A study on expression of programmed death ligand-1 in small cell lung carcinoma and correlation with clinicopathological parameters.

Small cell lung carcinoma (SCLC) is characterized by rapid growth and an aggressive clinical course. Standard therapy regimes have limited effects on disease course; therefore the prognosis of SCLC is poor. In the current study, the frequency of programmed death ligand 1 (PD-L1) expression in SCLC and its correlation with clinico-pathological features were evaluated. The study included 100 cases of SCLC wherein testing for PD-L1 was done with the SP263 clone on the Ventana benchmark XT system. Cases with > 1% PD-L1 expression in tumour cells or immune cells were categorized as positive. PD-L1 expression was identified in 14% of cases using the cut-off of ≥ 1%. The tumour proportion score was 10% and the immune proportion score was 9.78% using a cut-off of ≥ 1%. PD-L1 positive expression was more frequent in the male population with age > 40 years. All the patients with positive PD-L1 expression were smokers. In the PD-L1 positive group, presence of necrosis was identified in 71.4% of cases and when compared with the PD-L1 negative subgroup this finding was statistically significant (p = 0.010). Personalized targeted therapy for cases of SCLC is still under evaluation. The use of immunotherapeutic targets, such as PD-L1, may help to define a new treatment strategy for SCLC. Development of new treatment strategies may improve prognosis and survival.

Morphometric delineation of administrative boundaries and classification of threatened categories of small watersheds in transboundary rivers.

The ecological conservation of large rivers is impossible unless immediate attention is given to protecting their small tributaries at local levels. The natural boundaries of large river basins are shrinking because their tributaries and streams of different orders are disappearing at an unprecedented rate. Delineation of the fixed administrative boundaries (AB) to protect the natural boundary of small rivers and their classification into appropriate threatened categories, the present study was carried out on the 54.08 km long Banki River in the Ganga River basin. The > 70% irreversible loss in the number of streams (Nu), length of streams (Lu), and drainage density (Dd) resulted in the conversion of the 6th order Banki into the 4th order river. The extreme morphometric changes result in the Banki watershed being under the "Critically Endangered" category. The drainage density ratio (DdR) and mean stream width (Msw) were used to determine the width of AB (WAB). The "River Red List Categories and Criteria" are being proposed to strengthen global initiatives at the local levels to protect and conserve inland water bodies and transboundary rivers.

Programmed Death Ligand 1 Expression in Lung Adenocarcinoma: An Analysis of the Histomorphological Features.

Background: Immunotherapy is now a vital target therapy in the advanced cases of lung adenocarcinoma. The outstanding result of therapies with medications that inhibit the interaction of programmed death ligand 1 with programmed death protein 1 has revolutionarized prognostic treatment regimes. Aims: The study was undertaken with the objectives to analyze the detailed histomorphological features of adenocarcinoma lung with programmed death ligand-1 (PD-L1) expression in tumor cells and to compare the histomorphological features with PD-L1 negative group. Materials and Methods: The present study is a retrospective case series with 100 cases of non-small cell lung cancer-adenocarcinoma phenotype in which testing for PD-L1 had been done using immunohistochemistry. Detailed histomorphological analysis and comparison was performed for both the PD-L1 positive and negative phenotype. Results: Histomorphological features of 25 cases with positive PD-L1 positivity in the tumor cells and 75 cases that were negative for PD-L1 were analyzed. The most frequent pattern in the category that was PD-L1 positive was singly scattered cells or loose clusters present in 84% cases followed by solid nests that was identified in 60% cases. The presence of solid nests in the PD-L1 positive category was statistically significant (P = 0.018). Mucin was identified in 24% cases, and tumor necrosis was documented in 60% cases with PD-L1 positivity. In the cluster that was PD-L1 positive, 92% cases had moderate-to-severe nuclear pleomorphism. Conclusion: The identification of histomorphological patterns and characteristics may aid in triaging cases that have the likelihood to harbor PD-L1-positive phenotype, which has predictive and prognostic outcome.

Impact of glutathione S transferases P1 (Ile105Val) variants on the risk of GSTp, phosphorylated c-Jun kinase, and P53 phenotypic expression and their implications on overall survival outcomes in non-small cell lung cancer patients treated with chemotherapy.

AIM: This study focused on GST-M1, T1 null, and P1 Ile105Val variant genotypes associated with the risk of altered expression of GSTp, pJNK, and P53 in NSCLC patients. These markers and overall survival (OS) were correlated with a key set of clinicopathological characteristics. METHODS: Genotyping of GST- M1, T1 (+/-), and P1 (Ile105Val) was performed using PCR-RFLP.The expression of GSTp, pJNK, and P53 phenotypes was assessed by immunohistochemistry. The Spearman test was used to examine the correlation between GSTp, pJNK, and P53. Kaplan-Meier test was used for OS analysis. RESULTS: GSTP1 Val/Val and Ile/Val genotypes notably increased GSTp expression by 1.8 and 1.7 fold, respectively (p = 0.04,p = 0.06). GSTP1 Val/Val and Ile/Val genotypes considerably reduced P53 expression by 0.61 and 0.57 fold, respectively (p = 0.03& p = 0.05), respectively. GSTp, pJNK, and P53 were significantly co-expressed (p < 0.001). GSTp and pJNK expression showed a moderate negative correlation (ρ = -0.32, p = 0.046). In contrast, GSTp and P53 expression exhibited a strong negative correlation (ρ = -0.53, p < 0.0001). There was no correlation between P53 and pJNK expression(ρ = 0.07, p = 0.54). The patient's median OS was 8.9 months, and it was significantly related to pack-years, stage, metastasis, and GSTM1(-/-) genotypes (p > 0.05). SQCLC showed poor OS than ADC (5.7 months vs.9.1 months, p = 0.2). Stage IV and metastasis significantly reduced the OS (p = 0.001). The tumour size and lymph nodes reflected poor OS (p = 0.07&p = 0.06). Gemcitabine+Cisplatin and Gefitinib showed a slightly higher rate of survival (9.3 months and 8.1 months) than Pemtrexe+Cisplatin treatment (7.0 months,p = 0.8). Multivariate analysis revealed that pack-years and GSTp were independent predictors for OS (p = 0.03). CONCLUSION: GSTp, pJNK, and P53 showed interconnected cascading. Age, pack-year, stage, and GSTp were found to be significant predictive factors for OS.Pack-years, GSTp independent OS predictor.

Correlation between Programmed Death Ligand-1(PD-L1) Expression and Driver Gene Mutations in Non-Small Cell Lung Carcinoma- Adenocarcinoma Phenotype.

BACKGROUND: Targeted therapy in adenocarcinoma is recommended. The use of immune check point inhibitors for the treatment of Non-small cell lung carcinoma (NSCLC) is used as both first-line and the second-line treatment strategy. The current study was undertaken to assess the frequency of programmed cell death ligand-1 (PD-L1) expression with anaplastic lymphoma kinase (ALK), proto-oncogene 1, receptor tyrosine kinase (ROS), epidermal growth factor receptor (EGFR), Kirsten rat sarcoma (KRAS), and v-Raf murine sarcoma viral oncogene homolog B (BRAF)V600E driver gene mutations in NSCLC adenocarcinoma phenotype. It assesses the frequencies of all markers in the cases where both treatment strategies can be implemented. Expression of the all markers was further compared with demographic, clinical parameters, and overall survival rate. MATERIALS AND METHODS: The formalin-fixed paraffin-embedded (FFPE) tissue blocks were used in immunohistochemistry (IHC) staining and real-time polymerase chain reaction (RT-PCR) for determining the driver genes and PD-L1 expression in the 100 NSCLC-Adenocarcinoma cases. RESULTS: PD-L1 positivity was observed in 26.36% (n=29/110) cases in adenocarcinoma. No significant differences in PD-L1 expression were observed among patients harboring ALK, ROS1, EGFR, KRAS, and BRAF mutations EGFR mutations had significant association with smoking status. (p= 0.008), Thyroid transcription factor 1 (TTF1) (p=0.0005) and Napsin (p=0.002) expression. ALK gene re-arrangement was significantly related to age (p= 0.001), gender (p= 0.009) and smoking status (p= 0.043). The single versus multiple driver mutations were significantly correlated with smoking status (p=0.005). In the survival rate analysis, EGFR (p=0.058), KRAS (p=0.021), and PD-L1 (p=0.039) were significantly high with the positive versus negative group. CONCLUSIONS: The current study is a novel attempt to document the co-expression of multiple driver mutations in the NSCLC-adenocarcinoma phenotype. PD-L1 immunopositivity in NSCLC-adenocarcinoma was higher with EGFR mutation as compared to those of KRAS, ALK, ROS, and BRAF driver genes.

Primary Enteric Adenocarcinoma Lung with Mesenchymal Epithelial Transition Factor Amplification: A Case Report.

INTRODUCTION: Primary enteric adenocarcinoma (PEAC) of the lung is a rare variant characterized by the presence of colorectal adenocarcinoma like components. A rare case of PEAC of lung with over-expression of mesenchymal epithelial transition factor (MET) receptor tyrosine kinase protein is being presented. CASE REPORT: A 75-year old female presented with chest pain, cough, hemoptysis and fever for 2 months duration. Computerized tomography (CT) scan revealed a spiculated mass in left upper lobe of lung. A needle core biopsy was performed. The histopathological examination revealed a tumor composed of tall columnar cells with vesicular nuclei arranged in an acinar and glandular formation with luminal necrosis. On immunostaining, cytokeratin 7, thyroid transcription factor 1 and CDX-2 were positive. Based on the morphological and immunostaining profile a diagnosis of PEAC of the lung was rendered. Ancillary testing for driver mutations revealed over expression of c-MET while epidermal growth factor receptor (EGFR) mutations, anaplastic lymphoma kinase (ALK) rearrangement and C-ros oncogene 1, receptor tyrosine kinase (ROS 1) were negative. DISCUSSION: PEAC of the lung which was first described by Tsao and Fraser, is an uncommon variant and rarely harbors any driver mutations. MET over expression is present in 3-7% cases of lung adenocarcinoma and is indicative of aggressive tumor behavior. Colo-rectal adenocarcinomas with MET amplification have poor overall survival. The histogenesis of PEAC may be implicated to the dysregulation of MET pathway. Targeted therapy using MET inhibitors in cases that have MET amplification has predictive implications. CONCLUSION: This is a rare case of PEAC with c-MET over expression. The MET axis that is implicated in pathogenesis of colonic adenocarcinomas might also be the molecular pathway for the development of PEAC.

Kirsten rat sarcoma virus protein overexpression in adenocarcinoma lung: Association with clinicopathological and histomorphological features.

CONTEXT: Lung cancer is the leading cause of cancer-related deaths worldwide. The constitutive activation of multiple signaling pathways is the major cause of carcinogenesis. AIMS: The study evaluates the frequency of Kirsten rat sarcoma virus (KRAS) protein overexpression and correlates with clinicopathological and histomorphological features in non-small cell lung carcinoma (NSCLC)-adenocarcinoma. SETTINGS AND DESIGN: Tertiary hospital-based retrospective and prospective case series included 100 cases of NSCLC-adenocarcinoma. MATERIALS AND METHODS: The basic panel of Immunohistochemistry including Napsin-A, thyroid transcription factor-1 (TTF-1), and markers for squamous differentiation, p-40 was used in formalin-fixed paraffin-embedded tissue blocks. The KRAS monoclonal antibody (9.13, Thermo Fisher Scientific, USA) was used. STATISTICAL ANALYSIS USED: The IBM-Statistical Package for the Social Sciences (SPSS) (SPSS, International Business Machines Corporation, New York, NY, USA) analysis software, version 16 was used for all statistical calculations. RESULTS: KRAS protein expressed in 28.0% (28/100) cases. Cases were grouped as KRAS positive and negative. TTF-1 and Napsin-A were expressed in 89.25% (n = 25) and 92.86% (n = 26) cases, respectively. Stage IV clinical disease was identified in 55% of cases, and 36.84% of cases had a mean survival between 6 and 12 months. In KRAS positive group, the most common pattern of cellular arrangement was acinar/loose clusters pattern present in 64.29% (n = 21) and 75.0% (n = 18) cases followed by the solid pattern present in 42.86% of cases (n = 12), respectively. Necrosis was identified in 57.14% (n = 16) cases. Mucin pattern was present in 32.14% of cases (n = 9), which was significantly different when compared with the KRAS negative group (P = 0.036). CONCLUSIONS: This finding may imply that KRAS mutations may not be entirely triggered by alterations induced by carcinogens in smoke. KRAS gene is frequently mutated in pulmonary tumors. It should be tested in NSCLC owing to its predictive and prognostic effects.

Detection of Anaplastic Lymphoma Kinase Gene Re-Arrangement in Non-Small Cell Lung Carcinoma in the Indian Population: Comparison of Techniques and Immunohistochemistry Clones.

OBJECTIVE: Predictive and prognostic markers have revolutionized personalized therapy in non-small cell lung carcinoma (NSCLC). Crizotinib is now approved for locally advanced or metastatic NSCLC that is anaplastic lymphoma kinase (ALK) positive by either Fluorescence in situ hybridization (FISH) or immunohistochemistry (IHC). The current study aimed to detect the incidence of ALK gene re-arrangement in the Indian population, to compare the various IHC antibodies with FISH as a gold standard, and to analyze the morphology of cases with ALK phenotype. MATERIAL AND METHOD: A case series of 614 cases of NSCLC were included. IHC for detection of ALK phenotype was compared with FISH using 5A4 clone (Labvision, USA), ALK-1(Dako, Denmark) and D5F3 clone (Ventana, USA). RESULTS: ALK gene rearrangement was evident in 4.07% of the cases. Cases with ALK phenotype had unique histomorphology with presence of mucin or signet ring cells in association with necrosis, high tumour grade and poor differentiation. Comparison of various antibody clones used in IHC revealed that the sensitivity and specificity using the D5F3 clone (100%, 100%) and 5A4 clone (87.5%, 100%) were similar while the ALK-1 clone had the lowest sensitivity and specificity (50%, 95.5%). CONCLUSION: The incidence of ALK gene rearrangement in NSCLC in the current Indian study is within the worldwide reported range of 3-5%. This is the first study from the Indian subcontinent to compare various IHC antibodies used for detection of ALK phenotype. IHC using D5F3 clone and 5A4 clone may be considered as a rapid reliable and inexpensive method for detection of ALK gene rearrangement.