Marked increase in tumor transfection with a truncated branched polymer.
Marked increase in tumor transfection with a truncated branched polymer.
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DOI：
10.1002/jgm.3396
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发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
Mixson AJ
中科院分区：
文献类型：
--
作者：
Xu S;He J;Imtiyaz Z;Agrawal AK;Woodle MC;Mixson AJ
我们先前确定，由线性H2K肽形成的复合物在体内抑制肿瘤方面比由分支H2 K4 b-20肽形成的复合物更有效。基于胰蛋白酶消化和盐置换研究，线性H2K复合物的稳定性低于支链H2 K4 b-20复合物。由于聚合物和DNA从H2 K4 b-20多聚物中的结合和释放可能是无效的原因，我们研究了在其分支中具有不同数量的氨基酸的四分支组氨酸-赖氨酸（HK）肽是否在体内增加肿瘤中基因表达的能力方面更有效。通过固相合成法合成具有多个-KHHK-基序的线性和支链肽。分支H2 K4 b-20、-18、-14和12肽在其分支中分别具有20、18、14和12个氨基酸。在小鼠模型中检查这些肽携带荧光素酶表达质粒至人乳腺癌异种移植物的能力。凝胶阻滞和体内转染，掺入的靶向配体和内体裂解肽到这些复合物也进行了检查。在聚合复合物之前预注射阻断抗体，以确定神经纤毛蛋白1在肿瘤摄取这些聚合复合物中的作用。通过动态光散射测量聚合复合物的尺寸。在由分支载体形成的四种负表面电荷聚合复合物中，H2 K4 b-14聚合复合物被确定为最有效的质粒递送平台。将靶向配体和内体裂解肽掺入H2 K4 b-14多聚复合物中进一步增强了其体内抑制肿瘤的能力。此外，在给荷瘤小鼠预注射神经纤毛蛋白-1受体（NRP-1）的阻断抗体后，修饰的H2 K4 b-14复合物显著降低了肿瘤基因表达，表明NRP-1介导了它们转运到肿瘤中。本研究确定，长度中等的分支肽在体内将质粒递送至肿瘤中是非常有效的。为了利用肿瘤（包括其血管）中αvβ3整联蛋白和神经纤毛蛋白受体-1（NRP-1）转运系统表达的增加，设计了几种组氨酸-赖氨酸多聚复合物以靶向这些受体并将荧光素酶表达质粒携带至肿瘤。在最初靶向整联蛋白并随后靶向NRP-1受体后，具有中等长度分支载体的这些聚合复合物之一显示出对乳腺肿瘤异种移植物的显著高的转染和特异性。
We previously determined that polyplexes formed by linear H2K peptides were more effective in transfecting tumors in vivo than polyplexes formed by branched H2K4b‐20 peptides. Based on trypsin digest and salt displacement studies, the linear H2K polyplexes were less stable than the branched H2K4b‐20 polyplexes. Because binding and release of the polymer and DNA from the H2K4b‐20 polyplex may account for the ineffectiveness, we investigated whether four‐branched histidine‐lysine (HK) peptides with varying numbers of amino acids in their branches would be more effective in their ability to increase gene expression in tumors in vivo. Linear and branched peptides with multiple ‐KHHK‐ motifs were synthesized by solid‐phase synthesis. The branched H2K4b‐20, ‐18, ‐14 and 12 peptides had 20, 18, 14 and 12 amino acids in their branches, respectively. These peptides were examined for their ability to carry luciferase‐expressing plasmids to human breast cancer xenografts in a mouse model. With gel retardation and in vivo transfection, the incorporation of a targeting ligand and an endosomal lysis peptide into these polyplexes was also examined. A blocking antibody was pre‐injected prior to the polyplexes to determine the role of neuropilin 1 in the uptake of these polyplexes by the tumor. The size of the polyplexes was measured by dynamic light scattering. Of the four negative surface‐charge polyplexes formed by the branched carriers, the H2K4b‐14 polyplex was determined to be the most effective plasmid delivery platform to tumors. The incorporation of a targeting ligand and an endosomal lysis peptide into H2K4b‐14 polyplexes further enhanced their ability to transfect tumors in vivo. Furthermore, after pre‐injecting tumor‐bearing mice with a blocking antibody to the neuropilin‐1 receptor (NRP‐1), there was a marked reduction of tumor gene expression with the modified H2K4b‐14 polyplexes, suggesting that NRP‐1 mediated their transport into the tumor. The present study established that branched peptides intermediate in length were very efficient in delivering plasmids to tumors in vivo. To take advantage of increased expression of αvβ3 integrins and the neuropilin receptor‐1 (NRP‐1) transport system in tumors including their vessels, several histidine‐lysine polyplexes were designed to target these receptors and carry luciferase‐expressing plasmids to tumors. After initially targeting integrins and subsequently the NRP‐1 receptors, one of these polyplexes with an intermediate in length branched carrier showed remarkably high transfection and specificity for breast tumor xenografts.
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影响因子：
6.2
作者：
Chou, Szu-Ting;Leng, Qixin;Scaria, Puthupparampil;Kahn, Jason D.;Tricoli, Lucas J.;Woodle, Martin;Mixson, A. James
通讯作者：
Mixson, A. James
影响因子：
3.7
作者：
Karathanasis E;Chan L;Karumbaiah L;McNeeley K;D'Orsi CJ;Annapragada AV;Sechopoulos I;Bellamkonda RV
通讯作者：
Bellamkonda RV
影响因子：
8.8
作者：
Akashi, Y.;Oda, T.;Ohara, Y.;Miyamoto, R.;Kurokawa, T.;Hashimoto, S.;Enomoto, T.;Yamada, K.;Satake, M.;Ohkohchi, N.
通讯作者：
Ohkohchi, N.
DOI：
10.1016/j.nano.2013.07.008
发表时间：
2014-01-01
影响因子：
5.4
作者：
Laechelt, Ulrich;Kos, Petra;Wagner, Ernst
通讯作者：
Wagner, Ernst
DOI：
10.1002/jgm.3295
发表时间：
2021-03
期刊：
The journal of gene medicine
影响因子：
--
作者：
He J;Xu S;Leng Q;Mixson AJ
通讯作者：
Mixson AJ