bluuebunny/crossref_metadata_2025 huggingface dataset:
Metabolism of tumor cell in tumor micro environment assist immune escape.
Authors: He Ning , Wang Jianfei , Wang Lingyu , Jin Ge , Lin Rongbo , Yang Yadong
Published: May 2020

e15181

Background: Tumor cell’s metabolism has been identified assisting immune evasion in tumor micro environment (TME), and it is very different from that of immune cells. Previous studies mostly focus on bulk or cell lines transcriptome; we aim to find the specific metabolic model of tumor cell based on vast single cell RNA-seq data. Methods: Public transcriptome of 7186 cells (SMART-Seq2) from 33 melanoma tumors in Benjamin’s study were collected from Single Cell Portal of Broad Institute, and metabolic genes were obtained from Human Metabolome Database (HMDB). Raw count data was preprocessed and analyzed using Seurat R package. Statistical analysis and data visualization were all carried out using R software. Results: Metabolic signature genes of malignant cells (Mals), CD8+ T cells, CD4+ T cells, B cells, natural killer (NK) cells, macrophages, cancer associated fibroblasts (CAFs), and endothelial cells (Endos) were extracted using Seurat. Mals, CAFs, Endos and macrophages shared similar metabolism patterns (high-level glycolysis, pyruvate metabolism, purine metabolism and specific proteoglycans profile compared to immune cells, p < 0.01). Glutathione metabolism activity in Mal was higher than that in immune cell, especially GPX1, GPX4 and GSTO1 genes were highly expressed in Mal ( fc > 2, p < 0.01). Compared to immune cells, genes participate in glycolysis, PFKL, ADH5, ENO2, G6PC3, PGM1, ALDH1B1, PFKM, ALDH7A1, GAPDHS, TPI1, PGK1, LDHA, GPI, PKM, LDHB, ENO1, ALDOA and GAPDH were highly expressed in Mals, CAFs and Endos ( fc > 2, p < 0.01). Noteworthily, LGALS1, an inducer of T-cell apoptosis, was highly expressed in Mals and CAF ( fc > 2, p < 0.01). Otherwise, proteoglycans (PGs) in cancer, which contribute to cancer pathogenesis, exhibited high variation from immune cells. Conclusions: We firstly highlight the differential metabolism between tumor and immune cells in TME. Mals, CAFs, endothelial cells and macrophages can assist in evading immune surveillance through reshaping their metabolic patterns at cell level. This study is helpful for exploring new target of metabolic based anti-cancer immunotherapy.
Immune subtype of mismatch repair deficient colorectal carcinoma.
Authors: He Ning , Jia Qingzhu , Jin Ge , Zhu Bo
Published: May 2020

e15194

Background: Half of microsatellite instability-high (MSI-H) colorectal carcinoma (CRCs) patients do not response to checkpoint blockade (ICB) immunotherapy. We aimed to unleash the underlying mechanism of distinct efficacy to ICB by sophisticated depicting the immune microenvironment of MSI-H CRC. Methods: Public omics and clinical data of patients with MSI labels were collected from TCGA data portal (364 samples) and Gene Expression Omnibus (GSE39582, GSE13294, GSE13067, GSE35896, GSE24514, GSE25071, GSE79959, GSE103340, GSE29638, GSE33113 cohorts, 1119 samples). GEO datasets were used as discovering sets, and TCGA dataset was used as validating sets. The immune cell infiltrations and signaling pathways in the tumor microenvironment were quantified by single sample Gene Set Enrichment Analysis (ssGSEA), and the bio-similarities were estimated by t-Distributed Stochastic Neighbor Embedding (tSNE). Clustering was identified by k-mean unsupervised strategy with optimal k. Statistical analysis and data visualization were all carried out using the R. Results: Based on the whole transcriptome and immune cell infiltration profiling, both GEO and TCGA cohorts showed that MSI-H patients harbored unique pattern which significantly differed from the MSS counterpart. Unsupervised clustering further classified the MSI-H patients into “Hot” and “Cold” subgroups with remarkable different IFNG-related signatures, inflammatory response score, interferon-γ signaling score, and survival outcome. Screening assay identified 23 genes ( ADAMTSL3, DLGAP4, DNAH11, FFAR3, GABRA4, KCNA6, KCTD9, KIAA1033, KLHL6, LARGE, MORC1, NDST3, OR5H14, PAX5, PHF3, PIK3R5, RNF25, SPATA20, STAT4, TLR9, TMC3, TTC12, ZNF790) enriched and 5 genes ( CELSR3, DGKD, HECTD4, RECQL4, SLC9A4) spared in immune “hot” tumors. Moreover, “hot” tumor showed less WNT signaling pathway score, which was negatively correlated with the IFNG signaling and inflammatory response within the MSI-H CRC tumors. Conclusions: We firstly identified immune hot and cold groups from MSI-H CRCs, which may be related to different immunotherapy response. This result may help further understanding the anti-immunotherapy of MSI-H CRCs.
Pd-1 inhibitors plus nab-paclitaxel with S1 (AS) as first line in patients with advanced biliary tract cancer.
Authors: Gou Miaomiao , E Liu Tie , Yan Huan , Si Haiyan , Wang Zhikuan , Qian Niansong , Dai Guanghai
Published: May 2020

e15195

Background: The way to treat advanced biliary tract cancers (BTCs) including Intrahepatic cholangiocarcinoma, extrahepatic bile duct cancer, gallbladder cancer was disappointing. Hence, we explore the efficacy and safety of pd-1 inhibitors plus chemotherapy (nab-paclitaxel with S1, AS) as first line in patients with BTC. Methods: BTC patients from oncology department of Chinese PLA general hospital receiving PD-1 inhibitors with AS or AS alone were retrospectively analyzed. The primary endpoint was progression-free survival (PFS). The second endpoint was objective response rate (ORR), disease control rate (DCR) and safety. Results: 32 patients given pd-1 inhibitors (Pembrolizumab, Nivolumab, Sintilimab, Toripalimab) plus AS and 26 patients used AS alone were included in the study. The median PFS was 5.43m in combined group, significantly longer than the 2.82m with monotherapy alone group (HR 0.46, 95% CI 0.25–0.84, P = 0.01). OS was not matured in the pd-1 inhibitors group. PFS of Intrahepatic cholangiocarcinoma subgroup seemed better than extrahepatic group in the AS group (5.54m VS 2.57m, P = 0.004).however, there was no evidence of difference in intrahepatic group compared the other group in the pd-1 inhibitor group (5.07m vs 5.43m, P = 0.453). liver metastatic was poor predictor of PFS in both Pd-1 combined or not group (2.43m vs 7.63m, P = 0.01; 2.14m vs 5.54m, P = 0.04 ).ORR was 25% and DCR were 84.3% in Pd-1 inhibitor group and ORR was 15.3% and DCR was 69.2% in chemotherapy alone group respectively. No significant difference was seen for ORR and DCR compared between two groups (P = 0.364, P = 0.169). Grade 3 or 4 treatment-related adverse events(AE) in pd-1 inhibitor group were hypothyroidism (n = 3), rash (n = 2), hepatitis (n = 1). No AE related death. Myelosuppression was similar between the anti-PD-1 combination group and the chemotherapy alone group (12.5% and15.3 %, respectively). Conclusions: Anti-PD-1 therapy plus nab-paclitaxel with S1 prolonged the PFS and improved the ORR, DCR compared the nab-paclitaxel with S1 alone in advanced BTC with controllable AE. Further clinical trials were needed and we conduct the phrase II clinic trail of Toripalimab with AS (TSA-01 study) is ongoing (clinical registration number: NCT04027764).
Novel synthetic immune modulators for the activation of tumor antigen-specific T cells.
Authors: Zhang Di , Shi Lihua , Tam Susan , Fung Man-Cheong
Published: May 2020

e15203

Background: Although checkpoint inhibitor immunotherapy and adoptive T-cell therapy revolutionized cancer treatments, such approaches suffer either from lack of target specificity for checkpoint inhibitors or inability to target intracellular tumor-related antigens from CAR-T therapy. Here, we report the development of novel Tavo Immune Modulator (TIM) biologics molecules which can specifically recognize tumor antigen-specific T cells through an engineered pMHC complex with peptides derived from intracellular tumor-related antigens. These molecules can selectively activate such T cells through engineered T cell co-stimulatory modulators for enhanced tumor cell killing. Methods: NY-ESO-1 and MAGE-A10 TIM molecules were constructed as fusions of HLA-A*02:01 MHC complexed with either NY-ESO-1 (157-165) or MAGE-A10 (254-262) epitope peptides at the N-termini and various T cell costimulatory modulators at the C-termini of IgG heavy and light chains. TIM molecules were expressed in Expi293 cells and purified by Protein A affinity chromatography. Specific binding of TIM with cancer specific T cells was evaluated by immunostaining. The activation and proliferation of tumor specific CD8+ T cells were confirmed in T cell activation and recall assays. Results: Both NY-ESO-1 and MAGE-A10 specific TIM molecules were generated which recognized corresponding tumor specific T cells. NY-ESO-1 TIM engineered with IL2 could activate NY-ESO-1 specific CD8+ T cell exclusively. Engineering additional T cell costimulatory factors along with IL2 on NY-ESO-1 TIM molecule could further boost T cell proliferation and activation in T cell recall assays. Besides NY-ESO-1, combinations of T cell costimulatory factors with MAGE-A10 TIM molecules enhanced specific T cell activation. Additional in vitro and in vivo studies are ongoing to demonstrate efficacy of such novel TIM molecules in eliminating different types of NY-ESO-1 and MAGE-A10 which are over-expressed on tumor cells. Conclusions: This study demonstrates the utility of NY-ESO-1 and MAGE-A10 TIM molecules in the selective recognition and activation of tumor antigen-specific T cells. Such novel biologics molecules may provide target specificity in tumor treatment, and potential targeting of intracellular tumor-related antigens presented as peptides in MHC complexes on cell surfaces. Selective activation of tumor-specific T cells may provide a unique method for the treatment of various solid tumors and warrants further investigation.
The potential biomarkers of checkpoint inhibitor immunotherapy in solid malignancies.
Authors: Hakim Nausheen , Chi Jeffrey , Patel Rajvi , Quinn Ryann , Zhu Xinhua
Published: May 2020

e15226

Background: Tumor mutational burden (TMB), PDL1 status, microsatellite status (MS), tumor infiltrating lymphocyte, and the microbiome profile are commonly used biomarkers to predict which patients (pts) may benefit from immunotherapy (IO). However, they are imperfect and cannot be used to predict complete response (CR). Next Generation Sequencing (NGS) from Foundation Medicine has identified many gene mutations. We aim to identify potential biomarkers in pts who achieved CR with immunotherapies as opposed to in those who had upfront progression of disease (PD). Methods: We conducted a retrospective analysis of pts with metastatic solid tumors at our institution that received IO after NGS testing. Pts who achieved CR or PD per imaging studies and clinical settings were included. Gene alterations through NGS were identified in each group. Results: Eighty-one pts received IO who had NGS sent prior to treatment. Pembrolizumab, ipilimumab, nivolumab, durvalumab and atezolizumab were used as single agent or in combination with another IO or chemotherapy. 19 pts achieved CR, 18 pts had PD. In the CR group, melanoma and lung adenocarcinoma were most common tumor types (Table). 15 pts (78.9%) received IO as 1st line treatment. Most common gene mutations in CR group were ROS1 (52.6%), LRP1B (52.6%), GRIN2A (47.4%), and EPHA7 (42.1%) in 19 patients. 18 pts with various tumor types (Table) had PD. Most of IOs were given as 2nd or 3rd line setting in NR group. Most common gene alterations were MLL2 (33.3%), CDKN2A/B (27.8%), NTRK1 (27.8%), ARID1B (22.2%) in 18 patients. Conclusions: The cohort of patients who achieved CR with IO had the most common gene mutations of ROS1, LRP1B, GRIN2A, and EPHA7 which can potentially serve as a biomarker to predict response to IO. Increased frequencies of MLL2, CDKN2A/ B, NTRK1, ARID1B mutations were found in patients with PD. The gene mutations in CR and PD groups are mutually exclusive. Further explorations at tumor environment, RNA and proteins levels are needed to identify combined potential biomarkers to predict IO response, followed by validation in the prospective study. The limitation is that the study is retrospective. [Table: see text]
RAD51Bme as predictive biomarker for PD-1 blockade response in non-small cell lung cancer.
Authors: Guerreiro Inês Maria , Barros-Silva Daniela , Lopes Paula , Cunha Ana Luísa , Lobo João , Antunes Luís , Rodrigues Ana , Soares Marta Alexandra Silva , Henrique Rui , Jerónimo Carmen
Published: May 2020

e15235

Background: Lung cancer (LC) cells frequently express high levels of programmed death-ligand 1 (PD-L1). Although these levels grossly correlate with likelihood of response to specific checkpoint inhibitors, prediction of response is rather imperfect and, thus, more accurate predictive biomarkers are mandatory. We examined the methylation profile of RAD51B, a DNA-repair gene, as candidate predictive biomarker for efficacy of anti-PD-1 therapy in patients with non-small cell lung cancer (NSCLC), correlating with patients’ outcome. Methods: PD-L1 immunoexpression and RAD51Bme levels were analysed in samples of NSCLC obtained from patients not treated with anti-PD-1 blockade (testing cohort) and patients treated with anti-PD-1 (validation cohort). Results: Of a total of 127 patients assessed, 58.3% depicted positivity for PD-L1 (PD-L1+). RAD51Bme levels significantly associated with PD-L1 immunoexpression. Patients with clinical benefit from immunotherapy disclosed higher RAD51Bme levels (p = 0.04) and significantly higher median progression free survival (PFS) (p = 0.02). PD-L1+ was also associated with longer overall survival (p = 0.03). Conclusions: We demonstrated that RAD51Bme levels might be combined with validated predictive biomarker PD-L1 immunostaining to select patients who will most likely experience clinical benefit from PD-1 blockade. The predictive value of RAD51Bme should be confirmed in prospective studies.
CIMAC-CIDC CyTOF harmonization.
Authors: Rahman Adeeb , Sahaf Bita , Davila Melanie , Fernandez Nicholas , McWilliams Emily , Millerchip Karen , Bentebibel Salah E , Haymaker Cara L. , Sigal Natalia , Duault Caroline , Thrash Emily , Del Valle Diane , Espiridion Beatriz Sanchez , Pichavant Mina , Bernatchez Chantale , Wistuba Ignacio Ivan , Gnjatic Sacha , Bendall Sean , Maecker Holden ,   CIMAC-CIDC Network
Published: May 2020

e15242

Background: The Cancer Immune Monitoring and Analysis Centers – Cancer Immunology Data Commons (CIMAC-CIDC) network is a National Cancer Institute-funded initiative to identify biomarkers of mechanisms and response to cancer immunotherapy clinical trials, using state-of-the-art assay technologies. A primary platform for CIMAC-CIDC biomarker studies is CyTOF mass cytometry, which is performed at all four CIMAC laboratories. Methods: To test the ability to generate comparable data across labs, a cross-site harmonization effort was undertaken. We first harmonized SOPs between centers. Because of a new acquisition protocol introduced by the vendor (Fluidigm), we also tested this protocol across sites before finalizing the harmonized SOP. We then performed a cross-site assay harmonization experiment, using 5 shared cryopreserved PBMC samples and one lyophilized control cell preparation, along with a shared lyophilized antibody cocktail consisting of 14 markers, as validated in the HIPC consortium, plus CD45. These reagents and samples were distributed to the four sites, and FCS files were centrally analyzed by both manual gating and automated methods (Astrolabe). Results: Average CVs across sites for each cell population were reported and compared to a previous multisite CyTOF study. Once a cell recovery issue at two sites was resolved, this experiment resulted in inter-site reproducibility of under 20% CV for most cell subsets, very similar to the previous study. Conclusions: These results emphasize the ability to reproduce CyTOF across sites, and also highlights procedures, such as use of spike-in control samples, useful for tracking variability in this assay.
Comprehensive next-generation sequencing to portray distinctive molecular characteristics of hypermutant lung cancer.
Authors: Zhang Hongbin , Wang Yuan , Ji Qiaoxia , Cai Hongmei , Liang Xiangcun , Xie Jiong , Li Hua , Wang Jun , Zhu Guiyun , Tian Erpeng , Zhu Lingling , Cui Lingzhi , Su Wenzhong , Yuan Mingming , Chen Rongrong , Zhao Min
Published: May 2020

e15255

Background: Tumor mutation burden (TMB) has been confirmed to predict the sensitivity to immunotherapy across multiple tumor types. Multiple genetic factors have been confirmed to increase the level of TMB, such as mutations in DNA damage repair (DDR) genes, POLE/POLD1, and high microsatellite instability (MSI). However, the extent that these factors contribute to hypermutation in lung cancer has not been fully investigated. Methods: We retrospectively reviewed the genetic profiles of 1000 lung cancer patients (pts) who underwent 1021-panel matched tumor-normal next-generation sequencing using tumor tissue samples and peripheral blood. Their TMB status were analyzed to determine the threshold of hypermutation. The clinicopathological characteristics, genetic profiles and genetic factors related to hypermutation were investigated for the pts in hypermutant cohort. Results: The threshold of hypermutation was determined as 19 muts/MB (top 5% in 1000 pts). As a result, 53 pts were included in the hypermutant cohort. A total of 1725 nonsynonymous somatic variants in 506 genes were identified. The most frequently mutated genes included TP53 (88.7%), LRP1B (71.7%), MLL2 (35.8%), EPHA5 (34.0%), and FAT1 (34.0%). KRAS was mutated in 17% pts, whereas mutations in EGFR, BRAF, ERBB2, MET were identified less commonly. MSI-high was observed in 5 cases. A germline mutation in BRCA1 gene was identified in an adenocarcinoma patient. Compared to genetic profiles of non-small cell lung cancer from TCGA database, mutations in multiple DDR genes were enriched in the hypermutant cohort (Table). No known driver mutation in POLE/POLD1 was identified. Conclusions: MSI-high and mutations in DDR genes may be associated with high level of TMB, whereas POLE/POLD1 driver mutations may not be related to hypermutant lung cancer. Hypermutant lung cancer displays distinctive molecular features that may be used as complementary indicators to screening pts sensitive to immunotherapy. [Table: see text]
Clinical value of piR-36026 and piR-651 in esophageal squamous cell carcinoma.
Authors: Peng Wei , Zhu Jun , Liu Delin , Qian Pudong , Zhang Qin , Li Houhuai , Cao Guochun
Published: May 2020

e15265

Background: Piwi-interacting RNAs (piRNAs) are a novel type of small noncoding RNAs which have important biological functions in malignant diseases. However, little is known about the clinical value of piRNAs in esophageal squamous cell carcinoma (ESCC). Hence, we demonstrated that two representative piRNAs, piR-36026 and piR-651, to dig out their clinical value in ESCC. Methods: Quantitative real-time PCR was employed to detect the levels of piR-36026 and piR-651 expression in ESCC tissues. Furthermore, the correlation between them and ESCC patients' clinicopathological characteristics were explored. Receiver operating characteristic (ROC) curves were constructed for measuring significances of piR-36026 and piR-651 for ESCC diagnosis. Results: Both of piR-36026 and piR-651 were prominently increased in ESCC tissues compared with normal. Additionally, piR-36026 was related with ESCC patients' diameter (P = 0.013), TNM stage (P = 0.027), tumor differentiation (P = 0.008) and CEA (P = 0.013); and piR-651 was associated with diameter (P = 0.005), lymph node metastasis (P = 0.002), tumor differentiation (P = 0.008) and SCC (P = 0.002). The area under the ROC curve (AUC) of piR-36026 was up to 0.8634 (95% confidence interval: 0.8171-0.9089, P < 0.05). Similarly, the AUC of piR-651 was 0.8525 (95% confidence interval: 0.7804-0.9247, P < 0.05). Conclusions: These results demonstrated that both piR-36026 and piR-651 may play a crucial role in ESCC oncogenesis; and they may be diagnostic biomarker for ESCC.
Multipeptide vaccine to prevent epithelial ovarian cancer relapse by microenvironment immunomodulation.
Authors: Marquez-Manriquez Juan Pablo , Camacho-Hernandez Alejandro , Lucero-Diaz Pedro Alejandro , Gallardo-Rincon Dolores , Rosas-Delgado Martin Orlando , Barron Karla Ramirez , Durazo Alberto , Ruiz Jorge Alberto Cisneros , Meneses-Velazquez Diana Karen , Durazo-Acuna Alberto
Published: May 2020

e15268

Background: Typically cancer vaccines for relapse prevention in epithelial ovarian cancer (EOC) are administrated in distant anatomical sites away from the primary tumor location. Ideally for EOC relapse prevention vaccines we should explore different anatomical sites related with the original tumor location and nearby lymph nodes, in order to observe if the administration would have better clinical impact due to a potential access to the tumor microenvironment. Moreover most trials for EOC relapse prevention use one or few targets and is mainly unknown if they are relevant to promote EOC relapse. We treated EOC patients with a multi-peptide vaccine administered in lymph nodes (LN) and in the peritoneum area and were able to suppress cancer relapse with not significant adverse effects after eight years. Methods: We enrolled n = 25 EOC patients in our study and identified by systematic review and validated by siRNA six proteins involved in EOC relapse and fourteen peptides were predicted. Twelve peptides were immunogenic by Granzyme B ELISPOT, DTH and ELISA. Patients were immunized with the vaccine in axillary and inguinal LN subcutaneously every week for four weeks, every two-weeks four times and finally every month for a total of thirty-six months. Clinical and laboratory evaluations were performed every 2-months and at the end of the study. Results: 100% of the patients developed both antibodies against all the peptides by ELISA and CD8 specific cells. The peptides of the six proteins were able to induce T cell expansion in both peptides and purified protein (p = 0.0001) and (p = 0.05), respectively for Ape-1. The selected targets were all able to induce DTH and ELISA responses with statistical significance. Peptides predicted from Bcl-2, survivin, Ape-1, Sox2, EGFR and RCAS1/EBAG9 were able to increase and sustain the levels of Granzyme B ELISPOT. Bcl-2 A peptide (p = 0.001), Bcl-2 B (p = 0.005), Bcl-2 C (p.0002), survivin A (p = 0.001), Survivin C (0.05), Ape-1 A (0.0001), Ape-1 D (p = 0.001), Sox2 A (p = 0.003), Sox2 B (p = 0.0001), EGFR B (p = 0.0001) and EGFR D (p = 0.001). Conclusions: Treating EOC patients in remission in zones hardly explored previously in order to potentially improve the activation of antigen-specific Th1 and CD8 cells with specific peptides seems promising as a feasible and safe approach to prevent EOC relapse.
Enhancement of the therapeutic effects of oncolytic vaccinia virus by using autologous and allogeneic cell-based delivery platforms.
Authors: Minev Boris , Santidrian Antonio F. , Minev Ivelina , Nguyen Duong , Zamarin Dmitriy , Marincola Francesco , Draganov Dobrin Draganov
Published: May 2020

e15271

Background: Clinical trials with oncolytic viruses for cancer treatment have shown limited efficacy due to viruses’ rapid clearance by patients’ innate and adaptive immune systems. In a recent first-in-human clinical trial, we confirmed the safety and feasibility of our approach to enhance oncolytic vaccinia virus (OVV) delivery and improve tumor targeting by utilizing an autologous cell-based cell delivery system (auto-OVV). We also developed an allogeneic cell-based platform (SuperNova1c - SNV1c) aiming to protect and potentiate OVV’s antitumor effects in large patient populations. Methods: We evaluated the immunomodulatory potential of auto-OVV by an extensive time-course analysis of cytokines in patients’ plasma (Luminex profiling) and peripheral blood immune cells (flow cytometry). We also analyzed the ability of SNV1c to protect the OVV from antibody/complement inactivation in vitro and in vivo following intratumoral injection in various mouse tumors. The immune cell infiltrations of the injected tumors were also analyzed. Results: Therapy with auto-OVV induced a coordinated activation of cytokine, T cell and NK responses in patients as early as 1 day, peaking around 1-week and lasting for up to 1-month post treatment. Effective OVV amplification in cancer patients correlated with significant changes of multiple innate and adaptive immune parameters. Patient stratification into groups with transient versus persistent viral DNA was linked to opposing and mutually exclusive patterns of robust activation of NK versus T cell responses, respectively. SNV1c showed significantly enhanced protection of OVV in vitro and led to statistically significant tumor growth inhibition as compared to control non-treated tumors or to naked OVV-treated tumors. Importantly, local administration of SNV1c induced systemic therapeutic effects. Five days after SNV1c administration, tumor infiltrating lymphocytes from both treated and untreated tumors showed increased CD4 and CD8 T-cell infiltrations, decreased Tregs, and improved effector to Treg ratios, associated with tumor growth inhibition at both treated and untreated tumor sites. Conclusions: This study establishes the timeline of treatment-related immunological changes and identifies potential immunological correlates associated with the OVV persistence in vivo. We also demonstrate the ability of our cell-based platforms to protect and potentiate OVV by circumventing innate and adaptive immune barriers, resulting in enhanced oncolytic virotherapy.
Long-term survival rates in cancer patients achieved with metronomic chemotherapy (cyclophosphamide and methotrexate).
Authors: Chubenko Viacheslav , Zagorskaya Ludmila , Lukyanchikova Valentina , Moiseenko Fedor Vladimirovich , Abduloeva Nuriniso , Zhabina Albina , Levchenko Natalia , Kramchaninov Mikhail , Shelekhova Ksenia , Zykov Evgenii , Meldo Anna , Kudryavcev Alexey , Napolskaya Elena , Moiseyenko Vladimir
Published: May 2020

e15519

Background: Metronomic therapy (MT) is one of the most promising advances in cancer treatment strategy focused on both tumor cells and their microenvironment, specifically anti-angiogenic, immune and metabolic regulation. The treatment modality is a durable cytostatic drugs exposure with less toxicity. We investigated the efficacy of MT in point of prolonged duration of response (more than 6 months) in pts with the advanced resistant tumors. Methods: A total of 678 pts have been enrolled in the study and have been treated MT cyclophosphamide 50 mg (qd, PO) and methotrexate 2.5 mg (BID, PO) twice per week dosing schedule until progression. 508 pts (75%) had disease progression during first 6 months. Long-term efficacy (more than 6 months) was assessed in 170 pts (25%). The study population characteristics were female 122 (72%) vs male 48 (28%). Among the enrolled pts median age was 70 years (range from 37 to 91 years). ECOG status ranged from 0 to 4 (median - 1). The majority of pts received MT as a 2nd and 3rd line (57%) of the therapy (range from 1 to 11): 1-2 – 106 (62%), 3-4 - 52 (31%), 5 and more - 12 (7%). The distribution of pts by diagnosis is presented in the table 1. The majority of MT pts were represented by advanced clinical stage III-IV (97,6%): I - 1 (0,5%), II - 3 (2%), III - 7 (4%), IV - 159 (93,5%). MT commonly used in early-stages cancer in elderly patients with multiple comorbidities contraindicated to standard treatment. Most pts had two or multiple metastatic sites. Results: The ongoing long-term effect for MT indication was from 6 to 46 months. The main group consisted of the breast and colorectal cancer, 27% and 15%, respectively. The median time to progression was 9 months (16 months in CR was achieved (0,4%), 11 months and 9 months with a partial response (3,4%) and stable disease (21,2%), respectively). 12 (1,7%) pts are on ongoing long-term follow up more than 24 months. MT was well-tolerated. Drug related adverse events were described upon the study: hematological (10,2%, Gr 3-4 – 3,5%) vs non-hematological (6,6%, Gr 3-4 1,7%) toxicity. Conclusions: MT schedule demonstrated effectiveness in patients with advanced resistant tumors. The median duration of the effectiveness doesn’t depend on the line of therapy, tumor localization and objective response. The role of possible predictive markers of clinical benefit is question of further trials. [Table: see text]
Factors associated with low chemotherapy dosing in high risk breast cancer.
Authors: Hernandez Leilani , Fergestrom Nicole , Charlson John A. , Neuner Joan Marie
Published: May 2020

e15520

Background: Chemotherapy dose is often reduced or delayed due to adverse effects. However, a low relative dose intensity (RDI) is associated with reduced disease control and survival. There is little data among patients receiving modern breast cancer regimens regarding the risk of low RDI, or factors associated with it. Methods: We used an institutional registry at a large multidisciplinary cancer center to identify subjects with incident invasive, non-metastatic breast cancer diagnosed between 2009-2018. We focused on a subset of high-risk patients for whom chemotherapy is clearly recommended by guidelines: triple negative (estrogen receptor, progesterone receptor and HER2 negative) or HER2-positive subtypes, and either positive lymph node metastases or tumor size > 1.0 cm. We obtained administration dates and doses for the following agents: doxorubicin, cyclophosphamide, paclitaxel, docetaxel, and carboplatin. Our primary outcome was RDI < 85% of guideline-recommended doses of at least one of these agents. Other outcomes included dose of each agent (first cycle dose < 85% or total dose < 85) and timing (delay ≥7 days or early discontinuation). Variables used for analyses included BMI ≥25, race, HER2 status, age ≥55 years, marital status, stage, and Charlson co-morbidity score. Results: More than half of our cohort of high risk breast cancer patients experienced a low RDI of at least one chemotherapy agent. Out of the 237 patients who met our inclusion criteria, 60% had a low RDI, 10% received a low first cycle dose, 38% received a low total dose, and 46% experienced either a dose delay or discontinued treatment. Using logistic regression analyses, we found that being overweight (CI: 1.01-3.54; p = 0.046) and stage 3 breast cancer (CI:1.05-5.96; p = 0.038) were associated with increased odds of having a low RDI. Low total dose was associated with being overweight (p = 0.0181), unmarried (p = 0.0278), and stage 3 (p = 0.0201). HER2-positive status was also associated with a low first cycle dose (p = 0.001) and dose delays or discontinuation (p = 0.0002). Conclusions: Having a BMI ≥25, higher-stage, or being unmarried, which may serve as a proxy for low social support, are factors that may help identify patients who will have guideline-recommended chemotherapy doses reduced.
Taxane induced pulmonary toxicity in breast cancer.
Authors: Varayathu Hrishi , Mathew Hephzibah Sara , Sarathy Vinu , Thomas Beulah Elsa , Kumar Santosh Elluru , Reddy Nitheesha , Mufti Suhail Sayeed , Puii Lalnun , Sangi Lalram , Mathew Smruthy Ann , Koramati Samuel Luke , S Dinesh , Swamy Shivakumar , Naik Radheshyam , Ajaikumar Basavalinga Sadasivaiah
Published: May 2020

e15522

Background: Taxanes are an integral part of chemotherapy for a broad range of tumor types. The usual toxicities associated with Taxane use are peripheral neuropathy, myelosuppression and musculoskeletal adverse effects. Very few studies have reported taxane induced pulmonary toxicity objectively. In our study we explored Taxane induced pulmonary toxicity based on discrete CT findings. Methods: 40 non-smokers diagnosed with Her-2 negative breast cancer who received Taxane monotherapy from 2017 to 2018 were retrospectively analyzed for pre and post therapy CT changes. Following CT findings were considered significant as pulmonary toxicity; (a) Nonspecific area with ground-glass attenuation, (b) multifocal areas of airspace consolidation, (c) patchy distribution of ground-glass attenuation accompanied by interlobular septal thickening and (d) extensive bilateral ground-glass attenuation or airspace consolidations with traction bronchiectasis. The CT findings were assessed independently by two radiologists and one pulmonologist. Results: Our study showed that 5 patients (12.5%) developed pulmonary changes on chest CT post Taxane therapy as summarized in Table. Conclusions: Our study shows that taxane has more potential to induce pulmonary toxicity than reported in present literature. Given that taxanes are the most widely used chemotherapy, it is of utmost necessity to be cognizant of this under-reported potentially fatal adverse effect in a large population. Further studies should be conducted to better understand the true potential, mechanisms and patterns of taxanes induced pulmonary toxicity. [Table: see text]
Successful mutational profiling of single circulating epithelial tumor cells (CETCs) from patients with solid cancers for guiding therapy.
Authors: Schott Dorothea , Pizon Monika , Pachamnn Ulrich , Pachmann Katharina
Published: May 2020

e15527

Background: Circulating epithelial tumor cells (CETCs) are an important link between primary tumors and metastases and provide a readily accessible source of tumor material from patients with solid cancer. Therefore, they are representative for primary and metastatic lesions to evaluate treatment response in real-time and to guide therapeutic modifications. Beyond enumeration, information on mutations is extremely important in the clinical setting to monitor the development of the disease over time and to guide drug selection. Isolating single CETCs with high recovery and purity from the high background of blood cells is an indispensable prerequisite to investigate cell heterogeneity and provides a tool for a personalized medicine approach. Methods: Blood from patients with non-small cell lung cancer, colon cancer and malignant melanoma was analysed for CETCs using the maintrac approach. Between 8-20 EpCAM positive cells from each patient were isolated individually using a semiautomated capillary approach and deposited one by one into micro cups. For mutational profiling, EGFR, KRAS and BRAF hotspot mutations were analysed in CETCs after whole genome amplification. Results: DNA could be amplified from all individually isolated cells. EGFR mutations were detected in 17% of isolated tumor cells from patients with non-small cell lung cancer; KRAS mutations were detectable in 28% of cells from patients with colon cancer and BRAF mutations in 50% of evaluable cells. A high heterogeneity in mutation status was observed among CETCs within a patient. Conclusions: Individually isolating CETCs from the peripheral blood from patients with non-small cell lung cancer, colon cancer and melanoma allows not only to detect driver mutations but also to determine genotypic heterogeneity between cells and changes over time. Mutations in the CETCs resemble those detected in primary tumors present only in part of the cells. They can be used to monitor the development of disease and its status over time, particularly for its impact in guiding drug selection.
The application of dynamic monitoring of circulating tumor DNA for detecting minimal residual disease and predicting recurrence in colorectal cancer patients.
Authors: Chan Hiu Ting , Nagayama Satoshi , Chin Yoon Ming , Otaki Masumi , Hayashi Rie , Kiyotani Kazuma , Fukunaga Yosuke , Ueno Masashi , Nakamura Yusuke , Low Siew-Kee Kee
Published: May 2020

e15531

Background: Although the prognosis of colorectal cancer (CRC) has improved in the past decade, a subset of CRC patients may still suffer from relapse due to the progression from minimal residual disease (MRD) after surgical resection. A sensitive and non-invasive method to detect MRD and early diagnosis of recurrence disease is warranted to start therapeutic interventions at an earlier timing and improving the overall survival rate. In this study, we have evaluated the feasibility of circulating tumor DNA (ctDNA) analysis to detect MRD and early detection of recurrence in CRC patients. Methods: Plasma samples were collected prospectively from 38 CRC patients (stage I to IV), who underwent surgical resection. Preoperative blood samples were obtained just before surgery and post-operative samples were collected at multiple time-points to monitor the changes of tumor mutation profiles. Tumor-derived mutations were detected in preoperative blood samples as well as surgically resected-tumor tissues using ultradeep targeted next generation sequencing. Patient-paired peripheral blood cells (PBCs) were sequenced concurrently to exclude clonal hematopoiesis-related mutations. Results: Among the 38 patients, 74 non-synonymous mutations were identified in tumor tissues and 64 mutations in the preoperative plasma samples. Paired PBCs sequencing identified 11 mutations in plasma samples to be clonal hematopoiesis-related mutations. After the exclusion of clonal hematopoiesis-related mutations, 34 (89.5%) of the 38 patients harbors at least one somatic mutation either from tumor tissues or plasma samples to be monitored longitudinally. ctDNA was detectable in 5 of 14 (36%) post-surgical samples of patients who did not receive adjuvant chemotherapy and in 9 of 18 (50%) post-chemotherapy samples. Up to date, 6 patients have been detected with clinical recurrence and ctDNA analysis identified all 6 recurrences before imaging. Serial ctDNA analyses were able to detect disease recurrence up to 6 months before imaging tests. Furthermore, all patients that were ctDNA negative post-operative or post-chemotherapy showed no signs of clinical relapse. Conclusions: Our current results indicate that ctDNA analysis allows the detection of MRD in CRC patients. The integration of ctDNA analysis with current standard monitoring guidelines holds great promise in early detection of recurrence to allow clinical intervention to be applied promptly.
Focused ultrasound ablation of solid tumors: Feasibility of planning tissue-selective treatments.
Authors: Mancia Lauren , Vlaisavljevich Eli , Yousefi Nyousha , Maxwell Adam , Siegel Geoffrey W. , Xu Zhen , Johnsen Eric
Published: May 2020

e15600

Background: Focused ultrasound (FUS) is a noninvasive, nonionizing, repeatable local ablative therapy that induces mechanical fractionation or thermal necrosis of a variety of solid tumors including hepatocellular carcinoma, prostate cancer, and desmoid fibromatosis. Recent feasibility studies in animal models have demonstrated the possibility of designing focused ultrasound treatments that are selective (e.g. spare healthy tissue, nerves, and blood vessels) due to differences in tissue and tumor mechanical properties. Given wide variation in individual tumor and patient characteristics, mechanics-based predictions of ablation zone features in different tissues under a range of FUS device settings are needed to permit personalized treatment planning. Methods: A finite difference computational method is used to simulate FUS ablation of tissues with variable mechanical properties (shear moduli of 0.6 – 200 kPa) under different FUS sonication parameters (frequency and peak pressure). The model calculates strain fields contributing to tissue ablation in FUS treatments which are used to predict ablation zone radii and boundary characteristics. Simulation predictions in model tissues are then compared to histology obtained from FUS-treated porcine tissue samples with similar mechanical properties. Results: The mechanical properties of model tissues and FUS treatment parameters have distinct effects on predicted minimum ablation zone radii. For example, smaller ablation zone radii are achieved in stiffer vessel wall than liver under given FUS sonication parameters. In each tissue, lower frequency and higher peak pressure FUS sonication predict a larger ablation zone. Combined variation of sonication frequency and peak pressure are found to achieve wider variation in ablation zone radius than previously achieved with frequency variation alone. Predicted ablation zone radii and boundary characteristics are consistent with the observed histology of FUS-treated tissues. Conclusions: Results show that simulations accounting for tissue mechanical properties and device settings can predict tissue selectivity and ablation zone characteristics observed in FUS procedures. This study demonstrates the potential of using noninvasive measurements of tissue and tumor properties obtained, for example, via shear wave elastography, in combination with micromechanical tissue ablation simulations to develop personalized, selective focused ultrasound treatments for solid tumors.
Landscape of RAS variations in 17993 solid tumors: A pan-cancer analysis.
Authors: Wu Hao , Zhou Shaoqiang , Zhang Ding , Zhao Jing , Zhao Zhengyi , Wang Guoqiang , Zhang Yuzi , Cui Longgang , Cai Shangli , Bai Yuezong , Chen Dedian
Published: May 2020

e15615

Background: RAS family genes (HRAS, KRAS and NRAS) were frequently observed in several tumors. The expression of constitutively active RAS proteins mediated by RAS variations promote the development of tumors. KRAS is an important prognostic and drug resistance biomarker. It would also be a promising drug target. AMG 510 which targeting KRAS G12C showed an ORR of 48% in patients with NSCLC. Several trials which evaluating the efficacy of RAS G12C inhibitor in solid tumors are initiated. Herein, we analyzed the alterations status of KRAS/NRAS/HRAS across diverse solid tumors. Methods: The sing nucleotide variants (SNV) and copy number variants (CNV) data of 17993 Chinese patients from 22 types of cancer were obtained in 3DMed database. Genomic profiling of DNA was performed through a next-generation sequencing with 61 cancer-related genes on tissue. Only the pathogenic mutations and likely pathogenic mutations in clinical significance were rolled into our analysis. Results: Among 17993 pan-cancer patients, the total RAS variants frequency was 23.09%. KRAS was the most frequently altered (90.76% of RAS SNV, 84.70% of RAS CNV), followed by NRAS (6.56% of RAS SNV, 8.58% of RAS CNV) and HRAS (2.68% of RAS SNV, 6.72% of RAS CNV). For the SNV, KRAS were most commonly found in pancreas cancer (685/842, 81.35%), intestine cancer (85/175, 48.57%) and colorectal cancer (1609/3329, 48.33%). NRAS occurred most frequently in melanoma (31/196, 15.82%), colorectal cancer (116/3329, 3.48%) and thyroid cancer (2/60, 3.33%). HRAS were most often found in bladder/urinary tract cancer (46/367, 12.53%), prostate cancer (5/137, 3.65%) and head and neck cancer (7/314, 2.23%). Further analysis among KRAS SNV patients showed that the mutation frequency of KRAS G12C, G12D, G12R and G12V was 9.24%, 34.73%, 3.54% and 21.66%, respectively. A total of 21 in 22 types of solid tumors had KRAS G12C/D/R/V pathogenic or likely pathogenic mutation, which occurred most frequently in colorectal cancer, pancreas cancer and lung cancer. Conclusions: Our results suggested that a variety of solid tumors may harbor KRAS G12C/D/R/V mutation. These patients may benefit from KRAS inhibitors.
Repurposing antimuscarinics to treat lethal chemoresistant prostate and lung cancers.
Authors: Bland Tyler B , Wu Boyang
Published: May 2020

e15636

Background: Advanced non-small cell lung cancer (NSCLC) and prostate cancer (PCa) remain leading causes of cancer-related deaths in the United States and worldwide. The current study investigates the ability of antimuscarinics to resensitize chemotherapy resistant NSCLC and PCa to chemotherapy-induced apoptosis. Methods: The chemotherapy drug docetaxel (DTX) resistant human NSCLC and PCa cell cultures were established to recapitulate clinical emergence of chemoresistance. These cultures were used to harvest total RNA and protein for qPCR and Western blot analyses, respectively. DTX’s ability to induce apoptosis was measured with crystal violet survival assay and TUNEL assay. Results: DTX-resistant NSCLC (A549R) and PCa (DU145R) cells show increased expression of the muscarinic acetylcholine receptors 1 and 3 (CHRM1 / CHRM3). Treatment of these DTX-resistant cells with the CHRM1 antagonist dicyclomine (Dic) or the CHRM3 antagonist darifenacin (Dari) resensitize these cells to DTX-induced cell death. This treatment repurposes Dic and Dari, which are currently used in the clinic to treat irritable-bowel syndrome and overactive bladder, respectively. Decreased cell survival was observed with crystal violet survival assay, which led to increased cell death at a lower concentration of DTX compared to cells treated with DTX alone (A549R [DTX IC50]: DTX = No response, Dic + DTX = 5.2 nM, Dari + DTX = 2.2 nM / DU145R [DTX IC50]: DTX = No response, Dic + DTX = 4.2 nM, Dari + DTX = 1.4 nM). Furthermore, cells treated with Dic + DTX or Dari + DTX show increased apoptosis measured by TUNEL assay compared to cells treated with DTX alone (A549R [TUNEL positive %]: DTX = 0.9%, Dic + DTX = 22.2%, Dari + DTX = 88.2% / DU145R [TUNEL positive %]: DTX = 3.6%, Dic + DTX = 96.4%, Dari + DTX = 90.1%). Conclusions: We show that chemoresistant NSCLC and PCa have high expression of CHRM1 and CHRM3 and that antimuscarinics effectively resensitize these lethal forms of cancer to chemotherapeutics. Our data support the rationale of repurposing these therapies to treat chemoresistant NSCLC and PCa, allowing quick entry into a Phase II clinical trial and further translation into clinical practice, in a more cost-effective and less financially risky way, to benefit cancer patients.
X-ray crystal structure analysis of elacestrant (RAD1901), a novel selective estrogen receptor degrader (SERD), bound to estrogen receptor alpha ligand binding domain.
Authors: Fanning Sean W. , Greene Geoffrey , Conlan Maureen G.
Published: May 2020

e15647

Background: Antiestrogens are a mainstay of treatment for estrogen receptor positive (ER+) breast cancer in both the adjuvant and the advanced/metastatic settings. Elacestrant is a mixed activity selective estrogen receptor (SER) alpha (ERα) antagonist, acting as a SER modulator (SERM) at low doses and a SER degrader (SERD) at high doses. It has shown activity in hormone sensitive wild type (WT) ERα and insensitive estrogen receptor gene 1 (ESR1) mutation-harboring (Y537S and D538G) ERα breast cancer, both in preclinical models and in clinical studies. It also possesses a unique pharmacology compared to other competitive ER antagonists in its ability to cross the blood brain barrier. Competitive ERα antagonists are typically comprised of a core that sits in the ligand binding pocket and an arm that manipulates the structure to achieve SERM or SERD activities. In these molecules, the arm is attached in the same position as the triphenylethylene core of tamoxifen. However, elacestrant possesses a novel site of attachment. As such, we hypothesized that elacestrant adopts an alternative binding orientation in the ERα ligand binding pocket to achieve its unique pharmaceutical profiles. Methods: X-ray crystallography was used to solve a co-crystal structure of elacestrant in complex with WT ERα ligand binding domain to 2Å. Results: Overall, elacestrant promotes the formation of a canonical ERα ligand binding domain antagonist conformation, whereby helix 12 (H12) is docked into the activating function-2 cleft. However, elacestrant adopts a novel vector in the ERα ligand binding pocket that places it in close proximity to helix 12. As a result, it forms a bifurcated hydrogen bond that is not observed in other competitive antiestrogens and samples a chemical space known to increase H12 mobility and induce SERD activity. This novel vector also places it near positions 537 and 538, the two most common sites of somatic mutation. Conclusions: The high-resolution x-ray crystal structure of elacestrant suggests that the unique binding mode it adopts enables novel pharmacology and positions it to achieve potency in the WT and activating somatic ERα mutated breast cancer setting.