bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemical and pharmacological evaluation of manglicolous lichen Roccella montagnei Bel em. D. D. Awasthi
Authors: Tatipamula Vinay Bharadwaj , Vedula Girija Sastry , Sastry A. V. S. 
Published: November 2019

Abstract
Background
Lichen is a composite organism composed of fungus in association with algae or cyanobacteria. Particularly, lichen betide to mangroves are named as manglicolous lichens. From the folklore, lichen extracts were used in the management of many infections and diseases.

Results
The chemical investigation of acetone extract of manglicolous lichen Roccella montagnei (RM-Ac) yielded nine known metabolites namely divarinolmonomethylether (1), ethyl divaricatinate (2), divarinol (3), orcinol (4), methyl 2,6-dihydroxy-4-methylbenzoate (5), haematommic acid (6), atranol (7), ethyl haematommate (8) and ethyl orsellinate (9). Except 4, all are for the first reported from this species. The RM-Ac and its metabolites (1–9) were screened for antioxidant (DPPH, ABTS and superoxide free radical assays), anti-inflammatory, anticancer (SRB assay using A549, DLD-1, FADU, HeLa and MCF-7) and acute toxicity studies. The pharmacological results showed that compounds 6 and 8 depicted potent inhibitory profile against ABTS free radical (with IC50 value of 40.0 and 40.5 μg/mL, respectively) and protein denaturation (with IC50 value of 435 and 403 μg/mL, respectively). LD50 of RM-Ac was found to be above 2 g/Kg body weight. Moreover, the RM-Ac showed prominent inhibition of formalin-hind albino rat paw oedema at both the tested doses, i.e., 100 and 200 mg/Kg b.w than that of the standard drug (indomethacin). Furthermore, the compounds 6 and 8 exhibited significant degree of specificity against HeLa, FADU and A549; besides, they showed very little degree of specificity against NHME cell line specifying less toxicity to normal cells.

Conclusion
It can be concluded that the manglicolous lichen R. montagnei has an aptitude to act against free radicals, inflammation and cancer, and the main metabolites responsible for its biological activity are 6 and 8.
Comprehensive genomic analysis with immunological parameters to obtain the biomarkers for immune checkpoint inhibitors (ICIs) in non-small cell lung cancer (NSCLC).
Authors: Owada Yuki , Muto Satoshi , Takagi Hironori , Inoue Takuya , Watanabe Yuzuru , Yamaura Takumi , Fukuhara Mitsuro , Okabe Naoyuki , Matsumura Yuki , Hasegawa Takeo , Tanaka Daisuke , Ito Emi , Nanamiya Hideaki , Imai Jun-ichi , Isogai Takao , Watanabe Shinya , Suzuki Hiroyuki 
Published: November 2019

107 Background: Several kinds of predictors of ICIs efficacy in NSCLC have been analyzed. PD-L1 expression of tumor, frequency of tumor infiltrating lymphocytes (TIL) and microsatellite instability are cited as potential predictors of treatment efficacy of ICIs, but these still seem to be immature. Lately, mutation load in tumor, presumably involving to neo-antigens in tumor tissue, is also thought to be one of more probable predictors for ICIs. However, the hypothesis is still limited and it is very hard to analyze the mutation load routinely in clinic. We here describe more definitive and easily detecting factors by using Next generation sequencing (NGS) and by analyzing conventional immunological and molecular parameters. Methods: 94 patients with NSCLC who undergone resection in our institution were subjected for this study. Mutation load in tumor samples was analyzed by whole exome sequencing using NGS. The major gene alteration was also analyzed by NGS. TILs and PD-L1 expression in tumor were immunohistochemically assessed by using CD8 and SP142 (anti-PD-L1 antibody). Results: The median value of mutation load was 54 genes (10-363). The factors; squamous cell carcinoma, EGFR-mutation negative, and TP53 alteration positive indicated close connection with higher mutation load. PD-L1 expression, TILs, gender, age, and etc. were not associated with mutation load. Multiple regression analysis shows that EGFR-mutation negative and TP53 alteration positive contribute to higher mutation load (p = 0.018, p = 0.015). Conclusions: Mutation load in tumor could be associated with EGFR-mutation and TP53 alteration status. These profiles could potentially be more simple biomarkers in treating ICIs.
Tumor associated macrophages antagonize antitumor effect of chemotherapy.
Authors: Chen Fei , Su Shicheng , Song Erwei 
Published: November 2019

120 Background: Tumor associated macrophages play an essential role in tumor progression, promoting angiogenesis, immunosupression, invasion and metastasis. However, TAMs regulation of tumor responses to chemotherapy is complicated. In different type of treatment or tumor model, TAMs can either enhance or antagonize antitumor effect of chemotherapy. Methods: We isolated TAMs from breast cancer tissues, or PBMC stimulated with culture medium of mesenchymal breast cancer cells and cocultured with breast cancer cells MCF-7 for 6-10days. After cocultured with TAMs, we treated MCF-7 with docetaxel. In vivo, we use MCF-7 with or without TAMs to generate xenografts in NOD/SCID mice and treated with docetaxel. To testify if the antagonize was dependent on specific treatment, we also treated breast cancer cells with ciaplatin. Results: With coculture of TAMs, we obversed ruduced apoptosis of docetaxel treated breast cancer cells. In mice model, TAMs co-injection xenografts antagonized the antitumor effect of docetaxel.Consisted with previous results, TAMs coculture antagonize the antitumor effect of cisplatin, suggesting the antagonize of TAMs was independent on cell cycle. Conclusions: Collectively, our study indicated the antagonize of TAMs towards chemotherapy.
GRM1 to induce M-CSF and CCL2 expression to lead to an upregulation of M2 macrophages in melanoma.
Authors: Li Jiadong , Wen Yu , Jeong Byeong-Seon , Shah Raj , Isola Allison , Silk Ann W. , Chen Suzie , Goydos James Steven 
Published: November 2019

122 Background: GRM1 transgenic mice in an immune competent background (C57BL6) develope melanoma with 100% penetrance (TG-3 mice). Ectopic expression of GRM1 in immortalized mouse melanocytes (MelanA) produce cells (MASS20) that are tumorigenic in both SCID and immune competent C57BL6 mice. We have examined the downstream events of GRM1 signaling that allow transformed cells to evade immune destruction. Methods: Cytokine antibody arrays and Western blot were used to examine differences in cytokine expression between MASS20 cells and parental controls. Next, we cultured C57BL6 bone marrow cells in MASS20 conditioned medium for 24 and 48 hours and determined resultant M2 macrophage levels. Finally, we used oral weekly liposome clodronate (LC) treatment to deplete macrophages in murine models to determine the effect on tumor growth. Results: MASS20 cells produce higher levels of M-CSF and CCL2 as compared with control cells. These two cytokines recruit and induce the polarization of tumor associated macrophages (TAM) to increase the M2 fraction resulting in accelerated tumor progression. Culturing C57BL6 bone marrow cells in MASS20 conditioned medium increased expression of the M2 macrophage marker arginase1 at both time points. Weekly oral LC treatment to C57BL6 mice greatly inhibited the growth of MASS20 tumors as compared to growth in control animals. Single-cell suspensions of spleen and tumor cells from these animals were subjected to flow cytometry and showed that LC treatment depleted macrophages by 90% in the spleen, and increased the percentage of cd8+T and cd335+ NK cells in tumors from treated mice as compared to controls. Finally, we treated TG-3 transgenic mice with weekly oral LC and found that treated animals still developed tumors, but progression was greatly slowed, and survival greatly prolonged, compared to untreated animals. Within 1 week of stopping LC in the treated animals there was rapid growth of the tumors. These findings suggest a link between GRM1-signal transduction and TAM-induced suppression of tumor-immunity in melanoma. Conclusions: We conclude that GRM1-induced suppression of tumor immunity occurs through induction of M2 macrophages in the tumor microenvironment.
Effects of in vitro ATRA treatment on human MDSC expansion and function.
Authors: Tobin Richard P. , Jordan Kimberly R , Davis Dana , McCarter Martin 
Published: November 2019

125 Background: Myeloid derived suppressor cells (MDSCs) are a heterogeneous population of immature immunosuppressive myeloid cells that are expanded in tumor bearing hosts. Melanoma patients with high frequencies of MDSCs have decreased overall survival and an increased risk of death and disease progression, as well as impaired responses to ipilimumab. Targeting MDSCs to decease their frequency or suppressive activity may provide an effective means to improve the efficacy of anti-cancer therapies. All-trans retinoic acid (ATRA) is a vitamin A derivative currently used to treat APL that may target MDSCs. ATRA differentiates immature myeloid cells into macrophages, dendritic cells, or granulocytes. ATRA has been tested as an MDSC targeting agent in two completed caner clinical trials and is being investigated in several other open and completed trials. Methods: We purified myeloid cells from LRS chambers collected from normal donors. The purified cells were treated with GM-CSF+IL-6, GM-CSF+IL-4, or melanoma conditioned media with or without ATRA. After 5 days of incubation, PCR was used to determine the expression of immunoregulatory genes. Additionally, we used a MLR to determine the effect of ATRA on MDSC’s T cell suppressive function. Results: Here we report that in vitro ATRA treatment decreases the expression of ARG1, NOX1, INOS, and PD-L1. Further, we report that ATRA treatment improved T cell proliferation. Conclusions: Controlling MDSC suppressive function and accumulation may provide a mechanism to improve the efficacy of many current cancer therapies. Here we show that ATRA reduces the expression of the immunosuppressive genes ARG1, NOX1, INOS, and PD-L1 in MDSCs. Additionally, we show that treatment of MDSCs with ATRA decreases MDSC’s ability to suppress T cell proliferation. These results indicate that ATRA may improve the efficacy of conventional and immnunotherapeutic treatments.
Correlation between strength of T-cell response against HPV16 and survival after vaccination with HPV16 long peptides in combination with chemotherapy for late-stage cervical cancer.
Authors: Melief Cornelis Joseph , Gerritsen Winald R. , Welters Marij , Vergote Ignace , Kroep Judith R. , Kenter Gemma , Ottevanger P. B. , Tjalma Wiebren Age , Denys Hannelore , Nijman Hans , van Poelgeest Mariette IE , Reyners An K.L , Velu Thierry J. , Blumenstein Brent A. , Goffin Frederic , Lalisang Roy Iqbal , Stead Richard B. , van der Burg Sjoerd 
Published: November 2019

140 Background: Therapeutic vaccination with HPV type 16 synthetic long peptides (HPV16-SLP) results in T cell–mediated regression of HPV16-induced premalignant lesions but fails to install effective immunity in patients with advanced HPV16-positive cervical cancer. We showed that HPV16-SLP vaccination in mice and in patients with advanced cervical cancer patients fosters robust HPV16-specific T cell responses, when combined with chemotherapy. In this study we noted that a single dose of vaccine 2 weeks into the 2nd cycle of chemotherapy was optimal, because at this time the immunosuppressive myeloid cells were down. Methods: We now completed a chemo-immunotherapy study in a larger number of patients with late stage HPV16+ cervical cancer. Three HPV16-SLP vaccine doses were given 2 weeks after the second, third, and fourth cycles of standard chemotherapy. Cohorts of 12 patients each were vaccinated with each of 4 dose levels (20, 40, 100, and 300 µg/ per peptide) of 13 overlapping HPV16 synthetic long peptides (HPV16-SLP) together covering the length of the 2 E6 and E7 proteins. Results: Robust vaccine-induced HPV16-specific T cell responses as assessed by interferon-g Elispot were observed and were sustained throughout the cycles of chemotherapy. These T cell responses were substantially increased in all patients who received HPV16-SLP . In addition, the chemotherapy augmented recall responses to microbial antigens. Such robust T cell responses were not noted in previous trials when similar patients were vaccinated without timing of vaccination during chemotherapy. A marked correlation was observed between the strength of the vaccine-induced immune response and longer-term clinical outcomes such as overall survival. No such correlation exists between the strength of the T cell response against common recall antigens and survival. In addition, a remarkably high proportion of patients survived beyond 20 months after the start of therapy. Conclusions: These results indicate that the survival advantage is specifically related to the strength of the vaccine-induced T cell response and is not due to generally better immuno-competence. Clinical trial information: NCT02128126.
Chimeric antigen receptor (CAR) T cells genetically engineered to deliver IL-12 to the tumor microenvironment in ovarian cancer.
Authors: Yeku Oladapo O. , Purdon Terence , Spriggs David R. , Brentjens Renier J. 
Published: November 2019

141 Background: Chimeric antigen receptor (CAR) T cell therapy for solid tumor malignancies has not shown the same degree of clinical efficacy observed in hematologic malignancies such as B-ALL. The presence of an immunosuppressive cellular and cytokine microenvironment has been hypothesized as one reason for the failure of adoptive immunotherapy for solid tumors. In ovarian cancer, myeloid derived suppressor cells (MDSC) and immunosuppressive cytokines in the ascitic microenvironment have been reported. IL-12 is a proinflammatory cytokine produced by macrophages, dendritic cells (DC), and NK cells, and it has been shown to increase proliferation of T cells, induce differentiation of type 1 T helper cells, inhibit regulatory T cells, promote maturation of DCs, and enhance antigen presentation by macrophages. We hypothesize that CAR T cells genetically modified to constitutively secrete IL-12 will overcome a hostile tumor microenvironment in a peritoneal carcinomatosis model of ovarian cancer. Methods: CAR T cells were generated from retroviral transduction of second generation and IL-12 modified CAR’s directed to either an irrelevant CD-19 antigen or Muc16ecto. C57BL/6 mice were inoculated i.p with syngeneic ovarian cancer cells and treated with various CAR T cells. Results: Here we report increased production of IL-12, improved proliferation and cytotoxic activity of 4H1128ζ-IL12 CAR T cells. Further, we show increased levels of inflammatory cytokines at 24 and 48hrs after treatment of tumor-bearing mice, leading to increased survival at advanced stages of disease. Animals treated with 4H1128ζ-IL12 CAR T cells had decreased levels of F4/80+ CD11b+MDSCs. Genetic analysis of recovered MDSCs from the ascites of treated animals showed skewing towards an M1-phenotype via upregulation of cytokines, chemokines, MHC-II, and downregulation of Arg1. Furthermore, clodronate-mediated depletion of TAM’s further enhanced survival in mice treated with 4H1128ζ-IL12 CAR T cells. Conclusions: These results demonstrate the mechanisms of efficacy of localized delivery of IL-12 to the tumor microenvironment by 4H1128ζ-IL12 CAR T cells.
Modulating lncRNA NKILA in tumor-reactive T cells to enhance their trafficking to breast cancer by inhibition of FasL-induced T-cell apoptosis.
Authors: Huang Di , Su Shicheng , Song Erwei 
Published: November 2019

143 Background: Adoptive cell therapy (ACT) is a highly personalized cancer therapy that involves administration to the autologous tumor-reactive lymphocytes with direct anticancer activity. In addition to the effective application of ACT for cancer treatment to genetically engineer lymphocytes to express conventional T cell receptors or chimeric antigen receptors, many of the ACT therapy has failed because of the trafficking barrier of T cells. Methods: In vitro, we established tumor-reactive CTL by vaccination with autologous DCs pulsed with MUC1-derived peptides. Moreover, tumor-reactive T cells were transferred to the immunocompromised mice inoculated with autologous patient derived breast cancer xenografts (PDX). Results: In our study, we found that during the induction of tumor-reactive T cells, cytotoxic T lymphocytes (CTL) may undergo massive expansion and have the ability of specific lysis to tumor cells upon appropriate antigenic stimulation via NF-κB pathway activation. However, hyperactive tumor-specific lymphocytes were hypersensitivity to apoptosis signals in tumor milieu, such as Fas ligand. In this research, we showed that NKILA, an NF-κB interacting lncRNA, is up-regulated at the later phase of CTL activation. The up-regulated NKILA suppresses NF-κB activity and sensitizes T cells to apoptosis. In vivo, NKILA knockdown in tumor-reactive T cells upon transferring to the immunocompromised mice inoculated with autologous PDX enhances CTL in xenografts, and thus effectively inhibits tumor growth. Conclusions: Collectively, our findings suggested that NKILA upregulation is responsible for the apoptosis of tumor-responsive CTLs and targeting the lncRNA may provide a novel strategy to enhance the therapeutic efficacy of immune cell transferring for breast cancer.
Phase I study of the safety and activity of formulated IL-12 plasmid administered intraperitoneally in combination with standard neoadjuvant chemotherapy in patients with newly diagnosed advanced stage ovarian cancer.
Authors: Thaker Premal H. , Bradley William Hampton , Leath Charles A. , Gunderson Camille Catherine , Borys Nicholas , Anwer Khursheed , Musso Lauren , Alvarez Ronald David 
Published: November 2019

155 Background: This study evaluated weekly intraperitoneal GEN-1, an IL-12 plasmid formulated with polyethyleneglycol-polyethyleneimine cholesterol lipopolymer, in combination with dose-dense weekly taxane (T) and carboplatinum (C) every 3 weeks in newly diagnosed epithelial ovarian, fallopian tube, or primary peritoneal cancer (EOC) patients undergoing neoadjuvant therapy (NAC). The primary objective of this study was to evaluate the tolerability and safety of GEN-1 with NAC. Secondary objectives were objective clinical response and pathological complete response at interval debulking surgery (IDB). Methods: This protocol followed a standard 3+3 design. Patients received GEN-1 at escalating dose levels from 36 mg/m2, 47 mg/m2, 61 mg/m2, to 79 mg/m2 weekly for 8 treatments with concurrent T/C. After IDB, patients did not receive further GEN-1 but continued on T/C. Patients were assessed for dose limiting toxicities (DLT) and had tumor samples at a time of diagnostic laparoscopy and IDB. Translational endpoints are yet to be analyzed. Patients are being followed for every 3 months for 2 years. Results: Fifteen patients were enrolled and 9 patients received all 8 treatments with no DLTs. The 79 mg/m2GEN-1 cohort of patients are actively being treated. Two patients were screen failures; 1 patient discontinued prior to GEN-1 administration due to port site infection. Most common related toxicities were Gr 1 nausea, fatigue, abdominal pain, and vomiting. One patient experienced Gr 2 fevers associated with GEN-1 but responded to acetaminophen and fluids. One patient did not undergo IDB due to pulmonary embolism and severe deconditioning due to underlying cancer. Of the other 8 patients undergoing IDB: 3/8 had stable disease, 4/8 had partial responses (PR), and 1/8 had complete response (CR). There was 1 pathological CR, 3 micro PR, and 3 macro PR. All patients had a ≥ 89% drop in CA-125 following GEN-1. Conclusions: Adding GEN-1 to T/C is safe and appears to be active in NAC patients with newly diagnosed EOC. Updated results to be presented at meeting. Clinical trial information: NCT02480374.
Identification of T-cell-inflamed gastric adenocarcinoma in The Cancer Genome Atlas (TCGA).
Authors: Maron Steven Brad , Luke Jason John , Hovey Raymond , Bao Riyue , Gajewski Thomas , Ji Yuan , Catenacci Daniel V.T. 
Published: November 2019

16 Background: Gastroesophageal adenocarcinoma (GEC) is a significant global health problem. KEYNOTE-012 demonstrated a 22% objective response rate in patients with PD-L1 expressing GEC that received pembrolizumab. A subset of patients (pts) tumors express a T cell “inflamed” (TCI) phenotype, which can be measured using an IFN-γ-based immune signature and may prove more predictive of clinical benefit. Using a 160 gene RNA-Seq immune expression profile, we sought to characterize the molecular environments of TCI versus non-TCI GEC patients in The Cancer Genome Atlas (TCGA). Methods: 395 GEC pts with primary tumors in TCGA were clustered into TCI, non-TCI, and intermediate subtypes using both unsupervised hierarchical and K-means clustering (k = 3). Molecular characteristics were categorized using data acquired via CbioPortal and the UCSC Xena repository. Only non-silent somatic mutations and copy number variations (CNVs) reaching GISTIC2 -2 or +2 thresholds were considered. Statistical comparisons were performed using chi-square, ANOVA, and t-test. Results: The TCI subtype contained patients from all TCGA-defined subtypes - EBV-associated (56%), MSI-high (16%), chromosomal unstable (6%), and genomically stable (27%). No significant differences were seen between TCI and non-TCI for tumor site or stage. Mutations in PTEN, PIK3CA, CDH1, and RHOA were more frequent in TCI patients. ERBB2, CCNE1, and KRAS CNVs were infrequent in TCI patients as were PDE4D deletions ( p< 0.05 ). TCI tumors had higher expression of both co-inhibitory (PD-1, PD-L1/L2, CD28/80, BTLA, LAG3) and co-stimulatory (CD137/40/27, OX40, GITR, ICOS) checkpoint molecules ( p< 10-7). Total mutation burden was no different between TCI and non-TCI pts when excluding MSI-high pts nor when assessing MSI-high alone. Conclusions: The IFN immune phenotype encompassed GEC patients from all TCGA subsets. Correlation of clinical outcome with checkpoint blockade is necessary to confirm these molecular associations and the independent predictive utility of this immune-profile stratification.
Immunotherapy and survival in stage IV melanoma: A National Cancer Database (NCDB) analysis.
Authors: Ahmed Aabra , Krishnan Mridula , Siu Margaret , Walters Ryan W , Silberstein Peter T. 
Published: November 2019

160 Background: Interleukin-2 (IL-2) has been the predominant immunotherapy used for stage IV melanoma for the past decades, with newer immunotherapy drugs approved within the past five years. By using the National Cancer Database (NCDB), our study aims to be the largest to determine the impact of immunotherapy on patients with stage IV melanoma. To examine only IL-2 and interferon-alpha, only patients diagnosed prior to 2011 were used. Methods: 10,771 patients diagnosed with stage IV melanoma between 2004-2010 were identified with sufficient data. 1,009 of these patients were given immunotherapy. Chi-square analysis was used to determine demographic differences between those with and without immunotherapy. Between-therapy survival differences were estimated by the Kaplan-Meier method and associated log-rank tests; Tukey-Kramer adjusted p < .05 indicated statistical significance. Results: Patients with immunotherapy had a median survival of 17.4 months, while those without immunotherapy had a median survival of 7.1 months. 27% of patients given immunotherapy were alive three years post diagnosis, compared to only 14% of patients who were not given immunotherapy. Immunotherapy patients had a greater percentage of those with private insurance (65.3% vs. 37.7%, p<0.001), an income greater than $63,000 (40.5% vs. 31.9%, p<0.001), and no comorbidities (87.3% vs. 77.5%, p<0.001) compared to patients not given immunotherapy. Conclusions: Patients with immunotherapy had nearly a 2.5-fold increase in median survival compared to patients not provided immunotherapy. Compared to those without immunotherapy, immunotherapy patients lived in more wealthy and educated zip codes, and had more private insurance. [Table: see text]