Delivery of intact transcription factor by using self-assembled supramolecular nanoparticles.
Delivery of intact transcription factor by using self-assembled supramolecular nanoparticles.
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DOI：
10.1002/anie.201005740
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发表时间：
2011-03-21
影响因子：
16.6
通讯作者：
Tseng, Hsian-Rong
中科院分区：
文献类型：
--
作者：
Liu, Yang;Wang, Hao;Kamei, Ken-ichiro;Yan, Ming;Chen, Kuan-Ju;Yuan, Qinghua;Shi, Linqi;Lu, Yunfeng;Tseng, Hsian-Rong
蛋白质递送[1]被认为是调节细胞行为的最直接的策略，而没有与基因递送方法相关的安全性问题和表达性能问题。为了使其在生物学和医学中的实际应用成为可能，仍需克服两个主要挑战：1）如何促进蛋白质分子的细胞摄取; 2）如何在递送过程中保持其稳定性和功能。最近，人们尝试开发多种递送载体，包括脂质体、[3]聚合物胶束、[4]和纳米颗粒，[5]以增强靶细胞对蛋白质分子的吸收，同时稳定包裹的蛋白质。由于在优化运载材料方面采用的程序十分耗时，因此在寻求更好的运载系统方面作出了重大努力，尽管迄今为止在这一领域取得的进展有限。或者，重组技术[6]可用于将细胞穿透肽[7]（CPP）缀合到蛋白质分子上，这是最常用的蛋白质递送系统，具有改善的递送效率。在这种情况下，需要解决与产生重组蛋白的复杂程序和缺乏针对蛋白变性的保护机制相关的主要瓶颈。转录因子（TF）是一种负责调节细胞回路中基因转录的蛋白质。[8]一般来说，TF含有一个或多个DNA结合结构域（DBD），其识别与它们调节的基因相邻的匹配DNA序列。显然，TF的高效递送可以提供用于调节细胞行为的强大技术。需要高效TF递送的最重要的体外应用之一是产生人诱导多能干细胞（hiPSC），最近通过将CPP融合的重编程TF（即OCT 4、SOX 2、KLF 4和c-MYC）引入人体细胞中证明了这一点[9]。由此产生的hiPSC有可能彻底改变再生医学。[10]然而，四种重编程TF以其重组形式的高成本意味着这种方法不太可能在不进一步改善重编程蛋白的递送性能的情况下用于大规模hiPSC产生。因此，开发一种能够高效递送完整（未修饰）转录因子的新型载体是至关重要的。在此之前，我们展示了一种方便、灵活和模块化的自组装方法，用于通过基于金刚烷（Ad）和β-环糊精（CD）基序的多价分子识别从少量分子构建块制备超分子纳米颗粒（SNPs）。这种自组装合成策略能够控制所得SNP的大小，表面化学，zeta电位和有效载荷，这为生物医学应用开辟了许多有趣的机会，例如正电子发射断层扫描（PET）成像，[11]磁共振成像（MRI），[12]癌细胞的光热治疗，[13]和高效基因递送。[14]考虑到TF的独特作用，我们试图探索使用SNP作为一种新型的纳米级载体，用于以上级现有方法的效率递送完整的（未修饰的）TF。我们的想法是通过将阴离子特征引入TF来实现TF到阳离子SNP载体中的封装。具有TF特异性匹配识别序列的DNA质粒可用于形成阴离子TF· DNA复合物，该复合物随后可被封装到微囊中。
Protein delivery [1] has been considered as the most straightforward strategy for modulating cellular behavior without the safety concerns and expression performance issues associated with gene deliver approaches. Two major challenges remain to be overcome in order to enable practical applications in biology and medicine 1) how to foster cellular uptake of protein molecules and 2) how to retain their stabilities and functions [2] over the delivery process. Recently, attempts have been made to develop a variety of delivery vectors, including liposomes,[3] polymer micelles,[4] and nanoparticle,[5] to enhance the uptake of protein molecules in target cells, and at the same time, to stabilize the encapsulated proteins. Owing to the time-consuming procedures employed in optimization of delivery materials, significant endeavors have been made in search of better delivery systems, although there has been limited progress in the field to date. Alternatively, recombinant technology [6] can be utilized to conjugate cell-penetrating peptides [7](CPPs) onto protein molecules, this is the most commonly used protein delivery system with improved delivery efficiency. In this case, the major bottlenecks associated with the complicated procedure of generating recombinant proteins and the lack of protection mechanism against protein denature need to be solved. Transcription factor (TF) is a protein responsible for regulating gene transcription in cellular circuitry.[8] In general, TFs contain one or more DNA-binding domains (DBDs), which recognize matching DNA sequences adjacent to the genes they regulate. Apparently, highly efficient delivery of TFs can provide a powerful technology for modulating cellular behavior. One of the most important in-vitro applications that required highly efficient TF delivery is the generation of human induced pluripotent stem cells (hiPSCs) which has recently been demonstrated by introducing CPPsfused reprogramming TFs (ie, OCT4, SOX2, KLF4, and c-MYC)[9] into human somatic cells. The resulting hiPSCs have the potential to revolutionize regenerative medicine.[10] However, the high costs of the four reprogramming TFs in their recombinant forms, means it is unlikely that this approach can be used for large-scale hiPSCs generation without further improvement in the delivery performance of the reprogramming proteins. Therefore, it is crucial to develop a new type of vector capable of delivering intact (unmodified) TFs in a highly efficient manner.Previously, we demonstrated a convenient, flexible, and modular self-assembly approach for the preparation of supramolecular nanoparticles (SNPs) from a small collection of molecular building blocks through a multivalent molecular recognition based on adamantane (Ad) and β-cyclodextrin (CD) motifs. Such a self-assembly synthetic strategy enables control upon the sizes, surfaces chemistry, zeta potentials, and payloads of the resulting SNPs, which open up many interesting opportunities for biomedical applications, for example, positron emission tomography (PET) imaging,[11] magnetic resonance imaging (MRI),[12] photothermal treatment of cancer cells,[13] and highly efficient gene delivery.[14] Considering the unique role of TF, we attempted to explore the use of SNPs as a new type of nanoscale vector for delivering intact (unmodified) TFs with an efficiency superior to that of existing approaches. Our idea is to achieve the encapsulation of a TF into cationic SNP vectors by introducing anionic characteristics to the TF. A DNA plasmid with a matching recognition sequence specific to a TF can be employed to form an anionic TF· DNA complex, which can be subsequently encapsulated into …
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影响因子：
23.9
作者：
Kim D;Kim CH;Moon JI;Chung YG;Chang MY;Han BS;Ko S;Yang E;Cha KY;Lanza R;Kim KS
通讯作者：
Kim KS
影响因子：
16.6
作者：
Cao, Zehui;Tong, Rong;Lu, Yi
通讯作者：
Lu, Yi
影响因子：
4.8
作者：
Abbing, A;Blaschke, UK;Reiser, COA
通讯作者：
Reiser, COA
影响因子：
46.9
作者：
Kost, TA;Condreay, JP;Jarvis, DL
通讯作者：
Jarvis, DL
影响因子：
6.1
作者：
Kamei K;Ohashi M;Gschweng E;Ho Q;Suh J;Tang J;For Yu ZT;Clark AT;Pyle AD;Teitell MA;Lee KB;Witte ON;Tseng HR
通讯作者：
Tseng HR