A recombinant scFv antibody-based fusion protein that targets EGFR associated with IMPDH2 downregulation and its drug conjugate show therapeutic efficacy against esophageal cancer

Abstract

The present study aimed to evaluate the anti-tumor efficacy of the epidermal growth factor receptor (EGFR)-targeting recombinant fusion protein Fv-LDP-D3 and its antibody-drug conjugate Fv-LDP-D3-AE against esophageal cancer. Fv-LDP-D3, consisting of the fragment variable (Fv) of an anti-EGFR antibody, the apoprotein of lidamycin (LDP), and the third domain of human serum albumin (D3), exhibited a high binding affinity for EGFR-overexpressing esophageal cancer cells, inhibited EGFR phosphorylation and down-regulated inosine monophosphate dehydrogenase type II (IMPDH2) expression. Fv-LDP-D3 was taken up by cancer cells through intensive macropinocytosis; it inhibited the proliferation and induced the apoptosis of esophageal cancer cells. In vivo imaging revealed that Fv-LDP-D3 displayed specific tumor-site accumulation and a long-lasting retention over a 26-day period. Furthermore, Fv-LDP-D3-AE, a pertinent antibody-drug conjugate prepared by integrating the enediyne chromophore of lidamycin into the Fv-LDP-D3 molecule, displayed highly potent cytotoxicity, inhibited migration and invasion, induced apoptosis and DNA damage, arrested cells at G2/M phase, and caused mitochondrial damage in esophageal cancer cells. More importantly, both of Fv-LDP-D3 and Fv-LDP-D3-AE markedly inhibited the growth of esophageal cancer xenografts in athymic mice at well tolerated doses. The present results indicate that Fv-LDP-D3, and Fv-LDP-D3-AE exert prominent antitumor efficacy associated with targeting EGFR, suggesting their potential as promising candidates for targeted therapy against esophageal cancer.

Targeted degradation of oncogenic BCR-ABL by silencing the gene of NEDD8 E3 ligase RAPSYN

“…The experiments were performed according to a previous report [ 18 ] with minor modifications. Different concentrations of anti-CD79B-scFv ranging from 0.0001 to 1 mg/mL were added to each well in 96-well plates and incubated at 37 °C for 2 h. Horseradish peroxidase labeled anti-His monoclonal antibody (Proteintech, Wuhan, China) was used as the secondary antibody.…”

Drug Delivery Opportunities in Esophageal Cancer: Current Treatments and Future Prospects

“…253 Results from two other recent phase I trials with tisotumab vedotin 254 and sacituzumab govitecan 255 show serious adverse side effects during dosing, which has dampened enthusiasm for these particular agents. However, GPC1-ADC(MMAE), 256 enfortumab vedotin, 257 and Fv-LDP-D3-AE 258 all show in vitro and in vivo efficacy and merit their own clinical investigations, showing promise for use of ADCs against esophageal cancer.…”

Mechanisms of Action and Limitations of Monoclonal Antibodies and Single Chain Fragment Variable (scFv) in the Treatment of Cancer

“…Given the success of Moxetumomab, other scFvs coupled to toxins are currently under development, including an immunotoxin linked to an scFv with high binding affinity for glioblastoma multiforme (GBM) cells that express EGFRwt and EGFRvIII [ 133 ]. In addition to toxins, other antitumoral drugs have been used, such as lidamycin, composed of enediyne chromophore with extremely potent cytotoxicity, which is currently in phase II clinical trial [ 140 ].…”