Book chapter
Preparation and Characterization of a Liver Targeted, Poly(amidoamine) Based, Gene Delivery System
Non-Alcoholic Steatohepatitis, Vol.2455, pp.319-332
Methods in Molecular Biology, vol 2455, Humana
2022
DOI: 10.1007/978-1-0716-2128-8_24
PMCID: PMC9670859
PMID: 35213004
Abstract
Nonalcoholic steatohepatitis (NASH) is an aggressive liver disease that is considered a major cause of liver cirrhosis and hepatocellular carcinoma. NASH is characterized by multiple underlying genetic mutations, with no approved cure to date. Gene therapies that target those genetic mutations may play a major role in treating this disease, once delivered specifically to the hepatocytes. In this chapter we present, in detail, the synthesis and the characterization of an efficient gene delivery system capable of targeting hepatocytes by exploiting the overexpression of asialoglycoprotein receptors on their cell surface. The targeting ligand, galactose derivative, lactobionic acid (Gal), is first conjugated to bifunctional poly(ethylene glycol) (PEG), and then the formed PEG-Gal is further conjugated to the positively charged polymer, poly(amidoamine) (PAMAM) to form a PAMAM-PEG-Gal construct that can complex and deliver genetic material (e.g., pDNA, siRNA, mRNA) specifically to hepatocytes. We first synthesize PAMAM-PEG-Gal using carbodiimide click chemistry. The synthesized conjugate is characterized using 1H NMR spectroscopy and mass spectrometry. Next, nanoplexes are prepared by combining the positively charged conjugate and the negatively charged genetic material at different nitrogen to phosphate (N/P) ratios; then the size, charge, electrophoretic mobility, and surface morphology of those nanoplexes are estimated. The simplicity of complexing our conjugate with any type of genetic material, the ability of our delivery system to overcome the current limitations of delivering naked genetic material, and the efficiency of delivering its payload specifically to hepatocytes, makes our formulation a promising tool to treat any type of genetic abnormality that arises in hepatocytes, and specifically NASH.
Details
- Title: Subtitle
- Preparation and Characterization of a Liver Targeted, Poly(amidoamine) Based, Gene Delivery System
- Creators
- Kareem Ebeid - Department of Pharmaceutics, Faculty of Pharmacy and Pharmaceutical Manufacturing, Deraya University, New Minia, EgyptSean M Geary - Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, Iowa City, IA, USAAliasger K Salem - Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA, USA. aliasger-salem@uiowa.edu
- Resource Type
- Book chapter
- Publication Details
- Non-Alcoholic Steatohepatitis, Vol.2455, pp.319-332
- Series
- Methods in Molecular Biology; vol 2455
- DOI
- 10.1007/978-1-0716-2128-8_24
- PMID
- 35213004
- PMCID
- PMC9670859
- NLM abbreviation
- Methods Mol Biol
- eISSN
- 1940-6029
- Publisher
- Humana; New York, NY
- Language
- English
- Date published
- 2022
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Dental Research; Chemical and Biochemical Engineering
- Record Identifier
- 9984221630302771
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