A novel nanobody as therapeutics target for EGFR-positive colorectal cancer therapy: exploring the effects of the nanobody on SW480 cells using proteomics approach

Abstract

Background The epidermal growth factor receptor (EGFR) overexpression is found in metastatic colorectal cancer (mCRC). Targeted molecular therapies such as monoclonal antibodies (mAbs) and tyrosine kinase inhibitors (TKI) are becoming more precise, targeting specifically for cancer therapeutics. However, there are adverse effects of currently available anti-EGFR drugs, including drug-resistant and side effects. Nanobodies can overcome these limitations. Our previous study has found that cell-penetrable nanobodies targeted at EGFR-tyrosine kinase were significantly reduced EGFR-positive lung cancer cells viability and proliferation. The aim of the present study was to determine the effect of cell-penetrable nanobody (R9VH36) on cell viability and proteomic profile in EGFR-positive human colorectal cancer cell lines. Methods The human colorectal carcinoma cell line (SW480) was treated with R9VH36, compared with gefitinib. Cell viability was monitored using the MTT cell viability assay. The proteomic profiling was analyzed by LC–MS/MS . Results The half-maximal inhibitory concentration (IC50) values determined for R9VH36 and gefitinib against SW480 were 527 ± 0.03 nM and 13.31 ± 0.02 μM, respectively. Moreover, both the gefitinib-treated group and nanobody-treated group had completely different proteome profiles. A total 6626 differentially expressed proteins were identified. PCA analysis revealed different proteome profiling in R9VH36 experiment. There were 8 proteins in R9VH36 that significantly exhibited opposite expression directions when compared to gefitinib. These proteins are involved in DNA-damage checkpoint processes. Conclusion The proteomics explored those 6,626 proteins had different expressions between R9VH36 and gefitinib. There were 8 proteins in R9VH36 exhibited opposite expression direction when comparing to gefitinib. Our findings suggest that R9VH36 has the potential to be an alternative remedy for treating EGFR-positive colon cancer.

Molecular genotyping, histopathological and immunohistochemical studies of bovine papillomatosis

“…Suppression of binding sites of EGFR signaling at the level of extracellular receptor or tyrosine kinase intracellular activity may inhibit EGFR expression networks and thus improve patient outcomes ( Kitamura et al ., 2020 ; Wingert et al ., 2021 ). Several authors have suggested that the increased expression of EGFR has negative impacts ( Kwon et al ., 2018 ; Hashmi et al ., 2019 ; London and Gallo, 2020 ), while others maintain the opposite ( Pidugu et al ., 2019 ; Feng et al ., 2020 ; Lamtha et al ., 2022 ). However, meta-analysis data summarizing the results of various studies show that exposure to EGFR is not an independent predictor of survival in various diseases ( Yang et al ., 2019 ; Nastały et al ., 2020 ; Atef et al ., 2021 ).…”

New pyrazolyl-thiazolidinone/thiazole derivatives as celecoxib/dasatinib analogues with selective COX-2, HER-2 and EGFR inhibitory effects: design, synthesis, anti-inflammatory/anti-proliferative activities, apoptosis, molecular modelling and ADME studies

“…Since cancer is a compacted and multiple genes involved disease, drugs that act at multiple targets can improve efficacy and lower drug resistance and are considered as the future of worthy anticancer drug development 5 . One of the most efficient and known manner in cancer therapy, is targeting protein kinases due to their vital role in regulation of various cellular signal transductions and activities, such as, proliferation, differentiation, apoptosis, metabolism and transcription 6–8 .…”