Neuron-based high-content assay and screen for CNS active mitotherapeutics

Abstract

Impaired mitochondrial dynamics and function are hallmarks of many neurological and psychiatric disorders, but direct screens for mitotherapeutics using neurons have not been reported. We developed a multiplexed and high-content screening assay using primary neurons and identified 67 small-molecule modulators of neuronal mitostasis (MnMs). Most MnMs that increased mitochondrial content, length, and/or health also increased mitochondrial function without altering neurite outgrowth. A subset of MnMs protected mitochondria in primary neurons from Aβ(1–42) toxicity, glutamate toxicity, and increased oxidative stress. Some MnMs were shown to directly target mitochondria. The top MnM also increased the synaptic activity of hippocampal neurons and proved to be potent in vivo, increasing the respiration rate of brain mitochondria after administering the compound to mice. Our results offer a platform that directly queries mitostasis processes in neurons, a collection of small-molecule modulators of mitochondrial dynamics and function, and candidate molecules for mitotherapeutics.

Prevention of 7-Ketocholesterol-Induced Overproduction of Reactive Oxygen Species, Mitochondrial Dysfunction and Cell Death with Major Nutrients (Polyphenols, ω3 and ω9 Unsaturated Fatty Acids) of the Mediterranean Diet on N2a Neuronal Cells

“…As mitochondrial and ROS overproduction is a general feature of 7KC-induced cell death whatever the cells considered, it may be advisable to micro-or nano-encapsulate these nutrients in order to promote access to their therapeutic targets. In addition, the ability of polyphenols and fatty acids to strongly reduce the 7KC-induced drop in ∆Ψm indicates that these nutrients are potentially suitable molecules for mitotherapy [59]. Associated a targeted mitotherapy using functionalized nanoparticles (Targeted Organel Nano-therapy: TORN therapy) [60,61] containing polyphenols as well as ω3 and/or ω9 unsaturated fatty acids could thus be envisaged to treat diseases associated with increased 7KC-levels.…”

Sensory neurons derived from diabetic rats exhibit deficits in functional glycolysis and ATP that are ameliorated by IGF-1

“…Metabolically, mitochondria are tightly dependent on glycolysis, and this linkage is highlighted by the presence of hexokinase I binding to the mitochondrial outer membrane [ 56 , 57 ]. Mitochondria are mostly generated and refurbished in the neuronal perikarya and have a higher density in the cell body than axons [ [58] , [59] , [60] ]. They are distributed equally along the axons, excluding the axonal tips [ 56 ].…”

The Age-Dependent Decline in Neuron Growth Potential in the CNS is Associated with an Age-Related Dysfunction of Neuronal Mitochondria

“…Some examples of these that have begun to be explored include anti-apoptotic [100][101][102] as well as antioxidant strategies [103,104], boosting mitochondrial bioenergetics [105] and biogenesis [99,106], alternative biofuels [107][108][109], targeting the mPTP [30,[110][111][112][113], Ca2+ signaling and transmembrane channels. Of particular interest here, studies have even been conducted specifically to test pharmacological agents to target mitochondria in neurons [114]. In the case of SCI, 'MitoCeuticals' may prove to be an effective strategy to not only improve neuroprotection and create a more permissive lesion environment, but also potentially promote axon growth.…”