title: Intracellular delivery of peptide cargos using iron oxide based nanoparticles: studies on antitumor efficacy of a BCL-2 converting peptide, NuBCP-9.
abstract: Delivering peptides into cells targeting the undruggable oncoproteins is an emerging area in cancer therapeutics. Here we report a novel nanoparticle-based delivery system that can transport therapeutic cargos to the intracellular sites without the need for a cell transduction or penetration domain (CPP). In the present study, we have used iron oxide nanoparticles to deliver an oncopeptide, NuBCP-9, targeting the BCL-2 BH3 domain. Citric acid/2-bromo 2-methylpropanoic acid (CA/BMPA)-capped SPIONs were used to immobilize and deliver the NuBCP-9 peptide to the cancer cells without any noticeable off-target effects. Our results have demonstrated that NuBCP-9-SPIONs efficiently penetrate into cancer cells and bind to its intracellular target protein BCL-2. Moreover, significant inhibition of proliferation and substantial induction of cell death were observed when cancer cells were treated with NuBCP-9-SPIONs at different time intervals. Importantly, the IC50 values for killing of breast cancer cells with NuBCP-9-SPIONs were much lower compared to cells treated with the NuBCP-9 peptide linked with a CPP (Arg-8; NuBCP-9-R8). Molecular and biochemical analyses further supported that NuBCP-9-SPIONs killed breast cancer cells by apoptosis-mediated mechanisms. Furthermore, our data demonstrated that administration of NuBCP-9-SPIONs to mice bearing Ehrlich ascites tumors (EAT) was associated with loss of tumorigenicity and extensive apoptosis in tumor tissues. Taken together, these findings show that a non-CPP-tagged peptide can be successfully delivered to undruggable intracellular oncotargets using SPIONs.
authors.affiliation: Center for Biomedical Engineering, Indian Institute of Technology, Delhi,, Hauz Khas, New Delhi-110016, India. harpal2000@yahoo.com.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kumar
authors.forename: Gurpal
authors.initials: G
authors.lastname: Singh
authors.forename: Sapna
authors.initials: S
authors.lastname: Sharma
authors.forename: Dikshi
authors.initials: D
authors.lastname: Gupta
authors.forename: Vivek
authors.initials: V
authors.lastname: Bansal
authors.forename: Vikas
authors.initials: V
authors.lastname: Arora
authors.forename: Madhusudan
authors.initials: M
authors.lastname: Bhat
authors.forename: Sandeep K
authors.initials: SK
authors.lastname: Srivastava
authors.forename: Sameer
authors.initials: S
authors.lastname: Sapra
authors.forename: Surender
authors.initials: S
authors.lastname: Kharbanda
authors.forename: Amit K
authors.initials: AK
authors.lastname: Dinda
authors.forename: Harpal
authors.initials: H
authors.lastname: Singh
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002286:Carcinoma, Ehrlich Tumor / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects*; D004337:Drug Carriers; D005260:Female; D005290:Ferric Compounds / Q000737:chemistry / Q000494:pharmacology; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: C000595707:BCL2 protein, human; D004337:Drug Carriers; D005290:Ferric Compounds; D009842:Oligopeptides; D019253:Proto-Oncogene Proteins c-bcl-2; C000590397:phenylalanyl-seryl-arginyl-seryl-leucyl-histidyl-seryl-leucyl-leucine; C052617:Bcl2 protein, mouse; C000499:ferric oxide