Discovery of Peptide Ligands for Hepatic Stellate Cells Using Phage Display

被引:33
作者
Chen, Zhijin [1 ]
Jin, Wei [1 ]
Liu, Hao [1 ]
Zhao, Zhen [1 ]
Cheng, Kun [1 ]
机构
[1] Univ Missouri, Sch Pharm, Div Pharmaceut Sci, Kansas City, MO 64108 USA
基金
美国国家卫生研究院;
关键词
liver fibrosis; HSCs; IGF2R; peptide ligand; phage display; FACTOR-II RECEPTOR; TRIPLEX-FORMING OLIGONUCLEOTIDES; MANNOSE; 6-PHOSPHATE; TARGETED DELIVERY; LIVER FIBROSIS; GENE PROMOTER; CANCER CELLS; GROWTH; MANNOSE-6-PHOSPHATE; NANOPARTICLES;
D O I
10.1021/acs.molpharmaceut.5b00177
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Regardless of its cause, liver fibrosis is characterized by the excessive accumulation of extracellular matrix (ECM) in the liver. Hepatic stellate cells (HSCs) are the main producers responsible for the excessive production of ECM and profibrogenic cytokines in fibrotic liver. Therefore, development of HSC-specific delivery systems is essential for the success of antifibrotic agents. The objective of this study is to identify peptide ligands targeting the insulin-like growth factor 2 receptor (IGF2R), which is overexpressed on HSCs. We expect to use the peptide ligands for the future development of HSC-targeted drug delivery system. Protein- and whole cell-based phage display biopannings were conducted to identify phage/peptide candidates. Phage ELISA, cellular uptake, and cell viability assay were employed to evaluate the binding affinity and specificity of these peptide ligands to recombinant human IGF2R and HSCs. IGF2R siRNA was used to silence the IGF2R protein expression in human hepatic stellate cells (LX-2) to confirm the specificity of the identified peptide ligands. Among the identified peptide candidates, peptide-431 shows the highest binding affinity and specificity to recombinant human IGF2R protein and HSCs. The equilibrium dissociation constant (K-d) of peptide-431 is 6.19 mu M for LX-2 cells and 12.35 mu M for rat hepatic stellate cells HSC-T6. Cellular uptake of peptide-431 in LX-2 cells is significantly reduced after silencing IGF2R with siRNA. Peptide-431 also enhances the uptake of a proapoptotic peptide (KLA peptide) in LX-2 and HSC-T6 cells, indicating that peptide-431 can be used as a targeting ligand to deliver antifibrotic agents into not only rat but also human HSCs. Dimerization of peptide-431 further increase its binding affinity to LX-2 cells by approximately 9-fold.
引用
收藏
页码:2180 / 2188
页数:9
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