Withaferin A Suppresses Beta Amyloid in APP Expressing Cells: Studies for Tat and Cocaine Associated Neurological Dysfunctions

Abstract

Neurological disorders are the biggest concern globally. Out of ~36 million human immunodeficiency virus (HIV) positive people, about 30%–60% exhibit neurological disorders, including dementia and Alzheimer’s disease (AD) like pathology. In AD or AD like neurological disorders, the pathogenesis is mainly due to the abnormal accumulation of extracellular amyloid beta (Aβ). In this era of antiretroviral therapy, the life span of the HIV-infected individuals has increased leading towards increased neurocognitive dysfunction in nearly 30% of HIV-infected individuals, specifically older people. Deposition of the Aβ plaques in the CNS is one the major phenomenon happening in aging HIV patients. ART suppresses the viral replication, but the neurotoxic protein (Tat) is still produced and results in increased levels of Aβ. Furthermore, drugs of abuse like cocaine (coc) is known to induce the HIV associated neurocognitive disorders as well as the Aβ secretion. To target the Tat and coc induced Aβ secretion, we propose a potent bifunctional molecule Withaferin A (WA) which may act as a neuro-protectant against Aβ neurotoxicity. In this study, we show that WA reduces secreted Aβ and induced neurotoxicity in amyloid precursor protein (APP)-plasmid transfected SH-SY5Y cells (SH-APP). In this study, we show that in SH-APP cells, Aβ secretion is induced in the presence of HIV-1 Tat (neurotoxic) and drug of abuse coc. Our fluorescent microscopy studies show the increased concentration of Aβ40 in Tat (50 ng/ml) and coc (0.1 μM) treated SH-APP cells as compared to control. Our dose optimization study show, lower concentrations (0.5–2 μM) of WA significantly reduce the Aβ40 levels, without inducing cytotoxicity in the SH-APP cells. Additionally, WA reduces the Tat and cocaine induced Aβ levels. Therefore, we propose that Aβ aggregation is induced by the presence of Tat and coc and WA is potent in reducing the secreted Aβ and induced neurotoxicity. Our study provides new opportunities for exploring the pathophysiology and targeting the neurological disorders.

Inhibition of Amyloid-Beta Production, Associated Neuroinflammation, and Histone Deacetylase 2-Mediated Epigenetic Modifications Prevent Neuropathology in Alzheimer’s Disease in vitro Model

“…We observed these cells at 10× resolution. This study aligns with our prior results that WA has the ability to reduce Aβ in vitro (Tiwari et al, 2018).…”

“…In the brain of AD patients, NF-κB activation is predominantly found in neurons and glial cells present in the vicinity of Aβ plaque deposition (Terai et al, 1996;Boissière et al, 1997;Kaltschmidt et al, 1997;Lukiw and Bazan, 1998). In our earlier study, we have observed that withaferin A (WA), a steroidal lactone, derived from the plant Withania somnifera inhibits Aβ levels in amyloid overexpressing SH-SY5Y cells (SH-APP) (Tiwari et al, 2018). Based on these observations and reports on the role of WA in inhibiting the NF-κB-mediated neuroinflammation (Heyninck et al, 2014;Martorana et al, 2015), in this study we have utilized WA to study the Aβ levels and associated NF-κB-mediated neuroinflammation in SH-APP and microglial mixed cell culture.…”

Integrative genomics approach identifies conserved transcriptomic networks in Alzheimer’s disease

“…Previous studies have reported increased NFκB activation in AD and pointed to a relationship with Aβ pathology ( 108–111 ). Moreover, Withaferin A has been identified as an inhibitor of NFκB signaling ( 112 , 113 ) and has shown therapeutic potential to protect against Aβ toxicity ( 114 ), and we found Withaferin A is a modulator of CM9, CM4 and CM1 ( Supplementary Material, Fig. S11 ).…”

<p>Alzheimer’s disease: pathogenesis, diagnostics, and therapeutics</p>

“…There are promising drugs against Aβ toxicity,75 but in order to explore their maximum effect on CNS cells, there is a need of nanocarriers to be employed. Availability of drugs in the CNS is the major issue faced in the field of therapeutics against AD.…”