bluuebunny/crossref_metadata_2025 huggingface dataset:
Very low seroprevalence of sars-cov-2 among health care personnel (HCP) in a French northern comprehensive cancer center at the end of first national containment.
Authors: Doutrelant Philippe , Penel Nicolas , Renaudat Charlotte , Von Platen Cassandre , Lebitasy Marie Paule , Smis-Papillon Pauline , Hoen Bruno , Lartigau Eric , Robin Yves-Marie
Published: June 2021

e13604

Background: To limit SARS-Co-2 transmission in general population, French government had set up a first National containment from March 16, 2020 to June 2, 2020. Furthermore, General Direction of our hospital have implemented and organized physical distancing (telework, teleconsultation, virtual multi-disciplinary board), mask wearing, use of alcoholic ... to limit interpersonal contacts. To assess the impact of this policy, we have carried out a seroprevalence study and identified risk factors for SARS-Co-2 prevalence among HCP in May/June 2020. Methods: This is part of CORSER Study (“Etude séro-épidémiologique du virus SARS-CoV-2 en France d’une Cohorte CORSER-2d de personnels d’établissements de santé »), registered with ClinicalTrials.gov (NCT04325646). Two serological tests were applied in this cohort: S-Flow assay and Luciferase-linked immune-sorbent assay (LuLisa-N test). Results: Between 04 May to 26 June 2020, 392 Osar Lambret Cancer Center employees were included in the present study (about 40% of all staff). The most common jobs were: 92 nurses (23.4%), 80 radiology/radiotherapy manipulators (20.4%), 51 physicians (13.0%), 33 medical interns (8.4%) and 47 administrative staff members (11.9%). There were 98 men (25.0%) and 294 women (75.0%). The mean age was 38.7 (+/- 11.4). There were 7 seropositive cases; the seroprevalence was 1.8% (95%-CI: 0.7-3.6). Among the 7 positive cases, 5 were symptomatic (71.4%). In univariate analysis, factors associated with SARS-Co-2 seroprevalence: symptoms suggesting viral infection within 2 months (OR=5.33), dysgueusia (OR=37.00), anosmia (OR=66.29) and HCP exposed to COVID-19 patient outside work (OR=6.69). Gender, tobacco consumption, O blood group, HCP versus administrative staff, HCP working in different services, HCP providing care to suspected COVID-19 patients, HCP providing high-risk cares were not found to be associated with seropositivity. Of note, we have noticed that BMI≥ 24 kg/m² was associated with seroprevalence (OR=15.45), without biological rational. Conclusions: Our study suggests that strict implementation of protective measures was associated with low SARS-Cov-2 prevalence at the end of first National Containment, including among HCP treating COVID-19+ patients. HCPs seroprevalence seemed lower than seroprevalence of the general population at the same period (4-5%). Clinical trial information: NCT04325646.
A prospective data analysis of targeted therapy combined with concurrent radiation therapy for brain metastasis from NSCLC with driver gene mutation.
Authors: Duan Xiaotong , Zhu Xiaoxia , Wang Lijuan
Published: June 2021

e14006

Background: Previous studies have shown that brain metastases of non-small cell lung cancer (NSCLC) with positive driver genes have poor prognosis. There is still lack of prospective studies on the efficacy and safety of targeted therapy combined with concurrent radiotherapy for brain metastases(BM). Methods: NSCLC patients, with ECOG score 0-2, having MRI confirmed brain or meningeal metastases were eligible. Patients must have driver gene mutation and received corresponding targeted therapy. The intracranial radiotherapy regimen was SRS or whole brain radiotherapy. The primary objective was iPFS (intracranial progression-free survival); Secondary objectives were: iORR (intracranial objective response rate), PFS (progression-free survival), OS (overall survival). MMSE (Mini Mental State Examination) and FACT-Br was carried out before/after weekly radiotherapy and during systematic treatment. Treatment-related toxicities were assessed according RTOG/EORTC criteria. Tumor responses were evaluated using RECIST V1.1 criteria. Survival analysis was performed using the Graphprism version 6.0 by Kaplan-Meier method and log-rank test. Results: 23 NSCLC with BM was included. Among them, 10 patients were newly diagnosed with NSCLC BM. 2 patients’ BM progressed after targeted therapy. 11 NSCLC patients were newly diagnosed with BM after targeted therapy. 91.3% of patients presented an EGFR mutation, including primarily EGFR 19-exon deletion, EGFR 21-L8585R. 11.5% presented with c-MET mutation. Median age was 58.34 yrs(44-71yrs). Patients were mostly treated with Erolotinib and Gefitinib. All patients were adenocarcinoma. At last follow-up, for patients newly diagnosed with NSCLC BM, 8 patients had achieved intracranial progression, and 7 patients had reached OS, of which 1 died before completing WBRT. The median iPFS was 9.3m(95%CI:0.571-4.055) and the median OS was 11.9m (95%CI:0.2752 -2.732). As for patients who progressed after targeted therapy, one patient’s OS was 4.4m, iPFS of the other patient was 3.9m. Among NSCLC patients who were newly diagnosed with BM after targeted therapy, 8 patients had achieved intracranial progression and 5 patients had reached OS. The median iPFS was 6.13m (95%CI:0.247-1.751) and the mOS was 13.8m (95%CI:0.3660-3.634). Common adverse effects include dry skin, fatigue, dizziness, headache, anorexia, and grade I myelosuppression and no serious adverse events (SAEs); MMSE and FACT-Br scores were no significant differences at baseline and follow-up. Conclusions: In stage IV brain metastatic NSCLC with driver gene mutation, targeted therapy combined with concurrent radiotherapy for BM is tolerable, and there is no significant impact on the quality of life and cognitive function after radiotherapy. The evaluation of efficacy requires further follow-up. Support:LC2019ZD009,81972853 and 81572279.
Stem cell properties of GT101, a tumor infiltrating lymphocyte (TIL) product with multiple indications.
Authors: Liu Yarong , Zhao Deping , Shi Zixiao , Zhou Rong , Sun Jingwei , Han Yong , Zhao Peipei
Published: June 2021

e14506

Background: Tumor infiltrating lymphocyte (TIL) is a kind of adoptive T cell therapy that has demonstrated clinical benefits in several types of solid tumors. GT101 is an autologous TIL therapy for the treatment of multiple solid tumors. Previous studies showed that in vitro expansion resulted in over 80% terminally differentiated effector T cells in the final TIL products, which may show good cytotoxicity but weak capability of persistence and proliferation in vivo. Herein we present phenotype and function characterization of our TIL product GT101 with a focus on its stemness attributes. Methods: We have produced more than 30 batches of GT101 TIL samples from 10 different cancer indications and analyzed their proliferation, cytokine production and phenotype at preREP and/or REP stage. Results: GT101 products keep a consistent T cell phenotype and good proliferation capacity, and are capable of secreting large amounts of cytokines after stimulation by anti-CD3/CD28 antibody, reflecting its good functionality. The residues in the final products, such as feeder cells and tumor cells, are below the detection limit. In contrast to equivalent TIL products previously reported across the world, we find that GT101 features a manufacturing process that preserves the stem cell characteristics of T cells. Analysis of all the 30 GT101 TIL samples demonstrates that the average percentage of central memory T cells (Tcm) and naïve T cells is 40-50% and 10-20% respectively in both CD8+ and CD4+ populations at the end of preREP stage. Meanwhile, CD8+TCF1+ cells are close to 50%, mainly in the form of PD-1-. In the phase of REP, the stemness of T cell is still well maintained—the average proportion of both Tcm and naïve T cells only decreases by about 5%, and that of CD8+TCF1+ cells down by about 10%. Conclusions: In general, GT101 demonstrates good stemness with a higher percentage of Tcm and TCF1+ than equivalent products previously reported across the world, suggesting better performance in persistence and proliferation in clinic.
Effect of Wnt pathway inhibitor on ovarian cancer tumors.
Authors: Mindar M. V. , Goncharova Anna S. , Volkova A. V. , Verenikina Ekaterina V. , Menshenina Anna P. , Zaikina Ekaterina V. , Khodakova D. V. , Lukbanova Ekaterina A. , Ardzha Anna Yu. , Kurbanova L. Z. , Adamyan M. L.
Published: June 2021

e15010

Background: Ovarian cancer (OC) is the fourth most common cancer in women aged 40-54 years. Despite advances in cancer research, no existing methods provide effective early diagnosis and treatment of OC, so the search for new medications is an urgent task. The effectiveness of the XAV-939 pharmacological substance, which inhibits the Wnt signaling pathway, was studied in an orthotopic patient-derived xenograft of human ovarian cancer. Methods: A PDX model was created in 20 female immunodeficient Balb/c Nude mice. A tumor fragment was implanted into the ovary, and the ovary was transposed under the skin to better visualize the xenograft growth. When the average tumor volume was 67.4 mm3, animals were divided into 4 groups: group 1 – controls (n = 5); group 2 – animals receiving paclitaxel (n = 5); group 3 – animals with XAV-939 (n = 5); group 4 – animals with combined paclitaxel+XAV-939 (n = 5). The control group received the carrier substance, and the test substances were administered at 10 mg/kg; the substances were administered intraperitoneally twice a week for 22 days. The tumor growth was assessed throughout the experiment by measuring its linear dimensions (length and width), and the volume and tumor growth index were calculated. The data were statistically processed using Microsoft Excel and STATISTICA 10. The Mann-Whitney nonparametric test was used to assess the differences between the control and experimental groups. Results: No statistically significant differences were observed in the indices of tumor growth between groups 2 (paclitaxel) and 1 (control), and groups 3 (XAV-939) and 1 (control). However, the difference was statistically significant between groups 4 (paclitaxel+XAV-939) and 1 (control). The tumor growth inhibition index was calculated at the end of the experiment. It was maximal in group 4 (paclitaxel+XAV-939) – 67.78%; in groups 2 (paclitaxel) and 3 (XAV-939), it was 21.39% and 30.33%, respectively. Conclusions: An orthotopic PDX model of ovarian cancer was generated, and the activity of XAV-939 towards the OC xenograft was studied; the greatest efficacy of the preparation was noted in combination with paclitaxel.
Association of genes copy number in extracellular DNA in patients with prostate cancer and sensitivity to radiotherapy.
Authors: Kutilin Denis S. , Zinkovich Mikhail S. , Gusareva Marina A. , Faenson Aleksandr V. , Karnauhova Elena A. , Solntseva Anna A. , Fatkina Natalya B. , Krokhmal Julia N. , Vasilieva Ekaterina O. , Shevchenko Alexey N. , Filatova Elena V. , Khomutenko Irina A. , Rozenko Lyudmila Ya. , Udalenkova Irina A. , Gappoeva M. A. , Donskaya Aliya K.
Published: June 2021

e15014

Background: Radiotherapy (RT) is one of the main treatments for prostate cancer (PC). The effectiveness of such therapy depends on the initial radioresistance of tumor cells, which is ensured by their certain molecular features, which include the genes copy number variation (CNV). Model experiments on cell cultures (obtained from surgical material) have shown that CNVs have high potential as predictors of RT sensitivity. However, this potential is limited by the high level of invasiveness in obtaining biomaterials. A possible solution to this problem lies in the transition to CNV study in the extracellular DNA (cfDNA) of blood plasma. The aim of the study was to screen predictors of radioresistant PC based on the genes CNV in cfDNA. Methods: The study included 400 patients with diagnosed PC (T2a-3bN0M0, st. II-III), 40 of them after RT had a state of biochemical relapse (RT was performed on a Novalis TX linear accelerator (Varian, USA) (TFDisoeff = 75 Gr), mean time to biochemical relapse 7.5 months). Blood samples were separated into plasma and cell fraction by centrifugation. Isolation of cfDNA from blood plasma was performed using a set of reagents “DNA-Plasma-M” (Russia). Determination of the relative CNV of 13 genes (CDK1, CCND3, CDKN1B, TP53, PTEN, BCL2, XRCC4, BAX, RBBP8, H2AX, BRCA2, RAD50, EP300) was performed using the Real-Time qPCR method. Differences were assessed using the Mann-Whitney test; the Benjamin-Hochberg correction was used to correct multiple comparisons. Results: In the group with biochemical relapse (n = 40), the CNV of genes CDK1, CDKN1B, RBBP8, XRCC4, BRCA2 and RAD50 was statistically significantly (p < 0.05) higher by 2.0 times, 2.3 times, 2.1 times, 1.4 times, 2.4 times and 2.8 times, respectively, relative to the CNV of these genes in the cfDNA of the group without relapse (n = 360). Conclusions: Thus, it was found that the CNV of 6 genes (CDK1, CDKN1B, RBBP8, XRCC4, BRCA2 and RAD50) may be a potential molecular marker of radiosensitivity of prostate tumors. Based on the obtained data, a low invasive method for determining the prostate tumors sensitivity to RT has been developed.
Investigation of TLR4 and pSTAT3 expression on tumor and immune cells in the peripheral blood (PB) of patients with breast cancer (BC).
Authors: Papadaki Maria A , Monastirioti Alexia , Apostolopoulou Christina A , Aggouraki Despoina , Papadaki Chara , Michaelidou Kleita , Mavroudis Dimitrios , Agelaki Sofia
Published: June 2021

e15015

Background: Toll-like receptor 4 (TLR4) and phosphorylated signal transducer and activator of transcription protein 3 (pSTAT3) hold a key role in inflammatory response and tumor immune evasion. Their role in the peripheral immune response and in tumor immune interactions in circulation needs further investigation. In the current report we evaluated the incidence of TLR4 and pSTAT3 expression on tumor and immune compartments within the peripheral blood (PB) of patients with early (eBC) and metastatic breast cancer (mBC). Methods: PB was obtained from patients with eBC (n = 99) and mBC (n = 100), prior to adjuvant and first-line therapy, respectively. Triple immunofluorescence staining for cytokeratins (CK), TLR4 and pSTAT3 was performed on peripheral blood mononuclear cell (PBMC) cytospins. Single cell-level expression of TLR4 and pSTAT3 was assessed on CK+ circulating tumor cells (CTCs) and PBMCs using the Ariol microscopy system. Results: CK+ CTCs were detected in 6.1% and 19% of patients with eBC and mBC, respectively (p = 0.006) (number of CTCs: n = 34 and n = 28, respectively). Higher TLR4 expression rates on CTCs were observed in mBC as compared to eBC (53.6% vs 7.7% of CTCs, 13% vs 3% of patients; p = 0.016). pSTAT3+ CTCs also prevailed in mBC (60.7% vs 10.3% of CTCs; 14% vs 5.1% of patients; p = 0.032). In the metastatic patients, CTC detection was associated with shorter overall survival (median OS: 24.9 vs 36.5 months; p = 0.042; Kaplan Meier), whereas the detection of TLR4+ CTCs predicted for shorter progression free survival (median PFS: 11.4 vs 12.6 months; p = 0.036). At the PBMC level, TLR4 expression was more frequent among patients with mBC as compared to eBC (34% vs 20.2%, respectively; p = 0.029), whereas pSTAT3 expression prevailed in eBC (89.9% vs 77% of patients, respectively; p = 0.014). The detection of TLR4+ PBMCs was associated with reduced disease-free survival (median DFS: not reached; p = 0.007) and shorter OS (median OS: not reached; p = 0.028) in patients with eBC. Conclusions: TLR4 and pSTAT3 expression is demonstrated on CTCs and PBMCs and significantly varies among eBC and mBC, suggesting that the inflammatory signaling through TLR4 and pSTAT3 in PB could be associated with BC progression. Our results further imply that their assessment in PB may hold significant prognostic implications in BC.
Targeting DNAJC19 overcomes tumor growth and metastasis in NSCLC by regulating AKT1 signaling.
Authors: Ren Tao
Published: June 2021

e15024

Background: Some driver oncogenes are still unknown in non-small cell lung cancer (NSCLC). DNAJC19, known as a major component of the translocation machinery of mitochondrial membranes, is a disease-associated protein. Herein, we reported the role of DNAJC19 in NSCLC cell growth and metastasis. Methods: Immunohistochemistry (IHC) was performed to investigate DNAJC19 expression in NSCLC clinical samples. For knockdown or overexpression assays in A549 or NCI-H1299 lung cancer cells, lentiviral vectors were used. After assessment of cell functions, DNAJC19-knockdown A549 cells were further applied to establish mice xenograft and metastasis tumor models. STRING database, western, PCR, immunoprecipitation and IHC were performed for the mechanistic study. Results: Expression of DNAJC19 was higher in tumors than non-cancerous adjacent tissues. Survival analysis found that low DNAJC19 levels were correlated with increased progression-free survival rate. ShRNA-mediated knockdown of DNAJC19 markedly inhibited cell growth, viability, migration and invasion. Moreover, RNA-seq analysis explored that PI3K/AKT signaling pathway was involved in molecular events when A549 cells treated with shDNAJC19. And then DNAJC19 knockdown also decreased AKT1 expression in A549 cells. In addition, cellular functions were greatly rescued in DNAJC19-knockdown A549 cells by exotic overexpression of AKT1. Furthermore, tumor xenograft growth and metastasis were markedly repressed in the shDNAJC19 group than control group. As expected, DNAJC19 and AKT1 expression levels in xenograft mice samples were also lower in the shDNAJC19 group than in the shCtrl group. Conclusions: DNAJC19 greatly promotes NSCLC cell growth and metastasis via regulating AKT1 signaling, providing a novel therapeutic target for NSCLC patients.
Characterization of NTRK gene fusion events in solid tumors among Chinese patients.
Authors: Zhang Xuhui , Chen Dongsheng , Zhang Qin , Duan Qianqian , WANG Qing , Li Si , Xiao Mingzhe
Published: June 2021

e15040

Background: NTRK fusions are actionable genomic alterations detected across tumor types. NTRK gene fusions involving either NTRK1, NTRK2 or NTRK3 (encoding the neurotrophin receptors TRKA, TRKB and TRKC, respectively) are oncogenic drivers of various adult and paediatric tumour types. Here, we update the detection of NTRK gene fusions across tumor types and further describe fusion partner characteristics among Chinese patients. Methods: Samples submitted for clinical molecular profiling were retrospectively analyzed for NTRK fusion events. Method for identifying NTRK fusions was DNA-based next-generation sequencing that tumour DNA is extracted from formalin-fixed paraffin-embedded tissue. All NTRK fusion partners were identified for intact functional domains, domain prediction, breakpoints, frame retention and co-occurring alterations. Results: A total of 64 NTRK fusion events (0.26% of 24,451) were identified. NTRK fusions are characteristic in a few rare types of cancer, such as melanoma, glioma and carcinomas of the thyroid, lung and colon, but they are also infrequently seen in some uncommon cancers, such as secretory carcinoma of the breast or salivary gland and infantile fibrosarcoma. Among the fusions, NTRK1 (0.08% of 24,430), NTRK2 (0.02% of 24,445), NTRK3 (0.15% of 24,414) were identified. Twenty-six unique fusion partners were identified, the most common in NTRK1 fusion being TPM3 (23.8%), NTRK2 fusion being AGTPBP1 (33.3%), and NTRK3 fusion being TFG (13.5%). Almost 53.8 % (14 of 26) of all fusion events are expected to include the transmembrane domain contributed by the NTRK fusion partner. The most commonly identified breakpoints occur in exon 14 and exon 17 and in exon 15 and exon 20, in NTRK1, NTRK3, respectively. Conclusions: NTRK fusion products are diverse across tumor types, but the significance of these variations is not clear. The biological and clinical implications of retaining certain domains of NTRK and of fusion partners warrants further investigation.
Discovery of HBW-3-20, the first potent reversible inhibitor of Bruton’s tyrosine kinase (BTK) with high brain exposure.
Authors: Lee Ning , Li Yingfu , Yuan Chester , Liu Guanfeng , Yue Chunchao
Published: June 2021

e15068

Background: It has been an on-demand task to develop a BTK inhibitor of significant brain exposure, a critical property for extending its usages to treat Primary Central Nervous System Lymphoma (PCNSL) and autoimmune disorders. PCNSL is an aggressive extra nodal non-Hodgkin lymphoma that exclusively invades the central nervous system (CNS). Tirabrutinib, an irreversible BTK inhibitor with limited brain exposure, is the first BTK inhibitor approved for the treatment of recurrent or refractory primary central nervous system lymphoma recently. PRN2246 is another irreversible BTK inhibitor claimed to be of brain exposure, and is currently in clinical trials for the treatment of multiple sclerosis. Methods: New reversible BTK inhibitors were designed, synthesized and tested for enzymatic activities against wild-type and C481S-mutated BTK. Highly active compounds were confirmed for growth effects in diffuse large B-cell lymphoma derived TMD8 cells. Their microsomal stability and ADME properties were also assessed. Potent and bioavailable compounds were further measured for brain exposures in rats. Results: HBW-3-20 has excellent potency against both wild-type and C481S-mutated BTK, with IC50 of 2.5 and 3.8 nM, respectively. Its TMD8 cellular potency is 72 nM. In a head-to-head direct comparison of brain exposure experiment, HBW-3-20, tirabrutinib and PRN2246 were all dosed at 10mg/kg orally. The brain and plasma concentration were measured after 1 hour and the data are shown in the table below. The brain to plasma ratio for HBW-3-20, tirabrutinib and PRN2246 are 58%, 11.8% and 4.2% respectively. Our results show that HBW-3-20 has much greater brain permeability than tirabrutinib or PRN2246 in rats. Conclusions: HBW-3-20 is the first potent reversible BTK inhibitor that shows promisingly high brain permeability. HBW-3-20 provides a very valuable clinical candidate for treating B-cell malignancies in brain and autoimmune disorders![Table: see text]
Accurately estimating the homologous recombination deficiency based on low-pass whole-genome sequencing and target sequencing.
Authors: Lv Fang , Liu Yiqian , Zhang Yaxi , Liu Xiaoli , Zhihua Pei , Yu Jianing , Chen Weizhi , He Ji
Published: June 2021

e15077

Background: Homologous recombination deficiency (HRD) is a promising biomarker for poly ADP-ribose polymerase (PARP) inhibitors. In the past, the HRD status was mainly estimated by summing the loss of heterozygosity (LOH), telomeric allelic imbalance (TAI), and large-scale state transition (LST) based on target sequencing. However, the heterozygous loci coverage and probe number are closely correlated to B allele frequency and copy number variations (CNVs). Here we developed a novel method to combine the data from low-pass whole-genome sequencing (WGS) and target sequencing to accurately estimate the genome-wide CNVs and allelic imbalance. Methods: We first performed low-pass WGS and target sequencing on 30 healthy controls, formalin-fixed and paraffin-embedded (FFPE) samples, and matched blood cell samples for 29 patients with breast cancer or ovarian cancer. Two HRD positive samples were diluted to 40%, 30%, 20%, and 10% tumor purity with matched blood cell samples, respectively. Each diluted sample was sequenced 20 times. The baseline for CNVs was built based on the sequencing data of healthy donors. To eliminate the artifacts resulted from the sequencing platform, models for each candidate single nucleotide polymorphism (SNP) in target regions were trained using the data from healthy donors. The mutations of BRCA1/BRCA2 of 29 patients were used to define the threshold of the HRD score. The numbers of LOH, LST, and TAI were evaluated by CNVs and B allele frequency. Results: We defined a cutoff of 42 for the HRD status based on the low-pass WGS and target sequencing data of 29 patients. Specifically, the original HRD scores for two selected HRD positive samples were 68 and 89. The median HRD scores for 40%, 30%, 20%, and 10% tumor purity samples were 69 and 88, 66.5 and 90, 61 and 68, 52 and 34.5, respectively. Conclusions: We developed a novel method for robustly inferring the HRD status using low-pass WGS and target sequencing with a limit of detection of 20%.
Transformation from non-small cell lung cancer to small cell lung cancer: A report of clinicopathological characteristics and prognoses.
Authors: Wang Jun , Liu Qing , Shang Yanhong , Xiao Linlin , Huo Ran
Published: June 2021

e15084

Background: The transformation to small cell lung cancer (SCLC) is one of the mechanisms of drug resistance in non-small cell lung cancer (NSCLC). However, there are few reports on it. The study aims to analyze the clinicopathological characteristics of the patients, to explore the transformation factors and reasonable treatment strategy for them. Methods: A total of 9 patients with pathologically diagnosed NSCLC transformed into SCLC after first-line or multi-line treatment were involved in the study. Results: Among the 9 patients, there were 8 adenocarcinomas (ADC) and 1 squamous cell carcinoma (SCC), 7 of them were non-smokers. Epidermal growth factor receptor (EGFR) mutations were present in all ADC patients, including 7 with exon19 deletion (87.5%) and 1 with exon21 mutation (12.5%). All of them received EGFR tyrosine kinase inhibitors (TKIs) before the transformation, and the median transformation time was 19.5 months (95% CI, 14.1-32.7months). After first-line chemotherapy with paclitaxel and cisplatin, one SCC patient transformed to SCLC within 5 months. EGFR mutation was still maintained in 6 ADC patients when transformation occurred, of which 2 patients had a new TP53 mutation and 1 had RB1 gene mutation. Serum tumor markers were monitored in 6 patients, of them, progastrin releasing peptide (Pro-GRP) increased apparently at the time of disease progression in 5 patients, and neuron-specific enolase (NSE) also increased in 3 patients. This phenomenon occurred three to six months earlier than the transformation. The median survival time (MST) was 37 months from the initial diagnosis of NSCLC and 13 months after transformation. The median progression-free survival (mPFS) was only 2 months for those treated with chemotherapy (CT) after transformation, whereas 6 months treated with CT combined with TKIs as the first-line treatment. Three patients who received chemotherapy combined with programmed death receptor-1 (PD-1) inhibitor after multi-line therapy underwent progress within 2 months, nevertheless, 1 patient treated with the EP regimen combined with pembrolizumab as the first line treatment after transformation had a PFS of 12 months. After re-progression, the patients got another PFS of 5 months with a pemetrexed-platinum combined with atezolizumab. Conclusions: Phenotypic transformation is one of the mechanisms of drug resistance for NSCLC patients, especially for those with EFGR exon19 deletion received EGFR-TKIs. Tumor heterogeneity, TP53 gene and Rb gene mutation may be responsible for the transformation. The serum Pro-GRP over expression could as an early predictor of transformation. Classical chemotherapy is ineffective after SCLC transformation patients who still have sensitive mutation, it might be suitable to combine with EGFR-TKIs. The early application of immunotherapy could be attempted, but still needs to be explored further.
Profiling receptor tyrosine kinase fusions in Chinese breast cancers.
Authors: Tao Zhonghua , Hu Xichun , Cao Wen-Ming , Liu Jianxia , Li Ting , Xu Hong , Chen Kai , Zhang Jian , Zhou Hao , Sun Jie , Han Jinming , Yang Hua , Shao Lin , Xiang Jianxing , Guan Xiaoqian , Li Min
Published: June 2021

e15092

Background: Receptor tyrosine kinases (RTKs) are a class of tyrosine kinases that regulate cell-to-cell communication and control a variety of complex biological functions. Dysregulation of RTK signaling partly due to chromosomal rearrangements leads to novel tyrosine kinase fusion oncoproteins which are possibly driver alterations to cancers. Targeting some RTK fusions with specific tyrosine kinases inhibitors (TKIs) is an effective therapeutic strategy across a spectrum of RTK fusion-related cancers. However, there is still a paucity of extensive RTK fusion investigations in breast cancer. We aimed to characterize RTK fusions in Chinese breast cancer patients. Methods: An in-house sequencing database of 1440 Chinese breast cancer patients using a 520-gene NGS sequencing panel was thoroughly reviewed. RTK fusion was defined as an in-frame fusion with the tyrosine kinase domain of the RTK completely retained with the only exception of ERBB2 fusion which was not counted due to its unclear significance. Concomitant mutations and TMB were also analyzed and calculated. Patients’ clinical characteristics were retrieved from case records. Results: 27 RTK fusion-positive breast cancers (12 tissues + 15 plasmas) were identified, patients had a median age of 52 years. Triple-negative breast cancer subtype comprised 37% with luminal and HER2 positive subtypes being 40.8% and 22.2%, respectively. 77.8% of patients were at stage IV and 22.2% at stage I-III. Ten were treatment naïve. RTK fusions occurred in 2% of breast cancers in our database, compared with the prevalence of 0.6% and 1.3% in MSKCC and TCGA, respectively. In the subset of stage IV patients, our database showed a significantly higher RTK fusion frequency than that in MSKCC (3.2% vs. 0.6%, p = 0.013). FGFR2 fusions were seen most commonly (n = 7), followed by RET (n = 4), ROS1 (n = 3), NTRK3 (n = 3), BRAF (n = 2), and NTRK1 (n = 2). Other RTK fusions including ALK, EGFR, FGFR1, FGFR3, MET, and NTRK2 were identified in one patient each. Of note, the normalized abundance of RTK fusion (fusion AF/max AF) correlated negatively with TMB (r = -0.48, p = 0.017). Patients with TMB < 4 (Muts/Mb) displayed a higher fusion abundance than those with TMB ≥ 4 (Muts/Mb) (p = 0.018), suggesting a higher likelihood of subclonal nature for RTK fusions in TMB-high patients. Moreover, CREBBP mutation only co-occurred with FGFR2 fusion (p = 0.012), while NTRK3 fusion and TP53 mutation were mutually exclusive (p = 0.019). Conclusions: This is the first study comprehensively delineating the prevalence and spectrum of RTK fusions in Chinese breast cancers. Further study is ongoing to identify the enriched subpopulation which may benefit from RTK fusion inhibitors.
Treatment dependent improvements in survival of stage 2/3 rectal cancer patients treated with trimodality therapy between 2006-2016, an NCDB analysis.
Authors: Kennecke Hagen Fritz , Bahnson Henry T. , Lin Bruce Shih-Li , Kaplan Jennifer , Pham Huong , Suen Andrew , Simianu Val
Published: June 2021

e15543

Background: Trimodality therapy (TT) with chemo/radiation (C/RT), chemotherapy and total mesorectal excision (TME) surgery remains the standard for patients with stage 2/3 rectal cancer. Use of pre-operative (pre-op) C/RT is an important Commission on Cancer (CoC) quality benchmark but has not previously been shown to improve overall survival when compared to post-op C/RT. The objective of this study was to document the impact on survival of peri-op C/RT in stage 2/3 rectal cancer in a broad population. Methods: The National Cancer Database was used to identify all patients diagnosed with stage 2/3 rectal cancer from 2006-16. Included patients received true TT and were classified into groups A, Total Neoadjuvant Therapy (TNT) with pre-op C/RT + pre-op multi-agent (MA) chemotherapy (CT); group B, pre-op C/RT+ post-op single-agent CT; group C, pre-op C/RT + post-op MA CT; and group D, post-op C/RT and MA CT. Cox multivariate survival analysis were performed including demographics, peri-op C/RT, surgery type, stage, lymph node count, year of diagnosis and facility type: academic (Acad), Comprehensive (Comp)/Community (Comm), Integrated (Integ) and unknown (Unkn). Results: Of 110,372 stage 2/3 patients, 32,467 received TT (mean age 58, 61% male) and were included. Of these, 8883 (27%, group A) received TNT, 5967 (18%) were in group B, 12,928 (40%) in group C, while 4,689 (14%) were in group D. A reduction in use of post-operative C/RT (group D) was observed between 2006 (28%) and 2016 (8%), p < .001, accompanied by a reciprocal increase in patients receiving pre-op C/RT and post-op MA CT (Group C) between 2006 and 2016 (24 to 45%, respectively, p<0.001). Increasing use of pre-op C/RT led to a migration to lower pathologic stages 0/1/2/3 from 0.60/10/31/57% in 2006, to 2.8/22/29/45% in 2016, respectively (p < .001), while clinical stage 2/3 distribution remained unchanged. Receipt of pre-operative C/RT (Groups A/B/C) was associated with improved survival compared to post-op C/RT (group D) (table). Conclusions: Between 2006-2016 the proportion of patients with stage 2/3 rectal cancer treated with post-op C/RT declined dramatically and in 2016 accounted for 8% of all patients treated with TT. Multivariate analysis documented superior overall survival among patients treated with pre-operative C/RT, justifying the introduction of the CoC quality benchmark.[Table: see text]
Real-world time on treatment and overall survival in patients with metastatic colorectal cancer receiving cetuximab in second line after failing irinotecan or oxaliplatin-based regimens.
Authors: Gathirua-Mwangi Wambui , Yang Tony , Khan Taha , Wu Yixun , Afable Manuel
Published: June 2021

e15568

Background: Anti-epidermal growth factor receptor agents are increasingly used in later lines of therapy for the treatment of metastatic colorectal cancer (mCRC). However, the real-world time on treatment (TOT) and overall survival (OS) of patients with mCRC receiving cetuximab in second-line (2L) setting have not been described. Therefore, we sought to evaluate TOT, OS and identify factors associated with longer TOT and OS based on retrospective observational data. Methods: A total of 1,011 patients were selected from the nationwide Flatiron electronic health record database (January 2013-August 2020) who were: 1) diagnosed with mCRC, 2) received 2L treatment with cetuximab containing regimens, and 3) had failed oxaliplatin/irinotecan-based regimens in first-line (1L). TOT was defined as the time from initiation of cetuximab in 2L (index date), to the last date showing evidence of cetuximab administration. End of therapy was defined if patients progressed to third-line of therapy, or having a death record. OS was calculated from the index date to the date of death or censored to last visit date available. The Kaplan-Meier estimates, and stepwise Cox models were adapted to calculate hazard ratios (HR) and 95% confidence intervals (CI) for associated factors. Results: Majority of patients receiving 2L treatment with cetuximab containing regimens were: less than 65 years old (58%), male (60%), had a median body mass index (BMI) of 26.6 kg/m², received FOLFOX regimens in 1L (61%) and were treated in the community setting (96%). The most common 2L regimens were cetuximab+FOLFIRI (46%) and cetuximab+Irinotecan (29%). Overall, the median TOT (mTOT) for patients receiving 2L cetuximab was 3.94 months (median Interquartile range (mIQR) 3.51-4.40), and median OS (mOS) was 14.36 months (mIQR 13.01-15.70). Of all cetuximab containing regimens, receiving cetuximab+FOLFIRI in 2L (mTOT = 4.43 months, mIQR 3.71-5.36), KRAS wild-type (vs. KRAS+), and receiving 2L cetuximab after prior therapy with 1L CapeOX (vs. FOLFOX) were associated with a longer mTOT. Living in the South region (vs. Midwest) was associated with a shorter mTOT. Having a higher BMI (obese vs. underweight HR = 0.46, 95% CI 0.32-0.66) was associated with a longer mOS, while receiving cetuximab+FOLFOX in 2L (mOS = 10.97 months, mIQR 5.55-14.06) or being older (≥65 vs. < 65 years; HR = 1.24, 95% CI 1.05-1.46) were associated with a shorter mOS. Conclusions: In this real-world retrospective analysis we show TOT and OS overall in mCRC patients who received cetuximab containing regimens in 2L. These patients were mostly male, < 65 years, and majority received FOLFOX regimens in 1L therapy. Key factors associated with TOT and OS were treatment regimens received in 1L and 2L. In addition, KRAS status and region were associated with TOT, while BMI and age were associated with OS only.
Trop-2 inactivates E-cadherin and drives colon cancer metastasis.
Authors: Alberti Saverio , Guerra Emanuela , Altomare Donato F. , Depalo Raffaella , Trerotola Marco
Published: June 2021

e15576

Background: Metastatic diffusion is the biggest hurdle for colon cancer (CRC) cure and the identification of decisive drivers of CRC metastasis is an urgent need in CRC care. Methods: The expression of target molecules in primary tumors and metastases was systematically assessed by DNA array and IHC analysis. Cell-cell adhesion capacity was quantified in 2D cell cultures and in HCT116 CRC cell spheroids. Pro-metastatic impact of wtTrop-2 and activated, tail-less Trop-2 (Δcyto) was assessed in vivo in orthotopic diffusion models of KM12SM CRC cells. Primary CRC and metastasis transcriptomes were analyzed for differential induction of EMT determinants. Kaplan–Meier plots were used to illustrate survival and metastatic relapse in independent case series of CRC patients. Results: Trop-2 was identified as uniquely upregulated in CRC models by transcriptome analysis. wtTrop-2 and ΔcytoTrop-2 were shown to induce cell migration, wound-healing and resistance to apoptosis induction . wtTrop-2 increased the metastatic capacity of KM12SM cells, raising metastasis diffusion from 45% for control cells to 90% for wtTrop-2 transfectants. The constitutively-active ΔcytoTrop-2 further boosted metastatic spreading, with metastatic livers reaching up to four times their normal size. Constitutive high expression of E-cadherin was revealed in cancer metastases. No evidence was obtained for transcriptional down-regulation of epithelial differentiation biomarkers. No induction of EMT transcription factors was observed in Trop-2-activated cells. Trop-2 tightly bound E-cadherin and caused its release from the cytoskeleton, for loss of cell-cell adhesion and activation of β-catenin. The Trop-2/E-cadherin/β-catenin-driven pro-metastatic program was recapitulated in CRC patients and was shown to impact on CRC metastatic relapse and overall patient survival. Conclusions: We identify Trop-2-driven functional inactivation of E-cadherin as a key driver of metastatic diffusion in CRC. These findings may pave the way for novel multi-marker personalized diagnostics and anti-cancer therapies. [Table: see text]
Responsibility of immune microenvironment for poor response to immune checkpoint inhibitors in CRC patients with KRAS mutation.
Authors: Liu Qiyu , Zhang Hushan , Cai Shenggang , Xu Lin , Zhao Rong , Zhang Xiaolong , Yin Jianhui , Zhang Lijun , Liu Leyang , Gong Xiaolei , Huang Depei , Shen Xudong , Luo Shibi
Published: June 2021

e15588

Background: The clinical trial Keynote 177 is a breakthrough research in colon rectal cancer (CRC), however, the results of subgroups revealed that immunotherapy did not benefit more than chemotherapy in patients with KRAS mutation. Here we assessed characteristics of tumor immune microenvironment in patients with or without KRAS mutation, to find out the underlying mechanisms that prevent KRAS-mutated CRC patients from benefiting from immunotherapy. Methods: Tumor tissue samples of 77 CRC patients were collected from Janary 2019, several kinds of immune cells including CD8+ T cells, M1/M2 tumor associated macrophages (TAM), CD56bright NK cells and CD56dim NK cells in the tumor tissue were evaluated by multiple fluorescence immunohistochemistry (mIHC), and expression of PD-L1 were detected by using Dako PD-L1 IHC 22C3 pharmDx, Tumor Proportion Score (TPS) was used to determine expression of PD-L1. In addition, gene mutation was detected by means of next generation squesing (NGS). All these detections were performed in a College of American Pathologists (CAP)-certified and Clinical Laboratory Improvement Amendments (CLIA)-accredited lab for gene mutation analysis (3D Medicines Inc.,Shanghai, China). Statistical analysis was performed using GraphPad Prism (version 7.01) and SPSS version 21.0 (SPSS,Inc.). Results: According to whether KRAS is mutated, these 77 patients were divided into two groups, KRAS wild type (WT) (n = 49) and KRAS mutation group (n = 28). There was no significant difference in the CD8+T cells infiltrated in tumor tissue of KRAS WT and KRAS mutation samples, however, less CD8+ T cells were found in stroma of tumor tissue of KRAS WT samples than in KRAS mutation. What’s more, both the ratio of M1:M2 and CD56dim NK: CD56bright NK cells in the tumor tissues of KRAS WT were significantly higher than in KRAS mutation samples(p < 0.05). No significant difference was found between two groups, while higher MSI-H proportion was found in KRAS mutation group than in KRAS WT (33.4% and 12.5%, respectively). The proportion of positive PD-L1expression (TPS≥1) in both groups is very low, especially, proportion of strong positive (TPS≥50). Conclusions: The different response to immune checkpoint inhibitors therapy for CRC patient with or without KRAS mutation mainly associated with factors of TIME, not only CD8+T cells, but tumor associated macrophages and different sub-population of NK cells should be put into consideration.
Mutation profile of DNA damage response genes in Chinese colorectal cancer patients and its association with TMB and MSI.
Authors: Yang Meiling , Yao Jianfei , Liu Danni , Huang Tanxiao , Wang Zhizheng , Chen Shifu , Huang Haifu
Published: June 2021

e15608

Background: DNA damage response (DDR) genes are essential to maintain genomic integrity. Increasing evidence showed that mutations in DDR genes were related to tumor mutation load (TMB), microsatellite instability (MSI) and clinical benefit from immunotherapy. TMB and MSI are predictive biomarkers for immunotherapy. However, the mutation spectrum of genes involved in the DDR pathways and its association with TMB and MSI in Chinese colorectal cancer patients are still ambiguous. Methods: The genomic information of 728 Chinese colorectal cancer patients without germline mutation of Lynch syndrome related genes was obtained from HapLab database. HaploX WESPlus gene panel (an upgraded version of the standard WES) was performed to analyze the genomic data of patients. Results: 630 out of 728 patients (86.54%) had at least one mutation of the DDR genes. The frequently mutated DDR genes included TP53 (54.95%), BRCA2 (18.27%), PARP4 (13.46%), MSH2 (9.62%), XPC (7.55%), RAD54L2 (7.28%), ATM (6.73%), HMGB1 (5.77%), REV3L (5.49%) and POLD3 (5.36%). The TMB in patients with one, two, three and more than three mutations of DDR genes increased sequentially. The proportion of MSI-H in patients with more than three mutations of DDR genes (20.9%) was significantly higher than that in patients with less than 3 mutations of DDR genes (0.38%). Among the frequently mutated DDR genes, mutations of BRCA2, PARP4, MSH2, XPC, RAD54L2, ATM, REV3L or POLD3 were associated with higher TMB, mutations of MSH2, RAD54L2, ATM, REV3L or POLD3 were associated with higher proportion of MSI-H. The patients with mutation in each of eight DDR pathways showed significantly higher TMB and proportion of MSI-H than who without mutation of DDR genes. Conclusions: This study revealed the mutation profile of DDR genes in Chinese colorectal cancer patients. The mutation of DDR genes was closely related to TMB and MSI status. This result indicated the mutation of DDR genes may be a promising biomarker for immunotherapy in Chinese colorectal cancer patients.
Association of antibiotic exposure with the mortality in bevacizumab-treated metastatic colorectal cancer: A secondary analysis from Dryad database.
Authors: Sun Chenyu , Guo Xianwei , Kozma Kelly , Bhan Chandur , Kim Na Hyun
Published: June 2021

e15609

Background: Colorectal cancer (CRC) is one of the most diagnosed cancers and the second leading cause of cancer-related death worldwide. Globally, more than 1.8 million people are diagnosed of colorectal cancer (CRC) in 2018. In advanced CRC patients, bevacizumab plus 5-fluorouracil-based or platinum-based therapy has become one of the standard first-line chemotherapy regimen. A recent study also found that antibiotic exposure could be inversely associated with the mortality in metastatic colorectal cancer (mCRC) patients treated with bevacizumab. However, subgroup analysis of this study was no sufficient. Therefore, we conducted a secondary analysis based on the data of this study from Dryad database. Methods: In this retrospective cohort study, 147 mCRC patients treated with bevacizumab were included. All data was obtained from Dryad database (https://doi.org/10.5061/dryad.ft5sk66). Patients were divided into follow subgroups: (1) left colon vs right colon; (2) BMI < 18.5 vs BMI 18.5-24 vs BMI≥24;(3) Age < 45 vs Age≥45, (4) no surgery vs palliative surgery vs radical surgery, (5) bevacizumab plus FOLFIRI vs bevacizumab plus capeOX/FOLFOX vs bevacizumab plus other chemotherapy regimen. Specific survival of each subgroup was analyzed through the Kaplan-Meier curve, and the survival curves of the variables were compared using the log-rank test. Results: Survival analysis found no statistically significant differences of the cumulative survival rates between left colon cancer and right colon cancer, (58.9% vs 62.5%, respectively; χ2 = 0.043, P = 0.836), groups of different BMI (57.9% of BMI < 18.5 vs 55.2% of 18.5≤BMI < 24 vs 70.7% of BMI ≥24，χ2 = 3.026, P = 0.220), Age < 45 group and Age≥45 group (54.8% vs 61.2%, respectively; χ2 = 0.001, P = 0.976), surgical treatments (No surgery 59.0% vs Palliative surgery 57.6% vs Radical Surgery 61.3%; χ2 = 1.885, P = 0.390), as well as groups of different chemotherapy regimens (66.7% of FLOFIRI vs 52.0% of CapeOX/FOLFOX vs 59.9% of others; χ2 = 1.572, P = 0.456). Conclusions: Based on the data from this study, we found that different sites of colon cancer, age, BMI, different surgical treatments, and different chemotherapy regimens did not affect the survival outcome of patients with mCRC treated with bevacizumab after antibiotic exposure. Subsequent studies with larger sample size are still needed to further elaborate the effects of different antibiotics on survival outcomes.