Systematic evaluation of antibody-mediated siRNA delivery using an industrial platform of THIOMAB-siRNA conjugates.
Systematic evaluation of antibody-mediated siRNA delivery using an industrial platform of THIOMAB-siRNA conjugates.
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DOI：
10.1093/nar/gku1362
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发表时间：
2015-01
影响因子：
14.9
通讯作者：
Siebel CW
中科院分区：
文献类型：
--
作者：
Cuellar TL;Barnes D;Nelson C;Tanguay J;Yu SF;Wen X;Scales SJ;Gesch J;Davis D;van Brabant Smith A;Leake D;Vandlen R;Siebel CW
siRNA的递送是实现RNAi治疗前景的关键障碍。通过靶向内化细胞表面抗原，抗体-siRNA复合物提供了一种可能的解决方案。然而，抗体-siRNA复合物的最初报道依赖于非特异性带电相互作用，并且尚未广泛适用。为了评估和改进这种递送方法，我们建立了一个名为THIOMAB的治疗性抗体的工业平台，该平台被设计为能够实现siRNA的精确共价偶联。我们报告说，这种耦合产生单体抗体-siRNA缀合物（ARC），保留抗体和siRNA的活性。为了广泛地评估这项技术，我们针对使用多种内化途径的七个靶标生成了一组THIOMAB，从而能够系统地操纵影响递送的多个参数。我们鉴定了在体外诱导靶向沉默的ARC，并在小鼠模型中全身给药后将测试扩展到靶向前列腺癌细胞。然而，最佳沉默仅限于特定条件，并且仅使用ARC的子集观察到。运输研究指出，ARC截留在内吞隔室作为一个限制因素，独立的抗原内化的途径。我们使用治疗级缀合物技术对多个参数进行了广泛的表征，对这种递送技术进行了全面的评估，突出了成功的例子以及剩余的挑战。
Delivery of siRNA is a key hurdle to realizing the therapeutic promise of RNAi. By targeting internalizing cell surface antigens, antibody–siRNA complexes provide a possible solution. However, initial reports of antibody–siRNA complexes relied on non-specific charged interactions and have not been broadly applicable. To assess and improve this delivery method, we built on an industrial platform of therapeutic antibodies called THIOMABs, engineered to enable precise covalent coupling of siRNAs. We report that such coupling generates monomeric antibody–siRNA conjugates (ARCs) that retain antibody and siRNA activities. To broadly assess this technology, we generated a battery of THIOMABs against seven targets that use multiple internalization routes, enabling systematic manipulation of multiple parameters that impact delivery. We identify ARCs that induce targeted silencing in vitro and extend tests to target prostate carcinoma cells following systemic administration in mouse models. However, optimal silencing was restricted to specific conditions and only observed using a subset of ARCs. Trafficking studies point to ARC entrapment in endocytic compartments as a limiting factor, independent of the route of antigen internalization. Our broad characterization of multiple parameters using therapeutic-grade conjugate technology provides a thorough assessment of this delivery technology, highlighting both examples of success as well as remaining challenges.
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影响因子：
46.9
作者：
Jackson, AL;Bartz, SR;Linsley, PS
通讯作者：
Linsley, PS
影响因子：
11.2
作者：
Erickson, HK;Park, PU;Blättler, WA
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Blättler, WA
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Lee, Young Sik;Pressman, Sigal;Andress, Arlise P.;Kim, Kevin;White, Jamie L.;Cassidy, Justin J.;Li, Xin;Lubell, Kim;Lim, Do Hwan;Cho, Ik Sang;Nakahara, Kenji;Preall, Jonathan B.;Bellare, Priya;Sontheimer, Erik J.;Carthew, Richard W.
通讯作者：
Carthew, Richard W.
影响因子：
46.9
作者：
Morrissey, DV;Lockridge, JA;Polisky, B
通讯作者：
Polisky, B
影响因子：
2.2
作者：
Junutula, Jagath R.;Bhakta, Sunil;Lowman, Henry B.
通讯作者：
Lowman, Henry B.