Evaluation of cell penetrating peptides fused to elastin-like polypeptide for drug delivery
Evaluation of cell penetrating peptides fused to elastin-like polypeptide for drug delivery
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DOI：
10.1016/j.jconrel.2005.08.007
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发表时间：
2005-11-28
影响因子：
10.8
通讯作者：
Raucher, D
中科院分区：
文献类型：
--
作者：
Massodi, I;Bidwell, GL;Raucher, D
通过质膜的转运是大分子细胞递送的主要限制步骤。几种细胞穿透肽（CPP）已被证明可以有效地将各种分子货物内化到真核细胞内的靶点。在这项研究中，CPPs penetratin，达特，MTS融合弹性蛋白样多肽（CPP-ELP）的细胞摄取的效率和机制进行了评估。弹性蛋白样多肽是生物聚合物，其特征使得它们可用作用于递送治疗剂的聚合物载体。因此，通过将ELP融合到CPP来提高细胞摄取的效率并理解其细胞内化的机制可能有助于开发新的治疗方法。用流式细胞术和共聚焦荧光显微镜研究CPP-ELP的摄取机制。所有CPP-ELP的内化在4 ℃、ATP耗竭条件下和高渗蔗糖下显著受损，暗示CPP-ELP内化的内吞途径参与。Penetratin被确定为递送ELP的最有效CPP。最后，为了证明CPP-ELP用于药物递送的潜力，制备了含有穿透素、ELP和衍生自细胞周期蛋白依赖性激酶抑制剂p21的肽的融合多肽。该多肽显示通过减慢SKOV-3和HeLa细胞的生长速率来抑制它们的增殖。(c)2005 Elsevier B. V.保留所有权利。
Translocation through the plasma membrane is a major limiting step for the cellular delivery of macromolecules. Several cell penetrating peptides (CPP) have been demonstrated to efficiently internalize various molecular cargo to targets inside eukaryotic cells. In this study, the efficiency and mechanism of cellular uptake of the CPPs penetratin, Tat, and MTS fused to elastin-like polypeptides (CPP-ELP) were evaluated. Elastin-like polypeptides are biopolymers with features that make them useful as polymeric carriers for the delivery of therapeutics. Therefore, improving efficiency of cellular uptake by fusing ELPs to CPPs and understanding the mechanism of their cellular internalization could contribute to development of new therapeutic approaches. Flow cytometry and confocal fluorescence microscopy were used to elucidate the mechanism of CPP-ELP uptake. The internalization of all CPP-ELPs was impaired dramatically at 4 degrees C, under ATP depletion conditions, and with hyperosmolar sucrose, implicating involvement of an endocytic pathway for CPP-ELP internalization. Penetratin was identified as the most effective CPP for delivering ELP. Finally, in order to demonstrate the potential of CPP-ELP for drug delivery, a fusion polypeptide was made containing penetratin, ELP, and a peptide derived from the cyclin-dependent kinase inhibitor p21. This polypeptide was shown to inhibit proliferation of SKOV-3 and HeLa cells by slowing their growth rate. (c) 2005 Elsevier B.V. All rights reserved.