title: Glioma-targeted therapy using Cilengitide nanoparticles combined with UTMD enhanced delivery.
abstract: Malignant gliomas especially glioblastoma (GBM) are poorly responsive to the current treatments. Cilengitide (CGT) is a cyclic pentapeptide that demonstrated efficacy for GBM treatment by targeting the integrins avβ3 and avβ5 over-expressed on GBM cells. However, clinical translation of this therapy has been limited by issues including fast blood clearance, high kidney and liver uptake, poor blood-brain barrier (BBB) penetration, low tumor specificity and rapid washout from tumors. In this study, these issues were tackled in an integrated manner using a multi-stage strategy combining ultrasound-targeted microbubble destruction (UTMD) with CGT nanotherapy. CGT nanoparticles (CGT-NP) prepared using gelatin and Poloxamer 188-grafted heparin copolymer demonstrated significant apoptotic and cytotoxic effects in C6 GBM cells. Biodistribution study in a rat GBM model demonstrated buildup of high CGT level in tumors subjected to CGT-NP+UTMD combined therapy. The tumor CGT level in these animals was increased over 3-fold, tumor retention of CGT prolonged and renal clearance significantly reduced when compared with free CGT with or without UTMD. CGT-NP+UTMD treatment was further shown to extend the median survival period from less than 20days in the control and about 30days in free CGT group to about 80days. This was achieved with low CGT dosing level (2mg/kg twice weekly). In situ monitoring of GFAP, Ki67, caspase-3, Beclin-1, and LC-3 in the tumor samples together with TUNEL assay, transmission electron microscope imaging and Western blot assay all demonstrated high apoptotic and autophagy activities induced by the combined therapy. In conclusion, this study has provided extensive preclinical data supporting the use of this combined therapy to overcome the limitations of standard CGT treatment of gliomas.
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China; The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Ying-Zheng
authors.initials: YZ
authors.lastname: Zhao
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Lin
authors.affiliation: School of Pharmacy, Temple University School of Pharmacy, Philadelphia, PA 19140, USA.
authors.forename: Ho Lun
authors.initials: HL
authors.lastname: Wong
authors.affiliation: Department of Pharmacy, People's Hospital of Deqing County, Huzhou City, Zhejiang Province 313200, China.
authors.forename: Xiao-Tong
authors.initials: XT
authors.lastname: Shen
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: He-Lin
authors.initials: HL
authors.lastname: Xu
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Kai-Li
authors.initials: KL
authors.lastname: Mao
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Fu-Rong
authors.initials: FR
authors.lastname: Tian
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Jing-Jing
authors.initials: JJ
authors.lastname: Yang
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Xu
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Xiao
authors.affiliation: School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China; The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou City, Zhejiang Province 325035, China. Electronic address: lctuua@sina.com.
authors.forename: Cui-Tao
authors.initials: CT
authors.lastname: Lu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001812:Blood-Brain Barrier / Q000378:metabolism; D001932:Brain Neoplasms / Q000188:drug therapy*; D002470:Cell Survival / Q000187:drug effects; D016503:Drug Delivery Systems; D005910:Glioma / Q000188:drug therapy*; D006493:Heparin / Q000737:chemistry; D008297:Male; D045423:Microbubbles; D053758:Nanoparticles; D009363:Neoplasm Proteins / Q000378:metabolism; D020442:Poloxamer; D051381:Rats; D017207:Rats, Sprague-Dawley; D012910:Snake Venoms / Q000008:administration & dosage* / Q000627:therapeutic use*; D016019:Survival Analysis; D014018:Tissue Distribution; D014465:Ultrasonics
chemical_list: D009363:Neoplasm Proteins; D012910:Snake Venoms; D020442:Poloxamer; C422910:Cilengitide; D006493:Heparin
keywords: BBB opening technique; Cilengitide; Glioma; Nanoparticles; Ultrasound-targeted microbubble destruction