iPSC-Based Compound Screening and In Vitro Trials Identify a Synergistic Anti-amyloid β Combination for Alzheimer’s Disease

Kondo, Takayuki; Imamura, Keiko; Funayama, Misato; Tsukita, Kayoko; Miyake, Michiyo; Ohta, Akira; Woltjen, Knut; Nakagawa, Masato; Asada, Takashi; Arai, Tetsuaki; Kawakatsu, Shinobu; Izumi, Yuishin; Kaji, Ryuji; Iwata, Nobuhisa; Inoue, Haruhisa

doi:10.1016/j.celrep.2017.10.109

Cited by 182 publications

(171 citation statements)

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“…M.3243A>G heteroplasmy levels of iPS lines were sequenced, after which one iPS clone without-, and one iPS clone with heteroplasmy were selected. An additional, curated control cell line (409B2 (Kondo et al, 2017)) was included, and confirmed to have no m.3243A>G heteroplasmy. IPS cells were thawed and cultured at all times cultured on recombinant human laminin LN521 (Biolamina, 2021-21) kept in E8 basal medium (Thermo Fisher Scientific) supplemented with primocin (0.1 µg/ml, Invivogen), puromycin (0.5 µg/ml) and G418 (0.5 µg/ml), and uridine (50 µg/ml) and incubated at 37°C/5%CO2, with medium changes every 2-3 days and passages 1-2 times per week using ReLeSR (Stem Cell Technologies).…”

Section: Ips Cell Generation and Culturementioning

confidence: 99%

Mitochondrial dysfunction impairs human neuronal development and reduces neuronal network activity and synchronicity

Gunnewiek

Hugte

Frega

et al. 2019

Preprint

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Epilepsy, intellectual and cortical sensory deficits and psychiatric manifestations are among the most frequent manifestations of mitochondrial diseases. Yet, how mitochondrial dysfunction affects neural structure and function remains largely elusive. This is mostly due to the lack of a proper in vitro translational neuronal model system (s) with impaired energy metabolism. Leveraging the induced pluripotent stem cell technology, from a cohort of patients with the common pathogenic m.3243A>G variant of mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS), we differentiated excitatory cortical neurons (iNeurons) with normal (low heteroplasmy) and impaired (high heteroplasmy) mitochondrial function on an isogenic nuclear DNA background. iNeurons with high levels of heteroplasmy exhibited mitochondrial dysfunction, delayed neural maturation, reduced dendritic complexity and fewer functional excitatory synapses. Microelectrode array recordings of neuronal networks with high heteroplasmy displayed reduced network activity and decreased synchronous network bursting. The impaired neural energy metabolism of iNeurons compromising the structural and functional integrity of neurons and neural networks, could be the primary driver of increased susceptibility to neuropsychiatric manifestations of mitochondrial disease.

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Section: Ips Cell Generation and Culturementioning

confidence: 99%

Mitochondrial dysfunction impairs human neuronal development and reduces neuronal network activity and synchronicity

Gunnewiek

Hugte

Frega

et al. 2019

Preprint

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“…Recent studies also imply a large gap in the effectiveness of drug discovery studies between iPS-derived neurons and cultured cell lines [42]. Recently, Inoue and colleagues developed a reliable and robust iPS-based screening system for anti-Aβ drugs [43]. After incubating the differentiated neurons from iPS with PKC ligands for 24 h, Aβ42 and Aβ40 levels in the conditioned medium were calculated using electrochemiluminescence assays.…”

Section: Effects Of 1 On Aβ42/aβ40 Aβ Oligomerization and Neurotoximentioning

confidence: 99%

“…Human iPS cells of Alzheimer's patients were generated as described from skin fibroblasts [43,57] and maintained using StemFit AK02N medium (Ajinomoto, Tokyo, Japan) [64] and expanded for neural differentiation. To establish a robust and rapid differentiation method, we utilized direct conversion technology.…”

Section: Generation and Characterization Of Human Ips-derived Neuronsmentioning

confidence: 99%

Evaluation of Toxic Amyloid β42 Oligomers in Rat Primary Cerebral Cortex Cells and Human iPS-derived Neurons Treated with 10-Me-Aplog-1, a New PKC Activator

Murakami

Yoshimura

Nakagawa

et al. 2020

IJMS

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Amyloid β42 (Aβ42), a causative agent of Alzheimer's disease (AD), is derived extracellularly from Aβ precursor protein (APP) following the latter's cleavage by β-secretase, but not α-secretase. Protein kinase Cα (PKCα) activation is known to increase α-secretase activity, thereby suppressing Aβ production. Since Aβ42 oligomer formation causes potent neurotoxicity, APP modulation by PKC ligands is a promising strategy for AD treatment. Although bryostatin-1 (bryo-1) is a leading compound for this strategy, its limited natural availability and the difficulty of its total synthesis impedes further research. To address this limitation, Irie and colleagues have developed a new PKC activator with few side effects, 10-Me-Aplog-1, (1), which decreased Aβ42 in the conditioned medium of rat primary cerebral cortex cells. These results are associated with increased α-secretase but not PKCε-dependent Aβ-degrading enzyme. The amount of neuronal embryonic lethal abnormal vision (nELAV), a known β-secretase stabilizer, was reduced by treatment with 1. Notably, 1 prevented the formation of intracellular toxic oligomers. Furthermore, 1 suppressed toxic oligomerization within human iPS-derived neurons such as bryo-1. Given that 1 was not neurotoxic toward either cell line, these findings suggest that 1 is a potential drug lead for AD therapy. be more complex, given the recent discovery of APP proteolysis by ηand δ-secretases, for example, in [3]. The ability of Aβ42 to aggregate and exhibit neurotoxicity is higher than that of Aβ40 despite the lower in vivo amounts of Aβ42 [4]. Aβ42 oligomer formation causes synaptic dysfunction and neuronal death in AD pathology, whereas the contribution of end-stage mature fibrils of Aβ42 to AD is lower than that of oligomers [5]. Higher-order toxic oligomers that show potent synaptotoxicity and neurotoxicity have been reported, such as protofibrils (PFs), Aβ-derived diffusible ligands, and amylospheroids [6]. Therefore, suppressing toxic oligomerization of Aβ42 is a favorable strategy for developing AD therapies. This suppression can also be achieved by simultaneously decreasing Aβ production while inducing Aβ degradation.Protein kinase C (PKC) is a family of serine/threonine kinases that plays a pivotal role in various biological events such as signal transduction, proliferation, and apoptosis mediated by the second messenger 1,2-diacyl-sn-glycerol [7]. The PKC family, which contains at least 10 isozymes, is divided into three groups, namely conventional (α, βI, βII, and γ), novel (δ, ε, η, and θ), and atypical (µ, ξ, and ι) [7]. PKC activity is related to memory formation and learning [8], while PKC downregulation may induce cognitive impairment and memory loss in AD [9]. Regarding Aβ-driven molecular events, PKCα reportedly upregulates α-secretase activity either directly or indirectly through the mitogen-activated protein kinase (MAPK) pathway [10]. PKCα activation in a mouse model of AD has beneficial effects on AD pathology, including the disruption of Aβ production and reduction of toxi...

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“…Building on the strengths and relative maturity of adherent stem cell protocols, several PTS have provided proof-of-concept for iPSC-driven screening approaches (Kondo et al, 2017;Sterneckert et al, 2014) (also summarized in Table 1). At the same time, these approaches highlight the importance of careful hit validation strategies.…”

Section: Advances Of Stem Cell-based Pts In 2d Culturesmentioning

confidence: 99%

The Convergence of Stem Cell Technologies and Phenotypic Drug Discovery

Friese

Ursu

Hochheimer³

et al. 2019

Cell Chemical Biology

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Recent advances in induced pluripotent stem cell technologies and phenotypic screening shape the future of bioactive small-molecule discovery. In this review we analyze the impact of small-molecule phenotypic screens on drug discovery as well as on the investigation of human development and disease biology. We further examine the role of 3D spheroid/organoid structures, microfluidic systems, and miniaturized on-achip systems for future discovery strategies. In highlighting representative examples, we analyze how recent achievements can translate into future therapies. Finally, we discuss remaining challenges that need to be overcome for the adaptation of the next generation of screening approaches.

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iPSC-Based Compound Screening and In Vitro Trials Identify a Synergistic Anti-amyloid β Combination for Alzheimer’s Disease

Cited by 182 publications

References 39 publications

Mitochondrial dysfunction impairs human neuronal development and reduces neuronal network activity and synchronicity

Mitochondrial dysfunction impairs human neuronal development and reduces neuronal network activity and synchronicity

Evaluation of Toxic Amyloid β42 Oligomers in Rat Primary Cerebral Cortex Cells and Human iPS-derived Neurons Treated with 10-Me-Aplog-1, a New PKC Activator

The Convergence of Stem Cell Technologies and Phenotypic Drug Discovery

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