Project ID NS-MH2023_53


Co Supervisor 1A IoPPN/Basic and Clinical NeuroscienceWebsite

Co Supervisor 1B FoLSM/Institute of Pharmaceutical ScienceWebsite

Using human stem cell-derived neurons and TDP-43 knock-in mice to test therapies for ALS

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease characterised by misregulation of the RNA binding protein TDP-43. A major challenge has been the lack of good TDP-43 disease models, thereby limiting ‘bench to bedside’ success. However, the use of CRISPR genome engineering technology now means we can create ‘knock-in’ models, which recapitulate human disease more closely. The Sreedharan laboratory and others are using CRISPR to create both mouse and human stem cell models of ALS that harbour disease-linked TDP-43 mutations. Together with the Al-Jamal lab, who have created advanced therapies and intranasal delivery approaches, these models are being used to understand disease mechanism and develop treatments for ALS.

To derive TDP-43 knock-in human stem cell-derived neurons and test the efficacy of a range of drugs in vitro
To take the best candidate from the in vitro assays and test this in a TDP-43 knock-in mouse.

Skills the student will acquire:
Molecular biology, CRISPR
Biochemistry, RNA sequencing
Mouse husbandry, behavioural analysis, drug administration
Formulation and characterisation of nanomedicines
Drug quantification in tissues
Histology, microscopy

Proposed timeline:
Year 1 – Establish human stem cell lines with a novel TDP-43 mutation, derive and characterise neurons.
Year 2 – Treat neurons with candidate drugs. Identify best candidate, develop a formulation suitable for delivery to mice.
Year 3 – Expand colony of TDP-43 knock-in mutant mice. Administer drug. Analyse motor function. Harvest tissues.
Year 4 – Process tissues for histology, RNA sequencing to determine effect of drug on mitigating disease phenotypes.

Representative Publications

White MA, Kim E, Duffy A, Adalbert R, Phillips BU, Peters OM, Stephenson J, Yang S, Massenzio F, Lin Z, Andrews S, Segonds-Pichon A, Metterville J, Saksida LM, Mead R, Ribchester RR, Barhomi Y, Serre T, Coleman MP, Fallon J, Bussey TJ, Brown RH Jr, Sreedharan J*. TDP-43 gains function due to perturbed autoregulation in a Tardbp knock-in mouse model of ALS-FTD Nat Neurosci. 2018 Apr;21(4):552-563

Wang JT, Rodrigo AC, Patterson AK, Hawkins K, Aly MMS, Sun J, Al Jamal KT*, Smith DK*. (2021) Enhanced Delivery of Neuroactive Drugs via Nasal Delivery with a Self-Healing Supramolecular Gel. Advanced Science (Weinh). e2101058. doi: 10.1002/advs.202101058.