High-Throughput Small Molecule Screen Identifies Modulators of Mitochondrial Function in Neurons

Abstract

We developed a high-throughput assay for modulators of mitochondrial function in neurons measuring inner mitochondrial membrane potential (DJ m ) and ATP production. The assay was used to screen a library of small molecules, which led to the identification of structural/functional classes of mitochondrial modulators such as local anesthetics, isoflavones, COXII inhibitors, adrenergic receptor blockers, and neurotransmitter system effectors. Our results show that some of the isolated compounds promote mitochondrial health, enhance oxygen consumption rate, and protect neurons against toxic insults found in the cellular environment of Alzheimer disease. These studies offer a set of compounds that may provide efficacy in protecting the mitochondrial system in neurodegenerative disorders.

Neuronal cell-based high-throughput screen for enhancers of mitochondrial function reveals luteolin as a modulator of mitochondria-endoplasmic reticulum coupling

“…Most of these studies used cell lines derived from peripheral tissue such as mouse Hepa1-6 hepatoma cells [ 42 ] and murine myotubes [ 43 , 44 ]. Only recently a HTS using primary neurons was established to identify mitotherapeutics [ 45 ]. These large-scale screenings vary in their initial readouts, which include analysis of ΔΨ m , mitochondrial morphology, cellular respiration, mtDNA-encoded protein synthesis, NAD(P)H levels, and ATP content [ 46 ].…”

“…The recently published HTS using primary neurons have identified two isoflavone-derivates as mitotherapeutics. Of note, the use of ΔΨ m as a single readout in the primary screen may justify why luteolin was not identified among one of the primary 135 hits [ 45 ]. Biesemann and colleagues have also identified flavanone series as novel compounds that enhanced muscle performance in aged mice by increasing intracellular ATP levels and Nrf2 gene expression [ 44 ].…”

Assessment of Ethanol-Induced Toxicity on iPSC-Derived Human Neurons Using a Novel High-Throughput Mitochondrial Neuronal Health (MNH) Assay

“…Recent studies used MEA to investigate the effect of compounds on spontaneous neuronal activity and network activity ( Zwartsen et al, 2018 ). Various HT screening methods have also been established ( Saunders et al, 2014 ; Iannetti et al, 2015 ; Gelles and Chipuk, 2016 ; Daniele et al, 2017 ; Kepiro et al, 2018 ; Arias-Fuenzalida et al, 2019 ; Varkuti et al, 2020 ). Nonetheless, there is a need for additional in vitro systems centered on human iPSC derivatives in order to capture different aspects of human neurotoxicity ( Schmidt et al, 2017 ).…”

Assessment of ethanol-induced toxicity on iPSC-derived human dopaminergic neurons using a novel high-throughput mitochondrial neuronal health (MNH) assay

“…Recent studies used MEA to investigate the effect of compounds on spontaneous neuronal activity and network activity (34). Various HT screening methods have also been established (35)(36)(37)(38)(39)(40)(41). Nonetheless, there is a need for additional in vitro systems centered on human iPSC derivatives in order to capture different aspects of human neurotoxicity (42).…”