Can lowering mtHTT in human HD patient iPSC-derived neurons deliver potential benefit by alleviating HD-relevant neuronal disease phenotypes?

Academic Supervisor: Prof Paul J Kemp, Cardiff University
Industrial Supervisor: Dr George McAllister, Biofocus, Cambridge

In Huntington’s Disease (HD), expression of mutant huntingtin protein (mtHTT) leads to selective death of striatal medium spiny neurons (MSNs). This project will bring together expertise and two platform technologies from Cardiff and Biofocus, which together will enable high-content screening and target validation of drugs for lowering mtHTT/HTT in HD patient-derived cells. Biofocus have developed a FRET-based antibody triplex assay that detects the mtHTT:HTT ratio in non-excitable cells, and have lead compounds from a targeted library screen which lower mtHTT. This assay will now be translated for target validation, neuroprotection and further screening in HD neurons differentiated from patient-derived iPSCs. PJK and NDA (Cardiff) are partners in an international HD consortium that has generated an allelic series of patients induced pluripotent stem cells (iPSCs) with polyQ repeat lengths of 21, 33, 45, 50, 60, 109 and 180 and they have a patented a small molecule-based differentiation protocol that results in networked neurons with high synaptic activity, capable of initiating trains of fast action potentials. Critically, these neurons show many polyQ-dependent HD phenotypes, including differential excitotoxicity and mitochondrial dysfunction.

The project will test the hypothesis that lowering mtHTT in HD patient iPSC-derived neurons will rescue HD neuronal phenotypes.
Therefore, the student will develop expertise in iPSC culture, differentiation and CRISPR genome editing, to make reporter lines for functional studies and high-content assay development. S/he will translate assays for HTT lowering to iPSC-neurons and using the tools developed will be directly involved drug screen design and validation of target hits.

The student will meet the following milestones:

1. To develop a disease-relevant platform for screening new drug libraries;
2. To compare the efficacy of lead compounds with the current state-of-the-art antisense oligonucleotides;
3. To determine the effects of lead mtHTT lowering compounds on glutamate excitotoxicity in high throughput.

Lead compounds should be in pre-clinical trial by the end of the project.

The academic entry requirement is the equivalent of an upper second class Honours degree in a related science subject. If English is not your first language, you will need an overall score of 6.5 in IELTS with scores of 5.5. in each sub area or an equivalent qualification. For further information about English Language qualifications please see http://www.cardiff.ac.uk/for/prospective/international/english-language-requirements.html