Selective Intracellular Delivery of Recombinant Arginine Deiminase (ADI) Using pH-Sensitive Cell Penetrating Peptides To Overcome ADI Resistance in Hypoxic Breast Cancer Cells.
Selective Intracellular Delivery of Recombinant Arginine Deiminase (ADI) Using pH-Sensitive Cell Penetrating Peptides To Overcome ADI Resistance in Hypoxic Breast Cancer Cells.
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DOI：
10.1021/acs.molpharmaceut.5b00706
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发表时间：
2016-01-04
影响因子：
4.9
通讯作者：
Shen LJ
中科院分区：
文献类型：
--
作者：
Yeh TH;Chen YR;Chen SY;Shen WC;Ann DK;Zaro JL;Shen LJ
精氨酸消耗策略，如聚乙二醇化重组精氨酸脱亚胺酶（ADI-PEG 20），提供了一种有前途的抗癌治疗。许多肿瘤细胞抑制了一种关键酶的表达，即将瓜氨酸转化为精氨酸的氨基琥珀酸合成酶1（ASS 1）。这些肿瘤细胞成为精氨酸营养缺陷型，因为它们不再能在细胞内从瓜氨酸合成内源性精氨酸，因此对精氨酸耗竭疗法敏感。然而，由于ADI-PEG 20仅由于低内化而消耗细胞外精氨酸，表达ASS 1的细胞对治疗不敏感，因为它们可以在细胞内合成精氨酸。最近的研究发现，有几个因素影响ASS 1的表达。在这项研究中，我们评估了缺氧的影响，经常遇到的许多实体瘤，ASS 1的表达及其关系到ADI耐药的人MDA-MB-231乳腺癌细胞。结果发现，MDA-MB-231细胞在缺氧条件下产生ADI抗性，其中ASS 1表达增加。为了恢复ADI的敏感性以及实现缺氧条件下的肿瘤选择性递送，我们构建了一种基于pH敏感性细胞穿透肽（CPP）的递送系统，以在细胞内携带ADI，从而耗尽细胞内和细胞外精氨酸。递送系统被设计成仅在与缺氧肿瘤的微环境相关的弱酸性pH（6.5-7）下激活CPP介导的内化，从而实现对肿瘤细胞的更好选择性。CPP，HBHAc的pH敏感性通过与组氨酸-谷氨酰胺（HE）寡肽的重组融合来控制，产生HBHAc-HE-ADI。HBHAc-HE-ADI在人乳腺癌细胞的小鼠异种移植模型中的肿瘤分布与ADI-PEG 20相当。此外，与正常pH值（常氧条件）相比，在弱酸性pH值（缺氧条件）下孵育的细胞中，HBHAc-HE-ADI的体外细胞摄取增加，这与缺氧MDA-MB-231和人前列腺癌PC 3细胞中的pH敏感性体外细胞毒性相关。总之，我们得出结论，基于HBHAc-HE的肽递送提供了一种有用的手段来克服乳腺癌细胞中缺氧诱导的对ADI的抗性，并靶向弱酸性肿瘤微环境。使用pH敏感性细胞穿透肽选择性细胞内递送重组精氨酸脱亚胺酶（ADI）以克服缺氧乳腺癌细胞MDA-MB-231中的ADI抗性的示意图。
Arginine depletion strategies, such as pegylated recombinant arginine deiminase (ADI-PEG20), offer a promising anticancer treatment. Many tumor cells have suppressed expression of a key enzyme, argininosuccinate synthetase 1 (ASS1), which converts citrulline to arginine. These tumor cells become arginine auxotrophic, as they can no longer synthesize endogenous arginine intracellularly from citrulline, and are therefore sensitive to arginine depletion therapy. However, since ADI-PEG20 only depletes extracellular arginine due to low internalization, ASS1-expressing cells are not susceptible to treatment since they can synthesize arginine intracellularly. Recent studies have found that several factors influence ASS1 expression. In this study, we evaluated the effect of hypoxia, frequently encountered in many solid tumors, on ASS1 expression and its relationship to ADI-resistance in human MDA-MB-231 breast cancer cells. It was found that MDA-MB-231 cells developed ADI resistance in hypoxic conditions with increased ASS1 expression. To restore ADI sensitivity as well as achieve tumor-selective delivery under hypoxia, we constructed a pH-sensitive cell penetrating peptide (CPP)-based delivery system to carry ADI inside cells to deplete both intra- and extracellular arginine. The delivery system was designed to activate the CPP-mediated internalization only at the mildly acidic pH (6.5–7) associated with the microenvironment of hypoxic tumors, thus achieving better selectivity toward tumor cells. The pH sensitivity of the CPP, HBHAc, was controlled by recombinant fusion to a histidine-glutamine (HE) oligopeptide, generating HBHAc-HE-ADI. The tumor distribution of HBHAc-HE-ADI was comparable to ADI-PEG20 in a mouse xenograft model of human breast cancer cells in vivo. In addition, HBHAc-HE-ADI showed increased in vitro cellular uptake in cells incubated in a mildly acidic pH (hypoxic conditions) compared to normal pH (normoxic conditions), which correlated with pH-sensitive in vitro cytotoxicity in hypoxic MDA-MB-231 and human prostate cancer PC3 cells. Together, we conclude that the HBHAc-HE-based peptide delivery offers a useful means to overcome hypoxia-induced resistance to ADI in breast cancer cells, and to target the mildly acidic tumor microenvironment. Schematic presentation of the selective intracellular delivery of recombinant arginine deiminase (ADI) using pH-sensitive cell penetrating peptides to overcome ADI resistance in hypoxic breast cancer cells MDA-MB-231.
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影响因子：
14
作者：
Fei, Likun;Yap, Li-Peng;Conti, Peter S.;Shen, Wei-Chiang;Zaro, Jennica L.
通讯作者：
Zaro, Jennica L.
影响因子：
11.2
作者：
Kim, Randie H.;Coates, Jodi M.;Bowles, Tawnya L.;McNerney, Gregory P.;Sutcliffe, Julie;Jung, Jae U.;Gandour-Edwards, Regina;Chuang, Frank Y. S.;Bold, Richard J.;Kung, Hsing Jien
通讯作者：
Kung, Hsing Jien
影响因子：
13.8
作者：
Semenza GL
通讯作者：
Semenza GL
影响因子：
3.8
作者：
Belani CP;Yamamoto N;Bondarenko IM;Poltoratskiy A;Novello S;Tang J;Bycott P;Niethammer AG;Ingrosso A;Kim S;Scagliotti GV
通讯作者：
Scagliotti GV
DOI：
10.1073/pnas.1404171111
发表时间：
2014-09-30
影响因子：
11.1
作者：
Changou, Chun A.;Chen, Yun-Ru;Kung, Hsing-Jien
通讯作者：
Kung, Hsing-Jien