articles Targeted Delivery of Antisense Oligodeoxynucleotide by Transferrin Conjugated pH-Sensitive Lipopolyplex Nanoparticles: A Novel Oligonucleotide-Based Therapeutic Strategy in Acute Myeloid Leukemia Yan Jin,†,‡ Shujun Liu,|,⊥ Bo Yu,†,§ Sharon Golan,# Chee-Guan Koh,†,§ Jintao Yang,†,§ Lenguyen Huynh,|,⊥ Xiaojuan Yang,†,‡ Jiuxia Pang,|,⊥ Natarajan Muthusamy,|,⊥ Kenneth K. Chan,‡,⊥ John C. Byrd,|,⊥ Yeshayahu Talmon,# L. James Lee,*,†,§ Robert J. Lee,*,†,‡,| and Guido Marcucci*,|,⊥ NSF Nanoscale Science and Engineering Center, DiVision of Pharmaceutics, College of Pharmacy, Department of Chemical and Biomolecular Engineering, The ComprehensiVe Cancer Center, and DiVision of Hematology and Oncology, Department of Internal Medicine, The Ohio State UniVersity, Columbus, Ohio, and Technion-Israel Institute of Technology, Haifa, Israel Received August 21, 2009; Revised Manuscript Received October 13, 2009; Accepted October 23, 2009
Abstract: Therapeutic use of oligodeoxynucleotides (ODNs) that hybridize to and downregulate target mRNAs encoding proteins that contribute to malignant transformation has a sound rationale, but has had an overall limited clinical success in cancer due to insufficient intracellular delivery. Here we report a development of formulations capable of promoting targeted delivery and enhanced pharmacologic activity of ODNs in acute myeloid leukemia (AML) cell lines and patient primary cells. In this study, transferrin (Tf) conjugated pH-sensitive lipopolyplex nanoparticles (LPs) were prepared to deliver GTI-2040, an antisense ODN against the R2 subunit of ribonucleotide reductase that has been shown to contribute to chemoresistance in AML. LPs had an average particle size around 110 nm and a moderately positive zeta potential at ∼ 10 mV. The ODN encapsulation efficiency of LPs was >90%. These nanoparticles could release ODNs at acidic endosomal pH and facilitate the cytoplasmic delivery of ODNs after endocytosis. In addition, Tf-mediated targeted delivery of GTI2040 was achieved. R2 downregulation at both mRNA and protein levels was improved by 8-fold in Kasumi-1 cells and 2- to 20-fold in AML patient primary cells treated with GTI-2040-Tf-LPs, compared to free GTI-2040 treatment. Moreover, Tf-LPs were more effective than nontargeted LPs, with 10 to 100% improvement at various concentrations in Kasumi-1 cells and an average of 45% improvement at 3 µM concentration in AML patient primary cells. Treatment with 1 µM GTI-2040-Tf-LPs sensitized AML cells to the chemotherapy agent cytarabine, by decreasing its IC50 value from 47.69 nM to 9.05 nM. This study suggests that the combination of pH sensitive LP formulation and Tf mediated targeting is a promising strategy for antisense ODN delivery in leukemia therapy.
Keywords: Lipopolyplex nanoparticles; transferrin; GTI-2040; acute myeloid leukemia; antisense; oligodeoxynucleotides
1. Introduction Acute myeloid leukemia (AML) is a heterogeneous, malignant disease of the hematopoietic system characterized by clonal accumulation and expansion of immature myeloid cells in the bone marrow (BM) and blood. Mutations and 196
MOLECULAR PHARMACEUTICS VOL. 7, NO. 1, 196–206
overexpression of genes involved in normal patterns of cellular differentiation, proliferation and apoptosis have been discovered and correlated with the pathogenesis and prognosis of this disease. Despite the progress in the understanding of the mechanisms underlying the initiation and main10.1021/mp900205r 2010 American Chemical Society
Published on Web 10/23/2009
Lipopolyplex Nanoparticles for Antisense DeliVery in AML tenance of this disease, only approximately 40% of the treated younger patients (aged
