Anti-tau scFvs Targeted to the Cytoplasm or Secretory Pathway Variably Modify Pathology and Neurodegenerative Phenotypes

Marshall S Goodwin; Olga Sinyavskaya; Franklin Burg; Veronica O'Neal; Carolina Ceballos-Diaz; Pedro E Cruz; Jada Lewis; Benoit I Giasson; Peter Davies; Todd E Golde; Yona Levites

doi:10.1016/j.ymthe.2020.10.007

Anti-tau scFvs Targeted to the Cytoplasm or Secretory Pathway Variably Modify Pathology and Neurodegenerative Phenotypes

Mol Ther. 2021 Feb 3;29(2):859-872. doi: 10.1016/j.ymthe.2020.10.007. Epub 2020 Oct 14.

Affiliations

¹ Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
² Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
³ Litwin-Zucker Center for Research in Alzheimer's Disease, Feinstein Institute for Medical Research, North Shore/LIJ Health System, Manhasset, NY, USA.
⁴ Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA. Electronic address: tgolde@ufl.edu.
⁵ Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA. Electronic address: levites.yona@ufl.edu.

Abstract

Immunotherapies designed to treat neurodegenerative tauopathies that primarily engage extracellular tau may have limited efficacy as tau is primarily intracellular. We generated tau-targeting single-chain variable fragments (scFvs) and intrabodies (iBs) from the phosphorylated tau-specific antibodies CP13 and PHF1 and the pan-tau antibody Tau5. Recombinant adeno-associated virus (rAAV) was utilized to express these antibody fragments in homozygous JNPL3 P301L tau mice. Two iBs (CP13i, PHF1i) and one scFv (PHF1s) abrogated tau pathology and delayed time to severe hindlimb paralysis. In a second tauopathy model (rTg4510), CP13i and PHF1i reduced tau pathology, but cognate scFvs did not. These data demonstrate that (1) disease-modifying efficacy does not require antibody effector functions, (2) the intracellular targeting of tau with phosphorylated tau-specific iBs is more effective than extracellular targeting with the scFvs, and (3) robust effects on tau pathology before neurodegeneration only resulted in modest disease modification as assessed by delay of severe motor phenotype.

Keywords: Alzheimer's disease; adeno-associated virus; iB; immunotherapy; intrabody; neurodegeneration; phosphorylation; scFv; single-chain variable fragments; tauopathy.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Combined Modality Therapy
Dependovirus / genetics
Disease Models, Animal
Genetic Therapy
Genetic Vectors / administration & dosage
Genetic Vectors / genetics
Mice
Mice, Transgenic
Neurodegenerative Diseases / etiology
Neurodegenerative Diseases / metabolism*
Neurodegenerative Diseases / therapy*
Secretory Pathway / drug effects*
Single-Chain Antibodies / pharmacology*
Treatment Outcome
tau Proteins / antagonists & inhibitors*
tau Proteins / metabolism

Substances

Single-Chain Antibodies
tau Proteins

Grants and funding

R01 AG018454/AG/NIA NIH HHS/United States