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A dual AAV strategy for the treatment of Dravet Syndrome

Antinao Diaz, Juan Francisco; (2020) A dual AAV strategy for the treatment of Dravet Syndrome. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Dravet Syndrome (DS) is an inherited childhood epilepsy caused by a mutation in the SCN1A gene, which encodes the voltage-gated sodium channel NaV 1.1. It has an incidence rate of 1:16,000 live births; patients suffer from refractory and generalized seizures that can evolve into status epilepticus, which can result in premature death. The disease is also associated with significant cognitive impairments. DS remains untreatable by either medical or surgical means. Gene therapy for DS is challenging; we and others (Feldman and Lossin, 2014) have experienced difficulties in propagating wild-type SCN1A plasmids in E.coli competent cells. Furthermore, the coding sequence of SCN1A (6kb) prevents its incorporation into an adeno-associated virus vector (AAV). This thesis describes an approach to overcome these problems by splitting SCN1A into two complementary halves. The hypothesis is that the dual AAV8-SCN1A constructs would produce polypeptides which would reconstitute post-transcriptionally into a functional NaV 1.1 protein (Stühmer et al., 1989). Two AAV8 vectors were designed; the two halves of SCN1A were driven by a pan neuronal promoter, human Synapsin. In vitro, co-transfection appeared toxic for cells, in a subset of recordings a small sodium current was observed. A knock-out mouse model was established and characterised to test these vectors in vivo. Unfortunately the treatment did not modify the phenotype of the knockout mice. I propose this was the result of insufficient number of cells expressing both vectors or that the split channel does not have the same characteristics as its full-length counterpart.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: A dual AAV strategy for the treatment of Dravet Syndrome
Event: UCL
Language: English
Additional information: Copyright © The Author 2020. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request.
Keywords: Dravet Syndrome, Gene Therapy, SCN1A, AAV
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL EGA Institute for Womens Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL EGA Institute for Womens Health > Maternal and Fetal Medicine
URI: https://discovery.ucl.ac.uk/id/eprint/10110330
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