bluuebunny/crossref_metadata_2025_split huggingface dataset:
Correlation between tumor mutation burden and response to immunotherapy.
Authors: Bonta Ioana , Isac John Florin , Meiri Eyal , Bonta Dacian , Rich Patricia 
Published: September 2018

e14579 Background: The use of immunotherapy is exponentially increasing in treatment of patients with advanced solid tumors. However, the response rates vary significantly between different tumor types and even within same tumor type (e.g. in lung cancer approx. 1 in 4 patients respond to immunotherapy). In order to better identify patients that will respond to immunotherapy, several markers have been proposed. Tumor mutation burden (TMB) has emerged more recently as a quantitative marker that can help predict responses to immunotherapies across different cancers, including melanoma, lung cancer and bladder cancer. TMB is a measure of the overall number of somatic protein coding mutations occurring in the tumor specimen. Methods: We analyzed 54 consecutive patients treated with immunotherapy at our institution for which we had genomic sequencing (FoundationOne). There were 39 lung cases and 15 non-lung (GI, GU, sarcoma). For 30 cases we had TMB data. Favorable response was defined as stable disease or response to therapy at 3 months. The relationship between TMB and tumor response was explored using ROC analysis. Results: The probability of a favorable response to immunotherapy in our patient dataset was 57% (31/54 patients). Among the patients with known TMB 60% (18/30) had a favorable response (stable disease or response to therapy). The favorable response rate for tumors originating in the lung was 64% (25/39) and for non-lung primary tumors was 40% (6/15). The difference was not statistically significant, with p = 0.12. Higher TMB values were correlated with increased probability of a favorable response. ROC analysis demonstrated an Az of 74% for TMB values in differentiating between patients with and without a favorable response. A TMB cutoff value of 8 mutations/megabase yielded a sensitivity of 95% and a specificity of 58% for predicting favorable response. Conclusions: In our data base 57%of patients with different solid cancers had favorable response to immunotherapy, in second line and beyond .Higher TMB correlated with higher likelihood of response to immunotherapy, independent of the primary site of cancer.
Effect of high-dose corticosteroids on CD4+ or CD8+ lymphocyte proliferation or IL-2 production after stimulation with pembrolizumab.
Authors: Walker John WT , Xu Lai , Smylie Michael , Elahi Shokrollah 
Published: September 2018

e14587 Background: Immune checkpoint inhibitor (ICI) therapy has improved survival of patients with melanoma and other types of cancers. The effect of immunosuppresive corticosteroids on ICI efficacy is unknown, however treatment of immune-related adverse events with high-dose steroid therapy impairs neither efficacy nor time to failure of ICI. In certain clinical circumstances (for instance in the treatment of patients with symptomatic brain metastases) it may be advantageous to begin ICI therapy despite a requirement for steroid therapy. This study examined the effect of high-dose steroid therapy on lymphocyte proliferation/functionality after stimulation with anti-CD3 in the presence of pembrolizumab (PD-1 inhibitor) ex vivo. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from patients with either chronic hepatitis C infection or active malignant melanoma. CD4+ and CD8+ T cells were isolated and subjected to flow cytometry stating for the expression of PD-1. Isolated PBMCs were then pre-treated with prednisone (1 nM - 1 mM) before anti-CD3 stimulation. Anti-CD3-stimulated cells were then incubated with pembrolizumab (5 µg/mL) and their proliferative capacity and IL-2 secretion ability were measured by carboxyfluorescein succinimidyl ester (CFSE) staining and ELISA, respectively. Results: A dose-dependent inhibition of both lymphocyte proliferation and IL-2 production was observed, but at physiologically relevant nanomolar prednisone concentration the steroid effect was negligible. Millimolar prednisone concentrations resulted in > 50% relative reduction in lymphocyte proliferation, however even at this steroid concentration treatment with pembrolizumab was able to further stimulate proliferation and IL-2 secretion versus anti-CD3stimulation alone. Conclusions: Despite heavily immunosuppressive treatment, CD4+ and CD8+ lymphocytes demonstrate proliferative capacity and immunological activity following treatment with pembrolizumab. This suggests that treatment with pembrolizumab may be efficacious despite a requirement for steroid therapy, a scenario often encountered in patients with brain metastases.
The relative clinical value of combined antiPD-1 and antiCTLA-4 treatment, the NNT values of immuno-therapy in metastatic melanoma patients.
Authors: Porneczy Edit , Boncz Imre 
Published: September 2018

e14595 Background: The PD-1 inhibitor Nivo and antiCTLA-4 Ipi in combination was registered by FDA and EMA for the treatment of metastatic melanoma.The accelerated approval of this indication was based on PFS data.OS data in previously untreated melanoma pts was reported only in the Phase II CheckMate 069 study.In the Nivo+Ipi group the 1YOS% was 73,4% and 63,8% at 2Y, in the Ipi group 1YOS% was 64,8%, 2YOS% was 53,6% at median follow up of 24,5 months.The median OS had not been reached, the outcome measures could not assess the long-term benefit of antitumor immun response. Methods: In order to measure the relative clinical values, we analyzed the 1Y, 2Y survival data and calculated the ARR (absolut risk reduction) and NNT values (the number of patients who need to be treated to prevent one death) of the Phase II CheckMate 069 trial Nivo+Ipi vs Ipi group, and the overall survival data in subgroup of pts who discontinued the immun-therapy due to treatment related AEs. We used GraphPad Software QuickCalcs/NNT for calculating the results of 95% CI. Results: In the CheckMate 069, the Nivo+Ipi group vs Ipi group ARR was 10,93, the NNT was 10(95%CI 3,6-17,8) at 1Y.The ARR was 13,19, the NNT value was 8(95%CI 3,3-24,1) at 2Y. 1 in every 8-10 pts will benefit from the Nivo+Ipi combined immuno-therapy in order to prevent one death. In the subgroup of patients (37%) who discontinued treatment, the ARR was 21,16 at 1Y and 21,76 at 2Y. Despite of discontinuation, 1 in every 5 pts will benefit from immuno-therapy at 1Y and 2Y, the NNT was 5 (95%CI 2,5-40,4) and 2Y (95%CI 2,3-127,2). Conclusions: These results of NNT 5-10 represent the exceptional survival benefit of combined immuno-therapy. The limitations of NNT bias the pts demographic parameters and the value depends on comparator drug. The risk reduction was not constant over the follow up time and the long term benefit of immuno-therapy is not linear.The NNT data provide measurement of relative clinical value and clinical impact.
RNAi-mediated beta-catenin inhibition to promote T-cell infiltration and antitumor activity in combination with immune checkpoint blockade.
Authors: Ganesh Shanthi , Shui Serena , Lai Cheng , Dudek Hank , Wang Weimin , Brown Bob D , Abrams Marc 
Published: September 2018

e14608 Background: Recent research suggests an important role for Wnt/β-catenin signaling in mediating cancer immune evasion and resistance to immune checkpoint therapy. The mechnism is believed to involve blocking of specific cytokines which trigger immune cell recruitment to the tumor, resulting in the phenomenon of T-cell exclusion and rendering the tumor to a non-inflamed state. Inhibition of β-catenin may be an effective strategy for increasing the low response rate to these effective medicines in numerous cancer populations. DCR-BCAT is an advanced preclinical development candidate that has a potent and specific chemically-optimized RNA interference (RNAi) trigger targeting CTNNB1, the gene that encodes β-catenin, formulated in a tumor-selective lipid nanoparticle. Methods: Syngeneic murine models and transgenic MMTV-Wnt1 mouse models were used in this study. In both cases, a sequential dose regimen was employed where, in each dosing cycle, animals received DCR-BCAT, followed by a combination of anti-PD-1 and anti-CTLA-4 on subsequent days. Pharmacodynamic endpoints included CTNNB1 (β-catenin), CCL4, PD-1, PD-L1 mRNA measurement by quantitative PCR, as well as β-catenin, perforin, and granzyme B immunohistochemistry. Results: In syngeneic models, β-catenin inhibition with DCR-BCAT significantly improved the T-cell infiltration. The combination of DCR-BCAT and immune checkpoint blockade yielded significant tumor growth inhibition compared to monotherapy in B16F10, 4T1, Neuro2A and Renca tumors. The combination therapy was associated with high levels of granzyme B and perforin, strongly suggesting that the mechanism of sensitization to checkpoint therapy was a sharp increase in T-cell mediated cytotoxicity. Finally, when DCR-BCAT was combined with anti-PD-1/CTLA-4 antibodies in mice which develop spontaneous Wnt-driven mammary tumors, checkpoint therapy potentiation yielded complete tumor regressions. Conclusions: These data offer proof-of-concept for conversion of non-inflamed tumors to inflamed tumors by β-catenin inhibition, and support clinical evaluation of this combination approach using a first-in-class RNAi-based agent.
Upregulation of amphiregulin on CD14++CD16- (classical) monocytes in non-small cell lung cancer.
Authors: McCarthy Fiona Margaret , Candido Juliana , Rudd Robin , Steele Jeremy P. C. , Balkwill Frances 
Published: September 2018

e14612 Background: Amphiregulin is an EGFR ligand which contributes to many of the hallmarks of cancer including angiogenesis and metastasis. Recent studies demonstrate that Amphiregulin promotes an immunosuppressive tumour microenvironment by modulating the expression of T regulatory cells. Although its expression has been described on a variety of immune cells and tumour cells, there is no published data on Amphiregulin expression by monocytes to date. Previous work by this group has demonstrated that monocytes from non-small cell lung cancer patients are phenotypically and genomically different compared to healthy volunteers. This study aims to further investigate the differing genomic profile of monocytes in non-small cell lung cancer. Methods: Gene expression profiling was performed on CD14++CD16- (classical) monocytes of newly diagnosed patients with Stage IV non-small cell lung cancer (n = 6) and age-matched healthy volunteers (n = 6) using Affymetrix Human U133 Plus 2.0 array. Validation of differentially expressed genes of interest (including Amphiregulin) at mRNA level was performed using real-time PCR with TaqMan gene expression assays on independent patient (n = 6) and healthy volunteer cohorts (n = 6). Differential protein expression of Amphiregulin from the monocytes of healthy volunteers (n = 3) and non-small cell lung cancer patients (n = 3) was validated using ELISA technology. Results: Amphiregulin is one of the most upregulated genes on classical monocytes in non-small cell lung cancer patients compared to healthy volunteers (p = 0.001). The increased expression of Amphiregulin was confirmed on qPCR validation in an independent cohort of non-small cell lung cancer patients (p < 0.05) as well as in the original group (p < 0.001). Amphiregulin was also found to be increased at protein level on CD14++CD16- monocytes of non-small cell lung cancer compared to healthy volunteers. (p = 0.37). Conclusions: Amphiregulin is overexpressed by classical monocytes in non-small cell lung cancer and this over-expression is confirmed at mRNA and protein level. The production of Amphiregulin has not been ascribed to human monocytes in non-small cell lung cancer making its overexpression in this study a unique finding.
Association of an APOBEC mutational signature, mutational load, and BRCAness with inflammation and PD-L1 expression in HNSCC.
Authors: Rieke Damian Tobias , Messerschmidt Clemens , Ochsenreither Sebastian , Klinghammer Konrad Friedrich , Tinhofer Inge , Jöhrens Korinna , Klauschen Frederick , Beule Dieter , Keilholz Ulrich 
Published: September 2018

e14613 Background: Immune checkpoints are an emerging treatment target in head and neck squamous cell carcinoma (HNSCC). Expression of PD-L1 and an IFNG immune signature have been shown as biomarkers of response. In this study we analyzed mutational signatures to identify recurrent mutational patterns associated with inflammation. Methods: HNSCC mutation and gene expression data were downloaded (n = 496, TCGA). Inflamed tumors were identified through upregulation of a predefined 6-gene IFNG immune signature. Mutational load, base exchange motifs and a BRCAness signature were analyzed in inflamed and non-inflamed tumors. Results were validated in independent datasets of cervical, urothelial and lung cancer (TCGA). Results: Total mutational load did not correlate with tumor inflammation (r = -0.03, p = 0.5). An APOBEC-associated tCw (thymine – cytosine – adenine/thymine) mutational signature was significantly associated with inflammation (r = 0.26, p = 2.923e-09). The tCw signature was furthermore correlated with the expression of PD-L1 and other immune checkpoints (r = 0.19, p = 1.723e-05). The TCW mutational signature correlated with the expression of APOBEC3 genes. Expression of APOBEC3 genes also correlated with tumor inflammation (r = 0.62, p < 2.2e-16). As a second mutational signature, indels and an independent BRCAness signature were significantly enriched in non-inflamed tumors (p = 6.464e-14). Enrichment of an APOBEC mutational signature in inflamed tumors could be validated in cervical and urothelial carcinoma but not NSCLC. Conclusions: In HNSCC, overall mutational load is not associated with inflammation. BRCAness is significantly associated with immune-desertion whereas expression of APOBEC genes and an APOBEC mutational signature significantly correlates with the expression of PD-L1 and a T-cell inflamed signature. These findings suggest that a specific mutational pattern might serve as a biomarker for immune checkpoint inhibition in cancer.
Intra-tumour heterogeneity in the regulation of immune-tolerogenic pathways in primary and metastatic hepatocellular carcinoma (HCC).
Authors: Pinato David James , Victor Stephane , Spina Paolo , Trivedi Pritesh , Pirisi Mario , Burlone Michela , Black James M , Minisini Rosalba , Leutner Monica , Boldorini Renzo , Mauri Francesco A , Sharma Rohini 
Published: September 2018

e14614 Background: Anti-PD-1/PD-L1 checkpoint inhibitors have shown preliminary evidence of activity in HCC. PD-L1 status is a putative predictor of response, however PD ligands expression can be heterogeneous across sampled sites, with implications in the development of immunotherapy. Methods: We constructed a tissue microarray from 77 patients: 56 with surgically resected and 21 with intermediate/advanced HCC in whom 20 matching metastases were also available. Tumour and surrounding cirrhosis were analysed for PD-L1 and PD-L2 expression by immunohistochemistry. FFPE-extracted RNA was analyzed on the NanoString nCounter system using 12 paired primary/metastatic HCC cores from 6 patients, with validation in a subset of further 24 specimens. Results: Surgically resected cases were Child A (51/56, 91%), BCLC A (45/56, 80%). In resected tumour cores 11/56 (20%) were PD-L1 and 18/56 (32%) were PD-L2 positive. Surrounding cirrhotic liver revealed PD-L1 in the lymphocytic infiltrate in 8/56 samples (14%), whilst PD-L2 expression was found in the cirrhotic background in 29/56 cases (52%). PD-L2 expression in cirrhotic (p = 0.03) but not in tumour cores (p = 0.13) correlated with more advanced BCLC stage. In patients with intermediate-advanced HCC (n = 21), the prevalence of tumour PD-Ls expression was 40% (8/21) for both markers. Concordance between PD-L1 positive primary and metastases was 33% and 85% in PD-L1 negative tumours. In contrast, there was universal concordance between in PD-L2 expression between primary and metastatic HCC (p < 0.001). Targeted transcriptomic profiling revealed a subset of 40 genes involved in innate and adaptive immunity to be differentially regulated across primary and metastatic disease. Conclusions: Intra-tumour heterogeneity in the expression of PD-L1 is common in HCC, whilst PD-L2 is homogeneously distributed in primary and metastatic deposits. Unsupervised transcriptomic profiling confirms differential activation of innate and adaptive immune-related pathways in the metastatic dissemination of liver cancer. This has profound implications in the clinical development of immune response biomarkers in HCC.
Association of inflammatory cytokines with obesity and prevention with ASA or DHA/EPA supplementation.
Authors: Gruslova Aleksandra , Quach Duan , Lengfleder Lucy , Tiziani Stefano , Lodi Alessia , Kist Kenneth , Kaklamani Virginia G. , deGraffenried Linda A , Brenner Andrew Jacob 
Published: September 2018

e14619 Background: Obesity is associated with poor breast cancer outcomes. Our prior data suggested a positive feedback loop whereby circulating factors associated with obesity induce the COX-2 enzyme, resulting in increased pro-inflammatory cytokine prostaglandin E2 (PGE2), which can then increase aromatase levels. In order to establish a better understanding of baseline PGE2 levels, as well as select the proper COX2 inhibitor for prospective study, we performed a randomized study of aspirin (ASA), fish oil (DHA/EPA), and the combination in postmenopausal women of varying body habitus and assessed changes in cytokines. Methods: Women were recruited and randomly assigned into 3 interventions: ASA 81mg, docosahexaenoic acid (DHA, 1500mg) with eicosapentaenoic acid (EPA, 2500mg), or combination ASA and DHA/EPA (DUAL). Blood was collected before and after 28 days of supplementation, and 27 cytokines were assessed by Luminex array (Millipore); PGE2 was measured by ELISA. Investigators were blinded until analysis was complete. Statistical analyses were performed in R. Results: 110 patients were included: 90% participants white; of which 50% Hispanic; average BMI 31.5±0.67; and age: 60.1±0.58. Cytokines were not correlated with demographic variables. At baseline inflammatory cytokines IL-6 and Eotaxin were correlated with BMI ((p > 0.029 and p > 0.017, respectively). After 28 days of DHA/EPA BMI was correlated with Eotaxin, Il-2, IL-7 and GM-CSF (p < 0.05), in ASA group correlation was significant only with IL-2 (cor.coef= 0.58). Two sample t-test showed significant difference between DUAL and ASA or DHA/EPA for IL-6 (p = 0.008 and p = 0.034, respectively), and for IL-2 only between DUAL and ASA (p = 0.039). Furthermore, impact on PGE2 was observed with ASA with 81% obtaining a decrease from baseline (median change -28%); by comparison 55.1% (-1%) and 65.6% (-22%) of subjects showed decrease in the DUAL and DHA/EPA groups respectively. Level of macrophage inflammatory proteins (MIP-1b) were lower in the DUAL group compare to ASA (p < 0.014). Conclusions: Chronic inflammatory cytokines are correlated with obesity, and may be modified by use of ASA and DHE/EPA. This strategy may be useful in the secondary prevention setting. Clinical trial information: NCT02062255.
TLR9 agonist lefitolimod to improve antitumor effect of checkpoint inhibitors in vivo.
Authors: Schmidt Manuel , Kapp Kerstin , Volz Barbara , Oswald Detlef , Wittig Burghardt 
Published: September 2018

e14625 Background: TLR9 agonists are potent activators of the immune system via induction of cellular and humoral responses. Preclinical and ongoing clinical studies support the use of TLR9 agonists for immunotherapeutic approaches. Lefitolimod/MGN1703 is a covalently-closed dumbbell-like immune surveillance reactivator (ISR) with a broad immunomodulatory potential. Due to promising data from a phase 2 trial (IMPACT) as maintenance therapy after first-line chemotherapy in mCRC patients lefitolimod is recently evaluated in a phase 3 trial in mCRC patients (IMPALA). Furthermore, lefitolimod is currently investigated in a phase 2 trial in SCLC patients (IMPULSE) and in a phase 1 / 2 trial in HIV patients (TEACH). Methods: It was shown that lefitolimod reduces tumor growth in several murine tumor models. Since the mode-of-action of lefitolimod starts upstream of the initiation points of checkpoint inhibitors like anti-CTLA-4 or anti-PD-1/anti-PD-L1 a combinatory approach may result in an enhanced anti-tumor effect. Therefore, two syngeneic murine models were used – a colon carcinoma CT26 and a lymphoma A20 model – for evaluation of the anti-tumor effect in vivo. Results: In the CT26 model treatment with anti-PD-L1 (i.p.) had no effect on the tumor growth, whereas addition of lefitolimod (s.c.) to anti-PD-L1 led to an anti-tumor effect (tumor growth inhibition, TGI 48%) which consequently resulted in prolonged survival of the mice. This combinatory effect was even more pronounced in the A20 model where treatment with anti-PD-1 (i.p.) alone had a moderate anti-tumor effect which was vastly increased by the combination (TGI – anti-PD-1: 46%, anti-PD-1/lefitolimod 99%). Conclusions: In conclusion, we showed that lefitolimod, a member of dSLIM® family of TLR9 agonists and an ISR, can enhance the limited anti-tumor effects of checkpoint inhibitors in pilot studies in murine colon carcinoma and lymphoma tumor models in vivo. These data show the promising potential for the combination with checkpoint inhibitors. Notably, a clinical trial in cooperation with MD Anderson evaluating the benefit of lefitolimod in combination with ipilimumab is currently ongoing.