Molecular Imaging and Quantitation of EphA2 Expression in Xenograft Models with <sup>89</sup>Zr-DS-8895a

Burvenich, Ingrid; Parakh, Sagun; Gan, Hui; Lee, Fook-Thean; Guo, Nancy; Rigopoulos, Angela; Lee, Sze-Ting; Gong, Sylvia; O’Keefe, Graeme; Tochon-Danguy, Henri; Kotsuma, Masakatsu; Hasegawa, Jun; Senaldi, Giorgio; Scott, Andrew M.

doi:10.2967/jnumed.115.169839

Cited by 23 publications

(27 citation statements)

References 26 publications

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“…The biodistribution study demonstrated high uptake of 89 Zr-Df-Bz-NCS-DS-5573a in tumors, with levels rising by day 3 and maintained to day 7. The stable in vivo properties of 89 Zr-Df-Bz-NCS-DS-5573a were comparable to previously reported IgG1 humanized antibodies 32 , 44 , 45 . Uptake of 89 Zr-Df-Bz-NCS-DS-5573a was higher in the spleen of SCID mice versus BALB/c nu / nu mice.…”

Section: Discussionsupporting

confidence: 86%

“…As shown by Sharma et al, the interaction between Fc gamma receptors (FγcRs) expressed by immune effector cells of myeloid origin (such as monocytes/macrophages prevalent in the spleen and a subset of dendritic cells and neutrophils in the bone marrow) and the Fc portion of antibodies results in anomalous biodistribution of radioimmunoconjugates in non-target organs such as the spleen of mice that have a highly immunodeficient background. Furthermore, the glycosylation status of the DS-5573a, an afucosylated human IgG1 antibody with increased binding affinity for FcγRIIIa, could have contributed to the non-target uptake of 89 Zr-DS-5573a in organs such as bone and spleen when compared to previously reported 89 Zr-labeled humanized IgG1 antibodies 32 , 35 , 44 , 45 .…”

Section: Discussionmentioning

confidence: 94%

“…Analytical grade reagents, sterile technique and pyrogen-free plasticware were used in all labeling steps. DS-5573a was chelated with the bifunctional metal ion chelator p -isothiocyanatobenzyl-desferrioxamine (Df-Bz-NCS; Macrocyclics Inc Dallas, TX, USA) at a 3.0-fold molar excess as described before 32 . Df-Bz-NCS-DS-5573a conjugates prepared in 50 mM sodium acetate buffer (BDH/VWR Chemicals, Australia), pH 5.6, containing 5% w/v sorbitol (BDH/VWR Chemicals, Australia) and 0.01% w/v Tween 80 (Croda, Australia), were aliquoted in 1.0 mg aliquots and stored at -80 o C until required.…”

Section: Methodsmentioning

confidence: 99%

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Molecular imaging of T cell co-regulator factor B7-H3 with ⁸⁹Zr-DS-5573a

Burvenich¹,

Parakh²,

Lee³

et al. 2018

Theranostics

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B7-H3 is a transmembrane protein widely expressed in a variety of cancers and has been shown to play a role in anti-tumor immunity. This study aims to develop a molecular imaging probe to identify B7-H3 expression in tumors and to develop 89Zr-DS-5573a as a theranostic that could aid patient selection in clinical Phase I studies.Methods: The anti-B7-H3 humanised monoclonal antibody DS-5573a was labeled with zirconium-89 (89Zr-), and assessed for radiochemical purity, immunoreactivity (Lindmo analysis), antigen binding affinity (Scatchard analysis), and serum stability in vitro. In vivo biodistribution and imaging studies were performed with positron emission tomography and magnetic resonance imaging (PET/MRI) studies to identify and quantitate 89Zr-DS-5573a tumor uptake in a B7-H3-positive breast cancer model (MDA-MB-231) and a B7-H3-negative murine colon cancer model (CT26). Imaging and biodistribution studies were also performed in MDA-MB-231 tumor-bearing SCID mice in the absence and presence of therapeutic DS-5573a antibody dose (3 mg/kg DS-5573a).Results: 89Zr-DS-5573a showed high and specific binding to B7-H3-expressing MDA-MB-231 cells (immunoreactivity on day 0, 75.0 ± 2.9%), and low binding to B7-H3-negative CT26 cells (immunoreactivity on day 0, 10.85 ± 0.11%) in vitro. 89Zr-DS-5573a demonstrated good serum stability in vitro with 57.2 ± 2.0% of immunoreactivity remaining on day 7. In vivo biodistribution studies showed high uptake of 89Zr-DS-5573a in B7-H3-expressing MDA-MB-231 tumor-bearing mice, achieving 32.32 ± 6.55 %ID/g on day 7 post injection in BALB/c nu/nu mice and 25.76 ± 1.79 %ID/g in SCID mice, with minimal evidence of non-specific uptake in normal tissues, and excellent tumor localization on PET/MRI. In a combined imaging/therapy study, receptor saturation was demonstrated in tumors responding to therapy.Conclusion: 89Zr-DS-5573a demonstrates specific and prolonged targeting of B7-H3-expressing tumors in vivo. Saturation of binding sites was demonstrated in tumors responding to DS-5573a therapy. These results indicate that 89Zr-DS-5573a has potential to target B7-H3-expressing tumors in cancer patients. Furthermore 89Zr-DS-5573a has the potential to provide important insights into T cell biology through its specific binding to B7-H3.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

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Molecular imaging of T cell co-regulator factor B7-H3 with ⁸⁹Zr-DS-5573a

Burvenich¹,

Parakh²,

Lee³

et al. 2018

Theranostics

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“…As a member of RTK family, it is well known that EphA2 is an emerging target for cancer therapeutics, due to its increased expression in tumor tissues [ 12 – 14 ]. Downregulation of EphA2 expression through various approaches inhibits malignant behavior in vitro and in vivo [ 31 – 33 ]. To date, the knowledge of functional roles and regulatory mechanism of EphA2 in GC are remains unclear due to the lack of applicable and practicable method to target it.…”

Section: Discussionmentioning

confidence: 99%

miR-302b inhibits tumorigenesis by targeting EphA2 via Wnt/ β-catenin/EMT signaling cascade in gastric cancer

Huang¹,

He²,

McLeod³

et al. 2017

BMC Cancer

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BackgroundEphA2 is a crucial oncogene in gastric cancer (GC) development and metastasis, this study aims to identify microRNAs that target it and serve as key regulators of gastric carcinogenesis.MethodsWe identified several potential microRNAs targeting EphA2 by bioinformatics websites and then analyzed the role of miR-302b in modulating EphA2 in vitro and in vivo of GC, and it’s mechanism.ResultsOur analysis identified miR-302b, a novel regulator of EphA2, as one of the most significantly downregulated microRNA (miRNA) in GC tissues. Overexpression of miR-302b impaired GC cell migratory and invasive properties robustly and suppressed cell proliferation by arresting cells at G0–G1 phase in vitro. miR-302b exhibited anti-tumor activity by reversing EphA2 regulation, which relayed a signaling transduction cascade that attenuated the functions of N-cadherin, β-catenin, and Snail (markers of Wnt/β-catenin and epithelial-mesenchymal transition, EMT). This modulation of EphA2 also had distinct effects on cell proliferation and migration in GC in vivo.ConclusionsmiR-302b serves as a critical suppressor of GC cell tumorigenesis and metastasis by targeting the EphA2/Wnt/β-catenin/EMT pathway.Electronic supplementary materialThe online version of this article (10.1186/s12885-017-3875-3) contains supplementary material, which is available to authorized users.

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“…1C1 was also labeled with 64 Cu, after conjugation of the antibody with the chelating agent DOTA, and used for noninvasive PET of EphA2 levels in different tumor types (21). Recently, DS-8895a, an anti-EphA2 antibody, was labeled with appropriate radionuclides for PET and SPECT and applied to quantitative assessment of EphA2 expression and saturation in tumor models (22).…”

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confidence: 99%

PET-Guided Evaluation and Optimization of Internalized Antibody–Drug Conjugates Targeting Erythropoietin-Producing Hepatoma A2 Receptor

Jacobson

Chen

et al. 2017

J Nucl Med

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The erythropoietin-producing hepatoma A2 receptor (EphA2) is a tyrosine kinase overexpressed by tumor stroma and cancer cells. A high expression level of EphA2 predicts poor prognosis, correlating with disease progression and metastasis. Therefore, EphA2 is a relevant therapeutic target for human cancer. Antibodies, selectively bound to EphA2, can induce rapid receptor phosphorylation that results in antibody internalization and degradation. This internalization mechanism has been exploited with the development of antibody-drug conjugates (ADCs) for cancer chemotherapy. In this study, we used PET imaging to study the pharmacokinetics and tumor delivery of a panel of anti-EphA2 monoclonal antibodies (mAbs) with and without drug conjugates. A library of human anti-EphA2 mAbs were screened and evaluated for EphA2 internalization rate, binding affinity, epitope binding, and hydrophobicity. We chose 3 of these antibodies, denoted as 1C1, 3B10, and 2H7, which recognize different epitopes, for further evaluation. ADCs were generated by S239C mutation to give a ratio of 2 drug molecules per antibody. Native mAbs and ADCs were characterized, after conjugation to a DFO chelator andZr radiolabeling, in assays including cell uptake, internalization, hydrophobicity, and in vivo imaging using PET. All 3 mAbs had high affinities for EphA2 but exhibited different internalization rates following the order of 1C1> 3B10 > 2H7. Internalization rate is only 1 factor that affects in vitro cell uptake and in vivo tumor accumulation. Interestingly, the hydrophobicity of the mAbs, which followed the order of 2H7 > 1C1 > 3B10, had a strong correlation with in vivo tumor uptake measured by PET, with the least hydrophobic antibody, 3B10, showing the highest tumor uptake. ADC significantly reduced the in vivo uptake of all 3 mAbs. Tumor uptake of mAb is a complex process that is affected by multiple parameters, including internalization, hydrophobicity, and chemical modification. Our results suggest that the addition of drug molecules to mAb increases the clearance of the mAb presumably due to the increased hydrophobicity. Understanding the complexity of antibody-based tumor delivery may help improve ADC engineering for better tumor targeting and reduced side effects.

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Molecular Imaging and Quantitation of EphA2 Expression in Xenograft Models with ⁸⁹Zr-DS-8895a

Cited by 23 publications

References 26 publications

Molecular imaging of T cell co-regulator factor B7-H3 with ⁸⁹Zr-DS-5573a

Molecular imaging of T cell co-regulator factor B7-H3 with ⁸⁹Zr-DS-5573a

miR-302b inhibits tumorigenesis by targeting EphA2 via Wnt/ β-catenin/EMT signaling cascade in gastric cancer

PET-Guided Evaluation and Optimization of Internalized Antibody–Drug Conjugates Targeting Erythropoietin-Producing Hepatoma A2 Receptor

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Molecular Imaging and Quantitation of EphA2 Expression in Xenograft Models with 89Zr-DS-8895a

Cited by 23 publications

References 26 publications

Molecular imaging of T cell co-regulator factor B7-H3 with 89Zr-DS-5573a

Molecular imaging of T cell co-regulator factor B7-H3 with 89Zr-DS-5573a

miR-302b inhibits tumorigenesis by targeting EphA2 via Wnt/ β-catenin/EMT signaling cascade in gastric cancer

PET-Guided Evaluation and Optimization of Internalized Antibody–Drug Conjugates Targeting Erythropoietin-Producing Hepatoma A2 Receptor

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Product

Resources

About

Molecular Imaging and Quantitation of EphA2 Expression in Xenograft Models with ⁸⁹Zr-DS-8895a

Molecular imaging of T cell co-regulator factor B7-H3 with ⁸⁹Zr-DS-5573a

Molecular imaging of T cell co-regulator factor B7-H3 with ⁸⁹Zr-DS-5573a