. Methods: We conducted a phase I study for patients with metastatic solid tumors who progressed on standard treatment. The primary aim was to evaluate the safety of pembrolizumab Immunotherapy with Multi-Organ Site Ablative stereotactic body Radiation Therapy (iMOSART). Two to four metastases were chosen for each patient and not all sites of disease were targeted with SBRT. Metastases > 65cc were partially irradiated. Pembrolizumab was initiated within 7 days after the final SBRT treatment. Results: 79 patients were enrolled, three patients did not receive any treatment and three patients only received SBRT. The 73 patients included in the analysis were treated with SBRT and at least 1 cycle of pembrolizumab. 94.5% of patients had two lesions treated with SBRT. Median follow-up for toxicity was 5.5 months (IQR 3.3-8.1). Six patients experienced dose-limiting toxicity (DLT) with no radiation dose reductions. RECIST overall objective response rate was 13.5% in the 68 patients with imaging follow-up. Superior control of irradiated lesions was observed compared to non-irradiated target lesions for the 52 patients with paired data (p = 0·0005) with a mean percent tumor burden change -21·7% (SD 24·3%) for irradiated lesions vs. 1·7% (SD 46·3%) for non-irradiated lesions (p = 0·0008). While abscopal response defined by 30% reduction in any single non-irradiated measurable lesion was present in 26·9% of patients, abscopal response was 13·2% when defined by 30% reduction in aggregate diameter of non-irradiated measurable lesions. Conclusions: iMOSART was well tolerated with acceptable toxicity. Further randomized studies are warranted to investigate the potential clinical benefit of this combination approach. Clinical trial information: NCT02608385.
Association of the neutrophil-to-lymphocyte ratio prior to checkpoint blockade immunotherapy (CBI) or radiation plus CBI with overall survival in melanoma patients.
Authors: Rhodes Terence Duane , Fulde Gail , Romero Robin , Abraham Tyler , Moulton Bryce , Van Meter Margaret , Thota Ramya , Lewis Mark Andrew , Haslem Derrick S. , Nadauld Lincoln , Barker Tyler
Published: February 2018

200

Background: Studies in certain cancer types have indicated that radiation therapy prior to the use of CBI provides a survival benefit. This benefit has not been clearly defined for patients with metastatic melanoma. Additionally, the neutrophil-to-lymphocyte ratio (NLR) may be a potential biomarker. Methods: This retrospective study was performed in patients diagnosed with melanoma between January 2007 and August 2016 who received CBI with or without previous radiation treatment at Intermountain Healthcare (Utah, USA). Cases were identified from electronic medical records and data was manually extracted through August of 2017. The neutrophil-to-lymphocyte ratio (NLR) was calculated from the absolute neutrophil and lymphocyte counts of a complete blood cell count with differentials performed as a routine standard of care procedure in melanoma patients prior to therapy initiation. Overall survival was defined as the length of time (d) from start of CBI to death as of August, 2017. Results: Forty-six melanoma patients were initially identified. Of these, thirteen patients were excluded due to lack of follow-up data (n = 9), radiation performed after CBI (n = 3), or concurrent radiation and CBI (n = 1). The final analysis consisted of 33 subjects separated below (NLR < 3.12, n = 16) and above (NLR ≥ 3.12, n = 17) the NLR median. Age, height, body mass, and body mass index were not significantly different between groups (p-range: 0.11-0.60). Results from the Kaplan-Meier curve indicate that a NLR above the median associates with lower overall survival (Mantel, p = 0.04) in melanoma patients receiving CBI with or without previous radiation treatment. In a separate analysis of this cohort, overall survival was not significantly influenced by radiation therapy prior to CBI. Conclusions: Although prior radiation therapy offered no survival advantage for patients receiving CBI, NLR less than 3.12 was associated with an increase in overall survival. Further studies are need to explore NLR as a biomarker.