NSW-RPRN Research Pillar

Pilot research projects to seed the NSW RNA ecosystem.

Project 1 - Lung and nasal delivery of m/siRNA-LNPs

AIM:

This work aims to assess and optimise the aerosol performance of a series of lipid-nanoparticle siRNA delivery systems currently being developed at several collaborating institutions (the Kirby Institute, UNSW, USYD and CCI) to target the SARS-CoV-2 virus in the respiratory tract (nose and lung) using state of the art techniques developed at the Woolcock Institute and Macquarie University. 

METHODS:

The Woolcock/MQ Team is fully dedicated to formulating, delivering and assessing the aerosol and anti-viral performance and activity of the nano-system to the respiratory tract (using model SARS-CoV-2 virus that causes COVID-19). 

KEY OBJECTIVES: 
  1. To work in collaboration with the other project leaders in the Production Pillar (UNSW, USYD and CMRI at Westmead) to formulate the siRNA delivery system for nasal and lung delivery, assessing aerosols performances and bioactivity in vitro using physiologically relevant respiratory models developed in house. 

  1. Scale-up and manufacture of optimised, proof-of-principle mRNA lipid nanoparticles (NPs) to be assessed in vitro for their aerosol performance. 

  1. Test the efficacy of these mRNA and siRNA-loaded (lipid) NPs systems in vitro and in vivo (mouse) models. 

OUTCOMES:

Two optimised, inhalable anti-viral siRNA (lipid NPs suspended for nebulization; and lyophilized NPs for extemporaneous reconstitution prior to nebulization or dry powder) and generic mRNA NP systems for COVID treatments fully characterised in vitro, ready to progress for pre-clinical animal testing. 

KEY INSTITUTIONS:

Macquarie University, the Woolcock Institute of Medical Research, Kirby Institute, Children’s Cancer Institute, UNSW Sydney, Centenary Institute, University of Sydney, Children’s medical Research Institute Westmead 

 

Project 2 – Treatment of SARS-CoV-2 (COVID-19) infections with siRNA-LNPs 

AIM:  

The aim of this work is to upscale and optimise the performance of an siRNA delivery system targeting the SARS-CoV-2 virus that is currently being developed at the Kirby Institute. 

METHODS:  

The Kirby Institute has recently identified a number of antiviral siRNA’s that show a broad-range beta-Coronavirus antiviral activity, including a number of the key variants (alpha, delta…) of the SARS-CoV-2 virus that causes COVID-19. 

KEY OBJECTIVES:  
  1. Provide the project leaders with sufficient quantities of high-quality siRNA. Through the NSW-RPRN supported synthetic RNA facility at UNSW (Production Pillar), we will upscale the synthesis of these siRNA to enable a broader range of pre-clinically relevant siRNA sequences. 

  1. Optimise the formation of siRNA lipid nanoparticles. Initially the focus will be on commercially available lipid nanoparticle formulations, but other polymers and related materials will be sourced from a number of external collaborators and tested.  

  1. Test the efficacy of these siRNA-loaded (lipid) nanoparticle systems in vitro and in vivo (mouse) models. 

OUTCOMES:  

An optimised siRNA nanodelivery system that allows the researchers to carry out larger scale pre-clinical animal trials with their antiviral siRNA as a COVID-19 treatment. 

KEY INSTITUTIONS:  

Kirby Institute, Children’s Cancer Institute, UNSW Sydney, University of Technology Sydney, Centenary Institute 

Project 3 - CRISPR gene correction in a genetic disease mouse model 

AIM: 

Establish production and delivery of an mRNA encoding the CRISPR genome editing enzyme into human cells and demonstrate functional gene editing by reversing the deleterious phenotype in a genetic disease mouse model. 

METHODS: 

The Gene Therapy Research Unit (GTRU) at Children’s Medical Research Institute (Westmead) headed by Prof. Ian Alexander, has used CRISPR-based genome editing to correct patient-derived primary human hepatocytes in vivo. The patient hepatocytes were used to successfully repopulate the murine liver, generating a powerful system for evaluating human-specific therapies. The levels of correction in their study would be curative if recapitulated in the clinic and was achieved using elite vector-based gene delivery. The disease model, Ornithine transcarbamylase (OTC) deficiency, leads to hyperammonemia (elevated blood ammonia), which is neurotoxic and results in brain damage or death shortly after birth.  

KEY OBJECTIVES: 
  1. Generate and upscale sufficient quantities of high-quality mRNA for the preclinical studies 

  1. Develop a liposomal nanoparticle system capable of delivering the mRNA. 

  1. Encapsulate an mRNA encoding the CRISPR Cas9 editing enzyme together with the CRISPR guide RNA within a lipid nanoparticle for in vivo delivery to chimeric human-mouse livers. 

OUTCOMES: 

Development of gene therapy delivery for a rare disease that has broader application to other challenging diseases. An established mRNA-lipid nanoparticle platform that enables researchers within the NSW RPRN to apply across different disease models being researched across the network. 

KEY INSTITUTIONS:  

Children’s Medical Research Institute, Children’s Cancer Institute, University of New South Wales