Design of lipid based nanoparticles for gene delivery

Prof. Dr. Jörg Huwyler1)

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1.

Professor Dr. Jörg HuwylerUniversity of Basel, Department of Pharmaceutical Sciences, Division of Pharmaceutical Technology, Klingelbergstr. 50, CH-4056 Basel, Switzerland

People involved

Claudia Lotter (PhD fellow sponsored by the PRC) – University of Basel, Switzerland (claudia.lotter@unibas.ch)

Abstract

In this project, lipid nanoparticles (LNPs) will be designed, which are inspired by the lipid composition of enveloped viruses. They comprise synthetic ionized or ionizable lipids for DNA complexation and an array of natural and synthetic helper lipids for structural integrity, for steric stabilization, and to promote cellular uptake, membrane fusion, and intracellular processing. Preliminary experiments have indeed demonstrated that helper lipids enhance transfection efficiency of such particles by a factor of three.1)2)3) A design of experiment will be applied to identify optimal lipid compositions. LNPs will be combined with novel nanovector plasmid DNA. Such pDNA constructs are devoid of a bacterial backbone sequence and are therefore both smaller in size and less prone to epigenetic silencing as compared to conventional pDNA. These properties allow for a lifetime transfection of target cells. LNPs will be evaluated in vitro using, among others, physicochemical characterization and an array of hepatocyte-derived cell lines. For these experiments, marker expression plasmids will be designed encoding the green fluorescent protein (GFP) or luciferase to allow for convenient visualization and localization of gene products. Furthermore, mechanistic studies will be carried out to gain insight into the role of natural and synthetic phospholipids to highlight their importance as modifiers of gene transfection.

Benefit for the community

In the present work, we propose a novel lipid nanoparticle carrier, which utilizes multiple helper lipids found in viral envelopes to both enhance the binding and internalization of RNA (and eventually DNA) complexed LNPs by target cells. The proposed nanocarrier will include a lipid backbone, based on ionizable lipids in addition to previously unexplored helper lipids. The LNPs are designed to allow for a cost-effective production and a rapid transition from in vitro screening models to in vivo applications. If successful, these formulations might offer new treatment options for patients suffering from rare liver diseases.

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References:
1.
Witzigmann D, Grossen P, Quintavalle C, Lanzafame M, Schenk SH, Tran XT, Englinger B, Hauswirth P, Grünig D, van Schoonhoven S, Krähenbühl S, Terracciano LM, Berger W, Piscuoglio S, Quagliata L, Rommelaere J, Nüesch JPF, Huwyler J, 2021
Non-viral gene delivery of the oncotoxic protein NS1 for treatment of hepatocellular carcinoma
J. Controlled Release 334, 138-152
2.
Nicolle L, Casper J, Willimann M, Journot CMA, Detampel P, Einfalt T, Grisch-Chan HM, Thöny B, Gerber-Lemaire S, Huwyler J, 2021
Development of Covalent Chitosan-Polyethylenimine Derivatives as Gene Delivery Vehicle: Synthesis, Characterization, and Evaluation
Int. J. Mol. Sci. 22, 3828
3.
Witzigmann D, Wu D, Schenk SH, Balasubramanian V, Meier W, Huwyler J, 2015
Biocompatible polymer-Peptide hybrid-based DNA nanoparticles for gene delivery
ACS Appl. Mater. Interfaces 7, 10446-10456
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