A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer’s Disease and Frontotemporal Dementia Disease Modeling

Muñoz, Sonia Sanz; Engel, Martin; Balez, Rachelle; Do-Ha, Dzung; Cabral-da-Silva, Mauricio Castro; Hernández, Damián; Berg, Tracey; Fifita, Jennifer A.; Grima, Natalie; Yang, Shu; Blair, Ian P.; Nicholson, Garth A.; Cook, Anthony; Hewitt, Alex W.; Pébay, Alice; Ooi, Lezanne

doi:10.3390/cells9092018

Cited by 32 publications

(33 citation statements)

References 40 publications

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“…Human iPSCs can be differentiated in dopaminergic [ 87 ], glutamatergic [ 88 ], cholinergic [ 125 ], serotonergic [ 90 ] and GABAergic neurons [ 91 ] as well as several types of supportive cells. By mixing these different types of neurons and supportive cells in the right ratios, specific brain regions can be modeled thereby potentially increasing the predictive value.…”

Section: Future Directions and Challenges In In Vitro Seizure Liability Assessmentmentioning

confidence: 99%

Novel test strategies for in vitro seizure liability assessment

Tukker¹,

Westerink

2021

Expert Opinion on Drug Metabolism & Toxicology

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Introduction The increasing incidence of mental illnesses and neurodegenerative diseases results in a high demand for drugs targeting the central nervous system (CNS). These drugs easily reach the CNS, have a high affinity for CNS targets, and are prone to cause seizures as an adverse drug reaction. Current seizure liability assessment heavily depends on in vivo or ex vivo animal models and is therefore ethically debated, labor intensive, expensive, and not always predictive for human risk. Areas covered The demand for CNS drugs urges the development of alternative safety assessment strategies. Yet, the complexity of the CNS hampers reliable detection of compound-induced seizures. This review provides an overview of the requirements of in vitro seizure liability assays and highlights recent advances, including micro-electrode array (MEA) recordings using rodent and human cell models. Expert opinion Successful and cost-effective replacement of in vivo and ex vivo models for seizure liability screening can reduce animal use for drug development, while increasing the predictive value of the assays, particularly if human cell models are used. However, these novel test strategies require further validation and standardization as well as additional refinements to better mimic the human in vivo situation and increase their predictive value.

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Section: Future Directions and Challenges In In Vitro Seizure Liability Assessmentmentioning

confidence: 99%

Novel test strategies for in vitro seizure liability assessment

Tukker¹,

Westerink

2021

Expert Opinion on Drug Metabolism & Toxicology

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“…Approximately 40% of the resulting neurons express ChAT, the enzyme responsible for biosynthesis of the neurotransmitter acetylcholine and a mature BFCN marker ( Hu et al, 2016 ; Figure 2 ). Others have also successfully used small molecules to pattern BFCNs resulting in efficiencies ranging from 15 up to 80% ( Liu et al, 2013b ; Yue et al, 2015 ; Hu et al, 2016 ; Muñoz et al, 2020 ). Although the yields of BFCNs are good, the mixed neuronal cultures leave room for improvement in the established BFCN protocols.…”

Section: Modeling Ad In Ds and Bfcnsmentioning

confidence: 99%

Basal Forebrain Cholinergic Neurons: Linking Down Syndrome and Alzheimer’s Disease

Martínez

Zammit

West

et al. 2021

Front. Aging Neurosci.

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Down syndrome (DS, trisomy 21) is characterized by intellectual impairment at birth and Alzheimer’s disease (AD) pathology in middle age. As individuals with DS age, their cognitive functions decline as they develop AD pathology. The susceptibility to degeneration of a subset of neurons, known as basal forebrain cholinergic neurons (BFCNs), in DS and AD is a critical link between cognitive impairment and neurodegeneration in both disorders. BFCNs are the primary source of cholinergic innervation to the cerebral cortex and hippocampus, as well as the amygdala. They play a critical role in the processing of information related to cognitive function and are directly engaged in regulating circuits of attention and memory throughout the lifespan. Given the importance of BFCNs in attention and memory, it is not surprising that these neurons contribute to dysfunctional neuronal circuitry in DS and are vulnerable in adults with DS and AD, where their degeneration leads to memory loss and disturbance in language. BFCNs are thus a relevant cell target for therapeutics for both DS and AD but, despite some success, efforts in this area have waned. There are gaps in our knowledge of BFCN vulnerability that preclude our ability to effectively design interventions. Here, we review the role of BFCN function and degeneration in AD and DS and identify under-studied aspects of BFCN biology. The current gaps in BFCN relevant imaging studies, therapeutics, and human models limit our insight into the mechanistic vulnerability of BFCNs in individuals with DS and AD.

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“…In contrast to recent studies, our patterning regime omitted the use of non-adherent approaches, while simultaneously inhibiting the Wnt-pathway during neuralization and ventralization ( Ihnatovych et al, 2018 ; Muñoz et al, 2020 ). Plating cells under adherent conditions with constant numbers, increase the reproducibility and uniformity of the differentiation experiments in contrast to the formation of embryoid bodies, a process that is more difficult to control.…”

Section: Discussionmentioning

confidence: 99%

Ventral Telencephalic Patterning Protocols for Induced Pluripotent Stem Cells

Krajka

Naujock

Pauly

et al. 2021

Front. Cell Dev. Biol.

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The differentiation of human induced pluripotent stem cells (hiPSCs) into specific cell types for disease modeling and restorative therapies is a key research agenda and offers the possibility to obtain patient-specific cells of interest for a wide range of diseases. Basal forebrain cholinergic neurons (BFCNs) play a particular role in the pathophysiology of Alzheimer’s dementia and isolated dystonias. In this work, various directed differentiation protocols based on monolayer neural induction were tested for their effectiveness in promoting a ventral telencephalic phenotype and generating BFCN. Ventralizing factors [i.e., purmorphamine and Sonic hedgehog (SHH)] were applied at different time points, time intervals, and concentrations. In addition, caudal identity was prevented by the use of a small molecule XAV-939 that inhibits the Wnt-pathway. After patterning, gene expression profiles were analyzed by quantitative PCR (qPCR). Rostro-ventral patterning is most effective when initiated simultaneously with neural induction. The most promising combination of patterning factors was 0.5 μM of purmorphamine and 1 μM of XAV-939, which induces the highest expression of transcription factors specific for the medial ganglionic eminence, the source of GABAergic inter- and cholinergic neurons in the telencephalon. Upon maturation of cells, the immune phenotype, as well as electrophysiological properties were investigated showing the presence of marker proteins specific for BFCN (choline acetyltransferase, ISL1, p75, and NKX2.1) and GABAergic neurons. Moreover, a considerable fraction of measured cells displayed mature electrophysiological properties. Synaptic boutons containing the vesicular acetylcholine transporter (VACHT) could be observed in the vicinity of the cells. This work will help to generate basal forebrain interneurons from hiPSCs, providing a promising platform for modeling neurological diseases, such as Alzheimer’s disease or Dystonia.

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A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer’s Disease and Frontotemporal Dementia Disease Modeling

Cited by 32 publications

References 40 publications

Novel test strategies for in vitro seizure liability assessment

Novel test strategies for in vitro seizure liability assessment

Basal Forebrain Cholinergic Neurons: Linking Down Syndrome and Alzheimer’s Disease

Ventral Telencephalic Patterning Protocols for Induced Pluripotent Stem Cells

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