Ph.D position proposal: Microfluidic and biophysical modeling of human neurodegenerative diseases: Impact of RNA transport in the neuromuscular junction. Location : IGMM, Institut de Génétique Moléculaire de Montpellier. Montpellier, France Schedule : October 2018 Advisors : Florence RAGE, Benoit CHARLOT “NMJ-On-Chip” is a cooperative research project funded by the iSite MUSE that gathers several research teams within the University of Montpellier. The goal of this project is to reconstruct a neuro-muscular junction (NMJ) from human muscular cells and motor neurons derived from human induced Pluripotent Stem Cells (iPSC), on an instrumented microfluidic chip (Fig1) to understand the molecular mechanisms of motor neuron diseases such as Spinal Muscular Atrophy (SMA) or Amyotrophic Lateral Sclerosis (ALS, Charcot disease).

One of the major issues with neurodegenerative diseases is the difficulty to directly observe in-vivo biochemical and electrophysiological mechanisms involved in neuronal dysfunctions. One of the solutions is to reconstitute an in-vitro model recapitulating these pathological junctions using cultured cells. It will allow us to take advantage of advanced live cells imaging techniques to directly observe under the microscope the relationships between maturation, electrical activity, synapse formation, intracellular dynamics and vesicle transport. In particular, the project will aim at studying the impact of mRNA transport defect on the formation and stability of the NMJ.

The in vitro reconstruction of these neuromuscular junctions faces many challenges that microfluidics and micro-biotechnologies can manage. This is precisely what we intend to implement as part of a multidisciplinary and innovative research proposal. The study of neuromuscular junctions is usually the result of a biologists' consortium work using conventional molecular biology tools and techniques, either in genetic engineering or in microscopy. Indeed, the use of compartimentalized microfluidics coupled with microelectrode arrays (MEAs) and substrate microstructuration will allow reconstructing the spatial organisation, as well as the mechanical environment and electrical stimulations as close as possible to the in-vivo situation. In summary : - A public health problem: Degenerative diseases of the neuromuscular junction: Spinal Muscular Atrophy (SMA) and Amyotrophic Lateral Sclerosis (ALS: Charcot Disease). - A multidisciplinary international competent consortium of experts in biology, microtechnologies, microfluidics and biophysics already working together with preliminary results. - Advanced microfluidic technologies for in vitro neuromuscular junction reconstruction with microelectrode and electronics integration. - An entire-human model, derived from motor neuron-derived iPS cells and skeletal muscles to decipher molecular mechanisms for greater biological relevance. In the frame of the NMJ-on-chip project we are looking for a Ph.D student candidate that will take in charge parts of the project. The student will be located at IGMM laboratory but will also work in close cooperation with project partners such as IES and Phymedexp.

Figure 1. Schematic of the principle of the reconstruction of a neuromuscular junction in a microfluidic chip

Schematic of the NMJ on chip project Candidate profile : We are looking for a student with a background in biology and neurosciences with knowledges in microscopy. The Ph.D student will be in charge of: - Cell culture : iPS cells derived motoneurons and myocyte co-culture in microfluidic environment (in partnership with Phymedexp) - Microscopy and fluorescent labelling: confocal microscopy, time lapse, calcium imaging - Electrophysiology : Stimulation and recording of action potentials and electrical activities in co culture. (in partneship with IBMM) - Microfluidic devices preparation (in partneship with IES) - Transport analysis with videomicroscopy. Contact : Florence Rage (IGMM) florence.rage@igmm.cnrs.fr Benoit CHARLOT (IES) benoit.charlot@um2.fr