bluuebunny/crossref_metadata_2025 huggingface dataset:
Frequency of minimal residual disease as measured by ctDNA in mismatch repair deficient tumors following curative resection.
Authors: Lumish Melissa Amy , Jayakumaran Gowtham , Fox Maggie , Sabwa Shalom , Cercek Andrea , Stadler Zsofia Kinga , Ku Geoffrey Yuyat , Segal Neil Howard , Won Elizabeth , Maron Steven Brad , Troso-Sandoval Tiffany A. , Segal Michal Fani , Strong Vivian E. , Molena Daniela , Weiser Martin R. , Jones David Randolph , Zehir Ahmet , Berger Michael F. , Diaz Luis A. , Janjigian Yelena Y.
Published: June 2021

e14520

Background: Mismatch repair deficient (MMRd) tumors are highly sensitive to checkpoint blockade (CPB) in patients with metastatic disease regardless of tumor type. However, the efficacy of CPB in the adjuvant setting is unknown, especially since MMRd is considered a favorable biomarker for most resected tumor types. Circulating tumor DNA (ctDNA) could be used to screen for patients at high risk for recurrence following surgery or adjuvant chemotherapy and identify patients (pts) that would most benefit from CPB. Methods: To assess the frequency of ctDNA in the resected MMRd population, we prospectively screened pts with MMRd tumors who completed standard perioperative chemotherapy and surgery (NCT03832569). DNA from resected tumors and matched postoperative plasma was sequenced for the presence of somatic mutations. Patients were considered to have minimal residual disease (MRD) when mutations were identified in tumor and found to be identical to those in matched plasma DNA. Somatic tissue mutations were assessed using MSK-IMPACT and ctDNA was assessed using FoundationOne, Guardant360 or MSK-ACCESS. Results: A total of 86 pts were screened for the presence of MRD. These represented 7 tumor types with colorectal (63%), endometrial (16%) and esophagogastric (13%) being the most common. The majority of pts were stage III (49%). MRD was detected in 18% of cases (14 of 79). Among the MRD negative group (n=62), only one pt developed disease recurrence. Three samples failed ctDNA analysis for technical reasons. Conclusions: MRD was identified in 18% of resected MMRd tumors using ctDNA analysis, suggesting this to be a feasible tumor agnostic approach to test the efficacy of CPB in a pts at high-risk for recurrence. Future studies will assess the impact of CPB in MRD positive MMRd tumors.[Table: see text]
Artificial intelligence-powered spatial analysis of tumor infiltrating lymphocytes (TIL) to reflect target gene expressions of novel immuno-oncology agents.
Authors: Ock Chan-Young , Shin Seunghwan , Jung Wonkyung , Ahn Sangheon , Kim Haejoon , Lee Chunggi , Aum Jaehong , Tak Daim , Ryu Ahyoung , Chung Taiwon , Baek Eunji , Shin Jisoo , Lee Seungje , Shin Jiwon , Ma Minuk , Park Seonwook , Pereira Sergio , Kang Jeongseok , Yoo Donggeun , Paeng Kyunghyun
Published: June 2021

e14534

Background: Novel immuno-oncology (IO) agents are promising but showing their efficacy in early phase clinical trials has been challenging due to limited enrichment strategies using practical biomarker platforms. We hypothesize that an artificial intelligence (AI)-powered spatial analysis of TIL using practically feasible H&E slides, can reflect a specific target gene expression derived from RNA sequencing. This enhances its potential application in early development of novel IO agents. Methods: An AI-powered spatial TIL analyzer, namely Lunit SCOPE IO, was developed with data from 2.8 x 109 micrometer2 H&E-stained tissue regions and 5.9 x 106 TILs from 3,166 whole slide images of multiple cancer types, annotated by board-certified pathologists. Inflamed Score and Immune-Excluded Score was defined as the proportion of all tumor-containing 1 mm2-size tiles within a WSI classified as being of inflamed immune phenotype (high TIL density within cancer epithelium) and immune-excluded phenotype (low TIL density within cancer epithelium, but high TIL density within stroma), respectively. We used RNA sequencing data and H&E images from The Cancer Genome Atlas database, excluding those of mesenchymal origin (n = 7,467). Spearman's rank correlation between each gene expression and IS or IES, respectively, was calculated. Correlation coefficient > 0.2 and false discovery rate (FDR) < 1% was considered as a significant correlation. Results: In a total of 20,304 genes, 871 (4.3%) and 1,155 (5.7%) genes were significantly correlated with Inflamed Score (IS) and Immune-Excluded Score (IES), respectively. The IS was highly related to genes reflecting immune cytolytic activity and targets of approved immune checkpoint inhibitors (Table). Interestingly, it was also significantly correlated with target genes of novel IO such as TIGIT, LAG3, TIM3, IDO, Adenosine receptor A2A, OX40, ICOS, M-CSF, IL2, IL7, and IL12. Moreover, the IES was exclusively correlated with the target genes of CEACAM, TGFB, and IL1. Conclusions: Expression levels of novel I-O target genes are correlated with three scores derived from AI-powered TIL analysis using H&E slides, which can be easily applied to clinical research.[Table: see text]
MDNA11 is a long-acting IL-2 superkine that potentiates durable immune response in mouse tumor models and non-human primates.
Authors: To Minh D. , Merchant Fahar , Merchant Nina
Published: June 2021

e14546

Background: The efficacy and safety of recombinant human IL-2 (rhIL-2; Proleukin) to treat certain cancers is limited by a short half-life, marked toxicity and selective high affinity binding to IL2Ra over IL2Rb, resulting in preferential activation of suppressive Tregs. In contrast, MDNA11 has been engineered as a long-acting IL-2 superkine with high affinity IL2Rb receptor selectivity, resulting in preferential anti-cancer effector immune cell activation. Methods: MDNA11 was characterized in both in vitro and in vivo studies including assessment of receptor binding kinetics using BLI/Octet, receptor-mediated signaling in human PMBCs, efficacy in syngeneic mouse tumor models including memory response, as well as safety and PK/PD assessments in non-human primates (NHP). Results: Unlike rhIL-2, MDNA11 does not bind to human IL2Ra but demonstrates a 30-fold higher affinity binding to human IL2Rb. This selectivity resulted in enhanced in vitro STAT5 signaling in human NK and resting CD8 T cells with diminished signaling in Tregs; validation studies in humanized mice are ongoing. In CT26 and MC38 syngeneic tumor models, MDNA11 demonstrates potent and durable efficacy as monotherapy following a Q1W dose schedule for 2 weeks. Synergy with anti-PD1 and anti-CTLA4 immune checkpoint inhibitors (ICIs) was observed and a robust immune memory response developed in all mice with complete tumor clearance. These mice were protected against relapse and tumor re-challenges for up to 8 months without any further treatment, and showed the presence of antigen-specific CD8 T cells. In binding studies with IL-2 receptors of different species, MDNA11 showed highly similar affinity towards human and cynomolgus IL2Rb, confirming the latter as a highly relevant model for toxicology study. MDNA11 was well tolerated in cynomolgus monkeys up to 0.6 mg/kg, while inducing durable (≥10 days) proliferation and expansion of NK and CD8 T cells. Effects on Tregs were minimal and there was no eosinophilia and hypotension (associated with vascular leak syndrome). At high doses of MDNA11, the most common clinical observations were transient loss of appetite and diarrhea. There was modest increase in levels of IFNg and TNFa, but no sign of cytokine release syndrome. Dosing did not trigger development of anti-drug antibodies or histopathologic evidence of pulmonary edema (a major IL-2 induced toxicity). Conclusions: MDNA11 is a long-acting IL-2 superkine that exhibits robust efficacy in mouse tumor models as a single agent and was synergistic in combination with ICIs (anti-CTLA4 and anti-PD1). In NHP, MDNA11 demonstrates selective immune effector cell activation and a favorable safety profile. These data constitute a strong framework for the design of a pivotal GLP toxicology study to further support the planned clinical study of MDNA11 either as a single agent or in combination with ICIs.
Impact of cancer type on the incidence of cutaneous toxicities after immune checkpoint inhibitor therapy: A population-level analysis.
Authors: Pahalyants Vartan , Raval Neel Sanjiv , Ugwu-Dike Pearl Ojiugo , Le Thomas K. , Reynolds Kerry Lynn , Kwatra Shawn G. , Semenov Yevgeniy R.
Published: June 2021

e14553

Background: Cutaneous immune-related adverse events (cirAEs) from immune checkpoint inhibitor (ICI) therapy are increasing as these drugs are more widely used. Population-level studies are lacking. In this retrospective cohort study, we analyzed the primary tumor’s effect on cirAE incidence and downstream utilization of systemic immunosuppression in a national healthcare database (TriNetX). Methods: Index event was defined as day of ICI initiation; outcomes were restricted to 2 years from index. cirAEs were defined as 42 dermatoses identified in a comprehensive review of the cirAE literature (Sibaud 2018) and expert opinion. Systemic immunosuppression was classified as steroidal or non-steroidal. Primary outcomes included aggregate and cancer-specific incidence of cirAEs across the four most common cancer indications for ICI therapy (melanoma, lung, urinary, and gastrointestinal). Secondary outcomes included utilization of new steroidal or non-steroidal systemic immunosuppression. For each analysis, we excluded patients with an outcome prior to the index event. Risk ratios (RR) were calculated after 1-to-1 propensity score matching, adjusting for age at index, sex, race, ethnicity, and ICI target, with the lung cancer group as reference. Results: We identified 27,481 eligible subjects. Aggregate incidence of cirAEs across all cancer types was 23.41%, with non-specific rashes, pruritus, and drug eruptions as the most common diagnoses. Among all patients, new steroidal and non-steroidal immunosuppression use following ICI initiation was 16.1% and 4.1%, respectively. After adjusting for covariates, melanoma (RR 1.60, 95% CI 1.43-1.79, p < 0.001) was associated with a higher risk of cirAE, while urinary and gastrointestinal tract cancers were not significantly different from the reference. Melanoma (RR 0.64, 95% CI 0.55-0.74), urinary tract (RR 0.71, 95% CI 0.62-0.81) and gastrointestinal tract (RR 0.46, 95% CI 0.39-0.53) cancers were all less likely to require steroidal immunosuppression (all p < 0.001). However, patients with urinary tract cancer (RR 4.03, 95% CI 3.04-5.35) and melanoma (RR 2.44, 95% CI 1.75-3.40) were more likely to receive non-steroidal systemic immunosuppression (both p < 0.001). Conclusions: Our results reinforce that ICI recipients commonly develop skin toxicities and provide robust evidence that melanoma is independently associated with their incidence. Furthermore, we found that patients with lung cancers received steroidal systemic immunosuppression most frequently, an intervention associated with poorer overall survival (Riudavets 2020). In contrast, patients with melanoma were more likely to receive non-steroidal immunosuppression. This study is the first population-based analysis of an irAE across tumor types and is proof-of-concept for future investigations into clinical risk factors in a large dataset.
Efficacy of KD033, an anti-human-PD-L1-IL-15 bispecific protein, in human-PD-L1 positive and negative murine tumor models.
Authors: Martomo Stella , Lu Dan , Polonskaya Zhanna , Luna Xenia , Zhang Zhikai , Regev Galit , Schueller Olivier , Patel Jeegar
Published: June 2021

e14556

Background: KD033 is a clinical-stage bispecific fusion molecule consisting of a high-affinity anti-human-PD-L1 antibody and human IL-15. Preclinical studies have demonstrated that targeting IL-15 with anti-PD-L1 antibody resulted in increased efficacy, safety and maximal tolerated dose of the fusion protein compared to administration of free IL-15, as well as reduction of tumor growth in both PD-L1 positive and negative tumor models (1). The goal of the current study is to directly compare KD033 efficacy when PD-L1 is present or absent on the surface of the same tumor. Methods: KD033 was administered in the human-PD-L1/PD-1 transgenic C57/Bl6 mice subcutaneously transplanted with human-PD-L1 positive and negative MC38 colon carcinoma cells. This animal model allowed the evaluation of the impact of the presence or absence of human-PD-L1 expression on the tumor cell surface without altering human-PD-L1 expression on immune cells. Results: KD033 treatment resulted in significant tumor growth reduction in both human-PD-L1 positive and negative MC38 tumors. Analysis of peripheral immune cell populations showed similar increases of CD8 T and NK cells between human-PD-L1 positive and negative MC38- bearing mice after KD033 administration. Immunohistochemistry demonstrated a significant increase in CD8 T-cell infiltration into the human-PD-L1 positive MC38 tumors, whereas NK cells infiltration was more pronounced in the human-PD-L1 negative MC38 tumors. Analysis of tumor gene transcription after KD033 treatment highlighted differences in gene signatures between human-PD-L1 positive and negative MC38 tumors following KD033 treatment. Conclusions: These results showed that the efficacy of anti-PD-L1-IL-15 fusion protein is not limited to PD-L1 tumor expression as KD033 was efficacious in both PD-L1 positive and negative tumors. Mol Cancer Ther February 1 2021 (20) (2) 347-356; DOI: 10.1158/1535-7163.MCT-20-0457
HMBD-002 is a novel, neutralizing, anti-VISTA antibody exhibiting strong preclinical efficacy and safety, being developed as a monotherapy and in combination with pembrolizumab.
Authors: DiMascio Leah , Thakkar Dipti , Gandhi Namita , Guan Siyu , Rowinsky Eric Keith , Ingram Piers , Boyd-Kirkup Jerome Douglas
Published: June 2021

e14569

Background: Inhibition of immune checkpoints has been an effective strategy against several cancers, but efficacy gaps remain. VISTA is an immunoregulatory protein expressed on both hematopoietic and tumor cells and has been shown to induce an immunosuppressive environment by repressing T-cell activation and cytokine production. VISTA expression has been shown to be upregulated following treatment with immune checkpoint inhibitors. Hummingbird Bioscience is developing HMBD-002, an IgG4 antibody against VISTA with high binding specificity across species (human, cyno and rodent) and low pM affinity to the region of VISTA required for engagement with interaction partners, including VSIG3. As HMBD-002 is an IgG4 isotype, it uniquely blocks the inhibitory function of VISTA and releases immune suppression without depleting VISTA+ cells. Here we establish the preclinical efficacy and safety of HMBD-002 along with the rationale for development of HMBD-002 as a monotherapy and in combination with PD1 inhibitors in triple negative breast cancer (TNBC) and non-small cell lung cancer (NSCLC). Methods: The effect of HMBD-002 in an in vitro model of human immune activation was evaluated using an allogenic Mixed Lymphocyte Reaction (MLR) assay. Tumor growth inhibition studies were conducted in syngeneic murine cell-derived xenograft (CDX) models. The PK and tolerability of HMBD-002 was evaluated in rats and cynomolgus monkeys with a 21 or 28-day observation period. The expression profile of VISTA was examined in different cancers by IHC staining of FFPE TMAs of various solid cancers. Results: HMBD-002 induced a significant dose dependent increase in the levels of inflammatory cytokines, IFN-γ and TNF- α and suppressed the expansion of CD14+ monocytes. Treatment with HMBD-002 significantly inhibited tumor growth and improved survival in CDX models. HMBD-002 is well tolerated in rats and cynomolgus monkeys with acceptable serum half-life, no HMBD-002 related morbidity/mortality or clinical signs, and no HMBD-002 related changes were observed in body weight, food consumption, clinical chemistry, or hematology parameters. HMBD-002 did not induce cytokine release in human PBMCs or whole blood. TNBC and NSCLC patients were found to harbor the highest VISTA expression across cancer types tested, providing a rationale for exploring these indications in upcoming clinical studies. Conclusions: These studies provide the rationale for development of HMBD-002 as a monotherapy and in combination with pembrolizumab to treat patients with solid tumors expressing VISTA in the tumor microenvironment. A Phase 1 study will be initiated in 2021.
Myocarditis in patients treated with anti–programmed death-1/programmed death ligand 1 therapy.
Authors: Wang Hanping , Guo Xiaoxiao , Tian Ran , Si Xiaoyan , Zhang Xiaotong , Wang Jiangshan , Zhao Haitao , Zhang Li
Published: June 2021

e14579

Background: We retrospectively analysed the cases of immune checkpoint inhibitor(ICI) -associated myocarditis managed in PUMCH. We aim to investigate the clinical characteristics of myocarditis and the risk factors affecting the prognosis. Methods: Medical records of all checkpoint inhibitor-related myocarditis were reviewed retrospectively. Descriptive statistics were used for the report of clinical outcomes. Univariate and multivariate Logistic analyses were performed to establish a prognostic model. Results: The median age of 19 patients was 64 years old (range 39̃80 years). The median time of onset from the iniatial of ICIs was 21 days (ranging from 2 to 375 days). Seven cases were grade 4 (36.8%), 2 cases were grade 3 (10.5%), 8 cases were grade 2 (42.1%), and 2 cases were grade 1 (10.5%). Clinical symptoms included dyspnea (13/19, 68.4%), fatigue (11/19, 57.9%), myalgia/myasthenia (9/19, 47.4%), sweating (6/19, 31.6%) and palpitation (6/19, 31.6%) et al. 47.4% (9/19) of the patients showed cardiac insufficiency/hemodynamic instability at first, and 52.6% (10/19) of the patients showed symptoms related to myositis first. cTnI and CK increased in all patients. The level of cTnI was significantly higher in grade 3̃4 than that in grade 1-2. NTproBNP was elevated in all grade 4 cases and one grade 3 case. The LVEF decreased only in 4 patients with grade 4 myocarditis (4/19, 22.2%). The electrocardiogram showed ventricular arrhythmia in 4 cases and complete atrioventricular block (3°AVB) in 3 cases. All patients were treated with corticosteroids (GCS). Grade 4 (6 cases), grade 3 (1 case) and grade 2 (2 cases) were given GCS pulse dose. The median total duration of GCS treatment was 42 days (range: 14-60 days). Three patients were treated with tocilizumab and one patient with infliximab. All patients with grade 3-4 and 2 patients with grade 2 were also treated with IVIG. Pacemakers were implanted in 3 patients with 3°AVB. The mortality rate of 19 cases was 21.1% (4/19), and the mortality rate of grade 4 myocarditis was 42.9%(3/7). The longer course before GCS therapy (p = 0.023), the higher NTproBNP (p = 0.030) and the lower LVEF value (p = 0.014) were found in the death group compared to the survival group. Logistic regression analysis showed that LVEF is an independent risk factor. The myocarditis prognostic model was condstructed as follows: Y = 8.794-18.002*LVEF, the prognostic critical value of LVEF was 57.5%, the area under the ROC curve of the prognostic model was 0.894 (95%CI: 0.735̃1.000), the sensitivity was 0.769, and the specificity was 1.000. Conclusions: We describe the largest single-center immune myocarditis cohort to date. Paying attention to the symptoms related to myositis is helpful to find more grade 1-2 myocarditis. cTnI is sensitive to myocarditis, and NTproBNP is of great significance to grade 3 ̃ 4 cases. LVEF < 57% may be an independent risk factor for poor prognosis.
Genomic profiling by cell-free circulating tumor DNA in patients with recurrent adenoid cystic carcinoma (ACC) and the potential activity of selective fibroblast growth factor receptor (FGFR) inhibitors.
Authors: Perez Cesar Augusto , Dutcher Giselle , Sargi Zoukaa , Guevara Nelson , Tse David , Benedetto Pasquale W.
Published: June 2021

e15018

Background: Few effective systemic therapies are available for adenoid cystic carcinoma (ACC). Multitargeted Tyrosine Kinase inhibitors (TKIs) as Lenvatinib and Rivoceranib can provide disease control and limited radiographic response in unselected ACC patients. Next-generation sequencing in tissue (tNGS) has identified MYB gene fusions in [patients with ACC but currently, only Notch1 mutations have an available targeted agent. The fibroblast growth factor receptor (FGFR) pathway is a downstream target of MYB. Methods: We retrospectively reviewed adult patients with a diagnosis of recurrent ACC seen at the University of Miami Sylvester Comprehensive Cancer Center between January 1, 2019 and October 31, 2020. Commercially available cell free DNA panel of 73 genes was used for analyzing circulating tumor DNA (ctDNA). Patients included had radiographically measurable disease and both tNGS and ctDNA data available. Results: Twelve patients were included. Median age at diagnosis was 54 years (28 - 68), and 69% were males. 83% of the patients had genomic abnormalities reported by tNGS, with the same number by ctDNA sequencing. Seven patients (58%) had genomic abnormalities in ctDNA of potentially therapeutic significance which were not found on tNGS, including FGFR1 amplification as well as FGFR2, BRAF, KRAS and JAK2 mutations. Three patients with FGFR genomic abnormalities only seen by ctDNA had been previously treated with Lenvatinib. One patient with primary lacrimal gland ACC metastatic to the liver and tNGS positive for NOTCH 1 had disease progression despite treatment with Lenvatinib and a NOTCH γ-secretase inhibitor. The patient’s ctDNA reported a FGFR2 mutation. The patient was started on Erdafitinib, an oral selective FGFR inhibitor, achieving a partial response at the primary site and metastatic liver lesions and remained on therapy for 7 months. Follow-up ctDNA after 3 months of therapy noted resolution of both FGFR and NOTCH abnormalities. Conclusions: Patients treated with lenvantinib or other TKIs may develop acquired resistance mutations and other genomic abnormalities. ctDNA can reveal additional therapeutic targets not present on tNGS and can be repeated serially. FGFR genomic abnormalities can be a driver mutation in patients with refractory advanced ACC and the activity of Erdafinib should be evaluated in this subset of patients with few therapeutic options.
Correlation of SOX-2 and NOTCH1 copy numbers with vimentin expression in orthotopic patient-derived xenografts of cardioesophageal cancer in immunodeficient mice.
Authors: Goncharova Anna S. , Kolesnikov Evgeniy N. , Zaikina Ekaterina V. , Volkova A. V. , Mindar M. V. , Khodakova D. V. , Lukbanova Ekaterina A. , Kurbanova L. Z. , Kozhushko Mikhail A. , Anisimov Aleksandr E. , Myagkov Roman E. , Sanamyants Sergey V. , Katsieva Tanzila B.
Published: June 2021

e15034

Background: Cardioesophageal cancer is one of the most common tumors affecting the mucosa of the cardiac stomach and distal esophagus. Despite the variety of treatment strategies and chemotherapy agents, the prognosis for the patients remains poor. The purpose of the study was to analyze the relative copy numbers of SOX-2 and NOTCH1 and vimentin expression in orthotopic patient-derived xenografts of cardioesophageal cancer. Methods: The model of cardioesophageal cancer was created in Balb/c Nude mice with surgical bioptates of moderately differentiated adenocarcinoma obtained from a donor with infiltrative ulcerative cancer of the lower third of the esophagus with a transition to the cardiac stomach into the distal esophagus of Balb/c Nude mice. The index of proliferative activity in the bioptates was assessed by IHC. The relative copy numbers of SOX-2 and NOTCH1 and vimentin expression were determined by Real-Time qPCR. Results: Expression of vimentin was absent in tissues of the donor tumor. The levels of vimentin expression statistically significantly increased in xenografts (1+ and 3+). The SOX-2 and NOTCH1 relative copy numbers were statistically significantly increased in tissues of the donor tumor (0.9 and 0.7), compared to xenografts (1.5±0.03 and 1.7±0.03). Molecular genetic analysis demonstrated an association between an increased vimentin expression and changes in the relative copy numbers of SOX-2 and NOTCH1 (p = 0.013 and p = 0.0001). Conclusions: The relative copy numbers of SOX-2 and NOTCH1 genetic loci was associated with increased expression of the epithelial-mesenchymal transition marker vimentin in tumor tissue samples and changed with each new generation of orthotopic xenografts.
Organotin compounds as effective antimetastatic substances in the treatment of epidermoid lung carcinoma Lewis (LLC) in an experiment.
Authors: Dodokhova Margarita Avdeevna , Kotieva Inga Movlievna , Safronenko Andrej Vladimirovich , Shpakovsky Dmitriy Borisovich , Milaeva Elena Rudolfovna , Shtil Alexander Albertovich , Komarova Ekaterina Fedorovna , Alkhuseyn-Kulyaginova Margarita Stefanovna
Published: June 2021

e15054

Background: Lung cancer occupies one of the first places in the structure of oncological diseases in the world. Reducing the life expectancy of patients, as a rule, occurs as a result of metastatic complications, so the search for new antimetastatic agents is an urgent task for experimental pharmacology and oncology. The aim of our study is to assess the severity of metastatic lesion in the Lewis lung epidermoid carcinoma (LLC) model in the presence of an organotin compound. Methods: The study was conducted on experimental C57BL/6 mice (n = 24, each group contained 12 mice) with LLC (subcutaneous transplantation) in the presence of the cytotoxic organotin compound 2,6-bis (1,1-dimethylethyl) - 4 - [(triphenylstannyl)thio]phenol (Me5). All manipulations were carried out in accordance with the European Convention for the Protection of Vertebrates Used for Experimental and Other Scientific Purposes (ETS 123). Female mice (8 weeks of age, weighing 21-22 g) were intraperitoneally injected with a 1% aqueous gelatin solution of organotin Me5 daily for 5 days. The most effective total dose of Me5 was 250 mg / kg. The animals of the control group received saline solution in similar modes and volumes. In the experimental and control groups, the degree of metastatic lesion was assessed on the 21-day post-grafting period according to the scale proposed by D. Tarin and J. E. Price, which allows us to differentiate the severity of the lesion depending on the number of metastases and their size. The experiment describes three degrees of lesion: LCP-1 (metastases less than 10 pcs. with a diameter not exceeding 1 mm); LCP-2 (metastases from 10 to 30 pieces, some of them larger than 1 mm) and HCP-3 (metastases larger than 30 pieces of different sizes, but no drain). The pathohistological structure was studied by the light-optical method with hematoxylin and eosin staining. Results: When administered intraperitoneal to mice, Me5 did not inhibit the growth of the primary tumor site in the experimental group, but significantly reduced the severity of metastatic lesions in the lungs. In the control group, 75 % of mice with LLC had LCP - 2 (low colonization potential) and in 25% of HCP-3 (high colonization potential) metastatic lesions, in the experimental group only mild lesions were noted in all animals: 58% of mice had LCP – 1, 42% - LCP-2. Conclusions: It is concluded that the overall result of this study clearly demonstrates that the organotin compound 2,6-bis (1,1-dimethylethyl) - 4 - [(triphenylstannyl)thio] phenol (Me5) is an effective antimetastatic agent in the Lewis lung epidermoid carcinoma (LLC) model at a total dose of 250 mg/kg.
Mechanism of inhibitory effect of terpenoid compound from Achillea millefolium on human lung cancer cell proliferation.
Authors: Wang Siming , Wang Yu , Wu Yibing , Dong Mei , Ni Zhiyu
Published: June 2021

e15058

Background: A large number of reports have found that many components in traditional Chinese medicine preparations have obvious anti-tumor activities, and most of these effective components are realized by inducing tumor cell apoptosis. In this part of the experiment, we studied terpenoid compound (3β-acetoxy-1β,4α-dihydroxy-11αH-eudesman-12,6α-olide Achillea millefolium A) from Achillea millefolium anti-cancer mechanism, to determine whether this compound by inducing cell apoptosis plays its antitumor activity of cell proliferation. Methods: (1) The apoptosis of NCI-H292 cells treated with 1 μmol/L and 10 μmol/L Achillea millefolium A was detected by Annexinov-FITC /PI stain. (2) The apoptosis of NCI-H292 cells treated with 10 μmol/L Achillea millefolium A for 12 h and 24 h was detected by in situ end labeling. (3) The changes of p53, Bax, Caspase-3/8/12 and GRP78 in NCI-H292 cells were detected by Western blotting. Results: (1) Effects of Achillea millefolium A on apoptosis of NCI-H292 cells: The apoptosis rate of NCI-H292 cells treated with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 12 h was (15.31)% and (17.84)% respectively. The apoptosis rate of NCI-H292 cells treated with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 12 h was (15.37)% . (2), TUNEL method further verified the cell apoptosis: NCI-H292 cells were treated with 10 μmol/L Achillea millefolium A for 12 h and 24 h, and the positive cells accounted for (13.89)% and (28.37)% respectively, compared with blank control group (4.17)%. (3), Achillea millefolium A up-regulated the expression of p53 protein, Bax protein, Caspase-3/8/12 and GRP78 protein in NCI-H292 cells: After NCI-H292 cells were treated with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 24 h, the expressions of p53, Bax, Caspase-3 and Caspase-8 in NCI-H292 cells were significantly up-regulated. After treatment with 1 μmol/L and 10 μmol/ L Achillea millefolium A for 12 h, the expression of Caspase-12 and GRP78 increased significantly. Conclusions: Achillea millefolium A is a terpenoid compound with obvious apoptosis-inducing activity on human lung cancer cell line NCI-H292. Achillea millefolium A induced apoptosis of NCI-H292 cells was the result of multiple pathways. Achillea millefolium A can up-regulate p53 protein, and then up-regulate Bax, and then activate Caspase-3 protein expression. At the same time, the expression of caspase-3 protein was further increased by activating caspase-8, and apoptosis was induced by both exogenous and endogenous pathways. Notably, the complex of ER stress pathway GRP78 and caspase-12 may also be one of the mechanisms by which Achillea millefolium A induces apoptosis in NCI-H292 cells.
Frequency of homologous recombination deficiency gene mutations in melanoma and its relevance to the immunotherapeutic response.
Authors: Liu Huanhuan , Zhang Yanrui , Ding Feng , Zhang Yun , Liang Xiayuan , Lou Feng , Cao Shanbo , Wang Huina
Published: June 2021

e15073

Background: Homologous recombination deficiency (HRD) can regulate the tumor immune microenvironment based on increasing tumor-infiltrating lymphocytes and therefore might serve as a biomarker of immunotherapeutic response. The HRD phenotype has been defined as the presence of a non-silent somatic mutation in homologous recombination-related (HRR) genes. Here, we focused on frequency of HRR genes mutation and the relationship between HRR genes mutation and clinical immunotherapy responses in melanoma. Methods: We prospectively sequenced 130 Chinese melanoma patients using next-generation sequencing techniques with Acornmed panel with 808 cancer-related genes. 208 patients with WES data from the TCGA project were included. Correlations between HRR gene mutations and clinical outcomes were identified via bioinformatic analysis using TCGA datasets. Results: In the Chinese cohort, 40% (52/130) patients exhibited genomic alterations in HRR genes. The frequently mutated genes were ATM (14%), BRCA2 (13%), CHEK2 (6%), PALB2 (6%), BRIP1 (4.6%), BARD1 (3%), BRCA1 (2%), CDK12 (2%), CHEK1 (1%), FANCL (1%), and RAD54L (1%). In TCGA cohort, 26% (53/208) patients had at least one somatic mutation in an HRR gene. The frequently mutated genes were BRCA2 (9%), BRCA1 (7%), ATM (6%), BRIP1 (4%), PALB2 (4%), BRAD1 (1%), CHEK2 (1%), PPP2R2A (1%), RAD51B (1%), RAD51C (1%), RAD54L (1%), CHEK1 (1%), and RAD51D. Chinese melanoma patients had higher frequently mutated HRR genes compared to that in TCGA patients (40% vs 26%, p=0.005). In addition, compared with TCGA cohort, the Chinese cohort had higher frequency for ATM (14% vs 6%, p=0.019) and CHEK2 (6% vs 2%, p=0.006). The TCGA cohort had a higher frequency of BRCA1 mutations than the Chinese cohort, although the difference was not statistically significant. Unfortunately, we did not find a significant correlation between HRR mutation and immune cells. Interestingly, in the TCGA cohort, patients with HRR-mut had significantly better OS than patients with HRR-wt (log-rank p=0.027). Conclusions: Our data suggest that detection of somatic mutations in HRR genes might contribute to identify patients who might benefit from immune checkpoint blockade therapy. In addition, the molecular features of HRD provide new opportunities to predict the tumor response to multiple treatments.[Table: see text]
Gamma-interferon inducible lysosomal thiolreductase (GILT) effect on breast tumor microenvironment through regulating CXCR6/CXCL16 axis.
Authors: Fei YinJiao , Liang MingXing , Li Lei , Song Yuxin , Liu Zhen , Tang Jinhai
Published: June 2021

e15081

Background: Gamma-interferon Inducible Lysosomal Thiolreductase (GILT) is constitutively expressed in most antigens endocytosed by antigen presenting cells (APCs), and its function is to catalyze the reduction of disulfide (S-S) bonds in protein substrates. The cytokine CXCL16 is one of the only two known plasma membrane chemokines which induces chemotaxis of activated T cells and bone marrow plasma cells in tumor microenvironment. It contains a free end folded by two sulfur bonds and therefore could also be a zymolyte of GILT. Previous studies found that specific receptor of CXCL16, CXCR6, was significantly overexpressed in breast cancer tissues and metastatic axillary lymph nodes. We suppose whether CXCR6/CXCL16 axis is regulated by GILT and affect tumor microenvironment, thereby eliciting specific anti-tumor immune responses in breast cancer (BC). Methods: GILT expression in BC was evaluated using publicly available data from The Cancer Genome Atlas (TCGA). GILT gene was analyzed in UALCAN ( http://ualcan.path.uab.edu/analysis-prot.html ) . In vitro, Immunohistochemistry (IHC) was conducted to examine the location and relation of GILT and CXCR6. Gene Set Enrichment Analysis (GSEA) was performed to mine the biological pathways involved in BC related GILT regulatory network. The expression of GILT at protein and RNA levels were detected by Western Blot and RT-PCR assay. Overexpression and knockdown of GILT in BC cell lines was carried out to further analyzed the correlation between GILT and CXCL16/CXCR6. Results: TCGA database showed that GILT expression was increased in the stroma of BC compared with normal, and was correlated to shorter BC overall survival. GSEA suggested that the expression of GILT was associated with chemotactic factors. Pearson analysis and IHC showed GILT had a strong correlation with CXCL16/CXCR6 axis in the aspect of angiogenesis and immunity. qRT-PCR and Western Blot assay also revealed that GILT had high expression in BC. Besides, patients with high expression of GILT in IHC simultaneously showed high immunoreactive to macrophage markers, which was related to neovascularization and anti-tumor immune responses. Compared with the normal breast cell line MCF-10A, GILT protein had high expression in Hs578T and low expression in MDA-MB-231 cell line. GILT was overexpressed in MDA-MB-231 and knockdown in Hs578T. Result showed that high level GILT promoted the production of CXCL16/CXCR6,while GILT siRNA knockdown inhibited the production of CXCL16/CXCR6. Conclusions: GILT could catalyze CXCL16 in BC, function as a key mechanism to affect tumor microenvironment through CXCR6/CXCL16 pathway. GILT-activated CXCL16 levels showed a strong connection with unfavorable outcomes in BC, which could be a potential biomarker of prognosis and a novel therapeutic target.
Clinical application of next-generation sequencing in cancer patients.
Authors: Shamrikova Valentina , Shilo Polina , Stepanova Maria , Dekhanova Ksenia , Ledin Evgeny
Published: June 2021

e15090

Background: Next-generation sequencing (NGS) has been moving rapidly from pure cancer genomics research towards clinical oncology to provide a rigorous personalized approach to treating cancer. Still there is a number of limitations to overcome before we are ready to fully introduce NGS into everyday clinical decision-making. Here we are sharing our experience with using targeted NGS platforms in clinical practice. Methods: We retrospectively evaluated the NGS results and the treatments, provided to the patients after NGS testing between Marсh 2019 and January 2021. The patients’ clinical characteristics, response to treatment and genomic mutation profiles were reviewed. Coprimary endpoints were the percentage of patients with NGS revealed targeted therapy options and the percentage of patients who received mutation-driven therapy. Results: Samples from 95 patients were tested, most frequently GI tumors (n=25, 26%), hepatobiliary tumors (n=11, 12%), breast cancer (n=10, 11%) gynecological tumors (n=10, 11%), central nervous system tumors (n=7, 7%) and others (n=32, 33%). An average of two gene alterations were identified per patient (range 0-15). The 95% (n=90) of samples had at least one detected mutation, and in 42% (n=40) of cases gene mutations happened to be targetable with existing drugs, including off-label prescriptions. Mutation-driven therapy was performed in 9,5% cases (n=10) with clinically meaningful response in 5 patients (2 patients with ovarian cancer, 2 patients with colon cancer and 1 melanoma). Conclusion: Mutational profiling using a targeted NGS panel has identified potentially targetable alterations in a vast majority of advanced cancer patients. The assay has revealed additional therapeutic options in 42% of patients. However, due to the short-term follow-up, just a small number of patients have received mutation-driven therapy. Further study will be required to determine if the mutation-driven therapy will prove to be clinically meaningful in these patients.
Application of a colorectal cancer ctDNA methylation test in endocrine cancers.
Authors: Winter Jean M. , Sheehan-Hennessy Lorraine , Pedersen Susanne Kartin , Wassie Molla M. , Young Graeme P. , Symonds Erin L.
Published: June 2021

e15533

Background: Assays for circulating tumor DNA (ctDNA) have shown utility for cancer detection and management. It is important to demonstrate cancer specificity of targeted ctDNA biomarkers. Detection of methylated BCAT1 and IKZF1 ctDNA has shown high sensitivity for colorectal cancer (CRC). The aim of this study was to investigate whether hypermethylation of these two biomarkers is also present in prostate or breast adenocarcinoma (PCa or BCa) tissues, and if they are detectable as ctDNA from such cancer patients. Methods: A subset of TCGA data comprising matched tumor and normal samples from PRAD (n = 50), BRCA (n = 90) and COADREAD (n = 44) cohorts were accessed to interrogate DNA methylation (450K) at CpG probes covered by the BCAT1/ IKZF1 ctDNA assay (cg10764357 and cg07589773/cg18607529, respectively). Linear regression of BCAT1/ IKZF1 methylation and tumor staging was performed for cases with AJCC staging data (PRAD n = 503; BRCA n = 788; COADREAD n = 284). Blood was collected from BCa (n = 32) or PCa (n = 101) patients prior to starting treatment at Flinders Medical Centre (South Australia). DNA isolated from plasma was bisulphite-converted and assayed for BCAT1/ IKZF1 methylation. Age-adjusted comparisons of ctDNA positivity were made with blood results from N = 310 pre-treatment CRC patients (60% male), as well as male (N = 344) and female (N = 383) controls with a clear colonoscopy and no history of cancer. Results: BCAT1 and IKZF1 were significantly hypermethylated in PCa and CRC tumors compared to matched normal tissues (Table). Conversely, BCAT1/ IKZF1 CpG probes were hypomethylated in breast tumors compared to matched normal samples. There was no correlation between hypermethylation in IKZF1/ BCAT1 and stage of PCa or CRC. In contrast to PCa in silico findings of hypermethylation, BCAT1/IKZF1 ctDNA methylation was only positive in 7/101 PCa blood samples (6.9%; 95%CI 3.2-13.9%). Three of 32 BCa blood samples (9.4%; 95%CI 2.5-25%) were positive for BCAT1/IKZF1 methylation. These positivity rates were not significantly different to controls at 8.5% for females (p = 0.80 vs BCa) and 8.2% for males (p = 0.73 vs PCa), but all were significantly lower than that observed in CRC cases (59.0%, 183/310; p < 0.001). Conclusions: While interrogation of TCGA datasets revealed IKZF1 and BCAT1 hypermethylation in prostate tumors, detection rate of these biomarkers in blood from PCa patients was not higher than that of non-cancer controls. Likewise, the BCAT1/IKZF1 ctDNA assay positivity rate in BCa patients was also no different to non-cancer controls. These findings highlight the specificity of methylated BCAT1 and IKZF1 ctDNA for detection of CRC.[Table: see text]
Predicting biomarkers of hepatic metastasis in Chinese colorectal cancer patients.
Authors: Peng Honghua , Mu Tianhao , Sheng Yaping , Li Yingmei , Cao Peiguo
Published: June 2021

e15545

Background: Hepatic metastasis is the most common site of distant spread from colorectal cancer. About 15-25% patients with colorectal cancer harbors hepatic metastasis. The molecular mechanism and predicting biomarkers in colorectal cancer are still not fully understood. Methods: 57 Chinese colorectal cancer patients were enrolled in a cohort study. Samples of primary tumor were collected in these patients and underwent whole exome sequencing. Mutation profiles of primary tumors between the patients with metastasis and those without metastasis were analyzed and compared. Results: In the cohort, 54.4% (31/57) patients presented hepatic metastasis at the time of diagnosis, while 45.6% (26/57) did not. The patients were divided into 2 groups—with hepatic metastasis and without hepatic metastasis. The mutation landscape of primary tumor indicated that the Top 3 most frequently mutated genes of both groups were the same and presented mutated TP53, APC, and KRAS. 2. Interestingly, compared with the patients without hepatic metastasis, the patients with hepatic metastasis presented a higher frequency of mutated TCF7L2 (35.5% vs 3.85%) and TRIM77 (16.1% vs 0%). Moreover, in the patients with hepatic metastasis, the patients with TRIM77 mutation in primary tumor showed a worse overall survival (p < 0.0001). Conclusions: TCF7L2 and TRIM77 may be identified as potential candidate predicting biomarkers for hepatic metastasis in colorectal patients. In addition, mutated TRIM 77 predicted a poor overall survival in hepatic metastasis from colorectal cancer.
Update results from ALTER-C-001: Efficacy and safety of anlotinib plus XELOX regimen as first-line treatment followed by maintenance monotherapy of anlotinib for patients with mCRC-A single-arm, multicenter, phase Ⅱ clinical trial.
Authors: Yan Jin , Han Yunwei , Zhang Li , Jin Yongdong , Sun Hao
Published: June 2021

e15550

Background: The combination of anti-VEGF or anti-EGFR targeted drugs with chemotherapy is the standard first-line therapy for metastatic colorectal cancer (mCRC), and the followed maintenance treatment is an optional approach to balance the efficacy and toxicity. However, studies regarding the maintenance strategies based on antiangiogenic TKIs are limited currently. Anlotinib, a novel oral multi-target TKI which can inhibit both tumor angiogenesis and tumor cell proliferation simultaneously, substantially prolonged the PFS with manageable toxicity for refractory mCRC in the phase III ALTER0703 clinical trial. Here we report an update on the effectiveness and safety of anlotinib plus XELOX as first-line treatment followed by anlotinib monotherapy for mCRC. Methods: In this open label, single-arm, multicenter phase II clinical trial, 53 mCRC patients without prior systemic treated, aged 18-75 and an ECOG performance status of 0 or 1 were planned to recruit. Eligible patients received capecitabine (1000 mg/m2, po, d1-14, q3w) and oxaliplatin (130 mg/m2, iv, d1, q3w) plus anlotinib (10mg, po, d1̃14, q3w) treatment for 6 cycles. After 6 cycles of inducing therapy, patients would receive anlotinib (12mg, po, d1̃14, q3w) as maintenance therapy until disease progression or intolerable adverse events (AEs). The primary endpoint was PFS; Secondary endpoints included ORR, DCR, DOR and safety. Results: By the data analysis cutoff date of January 22, 2021, a total of 18 patients were enrolled, of which 12 patients were available for efficacy assessment. In best overall response assessment, there were 50.0% PR (6/12), 33.3% SD (4/12) and 16.7% PD (2/12). The ORR was 50.0% (95% CI, 21.1-78.9%) and DCR was 83.3% (95% CI, 51.5-97.9%). The longest duration of treatment was 8.8 months and the response was still ongoing. The median PFS was not reached. The most common treatment related adverse events (TRAEs) of any grade (≥20%) were leukopenia, hypertension, neutropenia, diarrhea, fatigue, hypertriglyceridemia. Grade 3/4 TRAEs included hypertension (22.2%), hypertriglyceridemia (11.1%), lipase elevated (11.1%) and neutropenia (5.6%). No grade 5 AEs occurred. Conclusions: The update results suggested that anlotinib combined with XELOX as first line regimen followed by anlotinib monotherapy showed promising anti-tumor activity and manageable safety for patients with mCRC. And the conclusions needed to be confirmed in trials continued subsequently. Clinical trial information: ChiCTR1900028417.
The temporal evaluation of RAS and BRAF mutation by liquid biopsy at progression after bevacizumab combinations in patients with metastatic colorectal cancer (mCRC).
Authors: Bilgetekin Irem , Dogan Mehmet , Karacin Cengiz , Basal Fatma Bugdayci , Esin Ece , Ucar Gokhan , Isak Ozlem Aydin , Imamoglu Goksen Inanc , Yildiran Keskin Gul Sema , Erturk Ismail , Aktas Burak Yasin , Kose Baytemur Naziyet , Aydin Kubra , Ozturk Selcuk Cemil , Aksel Bulent , Karadurmus Nuri , Kandemir Olcay , Oksuzoglu Berna , Demirci Umut
Published: June 2021

e15587

Background: Expanded RAS analysis is essential for the selection of biologic agents in mCRC. RAS mutations indicates anti-EGFR unresponsiveness. In this study, we aimed to investigate RAS and BRAF mutations by liquid biopsy at progression in patients with RAS mutant mCRC. Methods: Sixty patients with mCRC who harbored tissue RAS mutations were prospectively analyzed between July 2019 and April 2020. All the patients treated with chemotherapy plus bevacizumab combinations . The plasma samples of the patients were analyzed after progression of bevacizumab combinations. RAS mutation profile was evaluated in plasma using Idylla PCR-based molecular diagnostics method, which enables rapid detection of common mutations in RAS and BRAF genes in circulating tumor DNA (ctDNA). Kaplan-Meier method was used for survival analysis and log-rank test was performed for comparison of groups. Results: The median age of the patients was 60 years (IQR:35-83 years) and female was (n=23, 38.3%). Primary tumor was located in the left colon in 81.7% of all patients. There were 95.0% KRAS and 5% NRAS mutations in baseline tissue biopsy. As a result of liquid biopsy after progression, 55.0% of the patients had KRAS, 3.3% NRAS and 3.3% had BRAF mutations. The RAS mutation detected in 58.3% of the patients. While there was no significant difference in terms of clinicopathological features between wild type (RAS/BRAF) and mutant type (RAS/BRAF) determined by liquid biopsy, the overall survival (OS) of the wild type group was significantly longer than mutant group (43.8 vs. 20.4 months, p= 0.002). Conclusions: This study demonstrated that there may be changes in RAS/BRAF mutation from plasma analysis after progression in patients with mCRC. Since better survival in the patient group with wild type was detected compared to the RAS concordance group, the evaluation of RAS mutation status at the time of progression may be important in terms of disease prognosis and treatment options.