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

2019-07-23

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

Tom Dunkley1,*Veronica Costa2, Arno Friedlein1, Sebastian Lugert2, Stefan Aigner3, Martin Ebeling1,Meghan T. Miller2, Christoph Patsch4, Paolo Piraino5, Paul Cutler1 and Ravi Jagasia2,†

  1. Roche Pharmaceutical Research & Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel

  2. Roche Pharmaceutical Research & Early Development, Discovery Neuroscience, Roche Innovation Center Basel

  3. Dept. of Cellular and Molecular Medicine, University of California San Diego, School of Medicine

  4. Roche Pharmaceutical Research & Early Development, Small Molecule Research, Roche Innovation Center Basel

  5. Pvalue Research SRL

 PROTEOMICS – Clinical Applications (2015)

http://dx.doi.org/10.1002/prca.201400150

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

: 175 proteins were reliably quantified revealing a time-dependent pattern of protein regulation that reflected protein dynamics during in vivo brain development and which 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.