bluuebunny/crossref_metadata_2025_split huggingface dataset:
An effective and accurate algorithm for measuring the tumor mutation burden of cancer patients plasma.
Authors: Liu Bo , Zhou Shuang , Cai Yu Hang , Yang Ying , Ye Mingzhi , Shi Tai Ping 
Published: May 2019

e14177 Background: Tumor Mutation Burden (TMB) is deemed to be one of the effective genomic biomarkers that are used to predict the response to PD-1/PD-L1 blockade immunotherapy. As an exciting biomarker for diagnose and an alternative assess method for tissue TMB (tTMB), blood TMB (bTMB) is measured to further distinguish the patients who may benefit from immunotherapy. However, there is no recognized rule to calculate bTMB so far. Methods: In this study, we developed an in-house algorithm called PTAB to calculate bTMB of plasma, all credible single nucleotide variants and insertion-deletions whose allele frequency is greater than 1% are selected in PTAB without synonymous mutations and drive mutations. We also constructed a 636 genes panel which region size is 1.95Mb to identify all the associated mutations, due to the high costs and long turnaround time of WES. Results: Meanwhile, we assessed a cohort of 50 lung cancer plasma samples and paired tissues to ensure the consistency of different algorithm used for tissue and plasma. The results show that Pearson correlation coefficient (R2) between bTMB and tTMB in 50 patients is 0.7128. In addition, we found that the consistency of lung squamous carcinomas obviously higher than that of lung adenocarcinoma (0.8932 vs. 0.5947). We also described the distribution of bTMB and tTMB value of Chinese patients, including 594 tissue samples and 561 plasma samples from 8 types of cancers, including lung cancer, colorectal cancer, breast cancer, et al. The overall comparative analysis of 1155 previous data suggested that the distribution modes of bTMB and tTMB were almost same in certain cancer type, though the level of bTMB was generally lower than that of tTMB. Conclusions: Preliminary data highlight that PTAB is an effective and accurate algorithm for measure the TMB of plasma. The applicability of bTMB testing on liquid biopsy needs further investigation and may be clarified within the ongoing clinical trials.
Anti-HER2/Neu antibody therapy can reduce the immunosuppressive activity of MDSCs in breast tumor model.
Authors: Kim Il-Hwan 
Published: May 2019

e14181 Background: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous cell populations that play a critical role in tumor associated immune suppression. MDSCs consist of immature cells with myeloid lineage markers CD11b and Gr1 in mice and accumulate in large numbers in tumor bearing mice including HER2/neu+ breast cancers. This study was conducted to test whether anti-neu antibody treatment can affect on MDSC populations and immunosuppressive functions. Methods: Female BALB/c mice(5–6weeks of age) were used. The TUBO mammary carcinoma cell line was cloned from a spontaneous mammary tumor in a BALB Neu Tg mouse, as cell lines and reagents. Tumor-infiltrating lymphocyte (TIL) populations were evaluated by flow cytometry. The RT2 Profiler PCR array was used for cytokine expression analysis according to the manufacturer’s instructions. Results: Total number of MDSC in tumor and spleen were decreased within 3 days after anti-neu antibody treatment, but percentage of MDSC in tumor (day2) was reduced more quickly than percentage of MDSC in spleen (day3). The relative populations of MDSC in CD45.2+ cells were significantly decreased such as MO-MDSC. Immunosuppressive activity of MDSC was also reduced by anti-neu antibody treatment. MDSC reduced by anti-neu antibody was due to inhibition of the immunosuppressive factors Arg1 and IDO. In addition, combination therapy with 5-FU may enhance MDSC reduction. Conclusions: MDSC can be a potential therapeutic target in anti-neu antibody treated therapeutic model and the effect of anti-neu antibody could be further improved using other therapeutic treatment to target MDSC.
The influences of preoperative metformin on immunological factors in early breast cancer.
Authors: Takahiro Takahiro , Shien Tadahiko , Taira Naruto , Kochi Mariko , Iwamoto Takayuki , Doihara Hiroyoshi , Udono Heiichiro , Toyooka Shinichi 
Published: May 2019

e14182 Background: Metformin is one of the most commonly prescribed drugs for type 2 diabetes, and some reports have suggested that metformin may reduce cancer risk. Diabetics treated with metformin have a 23% reduction in the risk of cancer, including breast cancer. In addition, it is reported that the breast cancer patients with metformin treatment for diabetes showed favorable prognosis compared with those without metformin treatment. However, the mechanism underlying the positive effects of metformin on cancer treatment remains unclear. Methods: We conducted a prospective study to evaluate the effect of preoperative metformin on early breast cancer patients. The patients took a daily dose of metformin orally for 7 to 21 days before surgery. We evaluated the effects on immunological factors (TILs, CD4+, CD8+, PD-L1 and ALDH1) by comparing core needle biopsies (CNB) obtained before surgery with surgical specimens. Results: Seventeen breast cancer patients were enrolled in this prospective study and administered metformin before surgery, during the period from January to December 2016. We analyzed 59 patients who received surgery during the same period as a control group. In the control group, there was no significant difference in TILs between CNB and surgical specimens (Rs = 0.63). In the metformin group, TILs were negative in CNB and surgical specimens of 15 (88%) and 8 (48%) cases, low in 2 (12%) and 8 (48%), and intermediate in 0 and 1 (6%), respectively. These TILs increases were confirmed in 5 (29%) patients (p = 0.09), while a decrease was confirmed in 2 (12%). The expressions of CD4+ and CD8+ by TILs were increased in 41% and 18% of surgical specimens, respectively. (p = 0.02, p = 0.09) However, there was no statistically significant difference in these immunological factors and PD-L1 or ALDH1 expression between before and after metformin. Conclusions: In our small cohort, preoperative metformin administration shows positive impact on CD4 positive lymphocytes significantly and has tendency of increasing both TILs and CD8 positive lymphocytes. However, we should keep in mind that our sample size is small. Further study is necessary to uncover the mechanisms of favorable effects of metformin on breast cancer patients.
Use of PD-L1 and ICOSL to discriminate innate and adaptive dendritic cells in the head and neck cancer microenvironment.
Authors: Hoffmann Caroline , Surun Aurore , Noel Floriane , Michea Paula , Grandclaudon Maximilien , Faucheux Lilith , Sirven Philemon , Klijanienko Jerzy , Badois Nathalie , Lesnik Maria , Choussy Olivier , Kamal Maud , Lecerf Charlotte , Le Tourneau Christophe , Guillot-Delost Maude , Soumelis Vassili 
Published: May 2019

e14184 Background: Inflamed head and neck squamous cell carcinomas (HNSCC), have a better prognosis independently of the treatment modalities, and have an increased response rate to PD(L)1 blockade. Dendritic cells (DC) infiltration is associated to T cell infiltration and DC are critical in the immune response to cancer, but little is known on HNSCC infiltrating DC. The objectives were to determine the differences between DC infiltrating inflamed vs non inflamed tumors, to understand which mechanisms modulate DC phenotypes and which are the associated functions. Methods: 22 untreated HNSCC were characterized by flow cytometry for their immune infiltrate. RNA sequencing of sorted tumor infiltrating myeloid subsets and blood DC were analyzed for differentially expressed genes. The TMod database (Grandclaudon et al.), testing the effect of various stimuli on DC and subsequently on co-cultured T cells, was used to decipher DC and T cell modulation. Results: The level of CD3 infiltration was positively associated to the level of DC infiltration and PDL1 expression on myeloid cells, and negatively to the level neutrophils and of ICOSL expression on myeloid cells. PDL1high ICOSLneg DC from inflamed tumors were matched with the “innate DC” phenotype described in vitro, highly efficient at secreting cytokines and chemokines but poorly activating naïve CD4 T cells. DC from non-inflamed tumor resembled immature medium DC. A third type of DC, “adaptive DC”, expressing high levels of ICOSL and low of PDL1, poorly efficient at cytokine and chemokine secretion, but highly efficient at activating T cells is was described in vitro, but not found in tumors. RNAseq analysis confirmed this innate polarization and highlighted that those cells present a mature and NFkB activated phenotype. Conclusions: Inflamed HNSCC contain stimuli able to induce the recruitment and the differentiation of DC into mature, NFkB activated “innate” DC, highly efficient at secreting cytokines and chemokines but poorly activating naïve CD4 T cells. These data partly explain the superiority of CPS over TPS for the prediction of response to PD(L)1 blockade. More importantly, the stimuli identified here may be used as adjuvants to induce this DC « innate » maturation in non-inflamed tumors, but should only be used in combination with anti-PD(L)1 immunotherapy.
Demographic stratification of inflammatory signature in lung cancer patients in North Carolina: A prospective cohort study.
Authors: Sharma Nitika , Walker Paul R. , Stroud Chipman Robert Geoffrey , Cherry Cynthia R. 
Published: May 2019

e14200 Background: Lung Cancer remains the major cause of cancer related mortality in the state of North Carolina. There is a growing body of evidence that implicates inflammation as a mechanism of disease progression and reduced survival in patients with advanced cancer (Laird et al, Oncologist 2013). Smoldering inflammation in the tumor microenvironment regulates and escalates cancer invasion, angiogenesis and immune surveillance escape (Balkwill and Mantovani, Lancet 2001). We investigated the predictive value of inflammatory signature according to social stratification of cancer patients using Modified Glasgow Prognostic Score (mGPS). mGPS is a composite inflammatory score based on CRP and serum albumin with proven prognostic and predictive value in various tumor types. Methods: A prospective observational single institutional study was conducted whereby serum albumin and CRP were drawn at baseline for 333 patients with diagnosis of cancer regardless of stage from 30 counties in Eastern North Carolina. The mGPS score was compared according to rural urban divide and occupational regional exposure of various counties stratified per US Census Data. Results: Lung cancer was the predominant cancer type in 93% of patients. The mGPS of zero in Urban vs Rural counties was noted in 36% and 24% patients respectively. The mGPS score of two in Urban vs Rural counties was noted in 26% and 41% respectively. The mGPS of two in areas of hog farming, cattle farming and wet waste lands was seen in 41%, 38% and 43% respectively (p = 0.0019). The mGPS of zero was seen in 24%, 20% and 27% respectively (p = 0.0008). Conclusions: This study suggests a strikingly unfavorable inflammatory signature in rural population as well as areas of hog farm, cattle farm and wet waste lands. The hog and poultry operations heighten the harmful effect on waterways and can adversely affect the inflammatory signature, hence the tumor biology. This underscores additional interventions in these high risk populations that can have significant implications for quality of life and survival, especially in the era of immunotherapy.
The interplay of adiposity, metabolic factors, and tumor infiltrating lymphocytes in an East Asian breast cancer cohort.
Authors: Chen I Chun , Kuo Chun-Wei , Hsueh Tsu-Hsin , Lu Yen-Shen , Lin Ching-Hung 
Published: May 2019

e14207 Background: Breast cancer is rapidly emerging in East Asian countries in recent 2 decades. Unlike western countries in which obesity are quite prevalent and contributes to BC patients, the body mass index (BMI) of BC patients in East Asian countries are usually normal to no more than overweight. In our previous study, we noticed a 30% incidence of crown-like structure of breast (CLS-B) in our BC patients. Despite a lower BMI in this East Asian cohort than the US cohort, the body fat percentage (BF%) is significantly higher in the East Asian The roles of tumor infiltrating lymphocytes (TILs) in BC are inconslusive in different disease contexts. This study aims to decipher the interplay of adiposity, metabolic factors, and TILs in a BC cohort from an East Asian country. Methods: Breast cancer patients from National Taiwan University Hospital were prospectively enrolled into the protocol. Clinicopathologic factors were collected from electronic medical records. Fasting serum collected prior to surgery was used to quantify metabolic factors, including lipid and glucose profiles. Adiposity, including BF% and BMI were quantified by bioimpedance analysis method prior to surgery. Immunohistochemical staining from formalin-fixed, paraffin-embedded BC tumor samples were done to profile TILs by pathologist. CLS-B was stained and interpreted by pathologists. The correlation of TILs with different clinicopathologic factors, adiposity, or relapse-free survival (RFS) were analyzed. Results: 72 BC patients were included in this analysis. The density of CLS-B positively correlates with tumor infiltrating lymphocyte (TIL) percentage ( p = .042), higher BF% correlates with higher regulatory T cells ( p = .026) and tumor associated macrophages (TAM, p = .010), high glucose level is associated with higher CD56 ( p = .040) and higher TAMs ( p < 0.0001). Higher BMI is also associated with higher TAMs ( p= .005). In multivariate analysis, TAMx M2 macrophage is an independent predictor for RFS. Higher TAM level is associated with shorter RFS. Conclusions: Higher TAMs are associated with metabolic alterations such as higher adiposity, higher glucose level, and higher BMI. This result suggests a potential interplay between higher TAMs, altered metabolic factors, and worse RFS.
Possible viral oncolysis by members of the Reoviridae and Paramyxoviridae families (an in vitro study).
Authors: Sitkovskaya Anastasia O. , Zlatnik Elena Yu. , Kolpakov Sergey A. , Kolpakova Elena P. , Novikova Inna A. , Sagakyants Aleksandr B. , Dashkov Andrey , Kaymakchi Dmitry O. , Chupanov Gapiz M. , Gamzatova Patimat M. 
Published: May 2019

e14212 Background: Our purpose was to study possible oncolytic activity of Newcastle disease virus and reovirus in different tumor cell cultures in vitro. Methods: The study was performed on LaSota strain Newcastle disease virus (NDV) vaccine (OOO NPP AVIVAK, 5000 doses) and human reovirus strains R-92 (3 serotype) deposited in the State collection of viruses of the Research Institute of Virology named after. D.I.Ivanovsky. HeLa (cervical cancer), A549 (lung adenocarcinoma), BT20 (breast cancer), PC3 (prostate cancer), HT29 (colon cancer) cell lines were used; pig embryo kidney cell culture (PEVKC) was used as a positive control. Results: After Newcastle disease virus was placed in the medium with tumor cell lines, a 5-day visual microscopic control of the cell morphology did not show signs of the cytopathic action, while a complete lysis of cell biomass was observed in the positive control (PEKC) by the third day. After the incubation with reovirus, the oncolytic activity was registered already in 24 hours: partial or complete destruction of the monolayer, decreased cell adhesion (only solitary cellular conglomerates remained at the bottom of vials), changes in the typical cell appearance. The complete cytopathogenic effect was observed after 2 days. Conclusions: The results demonstrated a marked lytic effect of reovirus in vitro, as well as varying sensitivity of the studied tumor lines to it, which may depend on their molecular genetic characteristics, for example on the RAS mutation status. Despite ambiguous results, we consider it expedient to continue the studies of these and other viruses in tumor models both in vitro and in vivo.
Reovirus strain R-92 in in vitro generation of tumor-specific T-lymphocytes.
Authors: Sitkovskaya Anastasia O. , Zlatnik Elena Yu. , Kolpakov Sergey A. , Kolpakova Elena P. , Mezhevova Irina V. , Bondarenko Elena S. , Novikova Inna A. , Dashkov Andrey , Kaymakchi Dmitry O. , Chupanov Gapiz M. , Oganyan Nairi B. , Vladimirova Liubov Yu 
Published: May 2019

e14216 Background: The purpose of the study was to assess the potential of the reovirus strain R-92 for in vitro generation of tumor-specific T-lymphocytes. Methods: The human reovirus strain R-92 used as a master seed strain was previously isolated from a patient with infectious hepatitis and adapted to the growth on the pig embryo kidney cell culture (PEKC); it lost its pathogenicity after multiple passages and was characterized by electron microscopic, serological methods and PCR. HeLa lysate was prepared by the incubation with the R-92 reovirus in medium 199+L-glutamine during 5 days (37оC, 5% СО2), cytopathogenic effect was observed. Immature dendritic cells (DCs) were derived from blood monocytes cultivated for 7 days in presence of IL-4 and GM-CSF. For the DC loading during 48 hours we used: 1. HeLa lysate obtained by repeated freezing and thawing of cells (control); 2. HeLa lysate obtained by co-culturing with reovirus; 3. reovirus cultured in a standard PEKC. To assess the DC ability to activate lymphocytes, they were co-cultured for 5 days in a 3:1 ratio, 1.5x105 of target cells (HeLa) were added (1:1) for 48 hours; number of dead HeLa cells was count. At each stage of the generation of DCs and activated lymphocytes, they were phenotyped by flow cytometry. Results: DC samples 2 and 3 showed increased percentage of activated cells compared to the loaded with the control tumor lysate, CD86 and HLA-DR coexpression was 62 and 69% vs. 25%, respectively; CD83 expression was maximal after DC loading with reovirus (72.9%). After co-culturing of DC with autologous lymphocytes, the highest number of activated CD4+CD38+ and CD8+CD38+, was observed in sample 2, T-regs (CD4+CD25+CD127low) - in the control. The death of target cells cultured with activated lymphocytes obtained by DC samples 2 and 3, was 100%, control sample showed minimal cytotoxicity against HeLa. The number of T-lymphocytes in samples 2 and 3 cultured with HeLa exceeded the control values by 3-10 times. Conclusions: Reovirus R-92 used for the immature DC loading increases their presenting activity. DCs loaded with reovirus HeLa lysate, when co-cultured with lymphocytes, activate CD4+ and CD8+ and suppress T-regs, apparently leading to the total death of target tumor cells.
Correlation of radiosensitivity of lung carcinoma to CD137L endogenous expression levels.
Authors: Girdhani Swati , Rodriguez Andrew , Dua Rajiv , Parry Renate 
Published: May 2019

e14229 Background: Radiation therapy is a cornerstone in the management of cancer. Preclinical and clinical evidence indicate the therapeutic efficacy of radiation therapy can be augmented by immunomodulatory antibodies against targets such as CD137 and PD-1/PD-L1. Despite evidence of synergy, the mechanistic basis has yet to be fully explored. In this study, we investigated how the expression of CD137, CD137L, PD-1, and PD-L1 correlates with radiosensitivity and proliferation of lung tumor cell lines treated with radiation as well as anti-PD-L1 and agonist anti-CD137 antibodies. Methods: Expression of CD137, CD137L and PD-L1 was measured by RT-qPCR in eight NSCLC cell lines (HCC827, HCC2935, H358, H460, A549, H596, H1650, H520) treated with radiation in doses of 0-8 Gy. Clonogenic assays for radiosensitivity and the CyQUANT assay for cell proliferation were performed after treatment with radiation in combination with anti-PD-L1 (0.01-10µg/ml) and agonist anti-CD137 (0.01-10µg/ml) antibodies. Results: Out of the eight cell lines tested, four were found to be radioresistant – H460, A549, HCC827 and H358 had SF2 (survival at 2 Gy) = 0.76, 0.70, 0.67 and 0.62, respectively – two were moderately sensitive – H520 and HCC2935 (SF2 = 0.54 and 0.40) and two were sensitive to radiation – H596 and H1650 (SF2 = 0.38 and 0.27). Expression analysis by RT-qPCR showed high endogenous expression of CD137L in three out of four radioresistant cell lines while the fourth, H358, had low endogenous expression of CD137L but upon irradiation showed more than a 3-fold induction of CD137L expression. PD-L1 expression levels measured by RT-qPCR did not correlate with radiosensitivity. In vitro cell proliferation was found to be inhibited by 48-68% in all eight NSCLC cell lines after 4 Gy radiation, but combination with anti-PD-L1 and agonist anti-CD-137 antibodies did not contribute additional toxicity. Conclusions: Our results demonstrate that CD137L expression on NSCLC cell lines correlates with radioresistance. Further, addition of anti-PD-L1 and agonist anti-CD137 antibodies to radiation showed little synergistic effect in vitro on tumor cell proliferation. Additional studies to address the synergistic effect of radiation and immunomodulation on tumor cells co-cultured with immune cells is ongoing.
Preclinical development of combination therapy targeting the dominant cytokine interleukin-1β for renal cell carcinoma.
Authors: Aggen David Henry , Ghasemzadeh Ali , Mao Wendy , Chowdhury Nivi , Chaimowitz Matthew , Hawley Jessica , Wang Vinson , Dallos Matthew , Zhang Cheng , Li Hu , Guo Xinzheng Victor , Drake Charles G. 
Published: May 2019

e14237 Background: Targeting myeloid-derived suppressor cells (MDSCs), key mediators of intrinsic and adaptive immune resistance, remains a significant challenge in kidney cancer immunotherapy. One potential driver of MDSC mediated tumor immunosuppression is the cytokine interleukin-1 beta (IL-1β). Although large cancer prevention studies show that targeting IL1 decreases cancer risk, the exact mechanism of anti-IL-1 mediated anti-tumor efficacy is not yet defined. To understand the immunogenic effects of IL-1β blockade and effects on MDSCs, we used the RENCA model of RCC. Methods: We inoculated BALB/c mice with 1x106 RENCA cells. On Day +10 we treated mice (n = 8/group) with vehicle, αPD1, αIL1β, cabozantinib, combination αPD1/αIL1β, or cabozantinib/αIL1β. We harvested day +18 tumors to quantify tumor size and immune cells by flow cytometry. We then isolated immune cells for differential gene expression profiling by RNAseq. Results: Treatment with αIL1β significantly decreased tumor size compared to vehicle or αPD1 monotherapy. Combining αPD1/αIL1β or cabozantinib/αIL1β potentiated the anti-tumor effects of αPD1 or cabozantinib monotherapy respectively. Anti-IL1β therapy decreased polymorphonuclear (PMN)-MDSC infiltration (αIL1β: 0.76% +/- 0.21 vs. vehicle: 1.89% +/- 0.37, P = 0.014). Relative to PD1 monotherapy, combining αPD1 and αIL1β decreased PMN-MDSCs (αPD1: 2.14% +/- 0.56 vs αPD1/αIL1β: 0.91% +/- 0.15, P = 0.033) and increased M1-like tumor associated macrophages (TAM) (αPD1: 0.798% +/- 0.21 vs. αPD1/αIL1β: 12.04% +/- 2.9, P = 0.0008). Gene expression profiling from sorted immune cells from αIL1β treated tumors revealed multiple differentially regulated cytokines including interleukin-6, IFN-γ, and IL-10. Conclusions: Blockade of IL-1β depletes intratumoral PMN-MDSCs and increases M1-like TAM with minimal effects observed on the T cell compartment. Treatment with cabozantinib or αPD1 in combination with αIL1β led to sustained control of tumor growth. IL-1β represents a promising target for kidney cancer immunotherapy. Based on the preclinical data, a clinical trial of neoadjuvant αPD1 and αIL1β therapy is planned for patients with high risk RCC.