Application of Deacetylated Poly-N-Acetyl Glucosamine Nanoparticles for the Delivery of miR-126 for the Treatment of Cecal Ligation and Puncture-Induced Sepsis
Sepsis is an acute inflammatory syndrome in response to infection. In some cases, excessive inflammation from sepsis results in endothelial dysfunction and subsequent increased vascular permeability leading to organ failure. We previously showed that treatment with endothelial progenitor cells, which highly express microRNA-126 (miR-126), improved survival in mice subjected to cecal ligation and puncture (CLP) sepsis. miRNAs are important regulators of gene expression and cell function, play a major role in endothelial homeostasis, and may represent an emerging therapeutic modality. However, delivery of miRNAs to cells in vitro and in vivo is challenging due to rapid degradation by ubiquitous RNases. Herein, we developed a nanoparticle delivery system separately combining deacetylated poly-N-acetyl glucosamine (DEAC-pGlcNAc) polymers with miRNA-126-3p and miRNA-126-5p and testing these combinations in vitro and in vivo. Our results demonstrate that DEAC-pGlcNAc polymers have an appropriate size and zeta potential for cellular uptake and when complexed, DEAC-pGlcNAc protects miRNA from RNase A degradation. Further, DEAC-pGlcNAc efficiently encapsulates miRNAs as evidenced by preventing their migration in an agarose gel. The DEAC-pGlcNAc-miRNA complexes were taken up by multiple cell types and the delivered miRNAs had biological effects on their targets in vitro including pERK and DLK-1. In addition, we found that delivery of DEAC-pGlcNAc alone or DEAC-pGlcNAc:miRNA-126-5p nanoparticles to septic animals significantly improved survival, preserved vascular integrity, and modulated cytokine production. These composite studies support the concept that DEAC-pGlcNAc nanoparticles are an effective platform for delivering miRNAs and that they may provide therapeutic benefit in sepsis.
KEY WORDSEndothelial dysfunction Polymicrobial sepsis miRNA delivery Organ dysfunction Inflammation
We thank Ametria Harrison and Pengfei Li for their technical support throughout this project. Additionally, we thank Robin Muise-Helmericks and Amanda LaRue for their experimental expertise and project support.
This work was supported by the NIGMS 1R01GM113995 (HF). This work was also supported by grants NHLBI 5T32HL007260–39 (JJB), 1K23HL135263-01A1 (AG), and UL1 TR 001450 (PVH), and financial and technical supports were provided in part by Marine Polymer Sciences, Inc. Burlington, MA (JV and MD).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
Animal studies were conducted in accordance with the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health and were approved by the Institutional Animal Care and Use Committee at the Medical University of South Carolina.
- 1.Torio CM, Andrews RM. National inpatient hospital costs: the most expensive conditions by payer, 2011: Statistical Brief #160. Healthcare Cost and Utilization Project (HCUP) Statistical Briefs. Rockville (MD), 2006.Google Scholar
- 3.Torio CM, Moore BJ. National inpatient hospital costs: the most expensive conditions by payer, 2013: Statistical Brief #204. Healthcare Cost and Utilization Project (HCUP) Statistical Briefs. Rockville (MD), 2006.Google Scholar
- 12.Fan, H., A.J. Goodwin, E. Chang, B. Zingarelli, K. Borg, S. Guan, P.V. Halushka, and J.A. Cook. 2014. Endothelial progenitor cells and a stromal cell-derived factor-1alpha analogue synergistically improve survival in sepsis. American Journal of Respiratory and Critical Care Medicine 189: 1509–1519.CrossRefGoogle Scholar
- 15.Schober, A., M. Nazari-Jahantigh, Y. Wei, K. Bidzhekov, F. Gremse, J. Grommes, R.T.A. Megens, K. Heyll, H. Noels, M. Hristov, S. Wang, F. Kiessling, E.N. Olson, and C. Weber. 2014. MicroRNA-126-5p promotes endothelial proliferation and limits atherosclerosis by suppressing Dlk1. Nature Medicine 20: 368–376.CrossRefGoogle Scholar
- 17.Vournakis JN, Finkielsztein S, Pariser ER, Helton M. Bicompatible poly-β-1→4-N-acetylglucosamine. Google Patents, 2004.Google Scholar
- 26.Hao, D., W. Xiao, R. Liu, P. Kumar, Y. Li, P. Zhou, F. Guo, D.L. Farmer, K.S. Lam, F. Wang, and A. Wang. 2017. Discovery and characterization of a potent and specific peptide ligand targeting endothelial progenitor cells and endothelial cells for tissue regeneration. ACS Chemical Biology 12: 1075–1086.CrossRefGoogle Scholar