Aav-based modulation of gba1 and related compositions and uses thereof
AssigneeAspen Neuroscience, Inc.
InventorsAndres BRATT-LEAL, Ai ZHANG
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Abstract
The present disclosure provides adeno-associated vector (AAV)-based methods of increasing gene expression of a GBA1 in floor plate midbrain progenitor cells, determined dopamine (DA) neuron progenitor cells, and/or DA neurons, or glial cells, including those differentiated from pluripotent stem cells, and methods of lineage specific differentiation of the same. Also provided are compositions and uses thereof, such as for treating neurodegenerative diseases and conditions, including Parkinson's disease.
Claims111
A method of increasing expression of GBA1 in a cell, the method comprising:
- introducing, into a neurally differentiated cell, a recombinant adeno-associated viral (rAAV) vector comprising a promoter operably linked to a nucleic acid sequence encoding GBA1,
- wherein the introducing results in increased expression of GBA1 in the cell.
A method of increasing expression of GBA1 in a cell, the method comprising:
- introducing, into a neurally differentiated cell, a recombinant adeno-associated viral (rAAV) vector comprising a promoter operably linked to a nucleic acid sequence encoding GBA1,
- wherein the cell comprises a variant of GBA1 associated with Parkinson's Disease, and the introducing results in increased expression of GBA1 in the cell.
A method of differentiating neural cells, the method comprising:
- (a) performing a first incubation comprising culturing pluripotent stem cells (PSCs) in a non-adherent culture vessel under conditions to produce a cellular spheroid, wherein beginning at the initiation of the first incubation (day 0) the cells are exposed to (i) an inhibitor of TGF-β/activin-Nodal signaling;
- (ii) at least one activator of Sonic Hedgehog (SHH) signaling;
- (iii) an inhibitor of bone morphogenetic protein (BMP) signaling;
- and (iv) an inhibitor of glycogen synthase kinase 3β (GSK3β) signaling;
- (b) performing a second incubation comprising culturing cells of the spheroid in a substrate-coated culture vessel under conditions to neurally differentiate the cells;
- and
- (c) introducing into the neurally differentiated cells a recombinant adeno-associated viral (rAAV) vector comprising a promoter operably linked to a nucleic acid sequence encoding GBA1, wherein the introducing results in increased expression of GBA1 in the cell.
A modified neurally differentiated cell comprising an exogenous deoxyribonucleic acid (DNA) sequence encoding GBA1, wherein the DNA sequence is episomal in the cell.
A recombinant adeno-associated viral (rAAV) nucleic acid vector for increasing expression of GBA1 in a cell, the vector comprising a promoter operably linked to a nucleic acid sequence encoding GBA1, wherein the cell exhibits (i) reduced activity of the β-Glucocerebrosidase (GCase) enzyme encoded by GBA1 and/or (ii) reduced expression of GBA1 prior to being introduced with the rAAV vector, optionally as compared to a reference cell from a subject without Parkinson's Disease.
Dependencies are inferred from the claim text.