UPA-sensitive ACPP-conjugated nanoparticles for multi-targeting therapy of brain glioma Bo Zhang b , Yujie Zhang a , Ziwei Liao a , Ting Jiang b , Jingjing Zhao a , Yanyan Tuo a , Xiaojian She a , Shun Shen a , Jun Chen a , Qizhi Zhang a , Xinguo Jiang a , Yu Hu b, ** , Zhiqing Pang a, * a School of Pharmacy, Fudan University, Key Laboratory of Smart Drug Delivery, Ministry of Education, 826 Zhangheng Road, Shanghai 201203, PR China b Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, Hubei 430022, PR China article info Article history: Received 20 July 2014 Accepted 29 August 2014 Available online 13 October 2014 Keywords: Multi-targeting therapy Brain glioma Nanoparticles uPA Cell-penetrating peptide abstract Now it is well evidenced that tumor growth is a comprehensive result of multiple pathways, and glioma parenchyma cells and stroma cells are closely associated and mutually compensatory. Therefore, drug delivery strategies targeting both of them simultaneously might obtain more promising therapeutic benefits. In the present study, we developed a multi-targeting drug delivery system modified with uPA- activated cell-penetrating peptide (ACPP) for the treatment of brain glioma (ANP). In vitro experiments demonstrated nanoparticles (NP) decorated with cell-penetrating peptide (CPP) or ACPP could signifi- cantly improve nanoparticles uptake by C6 glioma cells and nanoparticles penetration into glioma spheroids as compared with traditional NP and thus enhanced the therapeutic effects of its payload when paclitaxel (PTX) was loaded. In vivo imaging experiment revealed that ANP accumulated more specifically in brain glioma site than NP decorated with or without CPP. Brain slides further showed that ACPP contributed to more nanoparticles accumulation in glioma site, and ANP could co-localize not only with glioma parenchyma cells, but also with stroma cells including neo-vascular cells and tumor- associated macrophages. The pharmacodynamics results demonstrated ACPP could significantly improve the therapeutic benefits of nanoparticles by significantly prolonging the survival time of glioma- bearing mice. In conclusion, the results suggested that nanoparticles modified with uPA-sensitive ACPP could reach multiple types of cells in glioma tissues and provide a novel strategy for glioma targeted therapy. © 2014 Elsevier Ltd. All rights reserved. 1. Introduction Glioma, as the most frequent brain tumor in adults, harbors a grim prognosis despite of multi-modality treatment due to its infiltrating property [1,2]. As a disease of the whole brain, surgical intervention is not sufficient to remove all the malignant cells [3], and the application of chemotherapeutics after surgery is neces- sary. Nanoparticle drug delivery systems achieve more chemo- therapeutic benefits over conventional small-molecule drugs to some extent, by the passive targeting strategy, namely, the enhanced permeability and retention (EPR) effect and the active targeting strategy as an extension [4]. However, most approaches utilize the interactions between targeting moieties and antigens highly expressed in glioma tissues to deliver therapeutic agents to a specific cell type in glioma, mainly glioma parenchyma cells [5,6], but neglect glioma stroma cells also as an important constitute of glioma [7]. It is well evidenced that tumor growth is a compre- hensive result of multiple pathways, and tumor parenchyma cells and stroma cells including neo-vascular cells and tumor-associated macrophages are closely associated and mutual compensatory [8e11]. As the “soil” of glioma parenchyma cells growth, glioma stroma cells play a critical role in the maintenance of glioma morphology, glioma cells differentiation, proliferation and invasion [12]. Thus, multi-targeting both glioma parenchyma cells and stroma cells and destroying glioma “seeds” and “soil” simulta- neously might benefit the treatment of glioma greatly. To the best of our knowledge, few studies in this filed has been reported, partly because targeting moieties capable of targeting all these cell types simultaneously are scarce. * Corresponding author. Tel./fax: þ86 21 51980069. ** Corresponding author. Tel.: þ86 27 85726335; fax: þ86 27 85776343. E-mail addresses: dr_huyu@126.com (Y. Hu), zqpang@fudan.edu.cn (Z. Pang). Contents lists available at ScienceDirect Biomaterials journal homepage: www.elsevier.com/locate/biomaterials http://dx.doi.org/10.1016/j.biomaterials.2014.09.008 0142-9612/© 2014 Elsevier Ltd. All rights reserved. Biomaterials 36 (2015) 98e109