Blocking EGFR Activation with Anti‐EGF Nanobodies via Two Distinct Molecular Recognition Mechanisms

Guardiola, Salvador; Varese, Monica; Sánchez‐Navarro, Macarena; Vincke, Cécile; Teixidó, Meritxell; García, Jesús; Giralt, Ernest

doi:10.1002/ange.201807736

Cited by 9 publications

(10 citation statements)

References 24 publications

(22 reference statements)

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“…After treatment with Nb@IC-NPs, the HIF-1α fluorescence signal was reduced significantly, demonstrating Nb@IC-NPs could effectively overcome hypoxic characteristic of cell-totumor, and this is the key to improving tumor hypoxia microenvironment and promoting PDT. Targeting of anti-EGFR Nb in Nb@IC-NPs to lung cancer promote the accumulation of IR1048MZ [34,35], resulting in precise and sufficient tumor oxygen production to retard tumor hypoxia and enhance PDT of lung cancer.…”

Section: Discussionmentioning

confidence: 99%

“…The superior stability, high affinity, high target specificity, and fast clearance of Nbs offer special advantages compared to conventional antibodies in application of noninvasive diagnostic imaging and tumor therapy [31][32][33]. Nb is able to mediate the endocytosis of specific receptors, such as EGFR, in tumor cells, promoting Nb as tumor-targeting ligands for drug nanocarrier designed in targeted cancer therapy and diagnosis [34][35][36]. In addition, anti-EGFR Nb also shows tumor suppressing properties, which further enhance the efficacy of drug delivery [37,38].…”

Section: Introductionmentioning

confidence: 99%

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Targeted nanobody complex enhanced photodynamic therapy for lung cancer by overcoming tumor microenvironment

Zhang

Liu

et al. 2020

Cancer Cell Int

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Background To investigate the efficacy of a PLGA-based nanobody complex in photodynamic therapy (PDT) and NIR-II imaging in A549 tumor hypoxic model. Method IR1048-MZ was firstly synthesized by conjugating a nitro imidazole group to IR1048. IR1048-MZ and Cat were then encapsulated in PLGA-SH solution. Anti-EGFR-Nanobody was also expressed and purified, and finally Anti-EGFR-Nanobody@PLGA-IR1048MZ-Cat (Nb@IC-NPs) nanobody complex was obtained based on the formation of desulfide bond between PLGA-SH and Anti-EGFR-Nanobody. Size distribution and morphology were characterized by TEM and DLS. Spectrum of Nb@IC-NPs towards NTR was measured by UV and fluorescence, while the particle’s selective response was studied using fluorescence. The uptake of Nb@IC-NPs in A549 cells was observed by flow cytometry and CLSM. In the meantime, its’ catalytic ability that decomposes H2O2 both extra-and intra-cellular was observed by fluorescence and CLSM. In vitro photodynamic toxicity of Nb@IC-NPs was examined by MTT, Live/Dead Cell Staining, Flow Cytometry and Apoptosis Assay. Tumor-bearing model was constructed to observe a semi-quantitative fluorescent distribution and the possibility of NIR-II fluorescence/photoacoustic (PA) imaging. Effect of Nb@IC-NPs on enhancing A549 tumor hypoxia and expression profile of HIF-1α was investigated in the presence of NIR. An A549 tumor metastasis model was also constructed to confirm the complex’ potential to destroy primary tumor, inhibit lung metastasis, and prolong mice’ survival. Lastly, impact of Nb@IC-NPs on mice’ main organs and blood indices was observed. Results Nb@IC-NPs was successfully fabricated with good homogeneity. The fluorescent absorbance of Nb@IC-NPs showed a linear relationship with the concentration of NTR, and a higher concentration of NTR corresponded to a stronger photoacoustic signal. In addition, Nb@IC-NPs showed a stable selectivity toward NTR. Our results also suggested a high efficient uptake of Nb@IC-NPs in A549 cells, which was more efficient than IC-NPs and IR1048-MZ alone. In vitro assays confirmed the effects of Nb@IC-NPs on catalytic O2 generation even in hypoxic cells. The cell viability was upregulated with the nanocomplex at the absence of the laser, whereas it was dramatically declined with laser treatment that excited at 980 nm. Nb@IC-NPs achieved tumor hypoxia NIR-II/PA imaging through assisting A549 gathering. When NIR was applied, Nb@IC-NPs can significantly relieve A549 cellular/tumor hypoxia by generating more reactive oxygen species (ROS), which in turn helps lower the expression level of HIF-1α. In summary, Nb@IC-NPs based PDT can efficiently decimate A549 primary tumor, inhibit metastatic lung cancer, and prolong the lifespan of the mice under tolerable dosage. At last, in vivo toxicity tests of the nanocomplex showed its biosafety to the main organs and normal blood indices values. Conclusion Nb@IC-NPs improves tumor hypoxia through catalytic reaction and lowers the expression level of HIF-1α. It achieves tumor PA imaging through intensified NIR-II fluorescence signal that caused by response of the complex to the lesion’s nitroreductase (NTR). Nb@IC-NPs based PDT can efficiently kill A549 primary tumor, inhibit a lung metastasis, as well as prolong mice’ survival cycle.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Targeted nanobody complex enhanced photodynamic therapy for lung cancer by overcoming tumor microenvironment

Zhang

Liu

et al. 2020

Cancer Cell Int

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“…We recently reported the discovery of the rst Nbs that directly target EGF and block EGFR phosphorylation and pathway activation through a new mechanism of action [17]. The most potent inhibitors (Nb1 and Nb6) showed remarkably different target recognition mechanisms, which were characterized, at the biophysical level, by distinct kinetics and thermodynamics ngerprints [18].…”

Section: Introductionmentioning

confidence: 99%

Anti-EGF nanobodies enhance the antitumoral effect of osimertinib and overcome resistance in non-small cell lung cancer (NSCLC) cellular models

et al. 2022

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Osimertinib is a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) that is effective against the EGFR T790M mutation in patients with advanced non-small-cell lung cancer (NSCLC). However, acquired resistance appears invariably due to several mechanisms. The strategy of using EGF-targeted nanobodies (Nbs) to block the initial step of the EGFR pathway constitutes a new research area. Nbs offer several advantages compared to traditional mAbs, such as their reduced size, increased stability and tissue penetration, which provide key advantages for targeting soluble tumoral growth factors. In this study we investigated the e cacy of anti-EGF Nbs to reduce Osimertinib resistance. Two anti-EGF Nbs, generated in our laboratory, were shown to inhibit cell viability and colony formation in PC9 and PC9-derived osimertinib-resistant cell lines. The combination of these Nbs with osimertinib improved the antitumoral e cacy of this EGFR-TKI in cell viability and colony formation experiments. In a mechanistic study of the EGFR pathway, the combination treatment dampened the activation of downstream proteins such as Akt and Erk1/2 MAP kinases. In addition, it increased cellular apoptosis and decreased the expression of Hes1, a cancer stem cell marker involved in metastasis and osimertinib resistance. We conclude that the addition of anti-EGF nanobodies enhances the antitumoral properties of osimertinib, thus representing a potentially effective strategy for NSCLC patients.

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“…In the years since their discovery, nanobodies have been engineered to serve a variety of roles, including fluorescent intracellular probes, 8–14 molecular tools for protein knockout experiments, 15 in vivo modulators of plant enzymes, 16 aids in protein crystallization, 17 diagnostic tools, 18,19 and potential human therapeutics. 20–23…”

Section: Introductionmentioning

confidence: 99%

Selection of Functional Intracellular Nanobodies

Woods

2019

SLAS Discovery

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Blocking EGFR Activation with Anti‐EGF Nanobodies via Two Distinct Molecular Recognition Mechanisms

Cited by 9 publications

References 24 publications

Targeted nanobody complex enhanced photodynamic therapy for lung cancer by overcoming tumor microenvironment

Targeted nanobody complex enhanced photodynamic therapy for lung cancer by overcoming tumor microenvironment

Anti-EGF nanobodies enhance the antitumoral effect of osimertinib and overcome resistance in non-small cell lung cancer (NSCLC) cellular models

Selection of Functional Intracellular Nanobodies

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