An optimized InCell Western screening technique identifies hexachlorophene as a novel potent TDP43 targeting drug

• Published in: Journal of biotechnology Vol. 207; pp. 34 - 38
• Main Authors: Narayan, Malathi, Peralta, Diego A., Gibson, Chelsea, Zitnyar, Ashley, Jinwal, Umesh K.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 10.08.2015
• Subjects: Alzheimer's disease; Amyotrophic lateral sclerosis; Animals; Binding; Biotechnology; Cell Line; Disorders; DNA-Binding Proteins - antagonists & inhibitors; DNA-Binding Proteins - metabolism; Drug Discovery - methods; Drugs; Fronto-temporal lobe dementia; HEK293 Cells; HeLa Cells; Hexachlorophene; Hexachlorophene - chemistry; Hexachlorophene - pharmacology; Humans; InCell Western; Mice; Molecular Targeted Therapy; Molecular weight; Proteins; Screening; TDP-43 Proteinopathies - drug therapy; TDP43 proteinopathies; Amyotrophic lateral sclerosis; TDP43 proteinopathies; Fronto-temporal lobe dementia; Alzheimer's disease; Hexachlorophene; InCell Western
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2015.04.012

•TDP43 is a protein implicated in Alzheimer's disease, ALS.•Development of TDP43 targeting InCell Western technique for drug screen.•Hexachlorophene (referred as B10) decreases levels of pathological TDP43.•Hexachlorophene is a novel drug molecule for modulating levels of TDP43. TAR DNA binding protein (TDP43) is a DNA- and RNA-binding protein that is implicated in several neurodegenerative disorders termed as “TDP43 proteinopathies” including Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS) and fronto-temporal lobe dementia (FTLD). We have developed an InCell Western (ICW) technique for screening TDP targeting drugs in 96 well plates. We tested 281 compounds and identified a novel compound hexachlorophene (referred to as B10) that showed potent reduction in TDP43 levels. The effect of B10 on TDP protein level was validated in two different cellular models: endogenous TDP43 expressing N9 microglial cells and TDP43-over-expressing HEK293 and HeLa cells. We also analyzed effect of B10 on various pathological forms of TDP such as the C25 cleaved fragment that localizes to the cytosol, insoluble high molecular weight species, and ALS-linked mutants. Our data suggest that B10 effectively reduces all forms of TDP. Overall, our data suggest that B10 could serve as a potential drug molecule for the treatment of AD, ALS and other TDP43 proteinopathies.