Epidermal Growth Factor Receptor Immunohistochemistry Research Articles

Detection of Early Esophageal Neoplastic Barrett Lesions with Quantified Fluorescence Molecular Endoscopy Using Cetuximab-800CW.

Esophageal adenocarcinoma causes 6% of cancer-related deaths worldwide. Near-infrared fluorescence molecular endoscopy (NIR-FME) uses a tracer that targets overexpressed proteins. In this study, we aimed to investigate the feasibility of an epidermal growth factor receptor (EGFR)-targeted tracer, cetuximab-800CW, to improve detection of early-stage esophageal adenocarcinoma. Methods: We validated EGFR expression in 73 esophageal tissue sections. Subsequently, we topically administered cetuximab-800CW and performed high-definition white-light endoscopy (HD-WLE), narrow-band imaging, and NIR-FME in 15 patients with Barrett esophagus (BE). Intrinsic fluorescence values were quantified using multidiameter single-fiber reflectance and single-fiber fluorescence spectroscopy. Back-table imaging, histopathologic examination, and EGFR immunohistochemistry on biopsy samples collected during NIR-FME procedures were performed and compared with invivo imaging results. Results: Immunohistochemical preanalysis showed high EGFR expression in 67% of dysplastic tissue sections. NIR-FME visualized all 12 HD-WLE-visible lesions and 5 HD-WLE-invisible dysplastic lesions, with increased fluorescence signal in visible dysplastic BE lesions compared with nondysplastic BE as shown by multidiameter single-fiber reflectance/single-fiber fluorescence, reflecting a target-to-background ratio of 1.5. Invisible dysplastic lesions also showed increased fluorescence, with a target-to-background ratio of 1.67. Immunohistochemistry analysis showed EGFR overexpression in 16 of 17 (94%) dysplastic BE lesions, which all showed fluorescence signal. Conclusion: This study has shown that NIR-FME using cetuximab-800CW can improve detection of dysplastic lesions missed by HD-WLE and narrow-band imaging.

STRIvE-01: Phase I study of EGFR806 CAR T-cell immunotherapy for recurrent/refractory solid tumors in children and young adults.

2541 Background: The epidermal growth factor receptor (EGFR) is a cell surface tyrosine kinase receptor associated with cell proliferation and differentiation. EGFR expression and activating mutations are associated with aggressive neoplastic disease, chemotherapy resistance, and increased metastatic potential. Published data and EGFR immunohistochemistry (IHC) performed on tissue microarrays indicate that 15-40% of pediatric solid tumors (ST) express EGFR. The unique EGFR monoclonal antibody (mAb) 806 selectively binds to an epitope that is conformationally hidden when EGFR is tethered but revealed when tethering is perturbed as occurs with EGFR overexpression, truncation, or through extra-cellular domain missense mutations. Methods: Children and young adults (CYA) with EGFR-expressing recurrent/refractory (R/R) ST were enrolled on a Phase 1 trial to examine the safety and feasibility of administering autologous chimeric antigen receptor (CAR) T cells derived from autologous T cells genetically modified to express a second generation EGFR806-specific scFV-IgG4hinge-CD28tm/cyto-4-1-BB-zeta and EGFRt tracking/suicide contract. All subjects received lymphodepleting chemotherapy with fludarabine and cyclophosphamide prior to the administration of cryopreserved CAR T cells a the prescribed dose level. The biologically effective dose (BED) or maximum tolerated dose was determined based upon observed toxicity through day 28 from initial CAR-T infusion and using a 3+3 statistical design. Results: Eleven subjects (n=10 evaluable, age range 9-25, median 18) were enrolled and received either dose level (DL) 1 (0.5 x 106 CAR-T/kg, n=4) or DL2 (1 x 106 CAR-T cells/kg, n=7). CAR T were manufactured successfully in all subjects. Most common toxicities were fatigue, tumor-related pain and cytokine release syndrome (n=2, maximum CTCAE grade 1). Dose limiting toxicity of CTCAE grade 4 transaminase level and hyperbilirubinemia occurred at DL2 (n=1). Maximum circulating CAR-T expansion was 29.66 cells/uL (range 0.05-29.66 cells/uL) with median persistence of 28 days (range 0-90). Two subjects on DL1 and one subject on DL2 demonstrated mixed response on day 28 and tolerated additional CAR T infusion without dose limiting toxicity. Conclusions: EGFR806 directed CAR-T cells have an acceptable toxicity profile in CYA with R/RST and demonstrate anti-tumor activity in some patients. Additional analyses are ongoing to identify biomarkers of response and toxicity. Clinical trial information: NCT03618381.

Biomarkers expression among paired serous ovarian cancer primary lesions and their peritoneal cavity metastases in treatment-naïve patients: A single-center study.

BackgroundHigh‐grade serous ovarian carcinoma (HGSOC), the most common histologic subtype of ovarian epithelial cancer, is associated with treatment resistance, enhanced recurrence rates, and poor prognosis. HGSOCs often metastasize to the peritoneal cavity, while fluid cytology examination could identify such metastases. This retrospective study aimed to identify potential biomarker discrepancies between paired HGSOC primary tissues and metastatic peritoneal fluid cytology samples, processed as cell blocks (CBs).MethodsTwenty‐four pairs of formalin‐fixed, paraffin‐embedded primary tissues and metastatic CBs from an equal number of treatment‐naïve patients were used, and immunohistochemistry (IHC) for epidermal growth factor receptor (EGFR), human epidermal growth factor receptor, programmed cell death‐1 ligand 1 (PD‐L1), and CD147 was applied.Results13/24 pairs showed discordant EGFR IHC results; in all these 13 patients, EGFR was positive (≥1+ membranous staining intensity found in at least 10% of the cancer cells) in the peritoneal, yet negative in the primary tissue samples. Notably, EGFR IHC was positive in 15/24 of the metastatic, whereas in just 2/24 of the primary HGSOC samples (p < 0.001). Although most PD‐L1 results were concordant, 5/24 and 6/24 pairs exhibited discordant results when stained with the E1L3N and 22C3 clones, respectively. Lastly, CD147 overexpression was found more often in the metastatic rather than the matched primary HGSOCs stained with CD147, though the difference was not significant.ConclusionsCytology from effusions could be considered for biomarker testing when present, even when tissue from the primary cancer is also available and adequately cellular, as it could provide additional information of potential clinical significance.

Visualizing surface marker expression and intratumoral heterogeneity with SERRS-NPs imaging.

Cell surface marker expression in tumors dictates the selection of therapeutics, therapy response, and survival. However, biopsies are invasive, sample only a small area of the tumor landscape and may miss significant areas of heterogeneous expression. Here, we investigated the potential of antibody-conjugated surface-enhanced resonance Raman scattering nanoparticles (SERRS-NPs) to depict and quantify high and low tumoral surface marker expression, focusing on the surface markers epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER2) in an intracerebral and peripheral setting with an inter- and intratumoral comparison of Raman signal intensities.Methods: ICR-Prkdc <scid> mice were injected with glioblastoma, epidermoid carcinoma, or breast tumor cell lines intracerebrally and peripherally. SERRS-NPs were functionalized with cetuximab or trastuzumab and administered via tail vein injection. Raman imaging was performed 18 hours post-injection in excised tumors and in vivo through the skull. Tumors were then fixed and processed for immunohistochemical evaluation.Results: Confirmed by MRI and immunohistochemistry for EGFR and HER2, our results demonstrate that antibody-conjugated SERRS-NPs go beyond the delineation of a tumor and offer clear and distinct Raman spectra that reflect the distribution of the targeted surface marker. The intensity of the SERRS-NP signal accurately discriminated high- versus low-expressing surface markers between tumors, and between different areas within tumors.Conclusion: Biopsies can be highly invasive procedures and provide a limited sample of molecular expression within a tumor. Our nanoparticle-based Raman imaging approach offers the potential to provide non-invasive and more comprehensive molecular imaging and an alternative to the current clinical gold standard of immunohistochemistry.

Immunohistochemical Detection and Prognostic Significance of p53, Epidermal Growth Factor Receptor, Murine Double Minute 2, and Isocitrate Dehydrogenase 1 in Glioblastoma Multiforme Patients of Pakistan.

Introduction:Glioblastoma multiforme (GBM) is one of the deadliest cranial tumors occurring in adults. Various biomarkers have been tested for their significance in diagnosis, prognosis, and treatment of GBM. Some well-studied markers in GBM are Isocitrate dehydrogenase 1 (IDH1), Murine double minute 2 (MDM2), Epidermal Growth Factor Receptor (EGFR), and p53. The aim of this study was to investigate the protein expression of these markers in GBM patients of Pakistan.Methods:A total of 102 surgically resected formalin-fixed paraffin-embedded specimens from patients diagnosed and treated at Aga Khan University Hospital were included in this study. Immunohistochemistry (IHC) for IDH1, MDM2, EGFR, and p53 was performed using Dako EnVision System and respective monoclonal antibodies. Survival analysis was performed to check association of markers protein expression with prognosis in GBM patients.Results:There were 73 males and 29 females in this study, with a median age of 49 years at the time diagnosis. Overexpression of molecular markers was as follows: 52% for EGFR, 26% for p53, 72% for IDH1, and 83% for MDM2. We did observe that EGFR was significantly associated with increased age of our patients and with worse survival. Age > 40 years was a predictor for worse prognosis as well.Conclusion:EGFR overexpression and advanced age were worse prognostic indicators.

Correlation between CD133 and EGFR Expression with Grading of Ovarian Carcinoma

BackgroundOvarian carcinoma account for 90% of all malignant ovarian tumors. Epidermal Growth Factor Receptor (EGFR), also known asErbB1/HER1, has an important role in tumor cell proliferation. CD133 is a marker for hematopoietic stem cell and it is involved in tumorcell proliferation, self renewal and cell differentiation. The objective to analyze the expression of CD 133 and EGFR in high grade andlow grade ovarian carcinoma, and to determine the correlation between the expression of CD133 and EGFR expression in ovariancarcinoma.MethodsThe study was conducted with observational and cross sectional method on paraffin blocks of patients diagnosed as ovariancarcinoma from 1 January until 31 December 2017 in Dr. Soetomo Hospital. Ovarian carcinoma was divided into 2 grade, low gradeand high grade. Immunohistochemistry for EGFR and CD133 was performed. Immuno-reactive Score (IRS) was applied to evaluatethe expression of EGFR and CD133. The difference expression of CD133 and EGFR in low grade and high grade ovarian carcinomawas analyzed with Mann-Whitney test. Correlation between CD133 with EGFR was analyzed with Spearman test.ResultsThere were significant difference in expression of CD133 (p=0.0001) and EGFR (p=0.0005) in high grade and low grade OvarianCarcinoma. There was significant positive correlation between CD133 and EGFR expression in Ovarian Carcinoma (p=0.035; r=0.37)ConclusionExpression of CD133 and EGFR were higher in high grade ovarian carcinoma. The higher expression of CD133 will increaseexpression of EGFR in ovarian carcinoma. Both markers may be used as prognostic factor.

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.

Congenital mesoblastic nephroma is characterised by kinase mutations including EGFR internal tandem duplications, the ETV6-NTRK3 fusion, and the rare KLHL7-BRAF fusion.

Congenital mesoblastic nephroma (CMN) is histologically classified into classic, cellular and mixed subtypes. The aims of this study were to characterise the clinical, pathological and molecular features of a series of CMNs, and to determine the utility of pan-Trk and epidermal growth factor receptor (EGFR) immunohistochemistry as surrogate markers for NTRK gene fusions and EGFR internal tandem duplications (ITDs). Twenty-two archival CMN cases (12 classic, five cellular, and five mixed) were tested for the ETV6-NTRK3 fusion and EGFR ITD transcripts by the use of reverse transcriptase polymerase chain reaction (PCR), and next-generation sequencing-based anchored multiplex PCR. All 12 classic CMNs had EGFR ITD. Of the five cellular CMNs, four had the ETV6-NTRK3 fusion and one had the KLHL7-BRAF fusion. Of the five mixed CMNs, four had EGFR ITD, and one had the ETV6-NTRK3 fusion. Pan-Trk immunoreactivity was 100% sensitive and 94.1% specific for the presence of NTRK rearrangement. However, EGFR staining was only 62.5% sensitive and 33.3% specific for EGFR ITD. EGFR ITD is a consistent genetic event in classic CMN. A majority of cellular CMNs have the ETV6-NTRK3 fusion. Rare cellular CMNs may harbour non-canonical mutations such as the KLHL7-BRAF fusion, which was found in one case. Mixed CMNs may have either EGFR ITD or the ETV6-NTRK3 fusion. Pan-Trk immunohistochemistry is a sensitive, albeit not perfectly specific, marker for NTRK rearrangement. EGFR immunohistochemistry is not helpful as a marker of EGFR ITD.

CD274 (PD-L1), CDKN2A (p16), TP53, and EGFR immunohistochemical profile in primary, recurrent and metastatic vulvar cancer.

Vulvar squamous cell carcinoma can be divided by human papillomaviruses (HPV) status into two distinct clinicopathological and molecular entities. New agents targeting the tumor surface expression of programmed cell death-1/programmed cell death-ligand-1 are becoming a therapeutic option in an increasing number of carcinomas. We evaluate CD274 (PD-L1), CDKN2A (p16), tumor protein p53 (TP53), and epidermal growth factor receptor (EGFR) immunoexpression in primary tumors, recurrences and lymph node metastases and its correlations with prognosis and HPV status. We report 93 cases of vulvar squamous cell carcinoma diagnosed between 2002 and 2016 with the description of their clinicopathological features and prognosis data. Immunohistochemistry for CD274, CDKN2A, TP53, and EGFR was performed on tissue microarrays collecting from primary tumor, recurrences and lymph node metastasis. Kaplan-Meier estimator and multivariable Cox regression analysis controlling for FIGO stage and age were used. Patients who underwent surgery had a superior overall survival (HR = 0.51, 95% CI = 0.26-0.99 p = 0.04). Lymph node metastasis size ≥5 mm was associated with an inferior overall survival (HR = 1.88, 95% CI = 1.22-2.92 p = 0.004). CDKN2A expression was correlated with an inferior rate of recurrent disease (p = 0.02). In high-risk HPV DNA+ vulvar squamous cell carcinomas patients with CDKN2A- carcinomas showed a significantly worse overall survival than women with CDKN2A+ tumors (56% vs.100%, p = 0.003). TP53 expression was associated with an increased rate of recurrent disease (p = 0.0005). CD274 expression was associated with lymph node metastasis (p = 0.04). In 16 patients the CD274, CDKN2A, TP53, and EGFR expression changed between primary tumors, recurrences and lymph node metastases during tumor progression. In conclusion, a significant percentage of vulvar squamous cell carcinoma has a heterogeneous biomarker expression during tumor progression. We highlight the importance of some of these markers to be used as prognostic biomarkers. This data brings new light to future treatment using targeted therapy to EGFR or CD274 to include retesting such biomarkers in recurrence and lymph nodes metastases.

Overexpression of EGFR, COX2 and p53 in oral squamous cell carcinoma patients of Pakistan and correlation with prognosis

Abstract Background Oral squamous cell carcinoma (OSCC), the sixth leading cancer worldwide, ranks as the most common cancer in males, and the third most common in females in Pakistan. It is influenced by risk factors which are widely consumed in our population. The epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor that is mutated and overexpressed in a variety of cancers. Cyclooxygenase 2 (COX2) is a rate-limiting enzyme that is upregulated in many cancers. It is involved in angiogenesis, apoptosis and metastasis of neoplasia. TP53 mutation and overexpression have been correlated with poor survival in many cancers including oral squamous cell carcinoma (OSCC). The aim of our study was to observe the expression of EGFR, COX2 and p53 in OSCC and correlate the expression with patient’s overall survival, and validate these markers for personalized therapy. Methods Formalin-fixed paraffin-embedded tissues of OSCC cases (n = 100) were used. Immunohistochemistry for EGFR, COX2 and p53 was performed using Dako monoclonal antibodies and EnVision system. Results There was a majority of males (55%) in our data set and the larger portion of patients was above 40 years of age (82%). Risk factor consumption was found in 73% of cases. EGFR overexpression was observed in 70%, COX2 in 55% and p53 in 67% of OSCC patients. Survival analysis revealed EGFR positive patients had markedly low overall survival (p = 0.098). Similarly, COX2 proved a good prognostic factor for patient survival (p = 0.013), while p53 overexpression was insignificant for survival (p = 0.107). Conclusions Our findings emphasize the role of EGFR, COX2 and p53 overexpression in OSCC patients. Molecular therapies targeted towards EGFR, COX2 and p53 may be used against Pakistani OSCC patients to improve prognosis. Legal entity responsible for the study Aga Khan University, Karachi, Pakistan. Funding Higher Education Commission, Pakistan. Disclosure All authors have declared no conflicts of interest.

Role of molecular markers in glioblastoma patients of Pakistan

Abstract Background Glioma is the most common primary brain malignancy with a largely unknown pathogenesis, although complex genetic abnormalities are thought to contribute to disease development. Several independent studies have investigated the expression of Epidermal Growth Factor Receptor (EGFR), Cyclooxygenase 2 (COX-2), p53 and Cyclin D1 in gliomas. However, conflicting evidence exists for expression and prognostic significance of these markers in glioma. Furthermore, no work relating to these genes has been performed on the glioma population of Pakistan. Our objective was to investigate the expression and prognostic significance of molecular markers p53, EGFR, COX-2 and Cyclin D1 in glioma in the Pakistani population. Methods Immunohistochemistry of markers p53, EGFR, COX-2 and Cyclin D1 was performed on 100 formalin-fixed paraffin-embedded specimens of patients diagnosed and treated atAga Khan University Hospital, Karachi, Pakistan. Expression was correlated with patient clinicopathological characteristics and overall survival. Results The prevalence of expression of Cyclin D1, EGFR, COX-2, and p53 was 57%, 52%, 54% and 39%, respectively. p53 and EGFR expression were positively correlated with younger and older age respectively. Chi squared test revealed a significant association between EGFR and Cyclin D1, with EGFR expression decreasing with increasing strength of Cyclin D1 staining. Only patient age was found to be a significant predictor for overall survival. Conclusions There is a relatively low prevalence of COX-2 and EGFR in the Pakistani population as compared to published literature. We also observed a correlation between EGFR and Cyclin D1 expression. The use of these biomarkers as targets for personalized therapies is worth investigating in the Pakistani population. Legal entity responsible for the study Aga Khan University, Karachi, Pakistan. Funding University Research Council, Aga Khan University, Karachi, Pakistan. Disclosure All authors have declared no conflicts of interest.

Immunohistochemical Expression (IHC) and Fluorescence In Situ Hybridization (FISH) of EGFR in Colorectal Carcinoma

Adenocarcinoma of the colon is the most common malignancy of the gastrointestinal tract and is a major cause of morbidity and mortality worldwide. Epidermal growth factor receptor (EGFR) is also known as HER-1 or erb-B1. The beginning of EGF to EGFR produces a biological signal to the cell that initiates several functions that promote tumor growth, including cell invasion and metastasis, repair and new blood vessel formation. Thus, EGFR is recognized as an important player in colorectal cancer (CRC) initiation and progression. In this study, total 61 cases of colorectal carcinoma were included and histological grading, immuno-histochemical (IHC) expression and fluorescence in situ hybridization of EGFR were conducted. For the grading, 11/61(18%) cases were well differentiated, 38/61(62%) cases were moderately differentiated and 12/61(20%) were poorly differentiated colorectal carcinoma. EGFR IHC immune expression was positive in 50/61(82%) cases and negative in 11/61(18%) cases. All 11 cases of well differentiated cases gave EGFR IHC positive immunoreaction. Among the 38 cases of moderately differentiated adenocarcinoma, 30 cases showed EGFR IHC positivity and 8 cases gave no reaction. Nine out of 12 cases of poorly differentiated adenocarcinoma showed EGFR IHC positive and 3 cases gave no reaction. Half (10/20, 50%) of the EGFR IHC highly positive cases showed FISH positive and other half cases give FISH negative reaction. Detection of EGFR is mainly for anti-EGFR targeted therapy. Therefore, this study aids in selection of patients for anti-EGFR targeted therapy and helpful in treatment options and disease management. Moreover, EGFR FISH can be tested together with KRAS mutation and can predict the treatment response and the disease outcome.

Genetic Alterations of Epidermal Growth Factor Receptor in Glioblastoma: The Usefulness of Immunohistochemistry.

Epidermal growth factor receptor (EGFR) amplification is one of the common alterations in IDH-wildtype glioblastoma. It is frequently associated with EGFRvIII mutation. To evaluate the correlation between EGFR overexpression, gene amplification, and EGFRvIII mutation, we performed immunohistochemical (IHC) analysis, fluorescence in situ hybridization by Vysis LSI EGFR/CEP7 dual color probe, and polymerase chain reaction studies in 76 patients diagnosed with glioblastomas (67 IDH-wildtype and 9 IDH-mutant). EGFR expression was scored ranging from 0 to 3+. Using formalin-fixed paraffin-embedded sections, real-time reverse transcription-polymerase chain reaction was carried out with primers specific for EGFRvIII and EGFR wildtype. In addition, we evaluated the impact of EGFR status on prognosis. EGFR gene amplifications and EGFRvIII mutations were identified in 30.3% and 15.5% of all cases, respectively. All the EGFR-amplified or EGFRvIII mutant cases were IDH-wildtype glioblastomas and tested positive with IHC. The sensitivity and specificity of EGFR IHC predicting EGFR gene amplification status were 100.0% and 46.5%, respectively. The EGFR-amplified cases tended to show more intense immunostaining (3+) in a considerable number of tumor cells (≥50%). Survival analyses of 37 IDH-wildtype glioblastoma patients revealed that none of the EGFR alterations significantly affected prognosis. EGFR IHC displayed high sensitivity and low specificity in predicting EGFR gene amplification, and interpretation of IHC results is a challenge. Therefore, EGFR IHC represents a possible screening tool for evaluation of EGFR gene amplification in clinical neuropathology, and both the intensity and proportion score facilitate interpretation of EGFR IHC.

Real‐world data on EGFR/ALK gene status and first‐line targeted therapy rate in newly diagnosed advanced non‐small cell lung cancer patients in Northern China: A prospective observational study

BackgroundTyrosine kinase inhibitors (TKIs) can significantly prolong overall survival for patients with advanced non‐small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR)‐mutation or anaplastic lymphoma kinase (ALK)‐rearrangement. However, the real‐world evaluation status of ALK/EGFR in China remains unclear.MethodsWe conducted a prospective study including 1134 patients with cytologically or histologically confirmed advanced NSCLC (stage IIIb–IV) at 12 Chinese hospitals.ResultsThe most common evaluation methods were amplification‐refractory mutation system for EGFR status and immunohistochemistry targeting D5F3 for ALK status. Among patients with non‐squamous, the EGFR mutation rate was 44.1% and the ALK rearrangement rate was 10.0%. Among patients with squamous cell carcinoma, the EGFR mutation rate was 8.3% and the ALK rearrangement rate was 3.7%. Among all patients, gender (HR = 1.7, 95%CI = 1.2–2.4, P = 0.006), smoking history (HR = 1.8, 95%CI = 1.3–2.7, P = 0.001), histology (HR = 5.0, 95%CI = 2.4–10.1, P < 0.001), and brain metastases (HR = 1.5, 95%CI = 1.1–2.2, P = 0.017) were independent predictors of EGFR mutation, while age (HR = 2.6, 95%CI = 1.7–4.1, P < 0.001) was an independent predictor of ALK rearrangement. The median time from tumor diagnosis to EGFR or ALK status confirmation was 7 and 5 days, respectively. Targeted therapy rate was 73.8% in EGFR‐positive patients and 51.4% in ALK‐positive patients. There was a negative correlation between the first‐line targeted therapy rate and the EGFR mutation detection period (r = −0.152, P = 0.02), while no significant correlation among patients with ALK rearrangement (r = −0.179, P = 0.076).ConclusionSquamous NSCLC patients should also be routinely tested to determine their EGFR/ALK statuses. The first‐line targeted therapy rate remains low in Chinese patients with NSCLC.

Paired-agent imaging for detection of head and neck cancers.

Head and neck cancers overwhelmingly overexpress epidermal growth factor receptor (EGFR). This overexpression has been utilized for head and neck cancers using molecular targeted agents for therapy and cancer cell detection. Significant progress has been made in using EGFR-targeted fluorescent antibody and Affibody molecule agents for fluorescent guided surgery in head and neck cancers. Although success in achieving tumor-to-background ratio of 3-5 have been achieved, the field is limited by the non-specific fluorescence in normal tissues as well as EGFR specific fluorescence in the oral cavity. We propose that paired-agent imaging (PAI) could improve the contrast between tumor and normal tissue by removing the fluorescent signal arising from non-specific binding. Here, ABY-029 - an anti-EGFR Affibody molecule labeled with IRDye 800CW - and IRDye 680RD conjugated to Affibody Control Imaging Agent molecule (IR680-Affctrl) are used as targeted and untargeted control agents, respectively, in a panel of head and neck squamous cell carcinomas (HNSCC) to test the ability of PAI to increase tumor detection. Initial results demonstrate that binding potential, a value proportional to receptor concentration, correlates well to EGFR expression but experimental limitations prevented pixel-by-pixel analysis that was desired. Although promising, a more rigorous and well-defined experimental protocol is required to align ex vivo EGFR immunohistochemistry with in vivo binding potential and fluorescence intensity. Additionally, a new set of paired-agents, ABY-029 and IRDye 700DX, are successfully tested in naïve mice and will be carried forward for clinical translation.

Epidermal growth factor receptor (EGFR) overexpression in triple-negative breast cancer: association with clinicopathologic features and prognostic parameters

IntroductionTriple-negative breast cancers are a poor prognostic group of breast cancers that don’t respond to conventional hormonal and her2neu targeted therapy. A subset of triple-negative breast cancer is known to overexpress epidermal growth factor receptor (EGFR); however prognostic significance of this biomarker has not been widely studied in our population. Therefore, we aimed to evaluate the frequency of EGFR overexpression in triple-negative breast cancer in our setup and its association with prognostic and predictive factors.MethodsWe performed EGFR immunohistochemistry on 150 cases of triple-negative breast cancers. Intensity and percentage of EGFR expression were combined to formulate an EGFR score, that was compared with prognostic features of breast cancer and recurrence status of patients.ResultsPositive EGFR expression was noted in 18.7% ( 28 cases); out of which 16% (24 cases) showed low EGFR expression, whereas high EGFR expression was seen in 2.7% ( 4 cases). No significant association of EGFR expression was noted when compared with various clinicopathological parameters and recurrence status of the patients.ConclusionWe found EGFR protein expression in 18.7% of cases while high expression was seen in only 2.7 % cases of triple-negative breast cancer which may harbor underlying genetic alterations like altered EGFR gene copy number, chromosome 7 copy number or average EGFR gene: chromosome 7 ratio; therefore we suggest that molecular tests like FISH to evaluate these EGFR molecular alterations should be performed in EGFR over expressing triple negative breast cancers in our setup to identify patients that can benefit from anti-EGFR targeted therapy. Moreover, regional difference in EGFR expression (high expression in chinese population compared to our population) are may be due to different underlying genetic alterations in triple-negative breast cancers, further necessitating a need of devising personalized therapeutic protocols for locoregional population.

Prognostic utility of epidermal growth factor receptor (EGFR) expression in prostatic acinar adenocarcinoma

BackgroundEpidermal growth factor receptor (EGFR) is potential prognostic biomarker expressed in many human cancers. Prognostic significance of EGFR immunohistochemical expression has not been established in prostatic acinar adenocarcinoma, therefore we aimed to evaluate the frequency of expression of EGFR in prostatic adenocarcinoma and its association with other prognostic parameters.MethodsThe study included 123 cases of biopsy proven prostatic acinar adenocarcinoma treated at Liaquat National hospital, Karachi from January 2013 till December 2017. Paraffin blocks of all cases were retrieved; sections were cut and stained with haematoxylin and eosin. Pathologic characteristics including tumor quantification, WHO grade group, gleason score, perineural and lymphovascular invasion were evaluated. EGFR immunohistochemistry (IHC) was performed on all tissue blocks.ResultsMean age of the patients included in the study was 69.05 ± 8.68 years. High gleason scores i.e. 8 & 9 were noted in 22% (27 cases) and 22.8% (28 cases) respectively. Similarly, 22.8% (28 cases) showed WHO grade group 5. 52.8% (65 cases) had > 50% tissue involvement by carcinoma and perineural invasion was seen in 37.4% (46 cases). Positive EGFR expression was noted in 18.7% (23 cases), while 81.3% (100 cases) showed negative EGFR expression. Significant association of EGFR expression was noted with gleason score (p-value = < 0.001), WHO grade (p = < 0.001), tumor quantification (p = 0.007) and perineural invasion (p = < 0.001). Moreover, significant association of EGFR expression was also seen with disease recurrence and Her2neu over expression. Patients with low gleason scores (score 6 and 7) and lower grade group (1, 2 & 3) were less likely to have positive EGFR expression as compared to patients with high gleason score (score 9) and higher grade group (5). Similarly, patients with perineural invasion were more likely to have positive EGFR expression.ConclusionWe found a relatively low EGFR expression in our patients with prostatic adenocarcinoma; however, its association with poor prognostic parameters like high gleason score, higher grade group, perineural invasion, higher tissue involvement by cancer and disease recurrence signifies its importance as a prognostic parameter in prostatic acinar adenocarcinoma.

A Comparative Study of Cell Block versus Biopsy for Detection of Epidermal Growth Factor Receptor Mutations and Anaplastic Lymphoma Kinase Rearrangement in Adenocarcinoma Lung.

Background:Lung cancer is a leading cause of deaths attributed to cancer worldwide. Epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) rearrangement are commonly found in patients of adenocarcinoma lung against, which targeted therapy is available. In this era of personalized medicine, it is a rationale to detect these molecular alterations in cases of lung carcinomas.Aims:The objectives were to compare the diagnostic efficacy of cytological samples for the detection of EGFR and ALK protein expression using immunocytochemistry in nonsmall cell lung carcinoma.Materials and Methods:We compared 22 cell blocks and biopsies for the detection of EGFR and ALK protein expression by immunohistochemistry (IHC). EGFR IHC was performed using EGFR Receptor (E746-A750 del Specific) (6B6) monoclonal antibody and ALK IHC was done using Ventana anti-ALK (D5F3) monoclonal primary antibody.Results:Two cases were found to be positive; 20 cases were negative for EGFR IHC both in biopsies and cell blocks. ALK IHC was positive in one case; negative in 21 cases. The results of IHC were also concordant for biopsies and cell blocks. The sensitivity and specificity were 100% for immunocytochemical detection of ALK and EGFR in cell blocks with respect to biopsies.Conclusion:We conclude that cell blocks can serve as a potential substitute for biopsies for detection of EGFR and ALK protein by immunocytochemistry, whenever patient presents with effusion and biopsy cannot be done or when tissue is not adequate.

A subset of esophageal squamous cell carcinoma patient-derived xenografts respond to cetuximab, which is predicted by high EGFR expression and amplification.

Epidermal growth factor receptor (EGFR) is reportedly overexpressed in most esophageal tumors, but most targeted therapies showed no efficacy in non-selected patients. This study aims at investigating the adaptive cetuximab subset in a cohort of esophageal squamous cell carcinoma (ESCC) patient-derived xenografts (PDXs). A large panel of ESCC PDXs has been established. The copy number, mRNA expression and immunohistochemistry (IHC) of key EGFR pathways have been examined along with cetuximab response. A preclinical trial on a randomly selected cohort of 16 ESCC PDXs was conducted, and the genomic annotations of these models were compared against the efficacy readout of the mouse trial. The trial identified that 7 of 16 (43.8%) responded to cetuximab (ΔT/ΔC <0 as responders). The gene amplification and expression analysis indicated that EGFR copy number ≥5 (P=0.035), high EGFR mRNA expression (P=0.001) and IHC score of 2-3 (P=0.034) are associated with tumor growth inhibition by cetuximab, suggesting EGFR may function as a single predictive biomarker for cetuximab response in ESCC. Overall, our results suggest that an ESCC subtype with EGFR amplification and overexpression benefits from cetuximab treatment, which warrants further clinical confirmation.

Expression of epidermal growth factor receptor (EGFR) in colorectal cancer: An immunohistochemical study

Background and study aimThe epidermal growth factor receptor (EGFR) plays an important role in tumourigenesis and tumour progression of colorectal cancer (CRC) and leads to the activation of intracellular signaling pathways. The use of anti-EGFR-targeted therapy has increased for patients with metastatic CRC. Today, the clinical utility of immunohistochemistry has remained somewhat inconclusive. It is based on EGFR screening methods using paraffin-embedded tumour specimen to select patients eligible for treatment. There is still lack of agreement on reproducible scoring criteria for EGFR immunohistochemistry has in various clinical trials. Patients and methodsWe retrospectively reviewed 36 CRC patients who underwent surgeries during 2011 in Habib Thameur hospital in Tunis. We analyzed the immunohistochemical overexpression of EGFR using a score based on immunostaining intensity. In addition, we analyzed the correlation between this overexpression and patients' clinicopathologic parameters. ResultsThe positive expression rate of EGFR was 78% (28/36). Using the immunoreactivity score, 21 cases were considered low grade expression and 15 tumours were high grade. Immunohistochemical expression of EGFR showed a significant difference with tumour’s location (p = 0.034) and vascular invasion (p = 0.03). This expression was not significantly associated with age, gender, tumour size, histological type, grade, TNM staging and perineural invasion. ConclusionsEGFR expression by immunohistochemistry in CRC is variably correlated with clinicopathological parameters. Its assessment by this method has still not proved its predictive value.