A New Optimized Version of a Colorectal Cancer-Targeted Immunotoxin Based on a Non-Immunogenic Variant of the Ribotoxin α-Sarcin

Narbona, Javier; Gordo, Rubén G.; Tomé-Amat, Jaime; Lacadena, Javier

doi:10.3390/cancers15041114

“…In this sense, the dual targeting of two different TAAs could be an effective tool for overcoming primary tumor heterogeneity, and subsequent tumor escape due to antigen loss, which makes tumors refractory to conventional treatments. In addition, the inclusion of αSDI allows a safer and more effective therapeutic tool since it has proven to be effective both in vitro and in vivo [76] and may prevent the formation of potentially neutralizing anti-drug antibodies (ADAs) by the patient s immune system.…”

Section: Discussionmentioning

confidence: 99%

Nanobody-Based EGFR-Targeting Immunotoxins for Colorectal Cancer Treatment

Narbona

¹

,

Hernández-Baraza

²

,

Gordo

³

et al. 2023

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Immunotoxins (ITXs) are chimeric molecules that combine the specificity of a targeting domain, usually derived from an antibody, and the cytotoxic potency of a toxin, leading to the selective death of tumor cells. However, several issues must be addressed and optimized in order to use ITXs as therapeutic tools, such as the selection of a suitable tumor-associated antigen (TAA), high tumor penetration and retention, low kidney elimination, or low immunogenicity of foreign proteins. To this end, we produced and characterized several ITX designs, using a nanobody against EGFR (VHH 7D12) as the targeting domain. First, we generated a nanoITX, combining VHH 7D12 and the fungal ribotoxin α-sarcin (αS) as the toxic moiety (VHHEGFRαS). Then, we incorporated a trimerization domain (TIEXVIII) into the construct, obtaining a trimeric nanoITX (TriVHHEGFRαS). Finally, we designed and characterized a bispecific ITX, combining the VHH 7D12 and the scFv against GPA33 as targeting domains, and a deimmunized (DI) variant of α-sarcin (BsITXαSDI). The results confirm the therapeutic potential of α-sarcin-based nanoITXs. The incorporation of nanobodies as target domains improves their therapeutic use due to their lower molecular size and binding features. The enhanced avidity and toxic load in the trimeric nanoITX and the combination of two different target domains in the bispecific nanoITX allow for increased antitumor effectiveness.

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“…However, these approaches still face limitations in achieving efficient cytosolic delivery at the single-cell level. A more recent strategy involves manipulating the dynamics of membrane proteins between the inside and outside of cell membranes 9,10 or harnessing the channeling properties of channel membrane proteins. 11 To leverage these uptake routes, binding to target membrane proteins is essential.…”

Section: Introductionmentioning

confidence: 99%

Target-selective cytosolic delivery of cargo proteins using the VHH-presented OLE-ZIP capsules

Takahashi,

Inoue,

Hida

et al. 2024

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In the pursuit of a new generation of protein pharmaceuticals, the efficient delivery of these therapeutics into cells stands out as a crucial challenge. In this study, we have developed a novel approach utilizing protein capsules modified with VHH antibodies as cytosolic carriers for protein pharmaceuticals. For the protein capsule component, we opted for the OLE-ZIP protein capsules, which can be prepared from the amphiphilic two-helix bundled protein OLE-ZIP using the water-in-oil (w/o) emulsion method. The spacious interior of the OLE-ZIP capsules allows for the stable encapsulation of over 200 molecules of protein pharmaceuticals, such as RNase A and Cre recombinase, in one capsule. By presenting the VHH antibody with an affinity for cell-type-specific receptors such as the epidermal growth factor receptor (EGFR) on the capsule surface, we achieved cell-type selective endocytic uptake in A431 cell lines (high expression level of EGFR) over NHDF and MCF-7 cells (normal expression level of EGFR). This selective uptake was followed by the subsequent release of the encapsulated protein pharmaceuticals into the cytosol of the target cells. Unlike our previous version of the OLE-ZIP protein capsules modified with IgG antibodies, cytosolic delivery of pharmaceutical proteins was little impacted by the presence of other IgGs, which are abundant in the bloodstream. This improved characteristic suggests potential advantages for practical applications, including intravenous administration.

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“…However, these approaches still face limitations in achieving efficient cytosolic delivery at the single-cell level. A more recent strategy involves manipulating the dynamics of membrane proteins between the inside and outside of cell membranes 9,10 or harnessing the channelling properties of channel membrane proteins. 11 To leverage these uptake routes, binding to target membrane proteins is essential.…”

Section: Introductionmentioning

confidence: 99%

Target-selective cytosolic delivery of cargo proteins using the VHH-presented OLE-ZIP capsules

Takahashi,

Inoue,

Hida

et al. 2024

Preprint

0

View full text Add to dashboard Cite

In the pursuit of a new generation of protein pharmaceuticals, the efficient delivery of these therapeutics into cells stands out as a crucial challenge. In this study, we have developed a novel approach utilizing protein capsules modified with VHH antibodies as cytosolic carriers for protein pharmaceuticals. For the protein capsule component, we opted for the OLE-ZIP protein capsules, which can be prepared from the amphiphilic two-helix bundled protein OLE-ZIP using the water-in-oil (w/o) emulsion method. The spacious interior of the OLE-ZIP capsules allows for the stable encapsulation of over 200 molecules of protein pharmaceuticals, such as RNase A and Cre recombinase, in one capsule. By presenting the VHH antibody with an affinity for cell-type-specific receptors such as the epidermal growth factor receptor (EGFR) on the capsule surface, we achieved cell-type selective endocytic uptake in A431 cell lines (high expression level of EGFR) over NHDF and MCF-7 cells (normal expression level of EGFR). This selective uptake was followed by the subsequent release of the encapsulated protein pharmaceuticals into the cytosol of the target cells. Unlike our previous version of the OLE-ZIP protein capsules modified with IgG antibodies, cytosolic delivery of pharmaceutical proteins was little impacted by the presence of other IgGs, which are abundant in the bloodstream. This improved characteristic suggests potential advantages for practical applications, including intravenous administration.

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A New Optimized Version of a Colorectal Cancer-Targeted Immunotoxin Based on a Non-Immunogenic Variant of the Ribotoxin α-Sarcin

Cited by 4 publications

References 54 publications

Nanobody-Based EGFR-Targeting Immunotoxins for Colorectal Cancer Treatment

Nanobody-Based EGFR-Targeting Immunotoxins for Colorectal Cancer Treatment

Target-selective cytosolic delivery of cargo proteins using the VHH-presented OLE-ZIP capsules

Target-selective cytosolic delivery of cargo proteins using the VHH-presented OLE-ZIP capsules

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