Project ID BE-MI2023_13

ThemeBE-MI

Co Supervisor 1A School of Biomedical Engineering & Imaging SciencesWebsite

Co Supervisor 1B Wolfson Centre For Age-related DiseasesWebsite

Gene therapy of traumatic brain injury using focused ultrasound

Background: Traumatic brain injury (TBI)is a leading cause of death and disability, with no effective restorative treatment to date. Current approaches include direct administration of viral vectors encoding proteins to promote neuroplasticity and neurogenesis. However, the blood-brain barrier (BBB) precludes systemic treatments. Here, we propose to develop focused ultrasound (FUS)-mediated gene therapyfor treating TBI. We will deliver adeno-associated virus (AAV)-encoded growth factors/enzymes using FUS andmicrobubbles. Our goal is to developtargeted gene therapy in brains with TBI, topromote neurorestoration in a non-invasive and translatablemanner.

Aims1) Deliver AAV-encoded growth factors into TBI lesions. AAVs encoding a reporter gene (e.g., mCherry) and brain-derived neurotrophic factors (BDNF)or chondroitinase ABC (Ch-ABC)will be delivered in wild-type and TBI miceusing FUS. We will use cortical contusion injury(CCI)as a TBI model. 2) Assess the neuroregenerative potential of FUS-mediated genedelivery. Neurorestoration will be evaluatedthrough BrdU and DCX staining. 3) Assess behavioural outcomes of FUS-mediated gene therapyin TBI mice. TBI mice will undergo behavioural tests,such as grasp-test, beam-walk, and rotarod, before/after TBI induction. We will sacrifice the mice and stain for neuronal density in the lesion/perilesional area.

Skills: In vivoexperiments, FUStechniques,TBI surgery,immunohistochemistry, microscopy, behavioural experiments

Objectives: Year 1 –Complete training on in vivoexperiments, FUS methods, CCI model, etc.Year 2 –Confirm gene delivery/expression in wild-type and TBI mice through fluorescence microscopy.Year 3 –Evaluate neurogenesis and neuroregeneration through immunohistochemistry.Year 4 –Measure behavioural improvement after gene therapy.

One representative publication from each co-supervisor:

Pouliopoulos A N, Kwon N, Jensen G, Meaney A, Niimi Y, Burgess M T, McLuckie A J, Munoz F, Ji R, Kamimura H A S, Teich A F, Ferrera V P, Konofagou E E 2021 Safety evaluation of a clinical focused ultrasound system for neuronavigation guided blood-brain barrier opening in non-human primates. Scientific Reports 11,15043.

Francos-Quijorna I, Sánchez-PetidierM, BurnsideER, BadeaSR, Torres-EspinA, MarshallL, de WinterF, VerhaagenJ, Moreno-Manzano V, BradburyEJ2022 Chondroitin sulfate proteoglycans prevent immune cell phenotypic conversion and inflammation resolution via TLR4 in rodent models of spinal cord injury. Nature Communications 13,2933

Ahmed A. I., Shtaya A. B., Zaben M. J., OwensE. V., Kiecker C., & Gray W. P. 2012Endogenous GFAP-positive neural stem/progenitor cells in the postnatal mouse cortex are activated following traumatic brain injury. Journal of Neurotrauma29,5,828–842.