Characterization of a human pluripotent stem cell‐derived model of neuronal development using multiplexed targeted proteomics. Issue 7 (26th May 2015)

• Record Type: Journal Article
• Authors: Dunkley, Tom; Costa, Veronica; Friedlein, Arno; Lugert, Sebastian; Aigner, Stefan; Ebeling, Martin; Miller, Meghan T.; Patsch, Christoph; Piraino, Paolo; Cutler, Paul; Jagasia, Ravi; Cutler, Paul; Voshol, Hans
• Is Part Of: Proteomics. Volume 9:Issue 7/8(2015)
• Page Start: 684
• Page End: 694
• Publication Date: 2015-05-26
• Subjects: Proteomics -- Periodicals; 572.605
• DOI: 10.1002/prca.201400150
• Languages: English
• ISSNs: 1862-8346

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="prca1652-sec-0010" sec-type="section"> <title>Purpose</title> <p>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.</p> </sec> <sec id="prca1652-sec-0020" sec-type="section"> <title>Experimental design</title> <p>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.</p> </sec> <sec id="prca1652-sec-0030" sec-type="section"> <title>Results</title> <p>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.</p> </sec> <sec id="prca1652-sec-0040" sec-type="section"> <title>Conclusions and clinical relevance</title> <p>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.</p> </sec> </abstract>