ABSTRACT
Developing vehicles that efficiently deliver genes throughout the human central nervous system (CNS) will broaden the range of treatable genetic diseases. We engineered an adeno-associated virus (AAV) capsid, BI-hTFR1, that binds human transferrin receptor (TfR1), a protein expressed on the blood-brain barrier. BI-hTFR1 was actively transported across human brain endothelial cells and, relative to AAV9, provided 40 to 50 times greater reporter expression in the CNS of human TFRC knockin mice. The enhanced tropism was CNS-specific and absent in wild-type mice. When used to deliver GBA1, mutations of which cause Gaucher disease and are linked to Parkinson's disease, BI-hTFR1 substantially increased brain and cerebrospinal fluid glucocerebrosidase activity compared with AAV9. These findings establish BI-hTFR1 as a potential vector for human CNS gene therapy.
Subject(s)
Antigens, CD , Brain , Capsid , Gene Transfer Techniques , Genetic Vectors , Glucosylceramidase , Receptors, Transferrin , Animals , Humans , Mice , Antigens, CD/metabolism , Antigens, CD/genetics , Blood-Brain Barrier/metabolism , Brain/metabolism , Capsid/metabolism , Capsid Proteins/metabolism , Capsid Proteins/genetics , Dependovirus , Endothelial Cells/metabolism , Gene Knock-In Techniques , Genetic Therapy , Receptors, Transferrin/metabolism , Receptors, Transferrin/genetics , Glucosylceramidase/genetics , Gaucher Disease/genetics , Gaucher Disease/therapy , Parkinson Disease/genetics , Parkinson Disease/therapyABSTRACT
Developing vehicles that efficiently deliver genes throughout the human central nervous system (CNS) will broaden the range of treatable genetic diseases. We engineered an AAV capsid, BI-hTFR1, that binds human Transferrin Receptor (TfR1), a protein expressed on the blood-brain barrier (BBB). BI-hTFR1 was actively transported across a human brain endothelial cell layer and, relative to AAV9, provided 40-50 times greater reporter expression in the CNS of human TFRC knock-in mice. The enhanced tropism was CNS-specific and absent in wild type mice. When used to deliver GBA1, mutations of which cause Gaucher disease and are linked to Parkinson's disease, BI-hTFR1 substantially increased brain and cerebrospinal fluid glucocerebrosidase activity compared to AAV9. These findings establish BI-hTFR1 as a promising vector for human CNS gene therapy.
