bluuebunny/crossref_metadata_2025_split huggingface dataset:
R-DOTAP (Versamune): A novel enantiospecific cationic lipid nanoparticle that induces CD4 and CD8 cellular immune responses to whole protein and tumor-specific peptide antigens.
Authors: Wood Lauren Virginia , Gandhapudi Siva K , Sundarapandiyan Karuna , Bedu-Addo Frank K , Conn Gregory , Woodward Jerold G 
Published: May 2020

e15211 Background: Immunotherapy approaches are limited in their ability to induce antigen-specific CD8+ T cells in vivo able to recognize and kill tumor cells. We developed a novel immunotherapy approach using enantiomerically pure, R-DOTAP cationic lipid nanoparticles and tumor-derived T cell antigens, and previously demonstrated that R-DOTAP formulations efficiently prime cytotoxic T cells through enhanced cross presentation and induction of type I interferons.[1] A phase I clinical trial of a R-DOTAP HPV16 peptide formulation confirmed induction of strong in vivo HPV-specific CD8+ cytolytic T-cells without associated systemic toxicities. In this study, we assessed R-DOTAP nanoparticle formulations containing whole protein (ovalbumin) or long multi-epitope peptides from the tumor antigen TARP (T-cell alternate reading frame protein): a 58-residue protein overexpressed in prostate and breast cancers, documented to be immunogenic in humans. Methods: R-DOTAP formulations were prepared containing ovalbumin (OVA) or TARP peptides. C57BL/6K mice were immunized with 10 μg/mouse of OVA plus R-DOTAP, CFA or sucrose on Days 0, 15 and 30. OVA-specific cellular and humoral responses following vaccination were assessed by measuring splenic CD4 and CD8 T cell IFN-γ production and circulating OVA-specific antibodies in serum. HLA-A2 transgenic mice (AAD mice) were vaccinated with long, multi-epitope TARP peptides delivered as an R-DOTAP admixture or with CFA or sucrose on Days 0 and 7. Antigen-specific T cell responses were measured by IFN-γ ELISpot assay. Results: OVA R-DOTAP formulations induced strong antigen-specific effector CD4 and CD8 immune and memory responses detected 7 and 30 days, respectively, following vaccination as well as OVA-specific antibody responses. In TARP peptide vaccinated mice, R-DOTAP formulations were able to present multiple CD8 T cell epitopes and stimulate responses that were superior to CFA. Conclusions: Our results suggest that R-DOTAP is a versatile immune activating therapy that can be formulated with long, multi-epitope tumor-derived peptides or whole proteins. R-DOTAP formulations induce quantitatively robust antigen-specific CD4 and CD8 T cells in vivo compared to well-established immune stimulants. Reference: 1.Gandhapudi SK, Ward M, Bush JP et al. Antigen Priming with Enantiospecific Cationic Lipid Nanoparticles Induces Potent Antitumor CTL Responses through Novel Induction of a Type I IFN Response. J Immunol 2019;202:3524-3536
Characteristics of carrier cells associated with clinical response in children treated with adenoviral virotherapy administered in autologous marrow-derived mesenchymal cells.
Authors: Ramirez Manuel , Franco-Luzón Lidia , González-Murillo África , Melen Gustavo , Ruano David , Lassaletta Álvaro , Madero Luis 
Published: May 2020

e15217 Background: We used oncolytic virotherapy carried by autologous marrow-derived mesenchymal cells to treat children with metastatic/refractory tumors (NCT01844661; 10.1016/j.ymthe.2020.01.019 and 10.1016/j.canlet.2015.11.036). Most patients did not show clinical response while a small group did. Methods: We have carried out a comprehensive study of the transcriptome of this new advanced therapy medicine (ATM) to find determinants that might be associated with clinical outcome. Results: The study of the ATM's transcriptome from a first cohort of patients identified signatures associated to biological processes in the infected carrier cells that distinguished responders from non-responders (viral infection and nucleic acid metabolism; immune regulatory routes; cell-cell adhesion; mitosis; cell metabolism; response to stress; and bone marrow fibrosis). Next, we used an approach based on systems biology and neural networks to identify candidates responsible for the differences found in gene expression signatures. We then validated the identified candidates using samples obtained from a second independent cohort of patients, by quantitative PCR and ELISA. Mesenchymal cells of the responding patients expressed significantly lower levels of the MAVS and NDRG1 genes compared with those of the non-responders after oncolytic adenoviral infection. Both genes are related to cellular antiviral responses and cellular metabolism. Accordingly, mesenchymal cells of responders produced significantly lower amounts of IL-6 and CCL-2 than non-responders’ cells. Conclusions: These results point to possible pretreatment biomarkers and the possibility of optimizing a universal version of the carrier cells for the oncolytic virus in this new ATM.
Predictive value of sonoelastography of secondary changed lymphous nodes in the assessment of effectiveness of immunotherapy.
Authors: Moiseenko Fedor Vladimirovich , Moiseyenko Vladimir , Chernobrivtceva Vera , Egorenkov Vitaliy , Zhegalina Arina , Artemeva Elizaveta 
Published: May 2020

e15225 Background: With the use of existing predictors of the effectiveness of immunotherapy it is not possible to identify patients who will have a response to treatment or a rapid progression. The mechanism of the antitumor effect of the modern generation of immunotherapeutic drugs leads to significant changes in the cellular structure of the tumor and, as a consequence of this, in its physical characteristics. Sonoelastography allows to evaluate the stiffness of the tumor. Methods: The study included 17 patients with malignant neoplastic disease with the presence of secondarily affected peripheral lymph nodes (l.n.). 11/17 patients had melanoma, 2/17 - NSCLC, 3/17 - neck cancer. 8/9 - M / F, average age 61.8 years. 5/17 patients received combined immunotherapy (Ipilimumab + Nivolumab), 12/17 - anti-PD-1 (8 people Nivolumab, 4 people Pembrolizumab). Results: An ultrasound examination of the l. n. in a multimodal mode with compression elastography was performed on a Philips Epic 7 device before the first administration of the antitumor drug and every 3 weeks. Before the first objective assessment of the effect was done, in 5/17 patients was registered an increase in the size of the studied lymph nodes by 2 or more times, in 7/17 patients the size of the l. n. did not change, in 5/17 patients the affected l. n. decreased (26% - 64%).The results of control examination after 2 months of treatment: PR - 2/17, CR - 4/17, SD- 9/17, PD- 4/17.In the group with an objective response (PR, CR, SD - 13/17), the average value. St. Ratio before treatment was 5.7 ± 0.4; during treatment decreased to an average of 3.2 ± 0.5 (p < 0.05). In the group with progression during treatment, the average St. Ratio value before treatment was 4.1 ± 0.4, and during treatment increased on average to 5.7 ± 0.5 (p < 0.05).A qualitative analysis of tumor blood flow was carried out, the Resistance Index (RI) was studied. The RI in the first group of patients was 0.65 ± 0.4; after treatment, the RI increased to 0.74. In the second group of patients, the RI before and after treatment did not significantly change. Conclusions: During the immunotherapy treatment, a decrease in the stiffness of the tumor is observed, regardless of the change in size and an increase in the indices of peripheral resistance to blood flow in case of further regression or stabilization of the process. An early change in the sonographic characteristics of tumor foci can identify patients with a subsequent treatment effect, and can also distinguish true progression from pseudoprogression, which requires confirmation in larger studies.
Crosstalk between the MSI status and tumor microenvironment in colorectal cancer.
Authors: Luo Peng , Lin Anqi , Zhang Jian 
Published: May 2020

e15236 Background: In recent years, cancer immunotherapy has been extensively studied, and colorectal cancer (CRC) patients have also derived clinical benefits from immunotherapy, especially CRC patients with mismatch repair deficiency (dMMR)/microsatellite instability-high (MSI-H), whose sensitivity to immune checkpoint inhibitors (ICIs) is significantly higher than that of patients with microsatellite-stable (MSS)/microsatellite instability-low (MSI-L) disease. This study suggests that patients with MSI-H CRC have a higher mutational burden and more immune cell infiltration than those with MSS/MSI-L disease. However, most studies have not systematically evaluated the immune characteristics and immune microenvironments of MSI-H and MSS/MSI-L CRC. Methods: A published CRC cohort with mutation and immunotherapy-related prognostic data was collected. We analyzed the relationship between the MSI status and prognosis of ICI treatment in an immunotherapy cohort. We then further used mutation data for the immunotherapy and The Cancer Genome Atlas (TCGA)-CRC (colon adenocarcinoma (COAD) + rectum adenocarcinoma (READ) cohorts. For mRNA expression, mutation data analysis of the immune microenvironment and immunogenicity under different MSI status was performed. Results: Compared with MSS/MSI-L CRC patients, patients with MSI-H CRC significantly benefited from ICI treatment. We found that MSI-H CRC had more immune cell infiltration, higher expression of immune-related genes and higher immunogenicity than MSS/MSI-L disease. The MANTIS score used to predict the MSI status was positively correlated with immune cells, immune-related genes, and immunogenicity. In addition, subtype analysis showed that COAD and READ might have different tumor immune microenvironments. Conclusions: MSI-H CRC may have an inflammatory tumor microenvironment and increased sensitivity to ICIs. Unlike those of MSI-H READ, the immune characteristics of MSI-H COAD may be consistent with those of MSI-H CRC. Furthermore, the possible mechanism underlying the prognostic differences among CRC patients receiving ICIs in relation to the immune microenvironment were elucidated to provide theoretical guidance for further improving the curative effect of ICIs treatment on MSI-H CRC patients in the future and solve the problems underlying why MSS/MSI-L CRC patients do not benefit from ICIs treatment.
Learning relevant H&E slide morphologies for prediction of colorectal cancer tumor mutation burden using weakly supervised deep learning.
Authors: Joshi Rohan P. , Kruger Andrew J. , Sha Lingdao , Kannan Madhavi , Khan Aly A. , Stumpe Martin 
Published: May 2020

e15244 Background: Tumor mutation burden (TMB) is the number of non-synonymous mutations present in a cancer exome. In colorectal cancer (CRC), high TMB is associated with microsatellite instability (MSI), POLE mutations, and response to immunotherapy. TMB prediction from whole-slide images (WSIs) could aid workflows that determine MSI and POLE status. Deep learning has previously been used to predict MSI status from WSIs. This approach assumed the morphologies of all regions within the tumor are equally associated with MSI. Here, we predict TMB using a weakly supervised deep learning framework that relaxes this assumption and automatically learns relevant regions within the tumor that are most associated with TMB, potentially uncovering morphological associations. Methods: Weakly supervised learning methods facilitate classification of samples that contain many individual instances, only some of which are related to the sample label. Here, a given WSI has a single TMB-high or -low label and contains individual regions that may or may not be associated with TMB status. We implemented a ResNet18 attention-based, multiple-instance learning (MIL), convolutional neural network to simultaneously learn which tiles are important for prediction of the slide-level TMB and the tile features that are associated TMB-high and -low. We determined performance through 8-fold cross-validation within a Tempus dataset using a 75%-12.5%-12.5% split of ~940 WSIs for training, validation, and testing folds. Results: In the cross-validation, we observed a receiver operating characteristic area under the curve of 0.854 (95% CI 0.776-0.932), an average precision of 0.723 (95% CI 0.580-0.865), and an accuracy of 0.889 (95% CI 0.833-0.945) in the held-out test sets. Morphologies predicted as irrelevant for TMB include adipose tissue and WSI artifacts. Visualizations of model weights show morphologies determined to be most associated with TMB-high and -low, such as high tumor/lymphocyte content and vasculature/red blood cells, respectively. Conclusions: Attention-MIL shows high performance for the prediction of TMB in CRC from H&E images and potentially reveals the morphologies of CRC that are most associated with TMB. Future directions include further investigation of morphological associations, generalizing this model beyond Tempus acquired data, and re-training on the entire Tempus dataset.
PD-L1 status and efficacy of immune check-point inhibitors (ICIs) in advanced cancer patients: A pooled analysis of randomized trials.
Authors: Biello Federica , Pinato David James , Borra Gloria , Genestroni Silvia , Siffredi Guido , Repetti Ilaria , Martini Veronica , Varughese Feba mariam , Vachanaram Ajay Ram , Mora Edoardo , Nardin Simone , Bruzzi Paolo , Sica Antonio , Gennari Alessandra 
Published: May 2020

e15263 Background: The introduction of immune check-point inhibitors (ICIs) in the treatment of a broad range of tumor types has led to a significant and clinically meaningful improvement in overall survival (OS) in advanced disease stages. However, the efficacy of these agents is not consistent across trials and in routine practice. The role of PD-L1 expression as a tumour-agnostic predictive correlate of response to ICIs remains unclear. We performed a pooled analysis of the efficacy of PD-1/PD-L1-targeted ICI regimens as compared to standard of care (SoC) therapy according to PD-L1 expression, based on landmark clinical studies. Methods: We searched literature databases to identify phase III randomized controlled trials that compared anti-PD-1/PD-L1 antibodies alone or in combination with chemotherapy or targeted agents against SoC therapy in the treatment of different tumor types. We reported efficacy data, in terms of OS, according to PD-L1 status. Log hazard ratios (HRs) were pooled across the studies overall and by PD-L1 status by inverse variance weighting. All statistical tests were two-sided. Results: Twenty-four studies including 17687 randomised patients with advanced lung, renal, urothelial, liver, breast, head and neck cancers and melanoma, were eligible for analysis. Efficacy according to PD-L1 immunohistochemical expression, with a 1% cutoff ( < 1% versus > 1%), was reported in 11 studies (7126 patients). Overall, ICI-containing regimes were significantly superior in terms of OS to SoC regimens (pooled HR = 0.64; 95% CI 0.60 to 0.68). When OS data were pooled according to PD-L1 expression, HR was 0.66; 95% CI 0.61 to 0.71 in PD-L1 < 1%, versus 0.62; 95% CI 0.56 to 0.68 in PD-L1 >1% (p = 0.7). Conclusions: The significant improvement in OS favoring the use of ICIs over SoC does not seem to be confined to patients with PD-L1-overexpressing tumours. The difference in efficacy according to PD-L1 status appears to be, at best, marginal.
Mechanisms of skin toxicity of paclitaxel: An in vitro preclinical assessment.
Authors: Perez-Fidalgo Jose Alejandro , Estornut Navarro Cristina , Sanz Garcia Celia , Perez-Leal Martin , Poveda Ferriols Jesus , Cervantes Andres , Milara Paya Javier , Cortijo Gimeno Julio 
Published: May 2020

e15511 Background: Paclitaxel skin toxicity is a frequent side effect extensively evaluated in the clinical setting. However little is known about the preclinical mechanisms that lead to this toxicity. The endpoint of this study was to analyse the cutaneous mechanisms that drive paclitaxel toxicity in a preclinical model. Methods: Primary human keratinocytes were co-cultured with human dermal fibroblast in collagen gel under air-liquid interface conditions to generate a multilayered 3D epidermis. Paclitaxel was added to 3D epidermis at 0.3 µM, 3 µM and 30 µM and total RNA and protein was extracted after 24h of incubation. Markers of cell senescence (p21 and p53), anti-apoptotic mediators (Bcl-2), skin elasticity (tropoelastin ELN, collagen type I and fibronectin), hidratation (aquaporin 3 AQP3), oxidative stress (NOX4), antioxidant (SOD1, Nrf2), and angiogenic markers (VEGFR, eNOS) were evaluated by RT-PCR and western blot. NfKB phosphorylation was measured by western blot and inflammatory citokines IL1α, IL-6 and IL-8 were measured in cultured supernatants by ELISA. Human primary melanocytes were cultured and stimulated with paclitaxel to measure melanogenesis through the expression of of TYR, TYRP1 and DCT genes. The effect of paclitaxel on skin endothelial cell angiogenesis was measured by endothelial tube formation. Results: In human 3D keratinocytesPaclitaxel inhibited the expression of Bcl-2, and increased the expression of p53 and p21. The angiogenic markers VEGF and eNOS were decreased. The expression of oxidative stress marker NOX4 was increased and the anti-oxidant mediators Nrf2 and SOD1 were decreased. Markers of elasticity, collagen type I, fibronectin and FN1 were decreased as occurs with AQP3 hidratation marker. Paclitaxel increased the phosphorylation of NfkB and elevated the secretion of IL1α, IL-6 and IL-8 cytokines. In skin endothelial cells, paclitaxel reduced the endothelial tube formation. In melanocytes, paclitaxel increased skin pigmentation represented by the increase of RNA expression of TYR, TYRP1 and DCT genes. Conclusions: This preclinical 3D model showed that paclitaxel impacts on the expression of proteins related with angiogenics, elasticity and senescence in human kerantinocytes. Moreover higher doses of paclitaxel increased inflammation parameters and confirmed phototoxic and anti-angiogenenic effects. This preclinical model could be a valuable tool to assess skin toxicity of new antineoplastic agents.
MicroRNAs involved in immune response as prognostic markers in early and metastatic breast cancer.
Authors: Papadaki Chara , Thomopoulou Konstantina , Koronakis George , Monastirioti Alexia A , Papadaki Maria A. , Kalbakis Kostas , Agelaki Sofia , Mavroudis Dimitrios 
Published: May 2020

e15528 Background: MicroRNAs (miRNAs) are involved in the regulation of immune response and have an important role in immune escape. We analyzed the expression levels of plasma miR-10b, miR-126, miR-19α, miR-20α and miR-155, which regulate tumor-immune interactions and investigated their prognostic implications in patients with early (eBC) and metastatic (mBC) breast cancer. Methods: Blood samples were obtained before treatment from 140 and 64 patients with eBC and mBC, respectively. Plasma miRNA expression levels were assessed by qRT-PCR and expression was classified as high or low according to the median values. Results: A panel of four miRNAs (miR-19a, miR-20a, miR-126 and miR-155) could discriminate eBC from mBC (AUC 0.802, p < 0.001). In early disease, miR-10b (p = 0.022) and miR-155 (p = 0.005) expression was lower in relapsed (n = 46) compared to non-relapsed (n = 94) patients; miR-155 expression along with lymph node infiltration and tumor grade had increased ability to predict relapse (AUC = 0.775; p = 0.003). In addition, miR-10b (p = 0.015), miR-19a (p = 0.003), miR-20a (p = 0.012), miR-126 (p = 0.001) and miR-155 (p < 0.001) expression levels were lower in patients with early relapse (relapse at ≤2 years). MiR-155 and miR-19a had the highest performance in discriminating early relapse (AUC 0.855; p < 0.001 and AUC = 0.729; p = 0.003, respectively), whereas, combined mir-155 and miR-19a expression further increased the accuracy of prediction (AUC = 0.867; p < 0.001). In eBC, the number of infiltrated lymph nodes and low miR-10b independently predicted for shorter DFS (p = 0.001 and p = 0.03, respectively) and axillary lymph nodes for shorter OS (p = 0.003). In the triple negative subgroup, low miR-155 strongly predicted for shorter DFS (p = 0.037). In mBC, recurrent disease and low miR-10b expression independently predicted for shorter PFS (p = 0.001 and p = 0.017, respectively), whereas performance status of 2 independently predicted for shorter OS (p = 0.03). Conclusions: Deregulated expression of circulating miRNAs involved in tumor-immune interactions can discriminate disease status in BC and independently predicts for patients’ outcome in early and mBC. Our results further support the notion that circulating miRNAs represent a useful prognostic tool in patients with BC.
Liquid biopsy of lung cancer by deep learning and spectroscopic analysis of circulating exosomes.
Authors: Shin Hyunku , Oh Seunghyun , Hong Soonwoo , Kang Minsung , Kang Daehyeon , Ji Yong-gu , Choi Byeong Hyeon , Kang Ka Won , Jeong Hyesun , Park Yong , Hong Sunghoi , Kim Hyun Koo , Choi Yeonho 
Published: May 2020

e15532 Background: Lung cancer has a high mortality rate because of belated diagnosis at advanced stages beyond the treatable condition. Early detection of lung cancer can improve the survival rate. A liquid biopsy that detects tumor-related biomarkers in body fluids has a great potential for the purpose. Particularly, tumor-derived exosomes in blood have been proposed as a promising biomarker. The tumor-derived exosomes carry molecules of their parental cells; thus, they provide information about the tumor in the body. Unfortunately, exosomal markers conducive to the early detection of lung cancer are still obscure. Therefore, using the molecular fingerprint of exosomes markers can be useful to detect the tumor exosomes. Raman spectroscopy is one of the representative methods for the purpose. However, because the exosomes have a heterogeneous composition in blood, interpreting their spectroscopic signals is hard. Thus, we utilized a deep learning approach to analyze the spectroscopic signal of the exosomes for liquid biopsy of lung cancer. Methods: The basic concept was to evaluate how much the exosomes in human plasma resemble cancer cell exosomes. As a proof of concept, exosomes of 43 non-small cell lung cancer (NSCLC) adenocarcinoma patients and 20 healthy controls were isolated from plasma of peripheral blood. Also, cell exosomes were isolated from culture media of adenocarcinoma cell lines and a human pulmonary alveolar epithelial cell line. Then, the spectroscopic signals were detected using surface-enhanced Raman spectroscopy (SERS). Further, the deep learning algorithm was employed to classify the signals. Then, we calculated the relative similarity to cancerous exosomes against human plasma exosomes. Results: Our method was able to classify cancer and normal cell exosomes with 95% accuracy. Also, Raman signals of cancer patients’ exosomes were more similar to the cancer cell exosomes than those of healthy controls. Notably, the similarity was proportional to cancer stages. Importantly, our method even detected stage I patients. The area under the curve (AUC) of receiver operating characteristic (ROC) curves was 0.912 for stage I and II, and 0.910 for stage I. Conclusions: We reported a novel diagnostic method using deep learning analysis against spectroscopic signals of circulating exosomes. Our method that evaluates the similarity to cancer exosomes accurately identified lung cancer patients, even stage I with high accuracy.