Virus-mediated delivery of antibody targeting TAR DNA-binding protein-43 mitigates associated neuropathology
Silvia Pozzi, … , Claude Gravel, Jean-Pierre Julien
Silvia Pozzi, … , Claude Gravel, Jean-Pierre Julien
Published January 22, 2019
Citation Information: J Clin Invest. 2019;129(4):1581-1595. https://doi.org/10.1172/JCI123931.
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Research Article Neuroscience

Virus-mediated delivery of antibody targeting TAR DNA-binding protein-43 mitigates associated neuropathology

Abstract

The cytoplasmic aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of degenerating neurons in amyotrophic lateral sclerosis (ALS) and subsets of frontotemporal dementia (FTD). In order to reduce TDP-43 pathology, we generated single-chain (scFv) antibodies against the RNA recognition motif 1 (RRM1) of TDP-43, which is involved in abnormal protein self-aggregation and interaction with p65 NF-κB. Virus-mediated delivery into the nervous system of a scFv antibody, named VH7Vk9, reduced microgliosis in a mouse model of acute neuroinflammation and mitigated cognitive impairment, motor defects, TDP-43 proteinopathy, and neuroinflammation in transgenic mice expressing ALS-linked TDP-43 mutations. These results suggest that antibodies targeting the RRM1 domain of TDP-43 might provide new therapeutic avenues for the treatment of ALS and FTD.

Authors

Silvia Pozzi, Sai Sampath Thammisetty, Philippe Codron, Reza Rahimian, Karine Valérie Plourde, Geneviève Soucy, Christine Bareil, Daniel Phaneuf, Jasna Kriz, Claude Gravel, Jean-Pierre Julien

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Figure 1

E6-derived scFv antibodies are able to recognize TDP-43.

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E6-derived scFv antibodies are able to recognize TDP-43. (A) Binding of ...
(A) Binding of p65 to TDP-43 was measured in the presence of PBS or anti–TDP-43 N-terminal antibody (Proteintech), monoclonal antibodies against RRM1 TDP-43 (named C10, G8, and E6), or BSA. n = 8 wells per condition (dots). One-way ANOVA P < 0.0001; *P < 0.05 and ***P < 0.001 versus PBS, by Tukey’s multiple comparisons test. Data represent the mean ± SEM. (B) Schematic representation of the pscFv9 plasmid used for scFv production and expression. (C) Representative Western blot of cytoplasmic and nuclear fractions of Hek293 cells. Anti-Myc antibody revealed scFv and laminin A/C or actin in the different fractions. (D) Representative image of media from transfected Hek293 cells probed with anti-Myc antibody. Ponceau staining was used as a reference. (E) Different concentrations of TDP-43 (1–206 aa, Proteintech) or BSA were loaded onto a dot blot membrane. Immunoblots were performed with media containing pscFv9-transfected Hek293 cells and E6 monoclonal antibody. Signals were revealed with anti–Myc-HRP antibody for scFv conditions or anti–mouse HRP for E6. Ponceau staining was used as a reference. (F) Representative blot of TDP-43 immunoprecipitation in pscFv9-transfected Hek293 cells. Experiments in C, D, and F were conducted more than 3 times. Empty, no scFv; CTR, control D1.3 scFv.