Engineered Multivalency Enhances Affibody-Based HER3 Inhibition and Downregulation in Cancer Cells

Schardt, John S.; Oubaid, Jinan M.; Williams, Sonya C.; Howard, James L.; Aloimonos, Chloe; Bookstaver, Michelle L.; Lamichhane, Tek N.; Sokic, Sonja; Liyasova, Mariya; O'Neill, Michael C.; Andrésson, Þorkell; Hussain, Arif; Lipkowitz, Stanley; Jay, Steven M.

doi:10.1021/acs.molpharmaceut.6b00919

Cited by 23 publications

(20 citation statements)

References 44 publications

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“…This is particularly interesting, because our findings contradict data published by Schardt et al [27,28], who investigated the therapeutic efficacy of multivalent anti-HER3 affibody constructs using the HER3-binding variant Z 05413 . The authors hypothesized that dimerization would improve affinity through avidity and potentially increase the inhibition of HER3 phosphorylation, downstream signaling, and tumor progression and therefore improve the therapeutic efficacy [27,28]. Indeed, the authors demonstrated a significant reduction in tumor progression compared to controls for groups treated with bivalent affibody constructs but not with monovalent constructs [27].…”

Section: Discussioncontrasting

confidence: 74%

“…While several HER3-targeting therapeutic agents are currently under preclinical and clinical development, no HER3-specific drugs or treatments have yet been approved [12]. Efforts by our group and others have demonstrated in preclinical models that anti-HER3 affibody molecules could be an alternative for HER3-targeted therapy [21,27,28]. For example, the affibody construct TAM-HER3 (an ABD flanked by two HER3-binding affibody molecules) delayed the growth of HER3 expressing tumors in a mouse model with equal potency as the HER3-targeting antibody MM-121 [21], indicating the main mode of action is based on receptor blocking and is not Fc-mediated.…”

Section: Discussionmentioning

confidence: 99%

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Evaluating the Therapeutic Efficacy of Mono- and Bivalent Affibody-Based Fusion Proteins Targeting HER3 in a Pancreatic Cancer Xenograft Model

et al. 2020

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Human epidermal growth factor receptor 3 (HER3) has been increasingly scrutinized as a potential drug target since the elucidation of its role in mediating tumor growth and acquired therapy resistance. Affibody molecules are so-called scaffold proteins with favorable biophysical properties, such as a small size for improved tissue penetration and extravasation, thermal and chemical stability, and a high tolerance to modifications. Additionally, affibody molecules are efficiently produced in prokaryotic hosts or by chemical peptide synthesis. We have previously evaluated the biodistribution profiles of five mono- and bivalent anti-HER3 affibody molecules (designated as 3) fused to an albumin-binding domain (designated as A), 3A, 33A, 3A3, A33, and A3, that inhibit ligand-dependent phosphorylation. In the present study, we examined the therapeutic efficacy of the three most promising variants, 3A, 33A, and 3A3, in a direct comparison with the HER3-targeting monoclonal antibody seribantumab (MM-121) in a preclinical BxPC-3 pancreatic cancer model. Xenografted mice were treated with either an affibody construct or MM-121 and the tumor growth was compared to a vehicle group. Receptor occupancy was estimated by positron emission tomography/computed tomography (PET/CT) imaging using a HER3-targeting affibody imaging agent [68Ga]Ga-(HE)3-Z08698-NODAGA. The affibody molecules could inhibit ligand-dependent phosphorylation and cell proliferation in vitro and demonstrated tumor growth inhibition in vivo comparable to that of MM-121. PET/CT imaging showed full receptor occupancy for all tested drug candidates. Treatment with 3A and 3A3 affibody constructs was more efficient than with 33A and similar to the anti-HER3 antibody seribantumab, showing that the molecular design of affibody-based therapeutics targeting HER3 in terms of the relative position of functional domains and valency has an impact on therapeutic effect.

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Section: Discussioncontrasting

confidence: 74%

Section: Discussionmentioning

confidence: 99%

Evaluating the Therapeutic Efficacy of Mono- and Bivalent Affibody-Based Fusion Proteins Targeting HER3 in a Pancreatic Cancer Xenograft Model

et al. 2020

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“…Because of their small size, affibody can be synthesized or recombinantly expressed. Since the first construct HER2-targeting affibody molecule Z HER2:342 was produced and confirmed to bind to HER2-positive cancer cell lines [24], several similar affibody molecules targeting EGFR [23,24], HER3 [27,28], IGF-1R [42], HIV-1-gp120 [43] and HPV16E7 [30,31] have been reported and also applied to image several tumour-associated molecular targets, such as HER2 [25,44], EGFR [45,46], HER3 [47]. The first clinical imaging study on patients with breast cancer was conducted using HER2-specific affibody, which Mice bearing C666-1tumor grafts were intravenously injected with 100 nmol/kg Z142X, Z142, or an equal amount of control agents or the same volume of PBS every two days for 15 times via tail vein.…”

Section: Discussionmentioning

confidence: 99%

“…To date, over 400 published studies show that affibody molecules have been selected for targeting more than 40 different proteins and served as affinity moieties in a variety of applications [22]. The affibody-targeted proteins include epidermal growth factor receptor (EGFR) [23,24], human epidermal growth factor receptor 2 (HER2) [25,26], human epidermal growth factor receptor 3 (HER3) [27,28], vascular endothelial growth factor (VEGF) [29], and human papillomavirus type 16 E7 (HPV16E7) [30,31]. In particularly, affibody molecules attached with a cytotoxin appear to be a straightforward and efficient way to direct the action of the toxin to desired cell targets [22].…”

Section: Introductionmentioning

confidence: 99%

Novel EBV LMP-2-affibody and affitoxin in molecular imaging and targeted therapy of nasopharyngeal carcinoma

et al. 2020

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Epstein-Barr virus (EBV) infection is closely linked to several human malignancies including endemic Burkitt's lymphoma, Hodgkin's lymphoma and nasopharyngeal carcinomas (NPC). Latent membrane protein 2 (LMP-2) of EBV plays a pivotal role in pathogenesis of EBVrelated tumors and thus, is a potential target for diagnosis and targeted therapy of EBV LMP-2 + malignant cancers. Affibody molecules are developing as imaging probes and tumor-targeted delivery of small molecules. In this study, four EBV LMP-2-binding affibodies (Z EBV LMP-2 12, Z EBV LMP-2 132, Z EBV LMP-2 137, and Z EBV LMP-2 142) were identified by screening a phage-displayed LMP-2 peptide library for molecular imaging and targeted therapy in EBV xenograft mice model. Z EBV LMP-2 affibody has high binding affinity for EBV LMP-2 and accumulates in mouse tumor derived from EBV LMP-2 + xenografts for 24 h after intravenous (IV) injection. Subsequent fusion of Pseudomonas exotoxin PE38KDEL to the Z EBV LMP-2 142 affibody led to production of Z142X affitoxin. This fused Z142X affitoxin exhibits high cytotoxicity specific for EBV + cells in vitro and significant antitumor effect in mice bearing EBV + tumor xenografts by IV injection. The data provide the proof of principle that EBV LMP-2-speicifc affibody molecules are useful for molecular imaging diagnosis and have potentials for targeted therapy of LMP-2-expressing EBV malignancies.

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“…It has been proposed that multivalent constructs could provide increased therapeutic potential compared to monomeric analogues. Indeed, in vitro the bi- and trivalent HER3-specific affibody molecules were significantly more efficient in blocking phosphorylation of HER3 and inhibiting cellular activity and proliferation compared to the monovalent control [ 20 , 26 ]. We demonstrated that a bivalent construct based on an anti-HER3 affibody molecule fused with ABD, 3A3, also efficiently inhibited growth of HER3-expressing cells in vitro, had prolonged in vivo half-life, delayed growth of HER3-expressing BxPC-3 xenografts in mice, and was equipotent to seribantumab (MM-121) [ 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

Influence of Molecular Design on the Targeting Properties of ABD-Fused Mono- and Bi-Valent Anti-HER3 Affibody Therapeutic Constructs

Altai

Leitao

Rinne

et al. 2018

Cells

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Overexpression of human epidermal growth factor receptor type 3 (HER3) is associated with tumour cell resistance to HER-targeted therapies. Monoclonal antibodies (mAbs) targeting HER3 are currently being investigated for treatment of various types of cancers. Cumulative evidence suggests that affibody molecules may be appropriate alternatives to mAbs. We previously reported a fusion construct (3A3) containing two HER3-targeting affibody molecules flanking an engineered albumin-binding domain (ABD035) included for the extension of half-life in circulation. The 3A3 fusion protein (19.7 kDa) was shown to delay tumour growth in mice bearing HER3-expressing xenografts and was equipotent to the mAb seribantumab. Here, we have designed and explored a series of novel formats of anti-HER3 affibody molecules fused to the ABD in different orientations. All constructs inhibited heregulin-induced phosphorylation in HER3-expressing BxPC-3 and DU-145 cell lines. Biodistribution studies demonstrated extended the half-life of all ABD-fused constructs, although at different levels. The capacity of our ABD-fused proteins to accumulate in HER3-expressing tumours was demonstrated in nude mice bearing BxPC-3 xenografts. Formats where the ABD was located on the C-terminus of affibody binding domains (3A, 33A, and 3A3) provided the best tumour targeting properties in vivo. Further development of these promising candidates for treatment of HER3-overexpressing tumours is therefore justified.

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Engineered Multivalency Enhances Affibody-Based HER3 Inhibition and Downregulation in Cancer Cells

Cited by 23 publications

References 44 publications

Evaluating the Therapeutic Efficacy of Mono- and Bivalent Affibody-Based Fusion Proteins Targeting HER3 in a Pancreatic Cancer Xenograft Model

Evaluating the Therapeutic Efficacy of Mono- and Bivalent Affibody-Based Fusion Proteins Targeting HER3 in a Pancreatic Cancer Xenograft Model

Novel EBV LMP-2-affibody and affitoxin in molecular imaging and targeted therapy of nasopharyngeal carcinoma

Influence of Molecular Design on the Targeting Properties of ABD-Fused Mono- and Bi-Valent Anti-HER3 Affibody Therapeutic Constructs

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