Development of Novel Cell Free Therapies to Prevent Age-related Motor Neuron loss

Addressing the adverse effects of ageing is a priority research area not only of the BBSRC but also of the Industrial sector. This project aims to develop new and novel cell free reagents to prevent age-induced motor neuron loss by forging an alliance between academia (University of Reading) and industry (NATURAL BIOSCIENCES LTD) to exploit a new finding made at the University of Reading. An extensive body of literature exists showing that as age increases beyond 60 years, human muscle undergoes denervation, due to a loss of MN that leads to a poor quality of life and often death. Permanently denervated muscle is ultimately lost and replaced by fat and fibrous tissue. This project will exploit our timely finding that deletion of the gene Myostatin in the mouse results in an unprecedented survival of MN, which are normally eliminated by programmed cell death during (Elashry et al 2010 Journal of Anatomy, submitted). Since the myostatin mutation only affects skeletal muscle, we have developed the central hypothesis 'that the skeletal muscle produces diffusible trophic factors that act as neuro-protective agents that prevent age related motor neuron death'. The hypothesis will be tested by the student. This study will take advantage of a strategic alliance between the groups of Professor Patel, who at the University of Reading have made this key discovery and Dr Steve Ray of NATURAL BIOSCIENCES LTD who brings to the research arena extensive (including expertise development of cell free therapies to promote neural regeneration) and research reagents (mouse lines displaying premature MN death, including SODG93A mouse line. Project aims: (1) To determine using in-vitro platforms whether MN that develop in the absence of myostatin are resistant to neuro-toxins. (2) To determine in-vivo whether the ageing resistant MN that form in the absence of myostatin display the survival characteristics of young MN. (3) To conduct in-vitro experiments to determine whether cell free reagents (conditioned media, micro vesicles, RNA) generated from myostatin null tissue harbour neuro-protective properties. (4) To determine whether cell free reagents developed in aim (3) offer neuro-protective properties in an in-vivo context. Timeline: 0-9 months. Isolation and culturing foetal MN from wild type and myostatin null mice by standard methods (Kume et al (2005) Neuroscience Letters 383:199-202). Training: cell culture, working with transgenic animals. Venue: University of Reading 10-18 months. Neuroprotective properties of MN from myostatin nulls to exposure to glutamate or nitric oxide agonists will be established. Training: cell culture, statistical analysis of data. Venue: NATURAL BIOSCIENCES LTD 12-18 months. Establish myostatin/SODG93A line. Training: animal handling, molecular biology. Venue: University of Reading 19-30 months. Histological examination of motor nerves in myostatin/ SODG93A line. Training: con-focal microscopy, electron microscopy. Venue: University of Reading 28-33 months. Isolation of cell free biologically active reagents (conditioned media, micro vesicles, RNA) from myostatin null muscle. Training: molecular biology, biochemistry. Venue: NATURAL BIOSCIENCES LTD 28-36 months. Confirm the neuroprotective properties of cell free reagents derived from myostatin null mice to prevent the toxic effects of glutamate or nitric oxide agonists. Training: toxicology, statistics. Venue: NATURAL BIOSCIENCES LTD 36-48 months. Quantify evidence of neuroprotection of cell free reagents from myostatin nulls in SODG93A mice. Training: histology. Venue: University of Reading The student will spend about each 24 months in the laboratory of Professor Patel and NATURAL BIOSCIENCES LTD. Natural Biosciences SA will provide £15,000 per year for animal, tissue culture, reagent and travel costs. Outputs from this project will be used for a future project grant application- details in later sections.