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Project title
Developing neuron-targeting lipid nanoparticles for translational applications: proof of concept in Niemann-Pick C disease.
Supervisors names
Julien Baruteau
Naresh Hanchate
Background
Among 3,000 cerebral genetic diseases, 75% of them affect children causing severe morbidity and early death. Cerebral inherited metabolic diseases (CIMD) are caused by the disruption of essential biochemical pathways, causing mostly diffuse neurological diseases. Less than 5% of these diseases have approved disease-changing therapies. Therefore, there is high unmet need for novel therapies.
Gene therapy applications mediated by viral vectors (GT-VV) have achieved clinical successes in various diseases but they have limitations including ongoing adverse events such as immunogenicity and insertional mutagenesis. Due to these potential adverse events, GT-VV-treated patients are often underdosed and do not have a full correction of the diseases and remain with severe neurodebilitating disease.
Messenger RNA (mRNA) replacement therapy is a novel gene therapy technology, which has shown its potent efficacy and high safety profile during the COVID-19 pandemic. mRNA therapy is actively being developed in many areas including rare genetic diseases. mRNA is encapsulated in lipid nanoparticles (LNPs) to prevent rapid degradation by RNases. Neither mRNA nor LNP trigger sustained immune response, enabling successful re-administration and adaptation of doses, overcoming GT-VV limitations. LNPs composition modify their biodistribution and improves efficacy. In vitro LNP screening does not correlate with in vivo findings.
Niemann Pick C disease (NPC) is a lysosomal storage disorder with 95% caused by NPC1 deficiency and subsequent deficient lipid export from endolysosomal compartment. This causes a severe neurodegenerative disease with early dementia, epilepsy and motor symptoms.
In this proposal, the applicant will create a library of barcoded mRNAs encapsulated in LNPs with different composition and addition of peptide-targeting ligands specific for neuronal cells and blood brain barrier crossing. In vivo screening using single cell RNA sequencing (scRNAseq) following intracerebroventricular and systemic injections, the latter testing blood brain barrier crossing. The applicant will then show proof of concept in NPC mice to test the therapeutic effect of this optimised LNP construct after mRNA NPC1 optimisation. This applicant will become proficient in 2 rapidly evolving key methods in Biological Sciences: LNP-mRNA and scRNAseq.
Aims/Objectives
mRNA-LNP has a transformative potential for cerebral monogenic diseases. The aim of this project is to screen for an optimal LNP able to target the brain and show proof of concept in NPC disease.
The objectives are
- Objective 1: Development and characterisation of LNP library
- Objective 2: Development and validation of neuronal-targeting scRNAseq-based screening
- Objective 3: Validation in CIMD mouse model, Niemann Pick C disease
Methods
- Objective 1: Molecular biology, cloning, cell culture, mRNA-LNP production and characterisation
- Objective 2: FACS, scRNAseq (from library preparation to post-processing and analysis), immunostaining.
- Objective 3: Animal work, genotyping, behaviour tests, immunostaining, mass spectrometry
Timelines
- OBJECTIVE 1: Months 1-12
- OBJECTIVE 2. Months 13-24
- OBJECTIVE 3. Months 18-30
- THESIS WRITING. Months 30-36
References
1. Hanchate et al. Science 2015
2. Hanchate et al. PNAS 2020
3. Seker et al, Int J Mol Sci 2020
4. Grant-Serroukh et al. J Control Release 2022
5. Gurung et al.2022, BioR
Contact
Julien Baruteau, j.baruteau@ucl.ac.uk