Characterization of a human pluripotent stem cell-derived model of neuronal development using multiplexed targeted proteomics

Proteomics Clin Appl. 2015 Aug;9(7-8):684-94. doi: 10.1002/prca.201400150. Epub 2015 May 26.

Abstract

Purpose: 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.

Experimental design: 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 and clinical relevance: 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.

Keywords: In vitro model; Neuronal development; Phenotypic screen; Pluripotent stem cell; SRM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation
  • Cell Line
  • Cluster Analysis
  • Humans
  • Models, Biological
  • Neurons / cytology*
  • Neurons / metabolism*
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism*
  • Principal Component Analysis
  • Proteomics / methods*
  • Time Factors