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Pharmacological evaluation in human iPSC-derived cortical and sensory neurons using high-throughput MEA system
Human iPSC-derived neurons are expected as a new toxicological evaluation assay to replace animal experiments in preclinical studies, improving the risk of accuracy.
MEAs have been adapted to drug discovery assays including cell-to-cell interaction, learning and memory, and network properties of neurons. High sensitivity consists key factor of their performance; it results in better signal to noise ratio leading to more reliable detection of action porentials. Michael Trujillo, Product Manager at Alpha Med Scientific, explains how low noise in engineered into MED64 systems.
Frontotemporal dementia and Parkinson's Disease cellular models: In vitro characterization of pathological phenotypes in gene-edited iPSC-derived neurons with MAPT and LRRK2 mutations
Mutations in MAPT and LRRK2 are reported to play a causative role in familial forms of frontotemporal dementia and Parkinson’s disease respectively. Human cell-based models in which to investigate the effect of these mutations on cellular functions such as microtubule assembly, oxidative stress, mitochondrial- and synaptic-functions, all of which could contribute to the onset of neurodegenerative conditions, are necessary to understand the disease mechanisms. Genome-edited human iPSC-derived neural stem cells (NSCs) offer a virtually unlimited source of physiologically relevant isogenic cell lines for use in disease modeling and drug discovery. Combining iPSC genome editing using CRISPR-Cas9 and directed differentiation, we have generated patient-relevant NSC disease models (axolGEMs) carrying heterozygous and homozygous combinations of missense mutations R406W, P301L and V337M in MAPT and G2019S in LRRK2. Genotype, karyotype, SNP frequency and copy number were assessed in genome-edited iPSC lines. Differentiated cells formed polarized neural tube-like rosettes in a monolayer culture. Immunocytochemistry confirmed the expression of typical NSC (PAX6, FOXG1, nestin, Ki67 and NCAD) and cerebral cortical neuronal markers (TUJ1, TBR1, MAP2 and CTIP). These cells enable a direct comparison of the variant effect on cellular phenotype between isogenic lines and offer a stable platform for drug screening and validation.
Abnormal phosphorylation of amyloid precursor protein tyrosine residues alters the APP trafficking in neurons from Alzheimer's Disease affected patients
Alzheimer’s Disease (AD) is the most common cause of dementia and is likely caused by defective amyloid precursor protein (APP) trafficking and processing in neurons leading to amyloid plaques containing the amyloid-β (Aβ) APP peptide byproduct. Understanding how APP is targeted and trafficked to selected destinations inside neurons and identifying the mechanisms responsible for the generation of Aβ are thus the key for the development of new therapies. We previously developed a mouse model showing that the Tyrosine (Tyr)682, in the C-terminus of APP, is essential for its binding to the coating protein clathrin and to the clathrin adaptor protein AP2 and for APP trafficking and sorting in neurons. In the present study, we investigated whether the Tyr682 residue of APP influences APP trafficking and sorting in neurons from differentiated neural stem cells (NSCs) of patients with AD carrying mutations on the PS1 gene (L286V; A246E; M146L), and on cortical tissues and fibroblasts from Göttingen minipigs engineered to develop AD (M146I mutant minipigs). Our results show that Tyrosine phosphorylation controls the APP binding to clathrin and AP2 in neurons carrying PS1 mutation and influences APP trafficking and sorting. Overall, our results provide a basis for the development of potential new therapies for AD.
Dr An Verheyen, Janssen R&D Discovery Services presented her work at ISSCR Basel, 2017. Dr Verheyen used human iPSC-derived neural cells generated using our custom differentiation service to model tauopathies linked to frontotemporal dementia with parkinsonism-17 and Alzheimer's disease.
Electrophysiological maturation and pharmacological responses of human induced pluripotent stem cell-derived cortical neuronal networks in long-term culture
Ikuro Suzuki, Associate Professor at Tohoku Institute of Technology, Japan who recently published his work with Axol Human iPSC-Derived Cerebral Cortical Neurons in Nature’s Scientific Reports (Odawara et al., 2016), presented our Innovation Showcase in collaboration with Alpha MED Scientific.
Prof Suzuki discussed the following results from his research:
- Morphology in long-term culture
- Development of spontaneous burst firings
- Pharmacological properties
- Induction of epileptiform activity & effects of anti-epilepsy drugs
- Induction of long-term potentiation and depression
Dr Alfredo Cabrera, Janssen Pharmaceutica, presented a robust, scalable and disease relevant model of tau aggregation using induced pluripotent stem cell (iPSC)-derived cortical neurons that can be applied to drug discovery programs in neurodegeneration. The resulting assay is highly reproducible across users and works in different commercially available iPSC-lines, providing a reliable tool for better understanding TAU pathophysiology and the identification of novel treatments against Alzheimer’s disease.
Currently, the majority of studies on Alzheimer's disease have used transgenic animal models or imaging studies of the human brain. It is difficult to validate these findings using human tissue. Whilst animal models have been central to our understanding of human physiology, human stem cell-based models may help us to further our understanding of human physiology and tackle devastating diseases such as Alzheimer's disease.