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 2

ScFv antibodies disrupt the interaction between TDP-43 and p65 and reduce NF-κB activity.

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ScFv antibodies disrupt the interaction between TDP-43 and p65 and reduc...
(A) Binding of p65 to TDP-43 was measured in the presence of equal volumes of PBS or media from pscFv9-transfected Hek293cells. n = 5–8 wells per condition (dots). One-way ANOVA P = 0.002; *P < 0.05 versus PBS; #P < 0.05 or ##P < 0.001 versus control scFv by Tukey’s multiple comparisons test. (B) BV2 cells were transfected with pscFv9, stimulated with LPS or PBS, and lysated for the luciferase assay. n = 4–5 individual experiments (dots). Two-way ANOVA P = 0.0002; §§§P < 0.001 versus LPS; ***P < 0.001 versus empty; and ###P < 0.001 versus control scFv, by Tukey’s multiple comparisons test. (C and D) BV2 p65-luc cells were treated or not for 6 hours or 10 hours with scFv antibodies, together with 4 hours of LPS treatment. (C) Representative Western blots of total lysates of Myc-tag scFv inside the cells and actin (n = 3 pooled experiments). The blot of the untreated cells at 6 hours was run on the same gel but was noncontiguous. (D) NF-κB activity assessed by luciferase assay. n = 4–6 replicates (dots) from 2 independent experiments (2–3 wells each). Two-way ANOVA P < 0.0001; *P < 0.05 and ***P < 0.001 versus untreated cells; ###P < 0.001 versus control scFv, by Tukey’s multiple comparisons test. Data represent the mean ± SEM. RLU, relative luminescence units; CTR, control D1.3 scFv.