Identification of a Novel Wnt Antagonist Based Therapeutic and Diagnostic Target for Alzheimer’s Disease Using a Stem Cell-Derived Model

Abstract

Currently, all the existing treatments for Alzheimer’s disease (AD) fail to stall progression due to longer duration of time between onset of the symptoms and diagnosis of the disease, raising the necessity of effective diagnostics and novel treatment. Specific molecular regulation of the onset and progression of disease is not yet elucidated. This warranted investigation of the role of Wnt signaling regulators which are thought to be involved in neurogenesis. The AD model was established using amyloid beta (Aβ) in human mesenchymal stem cells derived from amniotic membranes which were differentiated into neuronal cell types. In vivo studies were carried out with Aβ or a Wnt antagonist, AD201, belonging to the sFRP family. We further created an AD201-knockdown in vitro model to determine the role of Wnt antagonism. BACE1 upregulation, ChAT and α7nAChR downregulation with synapse and functionality loss with increases in ROS confirmed the neurodegeneration. Reduced β-catenin and increased AD201 expression indicated Wnt/canonical pathway inhibition. Similar results were exhibited in the in vivo study along with AD-associated behavioural and molecular changes. AD201-knockdown rescued neurons from Aβ-induced toxicity. We demonstrated for the first time a role of AD201 in Alzheimer’s disease manifestation, which indicates a promising disease target and biomarker.

Mesenchymal stem cell‐derived rapid drug screening system for Alzheimer's disease for the identification of novel drugs

“…Placenta was obtained after ethical clearance from Omega Multispecialty Hospital, Bangalore (OMHB/SORM/009). AM‐MSCs were isolated using the enzymatic method as described previously (Patil et al, 2023; Warrier et al, 2012). AM‐MSCs were cultured in Dulbecco's modified Eagle medium‐high glucose (DMEM‐HG) (Gibco), 10% fetal bovine serum (Gibco), 1x Antibiotic‐antimycotic solution (Gibco), and incubated at 37°C, 5% CO 2 .…”

“…AM‐MSCs were induced to differentiate into neuronal cells using a proprietary induction media containing DC304, an anti‐inflammatory glucocorticoid (1 µM), DC305, a phosphodiesterase inhibitor (0.5 mM), and DC306, a nonsteroidal anti‐inflammatory agent (100 µM) in serum‐free DMEM (Patil et al, 2023). Degeneration was induced by treating the differentiated cells with 2.5 µM oligomerized Aß 1‐42 for 24 h at 37°C.…”

“…Following treatments, the supernatant was discarded and cells were incubated with TMRE (0.3 µM) dissolved in PBS at 37°C for 30 min in the dark. The fluorescence intensity of TMRE was measured at Ex/Em = 549/574 nm (Patil et al, 2023).…”

“…dissolved in PBS at 37°C for 30 min in the dark. The fluorescence intensity of TMRE was measured at Ex/Em = 549/574 nm (Patil et al, 2023).…”

miRNA on the Battlefield of Cancer: Significance in Cancer Stem Cells, WNT Pathway, and Treatment

“…This involved a decrease in oxidative stress, improvement in mitochondrial function, and rescue of neuronal function. This emphasizes the importance of such a technology in therapy [ 54 ]. It has been reported in ovarian cancer that let-7d-5p targets CSC characteristics to subdue cancer progression.…”