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.