Epidermal Growth Factor Receptor Gene Mutations Research Articles

Molecular profiling and utility of cell-free DNA in nonsmall carcinoma of the lung: Study in a tertiary care hospital.

Lung carcinoma accounts to the most common cause of cancer globally. Optimal management of nonsmall cell lung carcinoma (NSCLC) requires prognostic biomarkers that help in targeted therapy and identification of tumor subsets with a distinctive molecular profile that can foretell response to therapy. Quantitative analysis of circulating cell-free DNA is considered as a possible aid for lung cancer screening. The main aim of our study was detection of the clinicopathological spectrum of NSCLC, immunohistochemical (IHC) study of lung adenocarcinoma with epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), and molecular expression of EGFR mutation using Formalin fixed paraffin embedded tissue (FFPE) and cell-free DNA (cfDNA) from blood samples. It was a prospective and observational study conducted in the Department of Pathology in association with the Department of Chest Medicine in a tertiary care hospital for 18 months, done on 50 patients. Histological subtyping of lung carcinomas was done, followed by IHC analysis using P40, thyroid transcription factor (TTF1), EGFR, and ALK. Molecular analysis for EGFR mutation was done using FFPE and cfDNA from the patient's blood samples. On histological subtyping, majority (66%) of the cases were found to be adenocarcinoma. All adenocarcinoma (66%) cases show TTF1 positivity and all squamous cell carcinoma (32%) cases show P40 positivity. All the ALK-positive (6%) cases were never smokers and histologically diagnosed as adenocarcinoma. About 58% of the NSCLC cases were found to be EGFR IHC positive. Formalin-fixed paraffin tissue (FFPE) showed EGFR mutation in 32% cases, of which majority were deletion (19, 28%) and rest (4%) of the cases involving mutation in exon 21. From cfDNA, mutations were noticed in 16% of the cases where majority involved deletion 19 (12%), whereas the rest of the cases were positive for missense mutation in exon 21 of the EGFR gene (2%) and compound heterozygous mutation involving deletion 19 and missense mutation for exon 21 (2%). On correlation of EGFR mutation studies from FFPE with that of cfDNA analysis, the study was statistically significant (P = 0.000). This study reports clinicopathological, immunochemical, and molecular analysis of EGFR among NSCLC cases. EGFR mutation detection from cfDNA has its advantage of being a noninvasive technique to avoid rebiopsy in cases of the progressive disease to detect resistance to a drug and emergence of a newer mutation. Mutation detection from FFPE samples still remains the gold standard for targeted therapy using EGFR tyrosine kinase inhibitors. ALK rearrangement detection using IHC serves as an adjunct to EGFR diagnosis.

P71.05 Use of a Multiscale NSCLC Tumor Heterogeneity Model to Predict Tumor Growth Under Gefitinib

Novel high-throughput techniques, advanced tumor micro-environment analyses and new insights in tumor cell heterogeneity have significantly increased understanding of regulatory processes in Non-small cell lung cancer (NSCLC): NSCLC treatment response to Gefitinib depends on the epidermal growth factor receptor (EGFR), but differs depending on EGFR gene mutations making clinical prognosis challenging. We therefore developed an in silico EGFR+ lung adenocarcinoma (LUAD) model to predict the effect of EGFR-related mutations on tumor size in advanced-stage adenocarcinoma patients (IIIb or higher), using a mechanistic representation of tumor evolution, including response to Gefitinib. Tumor heterogeneity, age, gender, initial clinical stage, and smoking status are included as covariates. 5-step in silico model development: 1. Model Building: Pathophysiology of EGFR+LUAD was characterized by extracting biological features and their functional relationships from literature and translating them into ordinary differential equations (ODEs). Mutational burden, EGFR downstream-pathways, tumor growth and heterogeneity, Gefitinib-PK/PD, treatment-induced resistance and clinical outcome were modeled in a computational simulation with 43 variables, 170 parameters and 18 to 83 ODEs reflecting intra-tumor heterogeneity. 2. Calibration: Published spheroid, xenograft and clinical data were used for stepwise calibration. 3. Virtual populations (VPOP): VPOPs were generated for validation and benchmarking respectively, adapting baseline characteristics of a real population. 4. Validation: A VPOP with comparable baseline characteristics was tested against published patient data[1]. 5. Benchmarking against a Bayesian reference model[2]: (1) coverage of experimental interquartile range (IQR) with simulated IQR (precision) assesses model fit with experimental data, (2) coverage of simulated IQR with experimental IQR (overlap) assesses model fit with experimental variability. Our model computed in silico data comparable to the reference model[2] without use of original data for calibration (Figure 1B.2: experimental vs. simulated, precision of 68%, overlap of 91%). The reference model reported precision of 72% and overlap of 86%. We simulated tumor growth and treatment response in advanced-stage adenocarcinoma patients and successfully validated results with a published study[2]. Access to patient-level data for calibration would have improved precision of our model.

Complete Response to Afatinib of an EGFR Exon 18 delE709_T710insD-Mutated Stage IV Lung Adenocarcinoma.

Epidermal growth factor receptor (EGFR) mutations are frequently found in patients with lung adenocarcinomas, 90% being deletions in exon 19 or point mutation in exon 21. Three generations of tyrosine kinase inhibitors (TKIs) targeting EGFR mutations are available and have changed patient prognosis but less data is available on exon 18 mutations and their sensitivity to TKI therapy. Exon 18 delE709_T710insD accounts for 0.06% (16/27,294) of all EGFR mutations and is an oncogenic driver. Several partial responses to afatinib have been described. We report the first case, to the best of our knowledge, of the complete response to afatinib of a 57-year-old patient with stage IV lung adenocarcinoma with a delE709_T710insD mutation in the EGFR gene detected by next-generation sequencing. Oral afatinib was prescribed and despite treatment interruptions and dosage tapering due to cutaneous adverse events, a complete response was achieved 12 months after treatment initiation and is currently maintained at 17 months. When EGFR mutation is suspected, complete DNA sequencing of exons 18 to 21 should be carried out and we suggest that afatinib should be the first-line treatment for exon 18 delE709_T710insD-mutated advanced lung adenocarcinomas. Rare EGFR gene mutations are not detected by standard diagnostic kits.DNA sequencing is required to diagnose rare mutations of the EGFR gene.delE709_T710insD-mutated stage IV lung adenocarcinomas respond to afatinib.

Abstract 120: Compartmentalized functions of epidermal growth factor receptor (EGFR) in tumorigenesis and malignant phenotypes

Abstract Previous studies demonstrated that nuclear EGFR is associated with malignancy and resistance to tyrosine kinase inhibitor (TKI). To decipher the cellular compartmentalized functions and underlying mechanism of EGFR, we generated EGFR mutants at nuclear translocation signal (NLS) and nuclear export signal (NES). The results showed that EGFR NLS mutant with decreased nuclear translocation sensitized the cancer cells to TKI and impaired invasiveness and stemness of the cancer cells. In contrast, EGFR NES mutant with increased nuclear accumulation promoted aggressiveness of the cancer cells including TKI resistance. Next, we generated genetically engineered mouse model (GEMM) to validate the resulting phenotypes of the EGFR mutants. To this end, CRISPR-Cas9-mediated genome editing approach was used for generating EGFR mutant knock-in mouse, which harbors an internalization-deficient mutation (EGFRLL/AA), pro-nuclear (NES) mutation (EGFRL749P(S)) and the dual mutations. Upon EGF stimulation nuclear EGFR (nEGFR) accumulation in the hepatocytes from EGFRL749P heterozygous mice was increased. Conversely, a significant reduction of nEGFR was observed in the hepatocyte from EGFTRLL/AA homozygous mice. We found that germline expression of EGFRL749P(S) gave rise to deficiency in breeding. However, EGFRLL/AA mutant mice displayed no gross abnormal even in homozygous offspring, suggesting that depletion of nEGFR has no significant phenotype. We failed to generate homozygotes of EGFR NES mutant mice due to embryonic lethality. Of note, the female and male founders and F1 EGFRL798P(S) mice developed tumors with huge spleen. The IHC staining results indicated that the infiltrating lymphocytes in liver, stomach and kidney were B cells. The latency of the B cell lymphomagenesis was from 17 to 23 months, and tumor incidence of F1 offspring of EGFRL749P(S) knock-in mice was 10 of 15 (66.7%). These findings indicated that single allele mutation in EGFR NES domain is capable of driving lymphomagenesis, suggesting that the EGFR NES mutant possesses a strong oncogenic activity. Surprisingly, by searching database of EGFR mutation in human tumors (Cosmic database https://cancer.sanger.ac.uk/cosmic ), we noticed that among the somatic mutations in EGFR gene the gene amplification represents only small part of the gene alternations in human cancers, and EGFR L747P and L747S mutation occur in the lung cancer patients with intrinsic TKI resistance. In addition, the majority of human B lymphocyte originated tumors harboring EGFR NES mutation are also heterozygotes. Cosmic database shows that somatic mutations at the EGFR NES region occur in the patients with plasma cell lymphoma. Taken together, our findings suggest that nEGFR contributes to malignant potential of the tumors and EGFR with NES mutation is a tumor driver. Citation Format: Lei Nie, Junwei Hou, Yu-Yi Chu, Ying-Nai Wnag, Li-Chuan Chan, Longfei Huo, Paul Chiao, Shao-Chun Wang, Mien-Chie Hung, Michael Wang. Compartmentalized functions of epidermal growth factor receptor (EGFR) in tumorigenesis and malignant phenotypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 120.

Epidermal Growth Factor Receptor Gene Mutation Detection in Histology and Cytology Specimens of Primary Lung Adenocarcinoma: Immunohistochemistry Versus the Molecular Method.

Background:Epidermal growth factor receptor (EGFR) gene in lung adenocarcinoma is associated with good clinical response to EGFR-tyrosine kinase therapy. The two most common EGFR gene mutations, representing 80 to 90%, are the E746-A750 deletion in exon 19 and the L858R point mutation in exon 21. Materials and Methods:We have conducted the study to evaluate immunohistochemistry’s performance in detecting the E746-A750 deletion in exon 19 of the EGFR gene in primary lung adenocarcinoma cases. This study examined 133 cases of primary lung adenocarcinoma for three years duration. The selected cases were tested for EGFR gene mutations by real-time PCR by a reference laboratory. Most cases (124) were diagnosed by tissue biopsy, though nine used cell block cytology. We performed an immunohistochemistry test on 75 cases that contained adequate diagnostic material in the paraffin block. Results:The test result was scored as 0 to 3+, based on the staining intensity and percentage of positive tumor cells. We evaluated the immunohistochemistry test’s sensitivity and specificity compared to the EGFR gene mutations by real-time PCR. There was a significant association between gender, smoking status, and the EGFR gene mutations (P < 0.001). The overall sensitivity and specificity of the immunohistochemistry test were 40% and 100%, respectively. The positive predictive value and negative predictive values were 100% and 76.9%, each. Conclusions:The immunohistochemistry has high specificity but low sensitivity in the detection of E746-A750 deletion in exon 19 of the EGFR gene. The mutation-specific antibody used in this study was unable to detect other uncommon variants of exon 19 deletions. With high specificity value, immunohistochemistry may provide an adjunct to molecular testing for detecting the most common EGFR gene mutations in cases of a low cellularity sample, financially-limited situations, or in critically ill cases where urgent targeted therapy is needed.

Analysis of the Relationship Between the Quality of Small Biopsy Specimens of Non-small Cell Lung Cancer and the Mutation Rate of EGFR Gene

背景与目的表皮生长因子受体（epidermal growth factor receptor, EGFR）是非小细胞肺癌（non-small cell lung cancer, NSCLC）患者中突变率最高的基因，准确检测出其突变类型有助于指导患者接受靶向药物治疗从而延长患者生存期。为了得到准确的检查结果，基因检测平台对标本质量有一定要求。已有文献报道标本的肿瘤细胞数量及其比例等会影响EGFR基因突变检出率，本研究旨在分析NSCLC小活检标本质量与突变扩增系统（amplification refractory mutation system, ARMS）法检测EGFR基因突变检出率的关系。方法收集299例小活检肺腺癌病例的临床特征、在显微镜下评估切片的肿瘤细胞数量及比例、标本提取的DNA浓度、EGFR基因突变检测结果，分析标本质量与EGFR基因突变检出率的关系。结果肿瘤细胞数≤500组与 > 500组EGFR基因突变阳性率分别为40.7%（11/27）、43.8%（119/272），两者间无统计学差异（P=0.764）。DNA浓度≤20.4 ng/μL组及 > 20.4 ng/μL组的基因突变阳性率分别为42.7%（64/150）、44.3%（66/149），两者间无统计学差异（P=0.776）。肿瘤细胞比例≤30%及 > 30%组的阳性率分别为29.4%（20/68）、47.6%（110/231），两者间存在统计学差异（P=0.008）。多因素Logistic分析显示男性、甲状腺转录因子1（thyroid transcription factor-1, TTF-1）阴性、有吸烟史及肿瘤细胞比例 < 30%是影响EGFR基因突变低检出率的主要因素。结论在达到检测的最低要求后，肿瘤细胞比例仍可影响EGFR基因突变检出率。有必要在基因检测切片后再次评估最后一张切片的肿瘤细胞比例，对于肿瘤细胞比例过低的标本，建议通过显微切割等方法富集肿瘤细胞，提高其肿瘤细胞比例，得出更准确的检测结果。对于无法进行肿瘤细胞富集的标本，可行循环肿瘤DNA（circulating tumor DNA, ctDNA）检测作为补充，若结果仍为阴性才需考虑再次活检获取足够的肿瘤标本进行分子检测。

Detection of circulating tumor cells with chromosomes 7 and 8 polysomy in non-small cell lung cancer and its correlation with epidermal growth factor receptor mutations in cancer tissue

Objective: To investigate the value of chromosomes 7 and 8 polysomy in circulating tumor cells (CTCs) for the diagnosis of non-small cell lung cancer, and the correlation of CTCs with clinical pathological characteristics and epidermal growth factor receptor (EGFR) mutations in cancer tissue. Methods: Fifty-seven patients with non-small cell lung cancer and 21 patients with benign lung diseases were enrolled at Beijing Chaoyang Hospital, Capital Medical University, Beijing, China from November 2017 to October 2020. Negative enrichment combined with immunofluorescence in situ hybridization (imFISH) was used to identify CTCs polysomy on chromosomes 7 and 8. EGFR mutations in 56 lung cancer patients was detected using ARMS-PCR. Results: CTCs were detected in 93.0% (53/57) of non-small cell lung cancers and 28.6% (6/21) benign lung lesions. The difference between lung cancer patients and the control cohort was statistically significant (P<0.01). Receive operator curve (ROC) analyses showed that, when the cut-off value was 1 cell/3.2 mL, Youden index had the highest sensitivity of 93.0% and specificity of 71.4% (AUC=0.906, 95%CI:0.833-0.980, P<0.01). The positive rate of CTCs in stage Ⅲ-Ⅳ cancers was significantly higher than that in stage Ⅰ-Ⅱ (P=0.023). No significant correlation was observed between positive rate of CTCs or chromosome polysomy and age, gender, smoking status, pathologic types and EGFR mutation status. The number of CTCs in EGFR mutated group was higher than that in the non-mutated group (6.5±1.1 vs. 3.7±0.7, P=0.045). The detection rate for CTCs ≥5 in the EGFR mutated group was also higher than the EGFR non-mutated group (52.0% vs. 19.4%,P=0.010). Conclusion: Detection of CTCs with chromosomes 7 and 8 polysomy has potential value in auxiliary diagnosis of non-small cell lung cancer, and the number of CTCs is correlated to TNM stage and EGFR gene mutation status.

PL01.06 Molecular Epidemiology of Asian Lung Cancer

Lung adenocarcinoma (LADC), the most frequent histological type of lung cancer, develops in both ever- and never-smokers, and has two distinct characteristics in Asian populations. First, the proportion of never-smoking patients is considerably higher than that among individuals of European descent (approximately 40% vs 10%), and the rates of development and death from lung cancer in never-smokers are significantly higher in Asians than those in individuals of European descent. Second, in Asian individuals LADC frequently (in approximately 50% of cases) carries oncogenic mutations of the epidermal growth factor receptor (EGFR) gene, in contrast to lower EGFR mutation rates (approximately 10%) in European/US patients. In Asian patients, EGFR mutation frequently occurs in both never- (60%) and ever- (40%) smokers, while it occurs almost exclusively in European/US ever-smokers. Notably, EGFR-mutated LADC does not respond well to immune checkpoint blockade therapy due to its non-inflamed (immune-cold) features, including low infiltration of CD4+ and CD8+ T cells. Therefore, it is highly likely that there are specific risk factors, independent from smoking, that promote LADC development in Asians and have immunogenic effects influencing tumour characteristics. By conducting genome-wide association study, we demonstrated that HLA loci influence lung adenocarcinoma risk independently from smoking and that the association of HLA-class II, but not class I, loci is more prominent for lung adenocarcinomas with EGFR gene mutations than those without; the HLA-DPB1 Asp57 and 76Val alleles are associated with elevated risk for EGFR-mutated lung adenocarcinoma. HLA variants might influence lung adenocarcinoma risk by modulating anti-tumour immunity.

P37.17 Prevalence of Epidermal Growth Factor Receptor (EGFR) Mutations in Non-small Cell Lung Cancer (NSCLC): A Global Meta-analysis

Identification of variable EGFR gene mutations in NSCLC is increasingly important for the selection of appropriate targeted therapies. In this timely, global overview of EGFR mutated NSCLC we aim to address this clinical data gap.

Our experience of lung resection in patients who decline blood transfusion for religious reasons

ObjectiveSurgical treatment for patients who refuse blood transfusion due to religious beliefs is an important issue related to medical safety. Few reports have examined pulmonary surgery for these patients, and we analyzed clinical characteristics in such cases.MethodsTen Jehovah’s Witness (JW) patients with lung tumor resection who declined blood transfusion for religious reasons between December 2013 and February 2020 at the Fukushima Medical University Hospital were included. Median total intraoperative blood loss was 17.5 mL (range 5–150 mL). Fibrin glue was used intraoperatively for 8 patients. Final pathological examination revealed pulmonary adenocarcinoma in 9 cases and metastasis of bladder cancer in 1 case. In 8 patients with pulmonary adenocarcinoma examined for epidermal growth factor receptor (EGFR) gene mutation, 6 cases showed mutation. No patients had serious complications, but 1 patient displayed temporary anemia due to postoperative hemorrhagic gastrointestinal ulcer.Result and conclusionsOur findings confirm that pulmonary resection is feasible and safe for JW patients if performed by experienced medical staff. However, awareness of complications associated with perioperative bleeding is important. Each JW patient should be interviewed individually and every available perioperative option aimed at blood-sparing management, including use of blood coagulation factors and fibrinogen concentrates, should be carefully discussed and clarified. In this study, the EGFR gene mutation rate was higher than usual for cases of lung adenocarcinoma. Further studies are necessary to assess clinical features in JW patients with lung cancer.

Clinical value of the detection of EGFR gene mutations in pleural effusion cell blocks among patients with non-small-cell lung cancer.

To evaluate the clinical value of epidermal growth factor receptor (EGFR) detection in pleural effusion cell blocks among patients with non-small-cell lung cancer (NSCLC). From July 2016 to September 2018, EGFR gene mutations in 40 lung tumor tissue samples and pleural fluid samples from NSCLC patients in Jinhua Municipal Central Hospital were assessed by the amplification refractory mutation system method. The EGFR results of the two types of samples were compared using the paired Chi-square test, and the mutation positive rates in EGFR exons 18, 19, 20 and 21 were compared between the two types of specimens using the four-grid Chi-square test. Among the 40 tissue samples and pleural effusion samples, 21 and 18 cases of EGFR mutations were detected, respectively, and the mutation positive rates were 52.5% and 45%, respectively. The κ value of the consistency test of the two specimens was 0.851. There were no significant differences in the mutation positive rates in EGFR exons 18, 19, 20, and 21 between the two types of specimens. The EGFR results of pleural fluid and tissue samples were in good agreement. Therefore, we can use pleural fluid samples to detect EGFR mutations to guide tyrosine kinase inhibitor treatment for NSCLC patients in whom tumor tissue samples cannot be obtained.

First-Line EGFR-TKIs Treatment in Stage I Non-Small-Cell Lung Cancer Patients Harboring EGFR Gene Mutations with Postoperative Intrapulmonary Recurrence.

BackgroundThe epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) is an effective treatment for advanced lung cancer harboring EGFR gene mutations, and has improved progression-free survival in several clinical trials.MethodsWe investigated 30 stage I non-small-cell lung cancer patients harboring EGFR gene mutations with postoperative intrapulmonary recurrence. Progression-free survival and response rate were analyzed.ResultsPartial response was achieved in 23 patients and stable disease was found in 7 patients. The objective response rate was 76.7% and disease control rate was 100%. The median progression-free survival (PFS) time was 24.5 months. The median PFS in patients with only intrapulmonary recurrence was significantly superior to patients with both intrapulmonary recurrence and metastasis (32.0 months vs 14.0 months, P = 0.003). The median PFS observed in patients who underwent icotinib treatment was significantly longer than in patients who underwent gefitinib treatment (30.5 months vs 12.0 months, p = 0.005). There were no statistical differences in median PFS between patients with tumors harboring exon 21 mutation and exon 19 deletion, age <65 and ≥65, male and female, smoker and non-smoker.ConclusionOur result reveals that first-line EGFR-TKIs treatment for stage I non-small-cell lung cancer patients harboring EGFR gene mutations with postoperative intrapulmonary recurrence is effective and could be a useful option in practical setting.

Humoral immune response to epidermal growth factor receptor in lung cancer.

The aim of this study was to explore the potential value of autoantibody to epidermal growth factor receptor (EGFR) in the diagnosis of lung cancer (LC) and its relation with EGFR mutations. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the level of autoantibody to EGFR in sera from 254 LC patients and 222 normal controls (NCs). Besides, the mRNA and protein levels of EGFR were investigated in Gene Expression Profiling Interactive Analysis (GEPIA) and Human Protein Atlas (HPA) database, respectively. The level of autoantibody to EGFR (anti-EGFR) in LC even different types of LC was obviously higher than that in NC (P < 0.05). The area under the curve (AUC) of anti-EGFR was 0.695 (95% CI 0.645-0.742) when comparing LC patients with NC, while the AUC of carcinoembryonic antigen (CEA) was 0.681 (95% CI 0.629-0.730). Moreover, by integrating anti-EGFR with CEA to diagnose LC, the AUC was up to 0.784 (95% CI 0.737-0.826). However, the expression level of autoantibody to EGFR had no difference between LC patients with and without EGFR gene mutation (P > 0.05). EGFR mRNA expression level was obviously upregulated in squamous cell carcinoma (SCC) tissues compared with normal tissues (P < 0.05), but not in adenocarcinoma (ADC) (P > 0.05). The study confirmed that anti-EGFR could be a potential biomarker for LC diagnosis; additionally, it could improve the diagnostic value of CEA in clinical work.

Analysis of Clinical Characteristics of Lung Cancer Combined with Multiple Primary Malignancies in Other Organs

背景和目的随着胸部计算机断层扫描（computed tomography, CT）肺癌早筛技术的普及，肺癌合并其他器官多原发癌（multiple primary malignancies, MPM）的检出比例不断提高。本研究就发病率、病理特征、诊疗特点和预后情况进行探讨分析，为完善此类疾病的临床诊疗提供研究依据。方法2011年9月-2015年9月江苏省肿瘤医院胸外科共收治5, 570例肺癌患者，回顾性分析其中61例肺癌合并其他器官MPM患者的临床病理特征。结果本研究中肺癌合并其他器官MPM的发病率为1.1%，其中同时多原发癌（synchronous MPM, SMPM）15例，异时多原发癌（metachronous MPM, MMPM）46例；结直肠癌、乳腺癌和甲状腺癌占肺癌合并其他器官MPM的前三位；患者总体5年生存率为39.3%，有71.4%的患者死于肺癌转移或复发，多因素分析发现肺癌患者的临床分期、肺癌与其他肿瘤发生的先后顺序、合并肿瘤的治疗状态以及是否存在表皮生长因子受体（epidermal growth factor receptor, EGFR）基因突变是影响患者生存期的重要因素。结论肺癌合并其他器官MPM的发病率并非罕见，相较于其他器官恶性肿瘤，肺癌是主要致死原因，中晚期肺癌、SMPM、肺癌先发、合并肿瘤仅获姑息治疗和无EGFR基因突变的患者预后较差。

Relationship between Epidermal Growth Factor Receptor Mutations and Adverse Events in Non-Small Cell Lung Cancer Patients treated with Afatinib.

Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors such as afatinib are used for non-small cell lung cancer (NSCLC) and show varying efficacy depending on EGFR gene mutation. Few studies have examined the relationship between EGFR gene mutations and the adverse events of afatinib in NSCLC. This retrospective study included 32 Japanese patients with NSCLC with EGFR gene mutation who were treated with afatinib between May 2014 and August 2018 at Kagawa University Hospital. Among the 32 Japanese patients with NSCLC treated with afatinib, 19 patients were positive for exon 19 deletion mutation (Del 19) and 13 patients were negative for Del 19. The incidence of grade ≥ 2 skin rash was slightly higher in patients positive for Del 19 (42.1% vs. 7.7%, P = 0.050). No significant differences were detected in other adverse events between the two patient groups. Patients positive for Del 19 also showed significantly longer median progression-free survival (288 vs. 84 days, P = 0.049). Our study indicates a higher incidence of skin rash associated with afatinib treatment in Japanese patients with NSCLC positive for Del 19 compared with patients without Del 19. The Del 19 positive patient group also showed better progression-free survival. J. Med. Invest. 68 : 125-128, February, 2021.

Development and validation of a multivariable predictive model for EGFR gene mutation status in patients with lung adenocarcinoma.

Detection of epidermal growth factor receptor (EGFR) is one real dilemma owing to the non-sufficient tissue for testing EGFR mutations in lung adenocarcinoma. A model for predicting EGFR mutations would be helpful for clinical decisions in those patients. A retrospective cohort of 1,196 patients diagnosed with lung adenocarcinoma was investigated between December 1, 2017, and December 31, 2019, in Renji Hospital, Shanghai, China. All patients were tested for EGFR mutations (amplification refractory mutation system, n=1,144; next-generation sequencing, n=52). Of 1,196 patients with lung adenocarcinoma, 944 met the inclusion criteria. A nomogram model was developed based on 567 patients and validated in 377 patients. Variables associated with EGFR mutations were age, sex, smoking history, lepidic predominant subtype, solid predominant subtype, mucinous adenocarcinoma, Ki67 expression, lobulation, solid texture in radiology, and pleural retraction. The nomogram based on the model performed well in the development group (c-index 0.789, 95% CI: 0.751-0.827), and the validation group (c-index 0.809, 95% CI: 0.771-0.847). At the probability cut-point of 0.7, the diagnostic efficiency was 82.7% in patients with NGS liquid biopsy. Decision curve analysis further confirmed the clinical usefulness of the nomogram, which showed that predicting the EGFR mutations probability applying this nomogram would be better than having all patients or none patients use this nomogram. A high probability group (>0.7) by nomogram model may suggest a high possibility of EGFR mutation, if tissue is limited, NGS-based ctDNA with liquid biopsy could be implemented effectively.

Results of EGFR Mutations Detected in Pleural Effusion and Its Clinical Significance in 132 Patients with Advanced Non-small Cell Lung Cancer: A Retrospective Study in A Single Center

背景与目的恶性胸腔积液表皮生长因子受体（epidermal growth factor receptor, EGFR）基因突变检测缺乏病理质控，导致对检测结果解释及指导临床EGFR酪氨酸激酶抑制剂（tyrosine kinase inhibitor, TKI）使用造成困惑。因此，提出质控标准，并按此标准进行胸水EGFR突变检测尤为重要。本研究拟回顾性分析按照严格病理质控标准进行的胸水沉渣切片EGFR基因突变检测结果以及据此结果指导EGFR-TKIs治疗的疗效。方法回顾性分析中国医学科学院北京协和医院病理科2012年1月-2018年6月收到的胸腔积液标本的患者临床资料，其中具有临床资料相对完整，且按照制定的质控标准进行了胸水沉渣石蜡包埋切片EGFR基因突变检测的患者132例。根据EGFR基因突变检测结果，分为阳性组和阴性组，比较不同组别使用EGFR-TKIs的疗效。结果胸腔积液经离心后，沉渣石蜡包埋、切片，HE染色后镜下观察，若肿瘤细胞数目≥100个，即满足病理质控标准，可用于后续EGFR基因突变检测。132例患者中，72例（54.5%）患者胸水中检出EGFR基因突变。72例突变阳性患者中，69例患者使用了EGFR-TKIs。60例EGFR突变阴性患者中，仅15例使用EGFR-TKIs。EGFR突变阳性组的疾病控制率（disease control rate, DCR）为95.8%，中位无疾病进展生存时间（progression-free survival, PFS）为11个月；EGFR突变阴性组的DCR为0%，中位PFS为1个月，两组患者DCR和PFS均有显著差异（P < 0.05）。结论经病理质控的胸水沉渣包埋切片可用于EGFR基因突变检测，其结果可指导临床EGFR-TKIs使用。

Value of serum tumor markers for predicting EGFR mutations in non-small cell lung cancer patients

ObjectivesWe investigated whether serum tumor markers (STMs) represent a valuable noninvasive tool to predict epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. MethodsA retrospective analysis was performed for 143 NSCLC patients at the Peking University International Hospital from December 2014 to December 2019. EGFR mutations in the tumor tissues were identified by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and next generation sequencing (NGS). The relationships between EGFR mutation and several clinicopathological features were analyzed. ResultEGFR mutation were found more frequently in female (56.67%, P = 0.01), never-smokers (55.26%, P = 0.004), and those with lung adenocarcinoma (ADC) (52.17%, P < 0.001). The positive mutation rate for the EGFR gene were higher in the squamous cell carcinoma antigen (SCCA)group (≤1.5 ng/ml) and in the gastrin-releasing peptide precursor (preGRP) increased group (≥69.2 pg/ml), and this difference was statistically significant (P < 0.05). Univariate logistic regression analysis demonstrated that females (Odd ratio [OR]: 2.435, 95% confidence interval [CI]: 1.232, 4.813, P = 0.01) and never-smokers (OR = 0.370; CI = 0.186, 0.734; P = 0.004), lung adenocarcinoma patients (OR = 9.091; CI = 2.599, 21.800; P = 0.001), the SCC group (≤1.5 ng/ml) (OR = 0.331, CI = 0.120, 0.914; P = 0.033), and the preGRP group (≥69.2 pg/ml) (OR = 5.478, CI = 1.462, 20.528; P = 0.012) patients were risk factors for EGFR gene mutation. Multivariate logistic regression analysis demonstrated that lung ADC and proGRP elevation were independent risk factors for predicting EGFR gene positivity (P < 0.05). ConclusionSTMs are associated with mutant EGFR status and could be integrated with other clinical factors to facilitate the classification of EGFR mutation status among NSCLC patients.

Sorting and gene mutation verification of circulating tumor cells of lung cancer with epidermal growth factor receptor peptide lipid magnetic spheres.

BackgroundThis study aimed to identify an efficient, simple, and specific method of detecting mutations in the epidermal growth factor receptor (EGFR) gene in isolated lung cancer circulating tumor cells (CTCs) and to improve the ability to obtain tumor tissue clinically.MethodsEGFR peptide lipid magnetic spheres (EG‐P‐LMB) were prepared by reverse evaporation, and characterization and cell capture efficiency assessed. The peripheral blood samples of 30 lung cancer patients were isolated and identified with the EG‐P‐LMB using 20 healthy volunteers as controls. Finally, the isolated CTCs were tested for EGFR gene mutations, and the tissue samples selected for comparison.ResultsThe prepared magnetic spheres had a smaller particle size and higher stability according to the particle size potential test. Their morphology was homogeneous by atomic force observation, and the UV test showed that there were peptides on the surface. The separation efficiency of EG‐P‐LMB was greater than 90% in PBS and greater than 80% in the blood simulation system. Compared with the tissue sample results, the positive rate of EGFR gene mutations was 94%. The CTC test results of 27 patients were consistent with the tissue test results of the corresponding patients, and the consistency with the tissue comparison test results was 90% (27/30).ConclusionsEG‐P‐LMB can effectively capture CTCs in the peripheral blood of patients with lung cancer. CTC detection can accurately identify mutations in the EGFR gene and improve the ability to obtain tumor tissue in clinical practice.Key pointsSignificant findings of the studyEG‐P‐LMB can effectively capture CTCs in the peripheral blood of patients with lung cancer. CTC detection can accurately identify mutations in the EGFR gene and improve the ability to obtain tumor tissue in clinical practice.What this study addsThis study added EGFR peptide lipid magnetic spheres to capture CTCs in the blood. Genetic testing was performed and compared with tissues. It solves the problem of clinically difficult tumor tissue sampling.

Assessment of BRAF V600E, KRAS, NRAS and EGFR mutations in Papillary Thyroid Carcinoma and Hashimoto Thyroiditis

Objective: The present study compared three study groups composed of patients with only papillary thyroid carcinoma (PTC), patients with only Hashimoto’s thyroiditis (HT) and patients with PTC+HT in terms of BRAF-V600E, KRAS, NRAS and epidermal growth factor receptor (EGFR) mutations. Also, the association between clinicopathological prognostic indicators including tumor multifocality, extrathyroidal extension (ETE), lymph node (LN) metastasis and recurrence and BRAF-V600E mutations were investigated in the PTC and PTC+HT groups. Methods: A total of 53 patients (two males and 51 females) who underwent a hemi/total thyroidectomy due to suspicion of malignancy or malignant lesion according to the thyroid cytopathology participated in the study. The study groups were composed of 19 patients with PTC, 18 with PTC+HT and 16 with HT according to the histopathological examination records. Histopathological sections from the paraffin blocks of the patients were investigated for BRAF-V600E, KRAS, NRAS and EGFR gene mutations using real-time polymerase chain reaction (PCR) assay. Results: There was no significant difference between the groups in terms of BRAF-V600E KRAS, NRAS and EGFR mutation rates. Also, presence of the BRAF-V600E mutation was not correlated with the prognostic indicators for the patients with PTC and PTC+HT. Conclusion: In the present study, no significant association was found between PTC and HT, and the BRAF-V600E, KRAS, NRAS and EGFR mutations. Further studies with a larger number of patients may help to clarify the clinicopathological and diagnostic importance of the BRAF-V600E, KRAS, NRAS and EGFR mutations in thyroid diseases.