title: Characterization of a human pluripotent stem cell-derived model of neuronal development using multiplexed targeted proteomics.
abstract: OBJECTIVE
  Human pluripotent stem cell (hPSC)-derived cellular models have great potential to enable drug discovery and improve translation of preclinical insights to the clinic. We have developed a hPSC-derived neural precursor cell model for studying early events in human brain development. We present protein-level characterization of this model, using a multiplexed SRM approach, to establish reproducibility and physiological relevance; essential prerequisites for utilization of the neuronal development model in phenotypic screening-based drug discovery.
  
  
  METHODS
  Profiles of 246 proteins across three key stages of in vitro neuron differentiation were analyzed by SRM. Three independently hPSC-derived isogenic neural stem cell (NSC) lines were analyzed across five to nine independent neuronal differentiations.
  
  
  RESULTS
  One hundred seventy-five proteins were reliably quantified revealing a time-dependent pattern of protein regulation that reflected protein dynamics during in vivo brain development and that was conserved across replicate differentiations and multiple cell lines.
  
  
  CONCLUSIONS
  SRM-based protein profiling enabled establishment of the reproducibility and physiological relevance of the hPSC-derived neuronal model. Combined with the successful quantification of proteins relevant to neurodevelopmental diseases, this validates the platform for use as a model to enable neuroscience drug discovery.
authors: Roche Pharmaceutical Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors: Tom
authors: T
authors: Dunkley
authors: Roche Pharmaceutical Research and Early Development, Discovery Neuroscience, Roche Innovation Center Basel, Basel, Switzerland.
authors: Veronica
authors: V
authors: Costa
authors: Roche Pharmaceutical Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors: Arno
authors: A
authors: Friedlein
authors: Roche Pharmaceutical Research and Early Development, Discovery Neuroscience, Roche Innovation Center Basel, Basel, Switzerland.
authors: Sebastian
authors: S
authors: Lugert
authors: Roche Pharmaceutical Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors: Stefan
authors: S
authors: Aigner
authors: Roche Pharmaceutical Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors: Martin
authors: M
authors: Ebeling
authors: Roche Pharmaceutical Research and Early Development, Discovery Neuroscience, Roche Innovation Center Basel, Basel, Switzerland.
authors: Meghan T
authors: MT
authors: Miller
authors: Roche Pharmaceutical Research and Early Development, Small Molecule Research, Roche Innovation Center Basel, Basel, Switzerland.
authors: Christoph
authors: C
authors: Patsch
authors: Pvalue Research S.R.L, Castel San Giovanni, Piacenza, Italy.
authors: Paolo
authors: P
authors: Piraino
authors: Roche Pharmaceutical Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors: Paul
authors: P
authors: Cutler
authors: Roche Pharmaceutical Research and Early Development, Discovery Neuroscience, Roche Innovation Center Basel, Basel, Switzerland.
authors: Ravi
authors: R
authors: Jagasia
mesh_terms: D002454:Cell Differentiation; D002460:Cell Line; D016000:Cluster Analysis; D006801:Humans; D008954:Models, Biological; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D025341:Principal Component Analysis; D040901:Proteomics / Q000379:methods*; D013997:Time Factors
keywords: In vitro model; Neuronal development; Phenotypic screen; Pluripotent stem cell; SRM