Gotowa bibliografia na temat „Eighteen Serum Cytokine Signature”

Abstract Abstract 4245 Introduction: Osteonecrosis of the jaw (ONJ) is a recognized risk of bisphosphonate infusion; the incidence, etiology, risk factors and pathophysiology remains largely undefined. In this prospective study we evaluated patients on long-term zoledronic acid infusions; the aims of the study were to provide a comprehensive clinical, oral/dental assessment of patients prospectively and sequentially measure changes in zoledronic acid PKs, cytokines, microbial signatures and bone turnover markers in saliva and serum in correlation with 2 major clinical end points development of ONJ and myeloma relapse. Methods and patients: Over 6 months 110 patients were enrolled and followed for 18 months. Patients were on zoledronic acid infusions: monthly (n=75) for patients with significant bone disease (relapsed or newly diagnosed) or every 3 months (n=35) for those in stable remission > 3years. During the study follow up therapy was changed to pamidronate (n=4) due to renal insufficiency or denosumab (n=1). Patients had clinical and oral evaluations and saliva and blood samples collected at baseline and every 3 months up to 18 months; saliva samples were obtained before and after (5 and 15 minutes) zoledronic acid infusions. Results: Median time from MM diagnosis was 3.7 years (range: 1.5–13) for 100 patients; 10 newly diagnosed patients were enrolled as control with a median of 7 months (range: 1–9). Median age was 60 yrs (range: 32–82); 60 were Caucasians, 49 African American, one Asian; 68 were males. At study entry 79% of the patients were in remission: CR (n=35, 32%) of them 15 were in CR for > 7 years, PR (n=52, 47%) and PD (n=23, 21%). Twenty-four patients were not receiving any MM therapy; most were on maintenance with lenalidomide (n=52) or thalidomide (n=11) and the rest on other regimens (n=23). Fifty patients progressed during the 18 month study period; they continued the study with various salvage therapies that included: bortezomib (n=14), carfilzomib (n=14), lenalidomide (n=6) and clinical trails (n=16). Thirteen patients had died: 9 from complications of relapsed MM and 4 from other causes. Eighteen patients withdrew consent for sample collection and dental evaluations but were followed clinically. Most patients had prior-to-study entry dental procedures (extractions, n=80). The prevalent oral pathology detected on dental evaluations included: gingivitis (n=41), moderate/severe periodontal disease (n=35) and gross dental caries (n=15). During the 18 months of observation; 13 patients developed ONJ. Median time from MM diagnosis to ONJ was 5.7 years (range: 1.9–12 years). Risk factors for ONJ were relapsed disease (n=11), monthly zoledronic acid (n=8), dental extractions (n=8), severe periodontal disease (n=9), older age [median age was 62 yrs] and prior ONJ (n=3). ONJ treatment was conservative with antibiotics and holding BP therapy: 9 patients had complete healing of the site of ONJ and 4 had persistent non-healing ONJ lesions. Sequential saliva and serum samples were assessed for various cytokines involved in immune/inflammatory process (IL-1β, IL-6, IL-17, TNF-α, IFN-α, TGF-β, MIP-1 α β, MMP-9); bone turnover (Osteopontin, ostiocalcin, RANKL, Osteoprotregerin) and angiogenesis factors (VEGF and EGF) in 42 patients with ONJ (n=13), severe periodontal disease (n=10) and normal control (n=19), the later included 10 patients in remission and 9 with relapsed MM. In conclusion: In this longitudinal observational study, the ONJ incidence was 11%, 70% eventually healed. Our cytokine analysis suggest that saliva may accurately detect changes reflective of mucosal inflammation as well as bone turnover markers; data correlating these changes with ONJ and MM status will be presented. Disclosures: No relevant conflicts of interest to declare.

2544 Background: CL of checkpoint inhibitors has been identified as a predictive covariate of overall survival (OS) in several tumors. Determination of CL requires post-treatment samples, which negates its utility as a baseline prognostic biomarker. This study aims to identify a baseline composite cytokine signature that correlates with NIVO CL in patients with RCC using translational PK/PD analysis. Methods: Peripheral serum PK (NIVO CL) and serum biomarkers to assess PD (Myriad Rules-Based Medicine customized inflammatory cytokine panel) were analyzed from 985 patients with RCC enrolled in 3 clinical trials. CheckMate (CM) 009 (NCT01358721) and CM 025 (NCT01668784) of NIVO (n = 481) were used for model development (training dataset). CM 010 (NCT01354431) of NIVO and a cohort treated with everolimus in CM 025 were included in model application (test dataset; n = 504). PK/PD analyses were conducted using a machine learning algorithm with performance assessed by receiver operating characteristic (ROC) curve and accuracy by confusion matrix. Results: The model selected the top-10 baseline inflammatory cytokines to form a composite cytokine signature, which predicted NIVO CL (high vs low) that was significantly associated with OS (p < 0.001) across all 3 studies (training and test datasets). The same cytokine features were associated with OS of everolimus (p < 0.01), suggesting the potential prognostic nature of the composite signature. The PK/PD analysis provided a robust description of the association between selected cytokines and CL (ROC = 0.71). Identified cytokines (eg, serum C-reactive protein known to reflect immune cell modulation) have been shown individually to be associated with RCC prognosis. A multivariable approach resulting in tumor-specific composite signatures may provide more accurate prognostic value. Conclusions: The baseline composite cytokine signature could serve as a clinically useful biomarker for patients with RCC, pending further evaluation, as it may provide improved prognostic accuracy for long-term clinical outcome compared to individual cytokines.

BackgroundType I interferon (IFN) plays a pivotal role in the pathogenesis of systemic lupus erythematosus (SLE) and dermatomyositis (DM)/polymyositis (PM) as typically evidenced by upregulation of type I IFN-stimulated genes (ISGs) in peripheral immune cells [1]. In addition, a distinct monocyte cytokine signature, which was induced by plasma from pediatric SLE patients and abrogated by Janus kinase (JAK) inhibition, was reported [2]. However, contribution of serum factors to expression of ISGs was not fully elucidated.ObjectivesTo investigate the cytokine induction profile in multiple myeloid lineages by serum from patients with SLE or DM/PM using the whole blood stimulation system and identify the signaling pathway relavant to the monocyte cytokine signature.MethodsSerum was collected from newly diagnosed, untreated, and clinically active adult patients with SLE (SLE group), anti-MDA5 antibody-positive DM (MDA5 group), and anti-aminoacyl-tRNA synthetase (ARS) antibody-positive DM/PM (ARS group), and healthy controls (HCs) (n= 10 for each group). Heparinized whole blood from healthy donors was incubated with control serum (10 v/v%), patient serum (10 v/v%), or IFN-β in the presence of protein transport inhibitor cocktail for 6 hours. Red blood cells were lysed, and then leucocytes were fixed at a single step. Intracellular staining was performed and expression of 11 cytokines in CD14+monocytes, CD1c+dendritic cells (DCs), and CD123+DCs were analyzed using flow cytometry. A cut-off level was determined by 2% positivity of each cytokine in unstimulated condition. For transcriptomic analyses, sorted CD14+monocytes from healthy donors were incubated with serum (SLE, MDA5, ARS, or HCs), or IFN-β (n= 3 for each group) for 4 hours, and bulk RNA-sequencing was performed using the Illumina NextSeq 500 platform. RNA-seq raw sequence reads analysis and pathway analysis were performed using the Strand NGS software. To evaluate significance of JAK-signal transducer and activator of transcription (STAT) pathway, whole blood from healthy donors was pre-incubated with upadacitinib, a JAK1 inhibitor, stimulated with patient serum for 6 hours, and analyzed for cytokine expression as described above.ResultsSerum from SLE and MDA5 groups induced significantly higher monocyte chemoattractant protein-1 (MCP1) and interleukin-1 receptor antagonist (IL-1RA) expression in CD14+monocytes than serum from HCs (Figure 1). These monocyte cytokine signatures were closely resembled that induced by IFN-β stimulation. Serum from ARS group did not induce any significant cytokine expression in CD14+monocytes. No significant cytokine expression was observed in CD1c+DCs or CD123+DCs. RNA-seq demonstrated that 612 and 462 genes were upregulated (fold change >2 relative to control) following stimulation with serum from SLE and MDA5 groups, respectively. In these upregulated genes, 383 genes were commonly upregulated across SLE and MDA5 groups and IFN-β. Pathway analysis revealed similar transcriptional profiles in SLE and MDA5 groups: upregulated genes were most frequently involved in IFN-αβ signaling pathway including multiple ISGs and STAT1/2. Upadacitinib significantly abrogated the monocyte cytokine signature, and MCP1 and IL-1RA expression induced by serum from SLE and MDA5 groups in a dose-dependent manner (p< 0.001 in all analyses).ConclusionThese results suggest that serum factors in patients with active SLE and anti-MDA5 antibody-positive DM can induce shared monocyte cytokine signature through type I IFN pathway. CD14+monocytes ‘primed’ by serum might contribute to the pathogenesis of these diseases.References[1]Rheumatology (Oxford).2017;56:1662[2]J Autoimmun. 2017;81:74Figure 1.MCP1 (A) and IL-1RA (B) expression in CD14+monocytes induced by serum from SLE and anti-MDA5 antibody-positive DM.Horizontal bars represent median values.P-values were calculated using Kruskal-Wallis test followed by Dunn’s multiple comparisons test.Acknowledgements:NIL.Disclosure of InterestsShohei Nakamura: None declared, Yuko Okamoto: None declared, Yasuhiro Katsumata Speakers bureau: GlaxoSmithKline K.K., AstraZeneca K.K., Sanofi K.K., Pfizer Japan Inc., Janssen Pharmaceutical K.K., Chugai Pharmaceutical Co., Ltd., Asahi Kasei Pharma, Astellas Pharma Inc., Mitsubishi Tanabe Pharma Corporation, Masayoshi Harigai: None declared.

Abstract Background: Chronic lymphocytic leukemia (CLL) is a common leukemia that is incurable. Survival curves for Rai stage 0/I and III/IV are distinct and a specific gene expression signature may be associated with stage progression (SP). However, distinct oncogenic pathways that differentiate stage progression are not well defined. Methods: Mononuclear cells from Rai stage 0/I (N=8) and III/IV (N=8) CLL patients were obtained through an IRB approved protocol and analyzed utilizing the HG-U133A 2.0 Affymetrix array after isolating and purifying total RNA (Qiagen, RNAeasy). Control RNA samples are from 4 normal volunteers’ peripheral blood (PB) B-cells (AllCell, CA) and a normal reactive lymph node. Statistical significance of individual genes and pathways was analyzed using ‘BioConductor’. Differential gene expression was analyzed using ‘Limma’. Pathways and cytogenetics regions were analyzed for enrichment using ‘Category’. Real time RT-PCR on specific genes was performed to validate the GEP based on stage. A serum cytokine profile (120 cytokines, RayBiotech) on 8 CLL patients (N=4 low risk; N=4 high risk) and 4 normal volunteers was evaluated to identify a signature for diagnosis and prognosis. Results: Data are presented as “robust” increases or decreases of relative gene expression common to 8 patients in the low or high risk stages. The Wnt/Ror-1 non-canonical, and TGFb pathways are over-expressed in Rai stage 0/I, implicating a program of cell survival and dysregulated migration. The Hedgehog, Proteasome, TNF/FAS/NF-kB, and MAPK pathways are over-expressed in Rai stage III/IV, suggestive of a tumor microenvironment driven program in SP. Genes located on chromosome 3q27.1 are also dysregulated in Rai stage III/IV. A distinct serum cytokine profile also differentiates low from high stage CLL. Conclusions: GEP of CLL with RT-PCR validation has identified distinct oncogenic pathways that distinguish Rai stage 0/I from III/IV. A serum cytokine profile also differentiates between low and high risk CLL. These data together provide a ‘molecular signature’ for SP in CLL.

Metastatic colorectal cancer is frequently associated with poor clinical conditions that may limit therapeutic options. Regorafenib is a small molecule approved for the treatment of metastatic colorectal cancer, but it is hampered by significative toxicities. Moreover, only a relatively limited number of patients benefit from the treatment. Therefore, the identification of reliable markers for response is an unmet need. Eighteen cytokines, selected based on their prevalent Th1 or Th2 effects, were collected. Peripheral blood samples were gathered at baseline in 25 metastatic colorectal cancer patients treated with regorafenib. Data extracted have been linked to progression-free survival. ROC identified the best cytokines associated with outcome. The relative value of the selected cytokines was determined by PCA. Data analysis identified 8 cytokines (TGF-β, TNF-α, CCL-2, IL-6, IL-8, IL-10, IL-13 and IL-21), used to create a signature (TGF-β, TNF-α high; CCL-2, IL-6, IL-8, IL-10, IL-13 and IL-21 low) corresponding to patients with a significantly longer progression-free survival. This report suggests that the analysis of multiple cytokines might identify a cytokine signature related to a patient’s outcome that is able to recognize patients who will benefit from treatment. If confirmed, future studies, also based on different drugs, using this approach and including larger patient populations, might identify a signature allowing the a priori identification of patients to be treated.

Although some signs of inflammation have been reported previously in patients with myalgic encephalomyelitis or chronic fatigue syndrome (ME/CFS), the data are limited and contradictory. High-throughput methods now allow us to interrogate the human immune system for multiple markers of inflammation at a scale that was not previously possible. To determine whether a signature of serum cytokines could be associated with ME/CFS and correlated with disease severity and fatigue duration, cytokines of 192 ME/CFS patients and 392 healthy controls were measured using a 51-multiplex array on a Luminex system. Each cytokine’s preprocessed data were regressed on ME/CFS severity plus covariates for age, sex, race, and an assay property of newly discovered importance: nonspecific binding. On average, TGF-β was elevated (P= 0.0052) and resistin was lower (P= 0.0052) in patients compared with controls. Seventeen cytokines had a statistically significant upward linear trend that correlated with ME/CFS severity: CCL11 (Eotaxin-1), CXCL1 (GROα), CXCL10 (IP-10), IFN-γ, IL-4, IL-5, IL-7, IL-12p70, IL-13, IL-17F, leptin, G-CSF, GM-CSF, LIF, NGF, SCF, and TGF-α. Of the 17 cytokines that correlated with severity, 13 are proinflammatory, likely contributing to many of the symptoms experienced by patients and establishing a strong immune system component of the disease. Only CXCL9 (MIG) inversely correlated with fatigue duration.

6081 Background: Human papillomavirus has been implicated in the development of head and neck squamous cell carcinoma and is associated with a more favorable clinical outcome. Previously, we identified a serum hypoxia signature associated with HNSCC progression (Byers et al, Proc ASCO. 2008). We investigated the association between HPV status, serum biomarkers, and clinical outcome in patients treated with induction chemotherapy. Methods: 47 previously untreated patients with locally advanced nodal disease (T0–4, N2b/c/3, M0) received 6 weekly cycles of paclitaxel (135 mg/m2), carboplatin (AUC 2), and cetuximab (400 mg/m2 week 1; 250 mg/m2 weeks 2–6) followed by definitive local therapy. 46 (98%) patients had a complete or partial response to induction chemotherapy (Kies et al, Proc ASCO. 2006), and 6 have had tumor progression (PD) after a minimum follow-up of two years. Formalin fixed biopsies were available for HPV testing by in situ hybridization with non-radioisotopic chromogen for 25 patients (including 5/6 with PD). 38 CAFs were measured by multiplex bead assay before and during treatment. Results: 12/25 patients were HPV-positive, all male and six were never smokers; of the 13 HPV-negative patients, four were male and three were never smokers. Among those with available data, all 5 patients with PD were HPV-negative (p = 0.02). There were four study deaths, all in the HPV-negative group. Overall survival was superior in HPV-positive patients (p = 0.04). There was no significant association between HPV status and serum CAF markers. However, among HPV-negative patients, PD was associated with the CAF hypoxia signature (5/8 patients with the hypoxia signature progressed versus 0/5 signature-negative). Of the individual CAFs, osteopontin was significantly elevated in all HPV-negative patients with PD. Conclusions: HPV positivity was associated with a longer progression-free survival and overall survival. Among HPV-negative patients, only those with a serum hypoxia signature had disease progression. These data suggest that the combination of HPV status and CAF profiling may identify patients at risk for relapse after sequential therapy. No significant financial relationships to disclose.

Autoinflammatory diseases (AIDs) are disorders characterised by recurrent inflammatory episodes in charge of different organs with no apparent involvement of autoantibodies or antigen-specific T lymphocytes. Few common clinical features have been identified among all monogenic AIDs (mAIDs), while the search for a common molecular pattern is still ongoing. The aim of this study was to increase knowledge on the inflammatory pathways in the development of mAIDs in order to identify possible predictive or diagnostic biomarkers for each disease and to develop future preventive and therapeutic strategies. Using protein array-based systems, we evaluated two signalling pathways known to be involved in inflammation and a wide range of inflammatory mediators (pro-inflammatory cytokines and chemokines) in a cohort of 23 patients affected by different mAIDs, as FMF, TRAPS, MKD, Blau syndrome (BS), and NLRP12D. Overall, we observed upregulation of multiple signalling pathway intermediates at protein levels in mAIDs patients’ PBMCs, compared with healthy controls, with significant differences also between patients. FMF, TRAPS, and BS presented also peculiar activations of inflammatory pathways that can distinguish them. MAPK pathway activation, however, seems to be a common feature. The serum level of cytokines and chemokines produced clear differences between patients with distinct diseases, which can help distinguish each autoinflammatory disease. The FMF cytokine production profile appears broader than that of TRAPS, which, in turn, has higher cytokine levels than BS. Our findings suggest an ongoing subclinical inflammation related to the abnormal and constitutive signalling pathways and define an elevated inflammatory cytokine signature. Moreover, the upregulation of Th17-related cytokines emphasises the important role for Th17 and/or Th17-like cells also in monogenic AIDs.

Gliomas are the neoplasia of glial cells present in the brain and comprises of more than 70% of all the neoplasms of the central nervous system. They consist of a family of primary brain tumors that are categorized based on the cell type of origin. Astrocytoma, a tumor of astrocytic glial cells has the highest frequency of occurrence as compared to other glioma types and often referred to as glioma. Based on the malignancy of the tumor,World Health Organization (WHO) has classified astrocytoma into Grade I/Pilocytic Astrocytoma (PA), Grade II/ Diffuse Astrocytoma (DA), Grade III/Anaplastic Astrocytoma (AA) and Grade IV/Glioblastoma (GBM). GBM is the most frequent and malignant primary brain tumor in adults. The standard treatment for GBM includes surgical resection of the tumor followed by radiation and temozolomide therapy. In spite of such mutlimodal treatment protocol followed, the median survival of the GBM remains as low as 15 months. While multiple markers with potential utility in diagnosis, prognosis and therapy of GBM have been reported based on profiling of gene expression, protein expression, miRNA and methylation pattern using tumor tissue, serum-based markers are scanty. Serum biomarkers have great potential in clinical decision making and management of glioma. Serum is an attractive source for biomarker mining since it can be obtained easily by less invasive method. Further, they have wide range of application which includes diagnosis, disease classification, prognosis, predicting risk and outcome of the disease. More importantly, serum can be obtained easily during follow-up of disease which could make it useful to detect tumor recurrence. This is particularly beneficial because glioma diagnosis and monitoring tumor recurrence are carried out by Magnetic Resonance Imaging (MRI) and Computed Tomography (CT) scan, which is time consuming and less cost effective. The current study was designed to identify serum biomarkers of glioma by using proteomic approaches and to understand the functional role of selected biomarkers in glioma pathogenesis. This study has been divided into three parts. In part I, we profiled cytokines using bead array method and by applying various statistical analyses we derived an 18-cytokine signature. In part II, Macrophage Colony Stimulating Factor (MCSF), one of the proteins elevated in GBM sera as identified by bead array method was investigated for its regulation and function in glioma. In part III, serum profiling by antibody microarray was performed and developed a serum based three-marker panel for distinguishing GBM from normal samples. Further, the role of C-Reactive Protein (CRP), a highest abundant serum protein in GBM, was investigated in detail.