This project will improve our understanding of ALS physiopathology and open new routes toward therapies. The student will be part of a dynamic team that collaborates with an international network (London, Washington DC, Montpellier, Paris).
Rationale: Studies in animal models and ALS patients show that motor neuron degeneration starts at the neuromuscular junction and that post-synaptic muscle changes may play an active role1,2. This axonopathy could be due to the secretion of toxic elements from the muscle. In our lab, we have already shown that ALS muscle cells release toxic exosomes. The purpose of the current project is to explore the in vivo capacity of the muscle exosomes to be transmitted to motor neurons.
Aim1 - Retrograde transport of murine exosomes in vivo: We will express exosomes tagged with a fluorescent marker in murine muscle and track them in vivo. At 5, 10, and 20 days post-injection, the muscles, sciatic nerves and central nervous system will be harvested for immunostaining. This experiment will allow us to determine the speed of diffusion of muscle exosomes in vivo.
Aim2 - Tracking ALS exosomes in vivo: We will track ALS muscle exosomes in vivo and determine whether they preserve their toxic properties.
Aim2a: Exosomal suspensions extracted from muscle cells of ALS patients and healthy subjects will be injected into muscles of immunodeficient mice. Similarly to aim1, various tissues will be harvested post-injection for immunostaining. This experiment will allow us to determine whether ALS muscle exosomes induce motor neuron death in vivo.
Aim2b: The sciatic nerves of mice of several ALS murine models will be harvested at pre-symptomatic ages and tested for muscle exosome markers. This experiment will allow us to determine whether ALS murine models are good models to study the retrograde transport of muscle exosomes. Aim3 - Correcting the toxicity of ALS exosomes in vivo: For this purpose, we will knockdown the expression of mutated SOD1 or FUS or TDP43 in vitro using an siRNA strategy. We will then extract the exosomes from these cells and inject them in vivo as described in Aim2a, to determine whether their toxic effect is abolished. This experiment will be a proof of concept to demonstrate whether targeting the secretion of toxic elements through exosomes could be a good therapeutic strategy.
Skills required: The successful candidate should have some familiarity with technical skills such as immunostaining and cell culture, should have good organizational and planning skills, a rigorous and methodical approach to lab-work, be capable of both independent and team work, and have good skills in writing and presenting data. Experience of the following would be advantageous: PCR and RT-qPCR, and RNA extractions.
References:
1.Fischer, L. R. et al. Exp. Neurol. 185, 232–40 (2004).
2.Tallon, C., Russell, K. A., Sakhalkar, S., Andrapallayal, N. & Farah, M. H. Neuroscience 312, 179–89 (2016).
3. Pradat, P.-F., Kabashi, E. & Desnuelle, C. Curr. Opin. Neurol. 28, 455–61 (2015).
4. Duguez, S. et al. Cell. Mol. Life Sci. (2013).
Applicants should hold, or expect to obtain, a First or Upper Second Class Honours Degree in a subject relevant to the proposed area of study.
We may also consider applications from those who hold equivalent qualifications, for example, a Lower Second Class Honours Degree plus a Master’s Degree with Distinction.
In exceptional circumstances, the University may consider a portfolio of evidence from applicants who have appropriate professional experience which is equivalent to the learning outcomes of an Honours degree in lieu of academic qualifications.
If the University receives a large number of applicants for the project, the following desirable criteria may be applied to shortlist applicants for interview.
The University offers the following levels of support:
The following scholarship options are available to applicants worldwide:
These scholarships will cover full-time PhD tuition fees for three years (subject to satisfactory academic performance) and will provide a £900 per annum research training support grant (RTSG) to help support the PhD researcher.
Applicants who already hold a doctoral degree or who have been registered on a programme of research leading to the award of a doctoral degree on a full-time basis for more than one year (or part-time equivalent) are NOT eligible to apply for an award.
Please note: you will automatically be entered into the competition for the Full Award, unless you state otherwise in your application.
The scholarship will cover tuition fees at the Home rate and a maintenance allowance of £19,000 (tbc) per annum for three years (subject to satisfactory academic performance).
This scholarship also comes with £900 per annum for three years as a research training support grant (RTSG) allocation to help support the PhD researcher.
Due consideration should be given to financing your studies. Further information on cost of living
Submission deadline
Monday 19 February 2018
12:00AM
Interview Date
6, 7 and 8 March 2018
Preferred student start date
Mid September 2018
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