HER2 monoclonal antibodies that do not interfere with receptor heterodimerization-mediated signaling induce effective internalization and represent valuable components for rational antibody-drug conjugate design

Goeij, Bart E.C.G. de; Peipp, Matthias; Haij, Simone de; Brink, Edward N. van den; Kellner, Christian; Riedl, Thilo; Jong, Rob N. de; Vink, Tom; Strumane, Kristin; Bleeker, Wim K.; Parren, Paul W.H.I.

doi:10.4161/mabs.27705

Cited by 28 publications

(33 citation statements)

References 47 publications

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“…IgG1-HER2 induced $40% downmodulation of total HER2 in HCC1954 cells that express high levels of HER2 (Fig. 3A), which was in line with previously published results (16). Despite the fact that the monovalent bsHER2xb12 showed dose-dependent binding to HER2-positive cells (Supplementary Fig.…”

Section: Generation Of a Low-affinity Cd63 Antibodysupporting

confidence: 91%

“…Cloning and production of HER2 antibody IgG1-153 has been described elsewhere (16 (20). Bispecific antibodies (Duobody) were made by controlled Fabarm exchange as described by Labrijn and colleagues (21).…”

Section: Cell Linesmentioning

confidence: 99%

“…HER2 antibody 153 (IgG1-HER2), which is known to internalize as bivalent IgG (16), was used to generate a bispecific IgG1 (bsHER2xCD63 his ) that can bind to both, HER2 and CD63. To validate the use of a monovalent HER2-specific IgG (bsHER2xb12) as a model for an IgG with poor internalization characteristics, we first investigated Ab-induced downmodulation of the monovalent bsHER2xb12.…”

Section: Generation Of a Low-affinity Cd63 Antibodymentioning

confidence: 99%

“…However, for internalization to be most effective HER2 requires cross-linking of HER2 molecules. Especially on tumor cells that highly overexpress HER2, Abinduced cross-linking has been described to improve internalization of HER2 (5,(16)(17)(18). The notion that monomeric HER2 does not internalize well suggested to us that a bsAb with monovalent HER2 binding characteristics may represent a suitable model system for testing whether internalization of cell surface molecules can be increased through binding to CD63.…”

Section: Introductionmentioning

confidence: 99%

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Efficient Payload Delivery by a Bispecific Antibody–Drug Conjugate Targeting HER2 and CD63

Goeij

Vink

Napel

et al. 2016

Molecular Cancer Therapeutics

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Antibody-drug conjugates (ADC) are designed to be stable in circulation and to release potent cytotoxic drugs intracellularly following antigen-specific binding, uptake, and degradation in tumor cells. Efficient internalization and routing to lysosomes where proteolysis can take place is therefore essential. For many cell surface proteins and carbohydrate structures on tumor cells, however, the magnitude of these processes is insufficient to allow for an effective ADC approach. We hypothesized that we could overcome this limitation by enhancing lysosomal ADC delivery via a bispecific antibody (bsAb) approach, in which one binding domain would provide tumor specificity, whereas the other binding domain would facilitate targeting to the lysosomal compartment. We therefore designed a bsAb in which one binding arm specifically targeted CD63, a protein that is described to shuttle between the plasma membrane and intracellular compartments, and combined it in a bsAb with a HER2 binding arm, which was selected as model antigen for tumor-specific binding. The resulting bsHER2xCD63 his demonstrated strong binding, internalization and lysosomal accumulation in HER2-positive tumor cells, and minimal internalization into HER2-negative cells. By conjugating bsHER2xCD63 his to the microtubule-disrupting agent duostatin-3, we were able to demonstrate potent cytotoxicity of bsHER2xCD63 his -ADC against HER2-positive tumors, which was not observed with monovalent HER2-and CD63-specific ADCs. Our data demonstrate, for the first time, that intracellular trafficking of ADCs can be improved using a bsAb approach that targets the lysosomal membrane protein CD63 and provide a rationale for the development of novel bsADCs that combine tumor-specific targeting with targeting of rapidly internalizing antigens. Mol Cancer Ther; 15(11); 2688-97. Ó2016 AACR.

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Section: Generation Of a Low-affinity Cd63 Antibodysupporting

confidence: 91%

Section: Cell Linesmentioning

confidence: 99%

Section: Generation Of a Low-affinity Cd63 Antibodymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Efficient Payload Delivery by a Bispecific Antibody–Drug Conjugate Targeting HER2 and CD63

Goeij

Vink

Napel

et al. 2016

Molecular Cancer Therapeutics

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“…TF-111 binds TF and partially interferes with FVIIa binding and ERK-phosphorylation. The HER2 mAbs 153 and 005 were described by (22). Both antibodies bound to epitopes distinct from those recognized by trastuzumab and pertuzumab.…”

Section: Antibody Generation and Conjugationmentioning

confidence: 99%

High Turnover of Tissue Factor Enables Efficient Intracellular Delivery of Antibody–Drug Conjugates

Goeij

Satijn

Freitag

et al. 2015

Molecular Cancer Therapeutics

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Antibody-drug conjugates (ADC) are emerging as powerful cancer treatments that combine antibody-mediated tumor targeting with the potent cytotoxic activity of toxins. We recently reported the development of a novel ADC that delivers the cytotoxic payload monomethyl auristatin E (MMAE) to tumor cells expressing tissue factor (TF). By carefully selecting a TFspecific antibody that interferes with TF:FVIIa-dependent intracellular signaling, but not with the procoagulant activity of TF, an ADC was developed (TF-011-MMAE/HuMax-TF-ADC) that efficiently kills tumor cells, with an acceptable toxicology profile. To gain more insight in the efficacy of TF-directed ADC treatment, we compared the internalization characteristics and intracellular routing of TF with the EGFR and HER2. Both in absence and presence of antibody, TF demonstrated more efficient internalization, lysosomal targeting, and degradation than EGFR and HER2. By conjugating TF, EGFR, and HER2-specific antibodies with duostatin-3, a toxin that induces potent cytotoxicity upon antibody-mediated internalization but lacks the ability to induce bystander killing, we were able to compare cytotoxicity of ADCs with different tumor specificities. TF-ADC demonstrated effective killing against tumor cell lines with variable levels of target expression. In xenograft models, TF-ADC was relatively potent in reducing tumor growth compared with EGFR-and HER2-ADCs. We hypothesize that the constant turnover of TF on tumor cells makes this protein specifically suitable for an ADC approach.

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In vitro incorporation of a cell‐binding protein to a lentiviral vector using an engineered split intein enables targeted delivery of genetic cargo

Chamoun‐Emanuelli

Wright

Roger

et al. 2015

Biotech & Bioengineering

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Gene therapy represents a promising therapeutic paradigm for addressing many disorders, but the absence of a vector that can be robustly and reproducibly functionalized with cell-homing functionality to mediate the delivery of genetic cargo specifically to target cells following systemic administration has stood as a major impediment. In this study, a high-affinity protein-protein pair comprising a splicing-deficient naturally split intein was used as molecular Velcro to append a HER2/neu-binding protein (DARPin) onto the surface of a binding-deficient, fusion-competent lentivirus. HER2/neu-specific lentiviruses created using this in vitro pseudotyping approach were able to deliver their genetic reporter cargo specifically to cells that express the target receptor at high levels in a co-culture. We envision that the described technology could provide a powerful, broadly applicable platform for the incorporation of cell-targeting functionality onto viral vectors.

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HER2 monoclonal antibodies that do not interfere with receptor heterodimerization-mediated signaling induce effective internalization and represent valuable components for rational antibody-drug conjugate design

Cited by 28 publications

References 47 publications

Efficient Payload Delivery by a Bispecific Antibody–Drug Conjugate Targeting HER2 and CD63

Efficient Payload Delivery by a Bispecific Antibody–Drug Conjugate Targeting HER2 and CD63

High Turnover of Tissue Factor Enables Efficient Intracellular Delivery of Antibody–Drug Conjugates

In vitro incorporation of a cell‐binding protein to a lentiviral vector using an engineered split intein enables targeted delivery of genetic cargo

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