title: Lentiviral arrays for live-cell dynamic monitoring of gene and pathway activity during stem cell differentiation.
abstract: Uncovering the complexity of mesenchymal stem cell (MSC) differentiation requires novel methods to capture the dynamics of the process in a quantitative and high-throughput manner. To this end, we developed a lentiviral array (LVA) of reporters to capture the dynamics of gene and pathway activity during MSC differentiation into adipogenic, chondrogenic, and osteogenic lineages. Our results identified signature promoters and pathways with unique activation profile for each MSC lineage. In combination with chemical inhibitors, lineage-specific reporters predicted the effects of signaling pathway perturbations on MSC differentiation. Interestingly, some pathways were critical for differentiation into all lineages, while others had differential effects on each lineage. Our study suggests that when combined with large chemical or siRNA libraries, the reporter LVA can be used to uncover novel genes and signaling pathways affecting complex biological processes such as stem cell differentiation or reprogramming.
authors.affiliation: Department of Chemical and Biological Engineering, Bioengineering Laboratory, University at Buffalo, The State University of New York, Amherst, New York, USA.
authors.forename: Roshan M
authors.initials: RM
authors.lastname: Padmashali
authors.affiliation: Department of Chemical and Biological Engineering, Bioengineering Laboratory, University at Buffalo, The State University of New York, Amherst, New York, USA.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Mistriotis
authors.affiliation: Department of Chemical and Biological Engineering, Bioengineering Laboratory, University at Buffalo, The State University of New York, Amherst, New York, USA.
authors.forename: Mao-shih
authors.initials: MS
authors.lastname: Liang
authors.affiliation: 1] Department of Chemical and Biological Engineering, Bioengineering Laboratory, University at Buffalo, The State University of New York, Amherst, New York, USA [2] Department of Biomedical Engineering, University at Buffalo, The State University of New York, New York, Amherst, USA [3] Center of Excellence in Bioinformatics and Life Sciences, Buffalo, New York, USA.
authors.forename: Stelios T
authors.initials: ST
authors.lastname: Andreadis
mesh_terms: D050156:Adipogenesis; D000328:Adult; D000368:Aged; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D020219:Chondrogenesis; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D005822:Genetic Vectors; D018859:Hair Follicle / Q000166:cytology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D020411:Oligonucleotide Array Sequence Analysis; D010012:Osteogenesis; D015398:Signal Transduction