Direct Cytosolic Delivery of Proteins through Coengineering of Proteins and Polymeric Delivery Vehicles
Direct Cytosolic Delivery of Proteins through Coengineering of Proteins and Polymeric Delivery Vehicles
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DOI：
10.1021/jacs.9b12759
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发表时间：
2020-03-04
影响因子：
15
通讯作者：
Rotello, Vincent M.
中科院分区：
文献类型：
--
作者：
Lee, Yi-Wei;Luther, David C.;Rotello, Vincent M.
纳米载体介导的蛋白质递送是基础研究和治疗应用的有前途的策略。然而，目前用于递送到细胞中的平台的功效受到递送的蛋白质货物的内体截留的限制，伴随着对胞质溶胶和其他细胞器（包括细胞核）的低效进入。我们在这里报告了一个强大的，多功能的聚合蛋白纳米复合材料（PPNC）平台，能够有效（>= 90%）的蛋白质传递到胞质溶胶。我们合成了一个库的胍官能化的聚氧杂降冰片烯酰亚胺（蓬特）的均聚物具有不同的分子量，以稳定和提供工程蛋白的末端寡谷氨酸“E-标签”。使用成像流式细胞术筛选聚合物的胞质递送效率，使用共聚焦显微镜验证胞质递送，并通过功能测定证明递送蛋白质的活性。这些研究表明，PPNC平台在广泛的制剂范围内提供了高度有效和可调的胞质递送，使其成为治疗性蛋白质递送的稳健试剂。
Nanocarrier-mediated protein delivery is a promising strategy for fundamental research and therapeutic applications. However, the efficacy of the current platforms for delivery into cells is limited by endosomal entrapment of delivered protein cargo with concomitantly inefficient access to the cytosol and other organelles, including the nucleus. We report here a robust, versatile polymeric protein nanocomposite (PPNC) platform capable of efficient (>= 90%) delivery of proteins to the cytosol. We synthesized a library of guanidinium-functionalized poly-(oxanorborneneimide) (PONT) homopolymers with varying molecular weights to stabilize and deliver engineered proteins featuring terminal oligoglutamate "E-tags". The polymers were screened for cytosolic delivery efficiency using imaging flow cytometry with cytosolic delivery validated using confocal microscopy and activity of the delivered proteins demonstrated through functional assays. These studies indicate that the PPNC platform provides highly effective and tunable cytosolic delivery over a wide range of formulations, making them robust agents for therapeutic protein delivery.