title: Derivation and expansion using only small molecules of human neural progenitors for neurodegenerative disease modeling.
abstract: Phenotypic drug discovery requires billions of cells for high-throughput screening (HTS) campaigns. Because up to several million different small molecules will be tested in a single HTS campaign, even small variability within the cell populations for screening could easily invalidate an entire campaign. Neurodegenerative assays are particularly challenging because neurons are post-mitotic and cannot be expanded for implementation in HTS. Therefore, HTS for neuroprotective compounds requires a cell type that is robustly expandable and able to differentiate into all of the neuronal subtypes involved in disease pathogenesis. Here, we report the derivation and propagation using only small molecules of human neural progenitor cells (small molecule neural precursor cells; smNPCs). smNPCs are robust, exhibit immortal expansion, and do not require cumbersome manual culture and selection steps. We demonstrate that smNPCs have the potential to clonally and efficiently differentiate into neural tube lineages, including motor neurons (MNs) and midbrain dopaminergic neurons (mDANs) as well as neural crest lineages, including peripheral neurons and mesenchymal cells. These properties are so far only matched by pluripotent stem cells. Finally, to demonstrate the usefulness of smNPCs we show that mDANs differentiated from smNPCs with LRRK2 G2019S are more susceptible to apoptosis in the presence of oxidative stress compared to wild-type. Therefore, smNPCs are a powerful biological tool with properties that are optimal for large-scale disease modeling, phenotypic screening, and studies of early human development.
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, North Rhine Westphalia, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Reinhardt
authors.forename: Michael
authors.initials: M
authors.lastname: Glatza
authors.forename: Kathrin
authors.initials: K
authors.lastname: Hemmer
authors.forename: Yaroslav
authors.initials: Y
authors.lastname: Tsytsyura
authors.forename: Cora S
authors.initials: CS
authors.lastname: Thiel
authors.forename: Susanne
authors.initials: S
authors.lastname: Höing
authors.forename: Sören
authors.initials: S
authors.lastname: Moritz
authors.forename: Juan A
authors.initials: JA
authors.lastname: Parga
authors.forename: Lydia
authors.initials: L
authors.lastname: Wagner
authors.forename: Jan M
authors.initials: JM
authors.lastname: Bruder
authors.forename: Guangming
authors.initials: G
authors.lastname: Wu
authors.forename: Benjamin
authors.initials: B
authors.lastname: Schmid
authors.forename: Albrecht
authors.initials: A
authors.lastname: Röpke
authors.forename: Jürgen
authors.initials: J
authors.lastname: Klingauf
authors.forename: Jens C
authors.initials: JC
authors.lastname: Schwamborn
authors.forename: Thomas
authors.initials: T
authors.lastname: Gasser
authors.forename: Hans R
authors.initials: HR
authors.lastname: Schöler
authors.forename: Jared
authors.initials: J
authors.lastname: Sterneckert
mesh_terms: D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D004594:Electrophysiology; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D009046:Motor Neurons / Q000166:cytology / Q000378:metabolism; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism*; D019636:Neurodegenerative Diseases / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases