title: Protein Kinase-A Inhibition Is Sufficient to Support Human Neural Stem Cells Self-Renewal.
abstract: Human pluripotent stem cell-derived neural stem cells offer unprecedented opportunities for producing specific types of neurons for several biomedical applications. However, to achieve it, protocols of production and amplification of human neural stem cells need to be standardized, cost effective, and safe. This means that small molecules should progressively replace the use of media containing cocktails of protein-based growth factors. Here we have conducted a phenotypical screening to identify pathways involved in the regulation of hNSC self-renewal. We analyzed 80 small molecules acting as kinase inhibitors and identified compounds of the 5-isoquinolinesulfonamide family, described as protein kinase A (PKA) and protein kinase G inhibitors, as candidates to support hNSC self-renewal. Investigating the mode of action of these compounds, we found that modulation of PKA activity was central in controlling the choice between self-renewal or terminal neuronal differentiation of hNSC. We finally demonstrated that the pharmacological inhibition of PKA using the small molecule HA1004 was sufficient to support the full derivation, propagation, and long-term maintenance of stable hNSC in absence of any other extrinsic signals. Our results indicated that tuning of PKA activity is a core mechanism regulating hNSC self-renewal and differentiation and delineate the minimal culture media requirement to maintain undifferentiated hNSC in vitro.
authors: CECS, 91030, Evry Cedex, France.
authors: Pauline
authors: P
authors: Georges
authors: CECS, 91030, Evry Cedex, France.
authors: Claire
authors: C
authors: Boissart
authors: CECS, 91030, Evry Cedex, France.
authors: Aurélie
authors: A
authors: Poulet
authors: INSERM UMR 861, 91030, Evry Cedex, France.|UEVE UMR 861, I-STEM, AFM, 91030, Evry Cedex, France.
authors: Marc
authors: M
authors: Peschanski
authors: CECS, 91030, Evry Cedex, France.
authors: Alexandra
authors: A
authors: Benchoua
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors*; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology / Q000201:enzymology*; D009474:Neurons / Q000166:cytology / Q000201:enzymology; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D047428:Protein Kinase Inhibitors; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: High content analysis; High throughput screening; Human neural stem cells; Pluripotent stem cells; Protein-kinase A; Self-renewal