Dual-Isotope Cryoimaging Quantitative Autoradiography: Investigating Antibody-Drug Conjugate Distribution and Payload Delivery Through Imaging
Dual-Isotope Cryoimaging Quantitative Autoradiography: Investigating Antibody-Drug Conjugate Distribution and Payload Delivery Through Imaging
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DOI：
10.2967/jnumed.118.207753
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发表时间：
2018-09-01
影响因子：
9.3
通讯作者：
Bradley, Daniel P.
中科院分区：
文献类型：
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作者：
Ilovich, Ohad;Qutaish, Mohammed;Bradley, Daniel P.
抗体-药物缀合物（ADC）的体外性质，例如结合、内化和细胞毒性，通常在体内研究之前充分表征。体内研究的解释可能会显着提高分子成像工具。我们在这里提出了一种双同位素冷冻成像定量放射自显影（CIQA）方法，结合先进的三维成像和分析，允许同时研究抗体和有效载荷分布在感兴趣的组织在临床前设置。方法：TAK-264是一种靶向抗鸟苷酸环化酶C（GCC）的研究性ADC，使用氚化单甲基澳瑞他汀E合成。然后将氚标记的ADC与二亚乙基三胺五乙酸缀合，用In-111标记，并在携带GCC阳性和GCC阴性肿瘤的动物中进行体内评价。结果如下：CIQA揭示了药物从ADC释放的时间过程及其分布到抗体较难接近的各种肿瘤区域。对于GCC阳性肿瘤，在示踪剂注射后96小时获得的代表性切片显示仅0.8%的体素具有共定位信号，而对于GCC阴性肿瘤切片，超过15%的体素具有共定位信号，这表明在GCC阳性病变中毒素的成功和特异性裂解。结论：成熟的放射自显影技术与先进的图像分析方法相结合，提供了一种实验工具，可以支持ADC肿瘤渗透和药代动力学的详细表征。
In vitro properties of antibody-drug conjugates (ADCs) such as binding, internalization, and cytotoxicity are often well characterized before in vivo studies. Interpretation of in vivo studies might be significantly enhanced by molecular imaging tools. We present here a dual-isotope cryoimaging quantitative autoradiography (CIQA) methodology combined with advanced 3-dimensional imaging and analysis allowing for the simultaneous study of both antibody and payload distribution in tissues of interest in a preclinical setting. Methods: TAK-264, an investigational ADC targeting anti-guanylyl cyclase C (GCC), was synthesized using tritiated monomethyl auristatin E. The tritiated ADC was then conjugated to diethylenetriaminepentaacetic acid, labeled with In-111, and evaluated in vivo in animals bearing GCC-positive and GCC-negative tumors. Results: CIQA revealed the time course of drug release from ADC and its distribution into various tumor regions that are less accessible to the antibody. For GCC-positive tumors, a representative section obtained 96 h after tracer injection showed only 0.8% of the voxels to have colocalized signal, versus over 15% of the voxels for a GCC-negative tumor section, suggesting successful and specific cleaving of the toxin in the GCC-positive lesions. Conclusion: The combination of a veteran established autoradiography technology with advanced image analysis methodologies affords an experimental tool that can support detailed characterization of ADC tumor penetration and pharmacokinetics.