Virus-mediated delivery of antibody targeting TAR DNA-binding protein-43 mitigates associated neuropathology
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
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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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 3

ScFv antibodies mediate TDP-43 degradation.

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ScFv antibodies mediate TDP-43 degradation. (A) Representative blot and ...
(A) Representative blot and quantification of TDP-43 levels in Hek293 cells transfected with pscFv9 plasmid. Graph shows quantification of TDP-43 normalized to actin and expressed as the fold change versus empty vector. n = 7–9 independent experiments (dots). One-way ANOVA P = 0.0003; *P < 0.05 and ***P < 0.0001 versus empty transfected, by Tukey’s multiple comparisons test. (B and C) Representative merged images of colocalization between scFv (Myc, green) and ubiquitin or LC3 (red). Arrows show cells with colocalization; yellow arrows indicate cells enlarged 2.5 times in the insets. Scale bars: 20 μm. The experiment was performed twice. (D) Representative blots and quantification of immunoprecipitation for all K48-polyubiquitinated (proteasome) or K63-polyubiquitinated (autophagy) proteins and Western blot for TDP-43. Graphs indicate the quantification of TDP-43 signal in immunoprecipitation experiments, expressed as the fold change versus empty vector. n = 3 independent experiment (dots). K63-ubiquitinated TDP-43: P = 0.0082, by 1-way ANOVA; **P < 0.01 versus empty vector–transfected cells, by Dunnett’s multiple comparisons test. K48-ubiquitinated TDP-43: P = 0.0128, by 1-way ANOVA; *P < 0.05 versus empty vector–transfected cells, by Dunnett’s multiple comparisons test. Data represent the mean ± SEM. Empty, no scFv (A and D); CTR, D1.3 scFv (B and C) or 8H11 anti-GFP scFv (A and D).