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.

One representative publication from each co-supervisor:

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.