Consistency comparison of programmed cell death 1-ligand 1 in different immuno-histochemical staining methods / 北京大学学报(医学版)

OBJECTIVE@#To compare the consistency of programmed cell death 1-ligand 1 (PD-L1, clone E1L3N, 22C3, SP263) in different immunohistochemical staining methods.@*METHODS@#The first step was to select the optimal process: The PD-L1(clone E1L3N) antibody recommended process, self-built process ①, self-built process ② and self-built process ③ were used to perform immunohistochemical staining in 5 cases of tonsil tissue. The quality of all slides was scored by expert pathologists (0-6 points). The process with the highest score was selected. The second step was to compare the consistency between the optimal procedure and the two standard procedures. Thirty-two cases of lung non-small cell carcinoma diagnosed by pathology in Peking University First Hospital in the past two years were randomly selected. The 32 cases were stained in parallel with the SP263 and 22C3 standard procedures, and all stained slides were scored by specialized pathologists for tumor proportion score (TPS). The scoring results were grouped according to < 1%, ≥1% to < 10%, ≥10% to < 50%, and ≥50%. The consistency of PD-L1 detection antibody clone E1L3N and 22C3, E1L3N and SP263 staining results was analyzed.@*RESULTS@#Tonsil stained slides scores (0-6 points) were as follows: The recommended protocol was 5, 5, 5, 5 and 5. The self-built process ① was 5, 6, 6, 5 and 6. The self-built process ② was 4, 4, 4, 4 and 4.The self-built process ③ was 3, 3, 3, 3 and 3. The self-built process ① was the best with the highest score. The TPSs of 32 non small cell lung carcinoma (NSCLC) cases were as follows: Of self-built process ①, 6 cases were lower than 1%, 5 cases were from 1% to 10%, 10 cases were from 10% to 50%, and 11 cases were higher than 50%; of 22C3 standard procedure, 5 cases were lower than 1%, 3 cases were from 1% to 10%, 13 cases were from 10% to 50%, 11 cases were higher than 50%; of SP263 standard procedure, 7 cases were lower than 1%, 4 cases were from 1% to 10%, 11 cases were from 10% to 50%, 10 cases were higher than 50%. The results of the consistency test were as follows: The κ value for self-built process ① and 22C3 standard procedure was 0.736 (P < 0.001), the agreement was good; the κ value for self-built process ① and SP263 standard procedure was 0.914 (P < 0.001), the agreement was very good.@*CONCLUSION@#The immunostaining using PD-L1(E1L3N) with validated self-built staining protocol ① by Ventana Benchmark GX platform can obtain high quality of slides, and the TPSs based on these slides are in good agreement with 22C3 and SP263 standard procedures.

The effect of PLK1 inhibitor in osimertinib resistant non-small cell lung carcinoma cells / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effects of PLK1 inhibitors on osimertinib-resistant non-small cell lung carcinoma (NSCLC) cells and the anti-tumor effect combined with osimertinib.@*METHODS@#An osimertinib resistant NCI-H1975 cell line was induced by exposure to gradually increasing drug concentrations. Osimertinib-resistant cells were co-treated with compounds from classical tumor pathway inhibitor library and osimertinib to screen for compounds with synergistic effects with osimertinib. The Gene Set Enrichment Analysis (GSEA) was used to investigate the activated signaling pathways in osimertinib-resistant cells; sulforhodamine B (SRB) staining was used to investigate the effect of PLK1 inhibitors on osimertinib-resistant cells and the synergistic effect of PLK1 inhibitors combined with osimertinib.@*RESULTS@#Osimertinib-resistance in NCI-H1975 cell (resistance index=43.45) was successfully established. The PLK1 inhibitors GSK 461364 and BI 2536 had synergistic effect with osimertinib. Compared with osimertinib-sensitive cells, PLK1 regulatory pathway and cell cycle pathway were significantly activated in osimertinib-resistant cells. In NSCLC patients with epidermal growth factor receptor mutations treated with osimertinib, PLK1 mRNA levels were negatively correlated with progression free survival of patients (R=－0.62, P<0.05), indicating that excessive activation of PLK1 in NSCLC cells may cause cell resistant to osimertinib. Further in vitro experiments showed that IC50 of PLK1 inhibitors BI 6727 and GSK 461364 in osimertinib-resistant cells were lower than those in sensitive ones. Compared with the mono treatment of osimertinib, PLK1 inhibitors combined with osimertinib behaved significantly stronger effect on the proliferation of osimertinib-resistant cells.@*CONCLUSIONS@#PLK1 inhibitors have a synergistic effect with osimertinib on osimertinib-resistant NSCLC cells which indicates that they may have potential clinical value in the treatment of NSCLC patients with osimertinib resistance.

Deubiquitinating enzyme JOSD2 affects susceptibility of non-small cell lung carcinoma cells to anti-cancer drugs through DNA damage repair / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effects and mechanisms of deubiquitinating enzyme Josephin domain containing 2 (JOSD2) on susceptibility of non-small cell lung carcinoma (NSCLC) cells to anti-cancer drugs.@*METHODS@#The transcriptome expression and clinical data of NSCLC were downloaded from the Gene Expression Omnibus. Principal component analysis and limma analysis were used to investigate the deubiquitinating enzymes up-regulated in NSCLC tissues. Kaplan-Meier analysis was used to investigate the relationship between the expression of deubiquitinating enzymes and overall survival of NSCLC patients. Gene ontology enrichment and gene set enrichment analysis (GSEA) were used to analyze the activation of signaling pathways in NSCLC patients with high expression of JOSD2. Gene set variation analysis and Pearson correlation were used to investigate the correlation between JOSD2 expression levels and DNA damage response (DDR) pathway. Western blotting was performed to examine the expression levels of JOSD2 and proteins associated with the DDR pathway. Immunofluorescence was used to detect the localization of JOSD2. Sulforhodamine B staining was used to examine the sensitivity of JOSD2-knock-down NSCLC cells to DNA damaging drugs.@*RESULTS@#Compared with adjacent tissues, the expression level of JOSD2 was significantly up-regulated in NSCLC tissues (P<0.05), and was significantly correlated with the prognosis in NSCLC patients (P<0.05). Compared with the tissues with low expression of JOSD2, the DDR-related pathways were significantly upregulated in NSCLC tissues with high expression of JOSD2 (all P<0.05). In addition, the expression of JOSD2 was positively correlated with the activation of DDR-related pathways (all P<0.01). Compared with the control group, overexpression of JOSD2 significantly promoted the DDR in NSCLC cells. In addition, DNA damaging agents significantly increase the nuclear localization of JOSD2, whereas depletion of JOSD2 significantly enhanced the sensitivity of NSCLC cells to DNA damaging agents (all P<0.05).@*CONCLUSIONS@#Deubiquitinating enzyme JOSD2 may regulate the malignant progression of NSCLC by promoting DNA damage repair pathway, and depletion of JOSD2 significantly enhances the sensitivity of NSCLC cells to DNA damaging agents.

Interpretation of CT images of pulmonary mucinous adenocarcinoma / 中国综合临床

Pulmonary mucinous adenocarcinoma (PMA) is relatively rare. On chest CT, it can be divided into two types: mass type and pneumonia type. Mass type PMA is more common and is difficult to distinguish from other nonsmall cell lung cancer. It is a solid or partial solid nodule or mass, predominantly located in the peripheral field of the lung with lobulation, spiculation, and more prone “vacuole sign”. Pneumonia type PMA has a poor prognosis and is more likely to develop into diffuse, multifocal and multilobular lesions similar to inflammatory manifestations, indicating dissemination along the airway. Typical signs include large areas of low density, low enhancement consolidation, and “dead tree sign”.

Osimertinibe comparado a Erlotinibe Gefitinibe ou quimioterapia para tratamento de carcinoma pulmonar de células não pequenas com mutação do gene do receptor do fator de crescimento epidérmico (EFGR): revisão rápida / Osimertinib compared to erlotinib, gefitinib or chemotherapy for the treatment of non-small cell pumonary carcinoma with epidermal growth factor receptor (EFGR) gene mutation: rapid review

Mesilato de osimertinibe, gefitinibe, erlotinibe, quimioterapia padrão. Indicação: Câncer de pulmão de células não pequenas com mutação do receptor do fator de crescimento epidérmico (EGFR). Pergunta: Mesilato de osimertinibe é mais eficaz e seguro que gefitinibe, erlotinibe ou quimioterapia para os desfechos de sobrevida global, sobrevida livre de progressão e de segurança no tratamento de carcinoma pulmonar de células não pequenas com mutação do EGFR? Métodos: Levantamento bibliográfico foi realizado na base de dados PUBMED e EPISTEMONIKOS, seguindo estratégias de buscas predefinidas. Foi feita avaliação da qualidade metodológica das revisões sistemáticas com a ferramenta AMSTAR-2 (Assessing the Methodological Quality of Systematic Reviews Version 2). Resultados: Foram selecionadas duas revisões sistemáticas que atenderam aos critérios de elegibilidade. Conclusão: Mesilato de osimertinibe é mais eficaz do que gefitinibe ou erlotinibe na melhora da sobrevida global e da sobrevida livre de progressão em pacientes virgens de tratamento. Em pacientes previamente tratados, o mesilato de osimertinibe não é superior à quimioterapia padrão à base de platina no prolongamento da sobrevida global, mas é mais eficaz no aumento da sobrevida livre de progressão. Para câncer avançado, mesilato de osimertinibe não é mais eficaz do que a quimioterapia com ou sem pemetrexede para prolongar a sobrevida global, mas é mais eficaz em melhorar a sobrevida livre de progressão. Gefitinibe combinado com quimioterapia à base de pemetrexede foi superior à quimioterapia com ou sem pemetrexede na melhora da sobrevida global e da sobrevida livre de progressão

Osimertinib mesylate, gefitinib, erlotinib, standard chemotherapy. Indication: Non-small cell lung cancer with epidermal growth factor receptor (EGFR) mutation. Question: Is osimertinib mesylate more effective and safer than gefitinib, erlotinib or chemotherapy for overall survival, progression-free survival and safety outcomes in the treatment of non-small cell lung cancer with EGFR mutation? Methods: A bibliographic search was done in the PUBMED and EPISTEMONIKOS database, following predefined search strategies. The methodological quality of systematic reviews was evaluated using the Assessing the Methodological Quality of Systematic Reviews Version 2 tool. Results: Two systematic reviews were selected because they met the eligibility criteria. Conclusion: Osimertinib mesylate is more effective than gefitinib or erlotinib in improving overall survival and progression-free survival in treatment-naive patients. In previously treated patients, osimertinib mesylate is not superior to standard platinum-based chemotherapy in prolonging overall survival, but it is more effective in increasing progression-free survival. For advanced cancer, osimertinib mesylate is not more effective than chemotherapy with or without pemetrexed in prolonging overall survival, but it is more effective in improving progression-free survival. Gefitinib combined with pemetrexed-based chemotherapy was superior to chemotherapy with or without pemetrexed in improving overall survival and progression-free survival

Evaluation of Enhancement Pattern of Malignant Lung Lesion During Contrast Enhanced Computed Tomography - Cross Sectional Study.

Background: At present, lung cancer is a leading cause of cancer death in the world. Most of the patients are asymptomatic at the early stages of the disease and manifest at the advanced stage. It is broadly classified into two types based on histopathology, Non-small cell lung cancer (NSCLC), the predominant type, and Small cell lung cancer (SCLC). Â Our study aimed to evaluate the enhancement pattern of malignant lung lesion during contrast-enhanced computed tomography. Methodology: A cross-sectional study was conducted on 34 patients with lung cancer after clearance from the Institutional Ethical Committee. Contrast enhanced computed tomography (CECT) Thorax was performed by 16 slice Multi-Detector computed tomography (MDCT )machine, and the tumor's pre and post-contrast Hounsfield Unit (HU) value was measured in the area of highest density. Results: Out of the 34 patients, Squamous cell carcinoma and adenocarcinoma were seen in 15 patients in each group, whereas small cell carcinoma was found in 4 patients. Pre and post-contrast mean HU value was 39.5 (SD= 2.95) HU and 72.13 (SD= 4.21) HU for adenocarcinoma, 34.6 (SD=2.61) HU and 63.8(SD=2.88) HU for squamous cell carcinoma and 31.00 (SD=1.41) HU and 55.7 (SD=2.75) HU for small cell carcinoma. The significant of density difference was measured using paired t-test (p<0.05). Conclusion: All histopathological subtypes of lung cancer showed significant post-contrast enhancement (>20HU) during the CECT study. Evaluation of enhancement pattern of lung mass by measuring pre and post-contrast HU value will help the radiologist to predict the histopathological type of lung cancer.

Interpretation of CT images of lung adenosquamous carcinoma / 中国综合临床

Lung adenosquamous carcinoma (ASC) is a rare histological subtype of lung cancer, which has a worse prognosis than simple lung adenocarcinoma and lung squamous cell carcinoma. ASC was predominantly located in the peripheral field of the lung with heterogeneous attenuation and the most common features included lobulation, spiculation, pleural tail sign, necrosis and cavitation on chest CT images. However, the imaging features of ASC are not specific. Therefore, acquring accurate pathological diagnosis and adequate treatment as soon as possible are very important.

Características de los pacientes con cáncer de pulmón de célula no pequeña en el Instituto Nacional de Cancerología de Colombia / Characteristics of patients with non-small cell lung cancer at the National Cancer Institute of Colombia

Resumen Objetivos: Describir las características demográficas, histopatológicas, biología molecular tumoral y estadificación de los pacientes con cáncer de pulmón de célula no pequeña atendidos entre diciembre de 2013 y diciembre de 2018 en el Instituto Nacional de Cancerología de Colombia. Métodos: Estudio descriptivo de corte transversal. Resultados: Se incluyeron 392 pacientes con cáncer de pulmón de célula no pequeña, la mediana de edad fue 65.9 años (rango, 28,9 a 88,9 años). 198 (50,5%) pacientes fueron mujeres, obteniendo una relación hombre mujer 1:1. El 90.6% de los casos eran mayores de 50 años. Antecedente de tabaquismo se presentó en 211 (53,8%) pacientes, 75.8% de la población masculina y 32,3% de la población femenina eran fumadores. El adenocarcinoma se encontró en 293 (74,7%) pacientes y el carcinoma escamocelular en 73 (18,6%) pacientes. La estadificación patológica fue: estadio I en 22 (5,6%) pacientes, estadio II en 18 (4,6%), estadio III en 40 (10,2%) pacientes, estadio IV en 311 (79,3%) pacientes y no hubo dato en un solo paciente. Se detectó la mutación del EGFR en 21,2% de los pacientes. Los reordenamientos de ALK se identificaron en 4,6% de los pacientes y el PDL 1 solo se midió en el 9% de la población. Conclusiones: este estudio nos muestra el panorama general del cáncer de pulmón de célula no pequeña en la población colombiana, en donde la mayoría de los pacientes se diagnostican en estadios avanzados de la enfermedad y nos expone la necesidad de nuevas estrategias para la detección temprana y el acceso oportuno de los pacientes con cáncer de pulmón.

Abstract Objectives: to describe the demographic and histopathologic characteristics, tumor molecular biology and staging of patients with non-small cell lung cancer treated between December 2013 and December 2018 at the National Cancer Institute of Colombia. Methods: a retrospective cohort study based on data from medical records. Results: 392 patients with non-small cell lung cancer were included. The median age was 65.9 years (range: 28.9 to 88.9 years); 198 (50.5%) patients were women, obtaining a 1:1 male-female ratio. 90.6% of the cases were older than 50 years. History of smoking occurred in 211 patients (53.8%), 75.8% of the male population and 32.3% of the female population were smokers. Adenocarcinoma was found in 293 (74.7%) patients, while squamous cell carcinoma was present in 73 (18.6%) patients. The pathological staging was: 22 (5.6%) patients were in stage I, 18 (4.6%) had stage II, 40 (10.2%) patients were in stage III, stage IV was found in 311 (79.3%) patients, and there was no data in one patient. EGFR mutation was detected in 21.2% of patients. ALK rearrangements were identified in 4.6% of patients and PDL-1 was only found in 9% of the population. Conclusions: this study presents a general panorama of non-small cell lung cancer in the Colombian population, where most patients are diagnosed in advanced stages of the disease, and highlights the need for new strategies for early detection and better access for patients with lung cancer.

Research status of radiotherapy combined with EGFR-TKI for brain metastases in patients with EGFR-mutant NSCLC / 中华放射肿瘤学杂志

The prognosis of patients with brain metastases from non-small cell lung cancer (NSCLC) is poor. Tyrosine kinase inhibitor (TKI) significantly improves the prognosis of patients with epidermal growth factor receptor (EGFR) sensitive mutation. EGFR sensitive mutations are associated with the incidence of brain metastases in NSCLC and may affect the efficacy of radiotherapy and TKI therapy. Both EGFR-TKI and radiotherapy are effective for EGFR-mutant NSCLC with brain metastases. Whether the combination of EGFR-TKI and radiotherapy may improve the prognosis compared with EGFR-TKI or radiotherapy alone has been studied. Retrospective studies have indicated that upfront radiotherapy, especially upfront stereotaxic radiosurgery combined with EGFR-TKI may be more advantageous in improving the prognosis, but it is still controversial. Therefore, clinical research progresses on the radiotherapy for EGFR-mutant NSCLC patients with brain metastases were reviewed.

Effectiveness of radiofrequency ablation therapy for patients with unresected Stage IA non-small cell lung cancer

Background: Approximately 20% of patients with resectable non-small cell lung cancer (NSCLC) are treated nonsurgically. To compare the clinical outcomes between nonsurgical patients receiving radiofrequency ablation (RFA) alone and those receiving no treatment (NT), we assessed RFA effectiveness in terms of survival using the surveillance, epidemiology, and end-results (SEER) database. Methods: Using the SEER registry process, we identified 5268 patients who were ineligible for the surgical treatment between 2004 and 2015. Overall survival (OS) and cancer-specific survival (CSS) were compared between the groups using propensity score matching (PSM), inverse probability of treatment weight (IPTW), and overlap weight analysis. In addition, an exploratory analysis was conducted to determine RFA treatment effectiveness based on clinically relevant patient subsets. Results: Of the 5268 patients, 189 (3.6%) received RFA. The OS and CSS in these patients were significantly better than those in the NT group (P < 0.0001). RFA was associated with a 16-month median OS improvement. Both OS and CSS improved in the nonsurgical patients (hazard ratio [HR], 0.695, 95% confidence interval [CI], 0.585–0.826, P < 0.0001; HR, 0.636; 95% CI, 0.505–0.800, P < 0.0001). The 1-, 3-, and 5-year OS in the unmatched RFA and NT groups were 84.2%, 49.0%, and 29.4% vs. 62.8%, 31.1%, and 17.1%, respectively (P < 0.001). PSM, IPTW, and overlap weight analysis showed comparable results. The odds of receiving RFA decreased with larger tumor size (>1, ≤2 cm, odds ratio [OR], 0.623, 95% CI, 0.402–0.966; >2, ≤3 cm, OR, 0.300, 95% CI, 0.186–0.483) compared to tumor size s1 cm (P < 0.05). Conclusion: RFA improves unresected stage IA NSCLC patient survival. Our results are limited by the retrospective nature of the study; however, we believe that our findings are noteworthy for recommending local ablative therapy

Effect of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell like cells / 解放军医学杂志

Objective To investigate the effect of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell-like cells of non-small cell lung cancer. Methods Non-small cell lung cancer H460 cell line was selected for serum-free suspension culture, and CD133 and CD44 positive H460 stem cells were screened by flow cytometry. qRT-PCR was performed to detect the expression levels of miR-30a, miR-148a, miR-148b, miR-152, miR-411, miR-497, miR-598, miR-424, miR-761, miR-1269a, miR-1280 and CD44, CD133 mRNA in H460 stem cell-like cells. The effects of miR-1269a expression on the radiotherapy sensitivity of H460 stem cell-like cells were analyzed using CCK-8 experiments, stem cell pelleting experiments, flow cytometry, and comet experiments. Results The expressions of CD133 and CD44 mRNA increased 1.30±0.03 times and 1.19±0.02 times, respectively, in H460 stem cell-like cells than in H460 cells (P<0.05). The expression level of miR-1269a was the highest in H460 stem cell-like cells, and was higher than that in H460 cells (P<0.05). After transfection of miR-1269a inhibitor and X-ray treatment, the growth inhibition rate of H460 stem cell-like cells increased significantly (72.11%±2.45%), the apoptosis rate reached to 17.70%±0.54%, and the fragmentation of DNA double-strand breaks in cells also increased significantly. After 2 Gy of X-ray radiation, the tail moment was 1.19±0.02 times of that before transfection, implying that transfection of miR-1269a inhibitor significantly increased the sensitivity of H460 stem cell-like cells to radiotherapy (P<0.05). After transfection of pcDNA3.1-SOX7, the effect of X-ray treatment on H460 stem cell-like cells was similar to that of miR-1269a inhibitor transfection. Simultaneous transfection of pcDNA3.1-SOX7 and miR-1269a mimics reversed the inhibitory effect of transfection of pcDNA3.1-SOX7 on the malignant biological behavior of H460 stem cell-like cells. Conclusion Inhibition of miR-1269a may enhance the radiotherapy sensitivity of non-small cell lung cancer, and the mechanism may be caused by up-regulation of SOX7 expression.

Charaoteristics and clinical significance of immune microenvironment in nonsmall cell lung cancer tissues / 中国肿瘤生物治疗杂志

@# Objective: To investigate the characteristics and clinical significance of the immunomicroenvironment typing based on the expression of programmed death-ligand 1 (PD-L1) and the infiltration of CD8+ T cells in the stroma in patients with non-small cell lung cancer (NSCLC). Methods: Paraffin tissue specimens and relevant clinicopathological data of 74 NSCLC patients admitted to our hospital from January 2016 to July 2018 were collected.All patients received EGFR gene test, and none received radiotherapy, chemotherapy or targeted therapy. Immunohistochemistry was used to detect the expression of PD-L1 in tissues and the infiltration of CD8+T cells in interstitium, and the relationship between PD-L1, CD8+T cells, and the immune microenvironment typing based on both, and the pathological parameters and the survival of patients was analyzed. Results: PD-L1 expression in the primary tumor of NSCLC patients showed statistical differences in gender, pathological type, smoking history, EFGR gene mutation status ( P <0.05). The infiltration of CD8+ T lymphocytes in tumor microenvironment showed statistically significant differences in different TNM stage and lymph node metastasis ( P <0.05), PD-L1 expression was significantly correlated with EGFR mutation ( P =0.000), while CD8+T lymphocyte infiltration was not correlated with EGFR mutation ( P =0.605). The immunomicroenvironment of EGFR wild-type patients was mainly (CD8+ PD-L1+) (type I), and the mutants were mainly (CD8-PD-L1-) (type II) and (CD8+PD-L1-) (type IV). The distribution of immune microenvironmental typing in each group with different EGFR mutation, smoking history and pathological differentiation degree was significantly different ( P <0.05) and significantly correlated with EGFR mutation ( P <0.05). Follow-up showed that the patients with disease free survival, recurrence and metastasis and death were the most in type I, type II and type I, respectively. Conclusions: In this study, the distribution of tumor immunomicroenvironmental typing in NSCLC patients was mainly the highest in type I and the lowest in type Ⅲ, which was related to EGFR mutation, smoking history and pathological differentiation. Patients with EGFR mutations were mainly of type Ⅱand type Ⅳ, and were associated with low expression of PD-L1. ··

Comparison of laboratory-developed test & validated assay of programmed death ligand-1 immunohistochemistry in non-small-cell lung carcinoma

Background & objectives: Inhibitors of immune checkpoint regulators, programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1), improve outcome in advanced non-small-cell lung carcinoma (NSCLC). Tumours expressing PD-L1 protein are more likely to benefit from this targeted therapy. Multiple concurrent clinical trials evaluating different anti-PD-1/PD-L1 therapies have validated five different immunohistochemistry (IHC) assays using varied antibody clones and staining conditions. This study was aimed at identification of a single harmonized PD-L1 assay for tumour tissue conservation and cost-effectiveness in patients with NSCLC. Methods: The performance of low-cost, manual, laboratory-developed technique (LDT) PD-L1 IHC assay using the easily available SP142 clone was compared with trial validated Ventana SP263 IHC performed on automated Ventana staining platform on tumour sections of NSCLCs. Results: Eighty cases of NSCLC were included. SP263 and SP142 stained both tumour cells and immune cells. The concordance rate of tumour cell staining was about 76 per cent, with SP263 detecting more tumour cells in 16 per cent of cases. The concordance rate of immune cell staining was only 61 per cent, with SP142 detecting more immune cells in 24 per cent of cases. The sensitivity, specificity, positive and negative predictive values of manual SP142 LDT assay against gold standard SP263 Ventana assay were 70, 94, 86 and 86 per cent, respectively, at positivity thresholds of ?1 per cent tumour cell staining. Interpretation & conclusions: The study findings suggested that LDT using SP142 clone showed only moderate concordance with SP263 Ventana assay, and the two assays were not interchangeable. More such validation studies need to be done to generate information that can complement patient therapy in cases of NSCLC.

Correlation between Ki-67 and PD-L1 expression in non-small cell lung cancer tissues and its effect on patients’prognosis / 中国肿瘤生物治疗杂志

@# Objective:To investigate the relationship between Ki-67 and PD-L1 in patients with non-small cell lung cancer (NSCLC) and their effects on prognosis. Methods: A total of 401patients, who were pathologically diagnosed as NSCLC in Changhai Hospital from January 2012 toAugust 2018, were enrolled as study subjects; and the patients were immunohistochemically tested for PD-L1 and Ki-67. The clinical and pathological data were collected, and the follow-up was performed regularly. The correlation between Ki-67 and PD-L1 and their effects on postoperative DFS and post-chemotherapy PFS were statistically analyzed. Results: Positive rates of PD-L1 and Ki-67 in NSCLC tissues were 37.9% (152/401) and 96.3% (386/401), respectively. Univariate analysis showed that Ki-67 was an influencing factor for PD-L1 expression (OR=0.33, 95%CI=0.28-0.39, P<0.0001); Curve Fitting analysis showed a positive correlation between Ki-67 and PD-L1; threshold effect analysis, segmentation multivariate logistic and ROC curve analysis showed 14% is a relatively suitable threshold for Ki-67 to be combined with PD-L1. Kaplan-Meier analysis showed that patients in Ki-67 high expression group had a significantly shorter post-operative DFS than those in Ki-67 low expression group ([21.88±11.25] vs [41.22±16.25]m, P< 0.0001), patients in PD-L1 positive group had a significantly shorter DFS than those in PD-L1 negative group ([24.75±14.59] vs [38.27± 16.75]m, P<0.0001)], and patients in Ki-67 high/PD-L1 positive group had the shortest DFS as compared to the other three groups ([20.57±11.33] vs [24.11±10.79], [36.00±16.79], [42.91±15.77]m, P<0.0001).As for post-chemotherapy PFS, patients in Ki-67 high expression group was significantly longer than those in Ki-67 low expression group [(7.70±3.01) vs (5.80±2.99)m, P=0.016), but there was no significant difference between PD-L1 positive group and PD-L1 negative group [(7.04±3.21) vs (6.33±3.06)m, P=0.22); for combined evaluation with Ki-67 and PD-L1, the PFS of two Ki-67 high expression groups was significantly longer than the other two Ki-67 low expression groups [(7.74±3.25) vs (7.43±2.38) vs (4.91±1.97) vs (6.02±3.19)m, P=0.041). Conclusion: Ki-67 is positively correlated with PD-L1 in NSCLC patients, and Ki-67 14% is a suitable threshold for combined use with PD-L1. Both Ki-67 and PD-L1 are predictors of poor prognosis. The combination of the two has an "additive effect" on the prediction of poor prognosis, and patients with high Ki-67 expression are more sensitive to chemotherapy.

Clinical Significance of Pleural Attachment and Indentation of Subsolid Nodule Lung Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Lung cancers presenting as subsolid nodule commonly have peripheral location, making the cancer-pleura relationship noteworthy. We aimed to evaluate the effect of pleural attachment and/or indentation on visceral pleural invasion (VPI) and recurrence-free survival. MATERIALS AND METHODS: Patients who underwent curative resection of lung cancer as subsolid nodules from April 2007 to January 2016 were retrospectively evaluated. They were divided into four groups according to their relationship with the pleura. Clinical, radiographical, and pathological findings were analyzed. RESULTS: Among 404 patients with malignant subsolid nodule, 120 (29.7%) had neither pleural attachment nor indentation, 26 (6.4%) had attachment only, 117 (29.0%) had indentation only, and 141 (34.9%) had both. VPI was observed in nodules of 36 patients (8.9%), but absent in nonsolid nodules and in those without pleural attachment and/or indentation. Compared to subsolid nodules with concurrent pleural attachment and indentation, those with attachment only (odds ratio, 0.12; 95% confidence interval [CI], 0.02 to 0.98) and indentation only (odds ratio, 0.10; 95% CI, 0.03 to 0.31) revealed lower odds of VPI. On subgroup analysis, the size of the solid portion was associated with VPI among those with pleural attachment and indentation (p=0.021). Such high-risk features for VPI were associated with earlier lung cancer recurrence (adjusted hazard ratio, 3.31; 95% CI, 1.58 to 6.91). CONCLUSION: Concurrent pleural attachment and indentation are risk factors for VPI, and the odds increase with larger solid portion in subsolid nodules. Considering the risk of recurrence, early surgical resection could be encouraged in these patients.

Acquired Resistance of MET-Amplified Non-small Cell Lung Cancer Cells to the MET Inhibitor Capmatinib / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Amplified mesenchymal-epithelial transition factor, MET, is a receptor tyrosine kinase (RTK) that has been considered a druggable target in non-small cell lung cancer (NSCLC). Although multiple MET tyrosine kinase inhibitors (TKIs) are being actively developed for MET-driven NSCLC, the mechanisms of acquired resistance to MET-TKIs have not been well elucidated. To understand the mechanisms of resistance and establish therapeutic strategies, we developed an in vitro model using the MET-amplified NSCLC cell line EBC-1. MATERIALS AND METHODS: We established capmatinib-resistant NSCLC cell lines and identified alternative signaling pathways using 3′ mRNA sequencing and human phospho-RTK arrays. Copy number alterations were evaluated by quantitative polymerase chain reaction and cell proliferation assay; activation of RTKs and downstream effectors were compared between the parental cell line EBC-1 and the resistant cell lines. RESULTS: We found that EBC-CR1 showed an epidermal growth factor receptor (EGFR)‒dependent growth and sensitivity to afatinib, an irreversible EGFR TKI. EBC-CR2 cells that had overexpression of EGFR-MET heterodimer dramatically responded to combined capmatinib with afatinib. In addition, EBC-CR3 cells derived from EBC-CR1 cells that activated EGFR with amplified phosphoinositide-3 kinase catalytic subunit α (PIK3CA) were sensitive to combined afatinib with BYL719, a phosphoinositide 3-kinase α (PI3Kα) inhibitor. CONCLUSION: Our in vitro studies suggested that activation of EGFR signaling and/or genetic alteration of downstream effectors like PIK3CA were alternative resistance mechanisms used by capmatinib-resistant NSCLC cell lines. In addition, combined treatments with MET, EGFR, and PI3Kα inhibitors may be effective therapeutic strategies in capmatinib-resistant NSCLC patients.

Effect of Platinum-Based Chemotherapy on PD-L1 Expression on Tumor Cells in Non-small Cell Lung Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Programmed death-1 (PD-1)/PD-1 ligand (PD-L1) axis blockades have revolutionized the treatment of advanced non-small cell lung cancer (NSCLC). We assessed the effect of platinum-based chemotherapy on tumor PD-L1 expression and its clinical implications. MATERIALS AND METHODS: We used immunohistochemistry to retrospectively evaluate the percentage of tumor cells with membranous PD-L1 staining (tumor proportion score) in paired tumor specimens obtained before and after platinum-based neoadjuvant chemotherapy (NACT) in 86 patients with NSCLC. We analyzed the correlation between the change in PD-L1 tumor proportion score and clinicopathologic characteristics, response to NACT, and survival. RESULTS: The PD-L1 tumor proportion score increased in a significant proportion of patients with NSCLC after platinum-based NACT (Wilcoxon signed-rank test, p=0.002). That pattern was consistent across clinically defined subgroups except for patients with partial response to NACT. Tumors from 26 patients (30.2%) were PD-L1‒negative before NACT but PD-L1-positive after NACT, whereas the reverse pattern occurred in six patients (7%) (McNemar’s test, p < 0.001). Increase in PD-L1 tumor proportion score was significantly associated with lack of response to NACT (Fisher exact test, p=0.015). There was a tendency, albeit not statistically significant, for patients with an increase in PD-L1 tumor proportion score to have shorter survival. CONCLUSION: Tumor PD-L1 expression increased after platinum-based NACT in a significant proportion of patients with NSCLC. Increase in tumor PD-L1 expression may predict poor clinical outcome.

Efficacy and Safety of Afatinib for EGFR-mutant Non-small Cell Lung Cancer, Compared with Gefitinib or Erlotinib / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: We tried to evaluate whether there are any specific features in treatment outcomes of firstline afatinib in patients with epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC), compared with gefitinib or erlotinib. MATERIALS AND METHODS: We analyzed patients treated with first-line afatinib, gefitinib, or erlotinib for advanced EGFR-mutant NSCLC at Samsung Medical Center between 2014 and 2016. RESULTS: In total, 467 patients received first-line afatinib (n=165), gefitinib (n=230), or erlotinib (n=72). Afatinib was used more often in patients with tumors harboring deletion in exon 19 (Del19), whereas the gefitinib group had more elderly, females, and never smokers. The median progression-free survival (PFS) time for afatinib, gefitinib, and erlotinib was 19.1 months, 13.7 months, and 14.0 months, respectively (p=0.001). The superior PFS of afatinib was more remarkable in subgroups of Del19 or uncommon EGFR mutations. Overall toxicity profiles of the three drugs were comparable, though more grade 3 or 4 toxicities were detected in afatinib (7.3%) compared with gefitinib (2.6%) or erlotinib (1.8%). The common grade 3 or 4 toxicities of afatinib included diarrhea (3.0%), paronychia (2.4%), and skin rash (1.8%). Dose modification was more frequently required in patients treated with afatinib (112/165, 68%), compared with gefitinib (5/230, 2%) and erlotinib (4/72, 6%). Interestingly, however, dose reduction in the afatinib group did not impair its efficacy in terms of PFS (dose reduction vs. no reduction group, 23.5 months vs. 12.4 months). CONCLUSION: First-line afatinib showed satisfactory efficacy data and manageable toxicity profiles.

A Randomized, Open-Label, Phase II Study Comparing Pemetrexed Plus Cisplatin Followed by Maintenance Pemetrexed versus Pemetrexed Alone in Patients with Epidermal Growth Factor Receptor (EGFR)-Mutant Non-small Cell Lung Cancer after Failure of First-Line EGFR Tyrosine Kinase Inhibitor: KCSG-LU12-13 / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The optimal cytotoxic regimens have not been established for patients with non-small cell lung cancer (NSCLC) who develop disease progression on first-line epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). MATERIALS AND METHODS: We conducted a multi-center randomized phase II trial to compare the clinical outcomes between pemetrexed plus cisplatin combination therapy followed by maintenance pemetrexed (PC) and pemetrexed monotherapy (P) after failure of first-line EGFR-TKI. The primary objective was progression-free survival (PFS), and secondary objectives included overall response rate (ORR), overall survival (OS), health-related quality of life (HRQOL), and safety and toxicity profiles. RESULTS: A total of 96 patientswere randomized, and 91 patientswere treated at 14 centers in Korea. The ORR was 34.8% (16/46) for the PC arm and 17.8% (8/45) for the P arm (p=0.066). With 23.4 months of follow-up, the median PFS was 5.4 months in the PC arm and 6.4 months in the P arm (p=0.114). The median OS was 17.9 months and 15.7 months in PC and P arms, respectively (p=0.787). Adverse events ≥ grade 3 were reported in 12 patients (26.1%) in the PC arm and nine patients (20.0%) in the P arm (p=0.491). The overall time trends of HRQOL were not significantly different between the two arms. CONCLUSION: The outcomes of pemetrexed therapy in NSCLC patients with disease progression after firstline EGFR-TKI might not be improved by adding cisplatin.

Salvage Concurrent Chemo-radiation Therapy for Loco-regional Recurrence Following Curative Surgery of Non-small Cell Lung Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: This study is to report clinical outcomes of salvage concurrent chemo-radiation therapy (CCRT) in treating patients with loco-regional recurrence (LRR) following initial complete resection of non-small cell lung cancer. MATERIALS AND METHODS: Between February 2004 and December 2016, 127 patients underwent salvage CCRT for LRR. The median radiation therapy (RT) dose was 66 Gy and clinical target volume was to cover recurrent lesion with margin without elective inclusion of regional lymphatics. Majority of patients (94.5%) received weekly platinum-based doublet chemotherapy during RT course. RESULTS: The median follow-up time from the start of CCRT was 25 months. The median survival duration was 49 months, and overall survival (OS) rates at 2 and 5 years were 72.9% and 43.9%. The 2- and 5-year rates of in-field failure-free survival, distant metastasis free survival, and progression free survival were 82.4% and 73.8%, 50.4% and 39.9%, and 34.6% and 22.3%, respectively. Grade ≥ 3 radiation-related esophagitis and pneumonitis occurred in 14 (11.0%) and six patients (4.7%), respectively. On both univariate and multivariate analysis, higher biologically equivalent dose (BED₁₀) (≥ 79.2 Gy₁₀ vs. 80 cm³; HR, 0.403), and longer disease-free interval (> 1 year vs. ≤ 1 year; HR, 0.489) were significantly favorable factors for OS. CONCLUSION: The current study has demonstrated that high dose salvage CCRT focused to the involved lesion only was highly effective and safe. In particular, higher BED₁₀, smaller CTV, and longer disease-free interval were favorable factors for improved survival.