Objective/Rationale:
The aim of this project is to create a model of Parkinson’s disease using skin cells from patients who have an inherited form of Parkinson’s disease. Using newly developed stem cell techniques, we can then convert these into a type similar to the brain cells that die in Parkinson’s disease, opening a window on what is actually happening within the brains of people who have this disorder.
Project Description:
After identifying patients carrying mutations in the LRRK2 gene, which causes Parkinson’s disease, we will take a small sample of skin from these individuals and grow the cells from this sample. By introducing a number of factors into the cells using viruses, we can convert them into a stem cell like state. Once they have been reverted to stem cells, they can be differentiated into any number of cell types. In the case of Parkinson’s disease we are interested in the specific cells within the substantia nigra in the brain that die in this disease, so we will use a further set of conditions and growth factors to induce the stem cells to adopt a neuronal identity.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
This project will, for the first time, allow us to look directly at the cells that die in Parkinson’s disease from patients who have a genetic form of the disease. By comparing these cells to those from individuals that do not have a genetic disposition to Parkinson’s disease, we can use a range of experimental techniques to investigate what makes these cells different, and more susceptible to a premature demise. This, in turn, will uncover novel pathways and targets for drug treatment in Parkinson’s disease.
Anticipated Outcome:
At the end of this project, we will have developed a novel model for Parkinson’s disease, from which we can learn more about the mechanism of this disorder - knowledge that is essential if we are to develop new treatments. In addition to providing a valuable resource for basic research into Parkinson’s, this model will also provide an important testing ground for therapies that seek to reverse any deficits that we uncover.
Final Outcome
Dr. Lewis obtained fibroblasts from both idiopathic and Lrrk2 PD patients as well as controls and has so far generated iPS cell lines from a patient carrying the R1441G mutation and one sibling control. Dr. Lewis is currently collaborating with Dr. Chandran of Cambridge to differentiate these lines into dopaminergic neurons.
Following up on the funding from the Fox Foundation, we have been awarded a clinical research fellowship from the UK Medical Research Council (to Dr Mike Devine, £213,825 over three years) and a post doctoral fellowship from the UK Alzheimer's research trust (to Dr Selina Wray, £161,951 over three years) to further extend the iPS experiments.
We have also received $642,790 from the Fox foundation to use the fibroblasts, in combination with LRRK2 mutation brain tissue in the Queen Square Brain Bank, to investigate signaling dysfunction due to mutations in LRRK2 and to examine potential substrates in a disease setting. Using mRNA isolated from LRRK2 mutation fibroblasts, we have generated whole genome expression data to investigate alterations due to mutation, and from differentiated embryonic stem cells and wild type iPS cells - which we are currently comparing to gene expression from post mortem human substantia nigra - to validate the experimental system.