Delivery of Raman active nanoparticles to dopaminergic neurons to investigate age-related protein aggregation

Supervisors: Prof. Alison Hulme, Dr Tilo Kunath, Dr Mathew Horrocks

Project Description:

This project will investigate age-associated protein aggregation within neurons using Raman active ‘loaded’ nanoparticles delivered both to human dopaminergic neurons in culture, and into the brain of a genetic rat model that has α-synuclein aggregation.

The first aim of the project will be to produce and test Raman active polymer analogues of poly(lactic-co-glycolic acid) to manufacture the nanoparticles. These particles will be functionalised with surface proteins, such as lactoferrin, to specifically target them to dopaminergic neurons (Hulme Group). The Raman active and standard nanoparticles loaded with fluorescent dyes will be used to treat a mixture of human stem cell-derived cortical and dopaminergic neurons to investigate neuronal specificity, and to determine the optimum imaging modality. The nanoparticles will further be used for intranasal delivery in rats to investigate regional brain uptake (Kunath Group).

The second aim of the project is to investigate the delivery of cargo with diagnostic potential. The PhD student will load the nanoparticles with validated DNA aptamers that specifically bind to α-synuclein oligomers (Horrocks Group). The DNA aptamers will be covalently linked to fluorescent dyes or to small active Raman tags with a different vibrational frequency to the nanoparticle polymer. The latter will be used for ‘dual colour’ imaging with Stimulated Raman Scattering microscopy [1]. This diagnostic α-synuclein tool will be used to detect α-synuclein aggregates in stem cell-derived dopaminergic neurons with familial PD mutations [2], and delivered intranasally to ageing rats with an α-synuclein mutation that causes protein aggregation. Finally, the functionalized nanoparticles will be loaded with small molecule modulators of α-synuclein aggregation to investigate their ability to alter the state of the protein in human neurons and in the rat α-synuclein model.

Working in world-leading laboratories at Edinburgh, the student will receive training across the fields of chemistry, cell biology and imaging. The ideal applicant will have a strong academic record, and a BSc or MChem/MSc degree in Chemistry, Biochemistry or related fields, with experience of working in an interdisciplinary environment.

[1] Tipping WJ, Lee M, Serrels A, Brunton VG, Hulme AN. (2017) Imaging drug uptake by biorthogonal stimulated Raman scattering microscopy. Chem Sci. 8:5606-5615.
[2] Devine MJ, Ryten M, Vodicka P, Thomson AJ, Burdon T, Houlden H, Cavaleri F, Nagano M, Drummond NJ, Taanman JW, Schapira AH, Gwinn K, Hardy J, Lewis PA, Kunath T. (2011) Parkinson's disease induced pluripotent stem cells with triplication of the α-synuclein locus. Nat Commun. 2:440.

To apply for this project, please go to this link.