Delivery of single-chain antibodies (scFvs) directed against the 37/67 kDa laminin receptor into mice via recombinant adeno-associated viral vectors for prion disease gene therapy.
The 37/67 kDa laminin receptor (LRP/LR) acts as a receptor for prions providing a promising target for the treatment of prion diseases. Recently, we selected anti-LRP/LR single-chain antibodies (scFvs) and proved a reduction of the peripheral PrP(Sc) propagation by passive immunotransfer into scrapie-infected mice. Here, we report the development of an in vivo gene delivery system based on adeno-associated virus (AAV) vectors expressing scFvs-S18 and -N3 directed against LRP/LR. Transduction of neuronal and non-neuronal cells with recombinant (r)AAV serotype 2 vectors encoding scFv-S18, -N3 and -C9 verified the efficient secretion of the antibodies. These vectors were administered via stereotactic intracerebral microinjection into the hippocampus of C57BL/6 mice, followed by intracerebral inoculation with 10 % RML at the same site 2 weeks post-injection of rAAV. After 90 days post-infection, scFv-S18 and -N3 expression resulted in the reduction of peripheral PrP(Sc) propagation by approximately 60 and 32 %, respectively, without a significant prolongation of incubation times and survival. Proof of rAAV vector DNA in spleen samples by real-time PCR strongly suggests a transport or trafficking of rAAV from the brain to the spleen, resulting in rAAV-mediated expression of scFv followed by reduced PrP(Sc) levels in the spleen most likely due to the blockage of the prion receptor LRP/LR by scFv.
Zuber
Chantal
C
Laboratorium für Molekulare Biologie - Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 München, Germany.
Mitteregger
Gerda
G
Zentrum für Neuropathologie und Prionforschung der LMU München, Feodor-Lynen-Str. 23, 81377 München, Germany.
Schuhmann
Natascha
N
Universität zu Köln, Klinik I für Innere Medizin, Kerpener Str. 62, 50937 Köln, Germany.
Rey
Clémence
C
Laboratorium für Molekulare Biologie - Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 München, Germany.
Knackmuss
Stefan
S
Affimed Therapeutics AG, Technologiepark, Im Neuenheimer Feld 582, 69120 Heidelberg, Germany.
Rupprecht
Wolfgang
W
Laboratorium für Molekulare Biologie - Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 München, Germany.
Reusch
Uwe
U
Affimed Therapeutics AG, Technologiepark, Im Neuenheimer Feld 582, 69120 Heidelberg, Germany.
Pace
Claudia
C
Zentrum für Neuropathologie und Prionforschung der LMU München, Feodor-Lynen-Str. 23, 81377 München, Germany.
Little
Melvyn
M
Affimed Therapeutics AG, Technologiepark, Im Neuenheimer Feld 582, 69120 Heidelberg, Germany.
Kretzschmar
Hans A
HA
Zentrum für Neuropathologie und Prionforschung der LMU München, Feodor-Lynen-Str. 23, 81377 München, Germany.
Hallek
Michael
M
Zentrum für Molekulare Medizin Köln, Universität zu Köln, Joseph-Stelzmann-Str. 52, 50931 Köln, Germany.|Universität zu Köln, Klinik I für Innere Medizin, Kerpener Str. 62, 50937 Köln, Germany.
Büning
Hildegard
H
Zentrum für Molekulare Medizin Köln, Universität zu Köln, Joseph-Stelzmann-Str. 52, 50931 Köln, Germany.|Universität zu Köln, Klinik I für Innere Medizin, Kerpener Str. 62, 50937 Köln, Germany.
Weiss
Stefan
S
Laboratorium für Molekulare Biologie - Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 München, Germany.
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006624:Hippocampus / Q000821:virology; D007141:Immunoglobulin Fc Fragments / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018620:PrPSc Proteins / Q000378:metabolism; D017096:Prion Diseases / Q000628:therapy*; D017474:Receptors, Laminin / Q000276:immunology*; D011995:Recombination, Genetic; D013154:Spleen / Q000378:metabolism / Q000821:virology; D014161:Transduction, Genetic
D000906:Antibodies; D007141:Immunoglobulin Fc Fragments; D018620:PrPSc Proteins; D017474:Receptors, Laminin