Mitochondrial dynamics and bioenergetics in Alzheimer's induced pluripotent stem cell-derived neurons

Abstract

Mitochondrial (MT) dysfunction is a hallmark of Alzheimer's Disease (AD), but the scope and severity of these specific deficits across forms of AD are not well characterized. We designed a high-throughput, longitudinal, phenotypic assay to track MT dynamics and bioenergetics in glutamatergic iPSC-derived human neurons possessing mutations in presenilin 1 (PSEN1), presenilin 2 (PSEN2) and the amyloid beta precursor protein (APP). Each gene set was comprised of iPSC-derived neurons from an AD patient as well as two to three engineered mutations with appropriate isogenic and age matched controls. These iPSC-derived neurons were imaged every other day beginning at DIV10 to assess how MT length and content change over a ten day time course using a mitochondrially targeted reporter. A second cytosolic reporter allowed for visualization of neurites. Bioenergetics assays, focusing on MT respiration and individual electron transport chain (ETC) complexes, were also surveyed over this time course. Mutations in all three genes altered MT function measured by basal, ATP-linked, and maximal oxygen consumption rate; and spare respiratory capacity, with PSEN1/PSEN2 alleles being more severe than APP mutations. Electron flow through Complexes I-IV was decreased in PSEN1/PSEN2 mutations but; in contrast, APP alleles had only modest impairments of Complexes I and II. We measured aspects of MT dynamics, including fragmentation, and neurite degeneration, both of which were dramatic in PSEN1/PSEN2 alleles, but essentially absent in APP alleles. The marked differences in MT pathology may occur from the distinct ways amyloids are processed into amyloid beta peptides (Aβ) and may correlate with the disease severity.

Keywords: allelic variants; early onset AD; high-content imaging; mitochondrial function.