Identification of Novel Ligands for Targeted Antifibrotic Therapy of Chronic Pancreatitis

Abstract

Jessica Hung,1 Rohni Awasthi,1 Alexander L Klibanov,2– 5 Kimberly A Kelly2 1Department of Biomedical Engineering, School of Engineering, University of Virginia, Charlottesville, Virginia, 22908, USA; 2Department of Biomedical Engineering, School of Medicine, University of Virginia, Charlottesville, Virginia, 22908, USA; 3Division of Cardiovascular Medicine, University of Virginia, Charlottesville, Virginia, 22908, USA; 4Robert M. Berne Cardiovascular Research Center, University of Virginia, Charlottesville, Virginia, 22908, USA; 5Department of Radiology, University of Virginia, Charlottesville, Virginia, 22908, USACorrespondence: Kimberly A KellyDepartment of Biomedical Engineering, School of Medicine, University of Virginia, Box 800759, Charlottesville, Virginia, 22908, USATel [phone]Fax [phone]Email [email protected]: Chronic pancreatitis (CP) is an inflammatory disorder of the pancreas that leads to impaired pancreatic function. The limited therapeutic options and the lack of molecular targeting ligands or non-serum-based biomarkers hinder the development of target-specific drugs. Thus, there is a need for an unbiased, comprehensive discovery and evaluation of pancreatitis-specific ligands.Methods: This study utilized a computational-guided in vivo phage display approach to select peptide ligands selective for cellular components in the caerulein-induced mouse model of CP. The identified peptides were conjugated to pegylated DOPC liposomes via the reverse-phase evaporation method, and the in vivo specificity and pharmacokinetics were determined. As proof of concept, CP-targeted liposomes were used to deliver an antifibrotic small molecular drug, apigenin. Antifibrotic effects determined by pancreatic histology, fibronectin expression, and collagen deposition were evaluated.Results: We have identified five peptides specific for chronic pancreatitis and demonstrated selectivity to activated pancreatic stellate cells, acinar cells, macrophages, and extracellular matrix, respectively. MDLSLKP-conjugated liposomes demonstrated an increased particle accumulation by 1.3-fold in the inflamed pancreas compared to the control liposomes. We also observed that targeted delivery of apigenin resulted in improved acini preservation, a 37.2% and 33.1% respective reduction in collagen and fibronectin expression compared to mice receiving the free drug, and reduced oxidative stress in the liver.Conclusion: In summary, we have developed a systematic approach to profile peptide ligands selective for cellular components of complex disease models and demonstrated the biomedical applications of the identified peptides to improve tissue remodeling in the inflamed pancreas.Keywords: phage display, next-generation sequencing, peptide ligands, targeted liposomes, drug delivery

Keywords

• phage display
• next-generation sequencing
• peptide ligands
• targeted liposomes
• drug delivery