title: Small molecules increase direct neural conversion of human fibroblasts.
abstract: The generation of human induced neurons (hiNs) via exogenous delivery of neural transcription factors represents a novel technique to obtain disease and patient specific neurons. These cells have the potential to be used for disease modeling, diagnostics and drug screening, and also to be further developed for brain repair. In the present study, we utilized hiNs to develop an unbiased screening assay for small molecules that increase the conversion efficiency. Using this assay, we screened 307 compounds from five annotated libraries and identified six compounds that were very potent in potentiating the reprogramming process. When combined in an optimal combination and dose, these compounds increased the reprogramming efficiency of human fibroblasts more than 6-fold. Global gene expression and CellNet analysis at different timepoints during the reprogramming process revealed that neuron-specific genes and gene regulatory networks (GRNs) became progressively more activated while converting cells shut down fibroblast-specific GRNs. Further bioinformatics analysis revealed that the addition of the six compound resulted in the accelerated upregulation of a subset of neuronal genes, and also increased expression of genes associated with transcriptional activity and mediation of cellular stress response.
authors.affiliation: Department of Experimental Medical Science, Wallenberg Neuroscience Center 221 84, Lund, Sweden.|Lund Stem Cell Center, Lund University, 221 84, Lund, Sweden.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Pfisterer
authors.affiliation: Chemical Biology &Therapeutics, Department of Experimental Medical Science, Lund University, 221 84, Lund, Sweden.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Ek
authors.affiliation: Lund Stem Cell Center, Lund University, 221 84, Lund, Sweden.|Division of Molecular Hematology, BMC, Lund University, Lund 22242, Sweden.
authors.forename: Stefan
authors.initials: S
authors.lastname: Lang
authors.affiliation: Lund Stem Cell Center, Lund University, 221 84, Lund, Sweden.|Division of Molecular Hematology, BMC, Lund University, Lund 22242, Sweden.
authors.forename: Shamit
authors.initials: S
authors.lastname: Soneji
authors.affiliation: Chemical Biology &Therapeutics, Department of Experimental Medical Science, Lund University, 221 84, Lund, Sweden.
authors.forename: Roger
authors.initials: R
authors.lastname: Olsson
authors.affiliation: Department of Experimental Medical Science, Wallenberg Neuroscience Center 221 84, Lund, Sweden.|Lund Stem Cell Center, Lund University, 221 84, Lund, Sweden.
authors.forename: Malin
authors.initials: M
authors.lastname: Parmar
mesh_terms: D000813:Anilides / Q000494:pharmacology; D001552:Benzazepines / Q000494:pharmacology; D002460:Cell Line; D065150:Cellular Reprogramming / Q000187:drug effects*; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000494:pharmacology; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D000077185:Resveratrol; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013267:Stilbenes / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: C412373:AG 1879; D000813:Anilides; D001552:Benzazepines; C473711:Chir 99021; D007211:Indoles; C514580:N1-(2-aminophenyl)-N8-phenyloctanediamide; D009419:Nerve Tissue Proteins; D011725:Pyridines; D011743:Pyrimidines; D054852:Small Molecule Libraries; D013267:Stilbenes; D014157:Transcription Factors; C119620:kenpaullone; D000242:Cyclic AMP; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D015232:Dinoprostone; D000077185:Resveratrol