Anthrax Protective Antigen Retargeted with Single-Chain Variable Fragments Delivers Enzymes to Pancreatic Cancer Cells.
Anthrax Protective Antigen Retargeted with Single-Chain Variable Fragments Delivers Enzymes to Pancreatic Cancer Cells.
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DOI：
10.1002/cbic.202000201
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发表时间：
2020-10-01
期刊：
影响因子：
--
通讯作者：
Pentelute BL
中科院分区：
文献类型：
--
作者：
Loftis AR;Santos MS;Truex NL;Biancucci M;Satchell KJF;Pentelute BL
无毒的炭疽保护性抗原/致死因子N-末端结构域（PA/LFN）复合物是将蛋白质转运到细胞胞质溶胶中的有效平台。突变型PA（mPA）最近与表皮生长因子（EGF）融合以将LFN重新靶向递送至携带EGF受体（EGFR）的细胞，但对已知同源配体的要求限制了该方法的适用性。在这里，我们提供实用的保护性抗原重靶向与mPA单链可变片段（scFv）融合构建体的各种受体。我们的设计能够靶向两种胰腺癌相关受体，EGFR和癌胚抗原。我们证明了与scFv的融合不会干扰mPA的基本功能。此外，mPA-scFv融合体能够将白喉毒素催化结构域和Ras/Rap 1特异性内肽酶细胞特异性递送至胰腺癌细胞。重要的是，基于mPA-scFv融合的治疗在体外显示出有效的细胞特异性毒性，从根本上开辟了工程化免疫毒素的新途径，并为靶向蛋白质递送至癌细胞胞质溶胶的挑战提供了潜在的解决方案。蛋白质的细胞特异性胞质递送是高度挑战性的。在此，我们证明了炭疽衍生的保护性抗原（mPA）的突变体可以表达为与scFv的融合物，用于将有效的蛋白质治疗剂靶向递送至细胞胞质溶胶。用这种策略靶向EGFR或癌胚抗原导致胰腺癌细胞的选择性递送和死亡。
The nontoxic, anthrax protective antigen/lethal factor N-terminal domain (PA/LFN) complex is an effective platform for translocating proteins into the cytosol of cells. Mutant PA (mPA) was recently fused to epidermal growth factor (EGF) to retarget delivery of LFN to cells bearing EGF receptors (EGFR), but the requirement for a known cognate ligand limits the applicability of this approach. Here, we render practical protective antigen retargeting to a variety of receptors with mPA single-chain variable fragment (scFv) fusion constructs. Our design enables the targeting of two pancreatic cancer-relevant receptors, EGFR and carcinoembryonic antigen. We demonstrate that fusion to scFvs does not disturb the basic functions of mPA. Moreover, mPA–scFv fusions enable cell-specific delivery of diphtheria toxin catalytic domain and Ras/Rap1-specific endopeptidase to pancreatic cancer cells. Importantly, mPA–scFv fusion-based treatments display potent cell-specific toxicity in vitro, opening fundamentally new routes toward engineered immunotoxins and providing a potential solution to the challenge of targeted protein delivery to the cytosol of cancer cells. Cell-specific cytosolic delivery of proteins is highly challenging. Herein, we demonstrate that mutants of anthrax-derived protective antigen (mPA) can be expressed as fusions to scFvs for targeted delivery of potent protein therapeutics to the cellular cytosol. Targeting of EGFR or carcinoembryonic antigen with this strategy led to selective delivery to and death of pancreatic cancer cells.
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影响因子：
4
作者：
Meng Q;Shi S;Liang C;Liang D;Xu W;Ji S;Zhang B;Ni Q;Xu J;Yu X
通讯作者：
Yu X
影响因子：
--
作者：
Ahmad ZA;Yeap SK;Ali AM;Ho WY;Alitheen NB;Hamid M
通讯作者：
Hamid M
影响因子：
15.9
作者：
Collins, Meredith A.;Bednar, Filip;di Magliano, Marina Pasca
通讯作者：
di Magliano, Marina Pasca
影响因子：
2.9
作者：
Antic, Irena;Biancucci, Marco;Satchell, Karla J. F.
通讯作者：
Satchell, Karla J. F.
DOI：
10.1083/jcb.200211018
发表时间：
2003-02-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Abrami L;Liu S;Cosson P;Leppla SH;van der Goot FG
通讯作者：
van der Goot FG