bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel TGF-beta targeted peptide HTPEP-001 enhances antitumor efficacy of anti-PD1 in mice models.
Authors: Li Xiaomei , Ding Yi , Xiao Ling , Chen Rui , Qing Ying , Jiang Qiu , Gao Yuanxue 
Published: May 2020

e15200 Background: With the emergence of immunotherapy in recent years, therapeutic antibodies against programmed death 1 (PD1) and its ligand (PDL1) have proven to be a huge success in multiple cancer types. However, these robust responses only occur in a subset of patients. Identifying mechanisms of responses and resistance is key to improve outcomes and setup new treatment strategies. Transforming Growth Factor-beta (TGF-beta) is an immunosuppressive cytokine and overexpression of TGF-beta in the tumor microenvironment is proven to be a key element of resistance to anti-tumor response. We have developed a novel peptide HTPEP-001 which blocks TGF-beta signaling in both cellular and fibrosis animal models (ATS-2020 abstract). In this study, we explored the antitumor effect and mechanism of HTPEP-001 when administered together with anti-PD1 antibody in mice models. Methods: Animal models: 1. C57BL/6 mice were inoculated subcutaneously with 5*105 MC38 cells. 2. BALB/c mice were inoculated on the right mammary fat pad with 1*105 EMT-6 cells. Treatment groups: A. HTPEP-001 (60 mg/kg, intravenously, qd); B. isotype IgG; C. rat anti-mouse PD1 (10 mg/kg, intraperitoneally, twice a week); D. Coadminstration of HTPEP-001 and anti-PD1 (combo). Treatments were initiated 4-5 days after inoculation and lasted for 10 days. ELISA, immunofluorescence and flow cytometry analysis of dissected tumors were performed. Results: Compared to anti-PD1 group, mice treated with both HTPEP-001 and anti-PD1 exhibited additional reductions in tumor size and 13% higher inhibition of tumor growth (59% in Anti-PD-1 vs 72% in combo) in MC38 model. In the EMT6 model, coadministration of HTPEP-001 and anti-PD1 also showed additional reduction in tumor burdens. Coadmistration of HTPEPE-001 and anti-PD-1 significantly decreased active TGF-beta1 and phosphorylated Smads expression in tumors in both MC38 and EMT6 models. Furthermore, immunofluorescence and flow cytometry analysis of CD8+ lymphocytes in tumor mass revealed markedly elevated number of cytotoxic tumor infiltrating lymphocytes in combo group compared to HTPEP-001 and anti-PD1 groups. Conclusions: This study showed that HTPEP-001 enhanced the anti-tumor activity of anti-PD1 via suppressing TGF-beta signaling and promoting infiltration of cytotoxic lymphocytes in two syngeneic tumor models. We will continue to develop HTPEP-001 as an add-on therapy to checkpoint inhibitors with a goal to improve the outcomes of current immunotherapy.
Intratumoral use of self-delivering RNAi to reprogram the tumor microenvironment and boost the antitumor response.
Authors: Dispersyn Gerrit , Lakshmikanthan Sribalaj , Yan Dingxue , Jedinak Andrej , Maxwell Melissa , Kadiyala Mani , Cardia James , Fricker Simon P 
Published: May 2020

e15206 Background: Immunotherapy targeting immune checkpoints has shown significant benefit in many cancers. However, systemic delivery can result in severe auto-immune toxicities, the risk increasing with the combination of checkpoint inhibiting antibodies. Local delivery of immunotherapies allows for multiple combination therapies, while potentially preventing significant systemic toxicities. One approach is to use self-delivering RNAi technology, INTASYL™, which has been shown to be safe and effective in clinical applications following local administration. Here we show the use of the INTASYL platform as a checkpoint inhibiting immunotherapy for intratumoral applications, through a series of preclinical in vivo studies. Methods: INTASYL compounds were made against mouse PD-1, TIGIT, PD-L1 and BRD4. INTASYL comprises an asymmetric duplex, a small duplex region (≤ 15 base pairs), a single-stranded phosphorothioate tail and chemical modifications to confer stability resulting in efficient cellular uptake allowing in situ delivery. The efficacy and immunomodulation of these INTASYL compounds was evaluated in the Hepa1-6 and CT26 murine syngeneic tumor models following intratumoral administration. Tumor volumes were recorded throughout the study. Tumor-Infiltrating-Lymphocytes (TILs) were isolated twenty-four hours post the last dose and examined for gene silencing by RT-qPCR. Tumor microenvironment (TME) and T cell activation were analyzed by flow cytometry. Results: Results showed dose-dependent attenuated tumor growth compared to control groups. Relevant changes in the TME included an increase of CD8+ T cells and an increase of activation markers on TILs. Tumor growth reduction correlated with silencing of target mRNA and immune effector cell levels and activation. Conclusions: These studies demonstrate the feasibility of using INTASYL compounds intratumorally to overcome the immunosuppressive TME, resulting in changes in T cell composition and activation, and inhibition of tumor growth. Together, these novel findings support using intratumoral INTASYL as a viable approach to immunotherapy and warrants further investigation in patients.
Modulation of CT-genes transcriptional activity in HT-29 cells by irradiation with a linear accelerator.
Authors: Kutilin Denis S. , Kosheleva Natalia G , Gusareva Marina A. , Legostaev Vladislav M. , Shlyakhova Olga V. , Dontsov Vladimir A. , Gaziev Umar Muhmadovich , Udalenkova Irina A. , Rozenko Lyudmila Y. , Donskaya Aliya K. , Rodionova Olga G. , Fatkina Natalya B. , Kit Oleg Ivanovich 
Published: May 2020

e15218 Background: Cancer-testis antigens (CTAs) are used as targets in immunotherapeutic approaches based on gene-cell vaccines (GCV). In some cases, it is possible to use experimental combinatorial therapy regimens that combine radiation and GCV. Despite numerous studies of the expression of CTA in tumors of various locations, including colon tumors, the transcriptional activity of CT-genes in tumor tissue of the colon after radiation therapy remains poorly studied. The aim of the study was to analyze changes in the expression of CT-genes in HT-29 cells after irradiation with a linear accelerator. Methods: The study used human cell culture HT-29. For the model experiment, 7 Gy fractionation option was used (irradiation was carried out 5 times every 24 hours). Irradiation was performed on a Novalis TX linear accelerator (Varian, USA). The ratio of living and dead cells was calculated in a Goryaev chamber using a 0.4% trypan blue dye solution. After microscopy on the 5th day of irradiation, the cell line was removed from the vial substrate by Trypsin/Versen solution. RNA was isolated by the Chomczynski&Sacchi method. For cDNA synthesis a set of "REVERTA-L" reagents used. Expression of 16 CT genes (MAGE-A1, -A2, -A3, -A4, MAGE-B1, -B2, GAGE-1, -3, -4, MAGEC1, BAGE, XAGE3, NYESO1, SSX2, SCP1, PRAME1) was determined using the Real-Time qPCR method (reference genes — GAPDH and B2M). Statistical analysis was performed using univariate analysis of variance (ANOVA) with repeated measurements. Results: After 5 days of exposure at a dose of 7 Gy, only 20% of the initial number of HT-29 cells remained viable. It was found that irradiation was statistically significantly (p < 0.05) influenced the increase in the expression of genes MAGEA1 by 1.8 times, MAGEB1 by 6.8 times, BAGE by 1.6 times, CTAGE1 by 3.3 times and SSX2 by 5.8 times relative to intact cells. Conclusions: Thus, tumor cells exposed to radiation at a dose of 7 Gy show an increase in the expression of 5 CT-genes, which, in turn, can increase their immunogenic potential.
Intratumor immunotherapy utilizing real time radiological image guidance: Early experience from a tertiary cancer center.
Authors: Murthy Ravi , Sheth Rahul , Subbiah Vivek , Janku Filip , Naing Aung , Rodon Jordi , Yap Timothy A , Diab Adi , Patel Sapna Pradyuman , Tam Alda , Gupta Sanjay , Meric-Bernstam Funda 
Published: May 2020

e15223 Background: Intratumoral (IT) delivery of immunotherapeutic agents is a compelling approach to overcoming the systemic barriers of toxicity and lack of efficacy associated with many novel agents. In this context, the application of IT delivery is growing rapidly, however the feasibility and safety profile of these interventions for lesions that require real time radiological image-guidance remains unknown. Methods: Patients who underwent IT injections of immunotherapeutic agents across several clinical trials over a 5 year period at a single tertiary cancer center were included in this analysis. Technical success & procedure related adverse events of the IT injection and the frequently accompanying concomitant or sequential image-guided biopsy of injected (adscopal) and non-injected (abscopal) sites were analyzed. Results: A total of 256 patients with metastatic &/or unresectable solid malignancies underwent a total of 1096 (median 5) image-guided IT investigational agent injections during the study period. There were no adverse events attributable to the technical component of the procedure, specifically during the needle insertions or 3862 concomitant biopsies. Soft tissue and nodal lesions in the extraparietal locations & visceral lesions in deeper, intraparietal locations and solid organs were also injected (adrenal, liver and lung). The median injected lesion tumor length was 3.3cm. Serious adverse events (NCI CTC AE ≥ 3) including dyspnea and severe flu-like symptoms developing within 24 hours of the injection & required hospitalization following 1.5% of injections. Conclusions: Intratumoral immunotherapeutic injections & tissue sampling with real time radiological image guidance are feasible across a wide range of regions and organs irrespective of agent and histology. Post-delivery anticipated & adverse events were rare & manageable.
Re-introducing immunotherapy in patients surviving immune checkpoint inhibitors-mediated myocarditis.
Authors: Peleg Hasson Shira , laufer-Perl Michal , Wolf Ido , Sivan Ayelet 
Published: May 2020

e15228 Background: Immune checkpoint inhibitors (ICI) have transformed the standard care in cancer treatment. Recent case reports describe ICI-mediated myocarditis with an atypical presentation and fatal potential which lead to permanent interruption of immunotherapy. We aim to characterize ICI-mediated myocarditis and re-introduction to immunotherapy. Methods: We retrospectively evaluated the presentation, severity, and prognosis of patients diagnosed with ICI-mediated myocarditis during 2019, and presented the clinical course and outcomes of patients that were chosen for re-introduction. Results: Among seven patients, only one patient had a history of cardiac disease. The majority were diagnosed with lung adenocarcinoma and treated with anti-programmed death-1 antibody (57%). All patients were treated with single agent ICI. Most patients presented with cardiac symptoms, elevated troponin and typical magnetic resonance imaging; however only 43% had reduced ejection fraction. Five patients were defined as grade I-II and two as grade III-IV. Overall, three patients were chosen for re-introduction with concomitant low dose steroids and weekly troponin follow-up. Two patients diagnosed with grade I/II renewed therapy successfully with no recurrence of symptoms and improvement in disease burden, while the one patient diagnosed with grade III developed worsening of cardiac symptoms after the 1st cycle and therefore therapy was interrupted permanently. Overall, survival was higher among the re-introduction patients (67% vs. 25%). Conclusions: ICI-mediated myocarditis is potentially fatal and leads to permanent interruption of life saving cancer therapy. We imply that re-introduction may be considered in low grade patients; however, better definition of the diagnosis and grading is needed.
Impact of corticosteroids use on efficacy of immune checkpoint inhibitors in cancer patients: A meta-analysis.
Authors: Li Jiarui , Yang Kaili , Zhao Lin , Bai Chunmei , Sun Zhao 
Published: May 2020

e15234 Background: Corticosteroids are commonly used for management of cancer-related symptoms or immune-related adverse events (irAEs) in cancer patients treated with immune checkpoint inhibitors (ICIs). However, given the inhibitory effects of corticosteroids on a broad range of immune responses, it is presumed that the use of these agents may affect the clinical outcomes of ICIs. This meta-analysis aims to explore the impact of corticosteroids use on the efficacy of ICIs in cancer patients. Methods: We conducted a search covering electronic databases (MEDLINE, EMBASE, and CENTRAL), conference abstracts (ASCO and ESMO) and reference lists to identify relevant studies. Studies that reported clinical outcomes of patients with corticosteroids administration before and/or after the initiation of ICIs treatment were eligible for evaluation. The primary outcomes included progression-free survival (PFS) and overall survival (OS). The random-effects model was utilized to synthesize the effect sizes of individual studies. Results: The initial literature search identified 1,900 records. After study selection, a total of 15 studies with 14,123 patients were included in the quantitative analysis. Corticosteroids use significantly reduced PFS (HR = 1.84; 95% CI: 1.30–2.61; P = 0.001) and OS (HR = 1.41; 95% CI: 1.18–1.68; P < 0.001) in cancer patients treated with ICIs. In subgroup analysis, corticosteroids use for cancer-related symptoms was associated with a shorter PFS (HR = 1.98; 95% CI: 1.40–2.78; P < 0.001) and OS (HR = 1.88; 95% CI: 1.25–2.83; P = 0.003), but their use for irAEs did not show a detrimental impact on OS (HR = 1.05; 95% CI: 0.77–1.44; P = 0.740). Conclusions: This meta-analysis indicated that corticosteroids use might hinder the efficacy of ICIs in cancer patients. The indications of corticosteroids use should be strictly controlled during the course of immunotherapy.
Immunohistochemical determination of PD-L1 expression with three different antibody clones and determination of SOX10 expression in triple negative breast cancer.
Authors: Guhl Margit Maria , Bohle R.M. , Ertz Martin , Kasoha Mariz , Hammadeh Mohammed Eid , Juhasz-Boess Ingolf , Solomayer Erich-Franz 
Published: May 2020

e15237 Background: In triple negative breast cancer (TBNC), checkpoint inhibitors directed against PD-L1/PD-1 show an improvement of therapy. For the immunohistochemical diagnosis of the predictive biomarker PD-L1, there are various antibodies and kits available. We examined the expression of PD-L1 in TNBC with different antibody clones to compare these results with regard to the staining of tumor cell membranes, staining of immune cells and cytoplasmic staining in order to find out whether the different clones or methods can be exchanged. It was also checked whether PD-L1 or SOX10 expression correlates with the existing clinical parameters. Methods: Breast cancer tissue of 60 patients with TNBC were examined for the expression of PD-L1 and SOX10 by immunohistochemistry. The detection kit used, was the ultraView universal alkaline phosphatase red detection kit for the antibodies anti-human PD-L1 clone 22C3 from Dako, the clone 28-8 from abcam and the SOX10 antibody clone SP267 from Cell Marque. The anti-PD-L1 antibody clone SP142 was detected with OptiView DAB. The cut-offs for the expression of PD-L1 at the tumor cell membrane were < 1%, > 1 to < 50% and > 50%. In the evaluation of SP142 the staining of the immune cells was evaluated with the score: percentage of PD-L1 positive immune cells to the tumor cells that were present. For SOX10, the nuclear staining was evaluated with the score: < 1, > 1 to 50, > 50 to > 100 and 100. The relationship between PD-L1 expression (TC and IC) and SOX10 expression was evaluated with the clinical parameters of the patients from the time of diagnosis until the end of data collection. Results: The antibodies clone 22C3 and clone 28-8 lead to the same results (22,0 % PD-L1 (TC) positive). Clone SP142 showed significantly (p < 0,001) more PD-L1 positive cases(40,7%). For the additional evaluation of the cytoplasmic staining with clone 22C3 and clone 28-8 it could be shown that the PD-L1 Expression is equivalent (40,7%) to the immune cell staining with clone SP142. With exception of Ki67 we were unable to demonstrate any correlation between PD-L1 (membrane and cytoplasmic), SOX10 and other clinical parameters in TBNC. Conclusions: The antibodies clone 22C3 and 28-8 can be used equivalently for PD-L1 determination in TBNC. Clone SP142 showed different results. The cytoplasmatic staining with 22C3 and 28-8 could gain importance in the future because the results were equivalent to the immune cell staining with clone SP142. There is no correlation between PD-L1 and SOX10 in TNBC.
Tumor lymphocytic infiltration impacts recurrence in endometrioid-type endometrial carcinoma.
Authors: Avila Monica , Fellman Bryan M. , Broaddus Russell 
Published: May 2020

e15239 Background: Endometrial cancers that have mismatch repair deficiency are associated with higher numbers of tumor-associated lymphocytes, but the clinical significance of this observation is unknown. Our objective was to quantify CD3+ and CD8+ tumor lymphocytes of MMR intact (MMRi) and MMRd endometrioid-type endometrial carcinomas and determine if there was an association with survival. Methods: MMRd was defined as endometrial carcinomas with loss of MLH1 expression due to MLH1 gene methylation and determined by immunohistochemistry. MMRi was defined as positive expression of MLH1, MSH2, MSH6, and PMS2. This was followed by Aperio image-based quantification was used to assess CD3+ and CD8+ lymphocyte populations in different regions of the primary endometrial carcinomas, including tumor periphery (tumor-myometrial interface), tumor center (bounded on all sides by tumor), and tumor hotspot (area with highest number of lymphocytes). Recurrence-free survival was estimated using Kaplan Meier and Cox regression. Median follow up time was 44 months. Results: 180 patients with endometrial carcinoma were analyzed of which 132 were MMRi and 48MMRd. The MMRd group had significantly higher levels of CD3+ and CD8+ lymphocytes regardless of which tumor region was assessed (Figure 1a, P < 0.001). Lymphocyte counts in both MMRd and MMRi groups had wide standard deviations such that there was some overlap in counts between the groups (Figure 1). Both MMRd and higher CD3+ counts were associated with worse recurrence-free survival. CD3+ quantification in the tumor periphery captured 21/23 recurrences (Figure 1b, HR = 8.04; 95% CI: 1.88 -34.31; p = 0.005); this included all of the MMRd cases that recurred and 7 MMRi cases with higher numbers of CD3+ lymphocytes that also recurred. Conclusions: MMRd endometrial cancers have increased numbers of CD3+ lymphocytic infiltrates within the primary tumor. Higher CD3+ infiltration is associated with greater risk of recurrence regardless of tumor location. In predicting tumor recurrence, lymphocytic counts performed better than assessment of MMR. Thus, quantification of CD3+ lymphocytes should be explored as a predictive biomarker.
Effect of CCND1 amplification on immunosuppression and the association with a poor prognosis to immune checkpoint inhibitors in solid tumors.
Authors: Huang Yingying , Chen Chuan-ben , Chen Yu , Gao Xuan , Li Yi , Lin Jing , Guan Yan-Fang , Xia Xuefeng 
Published: May 2020

e15249 Background: CCND1 amplification relevant to malignant biological behavior exist in solid tumors. The prevalence and utility of CCND1 amplification as a biomarker for the clinical response to immune checkpoint inhibitors (ICIs) therapy is unknown. Methods: This study included three cohorts: Geneplus Institute ( n= 6536), The Cancer Genome Atlas ( n= 10562) and Memorial Sloan Kettering Cancer Center ( n= 106109). Comprehensive profiling was performed to determine the prevalence of CCND1 amplification and their correlation with the prognosis and the response to ICIs. Results: CCND1 amplification occurs in many cancer types, correlates with shorter overall survival and inferior outcomes with ICIs therapy. Transcriptomic analysis showed various degrees of immune cells exclusion, including cytotoxic cells, T cells, CD8+ T cells, DC cells, B cells in the tumor microenvironment (TME) in a CCND1 amplification population. The gene set enrichment analysis suggested that CCND1 amplification correlates with multiple aggressive, immunosuppressive hallmarks including epithelial mesenchymal transition, TGF-β signaling, KRAS signaling, PI3K/AKT/mTOR signaling, p53 pathway and hypoxia signaling in solid tumors. Conclusions: Our study indicated that CCND1 amplification may be a key point related to immunosuppression in TME and multiple malignancy hallmarks, and it hinders not only the natural host immune responses but also the efficacy of ICIs in solid tumors.
Prevalence of prkdc mutations and association with response to immune checkpoint inhibitors in solid tumors.
Authors: Li Yi , Chen Chuan-ben , Chen Yu , Guan Yan-Fang , Huang Yingying , Lin Jing , Chen Gang , Xia Xuefeng , Chang Lianpeng , Yi Xin 
Published: May 2020

e15250 Background: Predictive biomarkers of response to immune checkpoint inhibitors (ICIs) help to identify cancer patients who will benefit from immunotherapy. PRKDC is an important gene relates to DNA double-strand break (DSB) repair and central T-cell tolerance. We aim to investigate the association between PRKDC mutations and tumor mutation burden (TMB), tumor microenvironments (TME), and response to ICIs. Methods: The whole exome sequencing data of 4022 solid tumor samples from the Cancer Genome Atlas (TCGA) and the panel-based sequencing data of 4652 solid tumor samples from GenePlus-Beijing, China were used to analyze TMB. The mRNA expression data of 3541 solid tumor samples from TCGA was used to explore the effect of PRKDC mutation on TME. Four ICIs-treated cohorts were analyzed for verifying the correlation between PRKDC mutation and the response to ICIs. Results: In TCGA datasets and GenePlus datasets, we both found that the TMB in PRKDC mutation samples were significantly higher than PRKDC wild-type samples(P < 0.05, P<0.0001 respectively). When compare to other pivotal DNA damage response (DDR) genes, it showed an identical effect of the degree in TMB by PRKDC mutation with MMR ( PMS2/ MLH1/ MSH2/ MSH6) genes, POLE/D1 and BRCA1/2 mutation (P > 0.05). Further, in TCGA datasets, it showed that PRKDC mutation samples were associated with significant expression increase of CD8+ T cells, NK cells, immune-checkpoint, chemokines etc compared to PRKDC wild-type samples (P < 0.05). In ICIs-treated cohorts, we also found the PRKDC mutation was associated with superior survival(median PFS, not reached (NR) vs. 6.8 months HR, 0.2893 95% CI, 0.1255-0.6672 P = 0.0650, Hellmann cohort; median OS, 1184 days vs. 250 days HR, 0.5126 95% CI, 0.2715-0.9679 P = 0.1020, Allen cohort), and the superiority was significantly in multivariate Cox regression analyses (HR, 0.361 95% CI,0.155-0.841; P = 0.018, Allen cohort; HR, 0.240 95% CI,0.058-0.998; P = 0.050, Hellmann cohort). Conclusions: PRKDC mutations are associated with an increased TMB, an inflamed TME and a better response to ICIs, it may be a potential predictive biomarker for ICIs-immunotherapy.