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Lineage Reprogramming of mouse fibroblasts into induced cardiac progenitor cells by defined factors

ABSTRACT: Mouse adult cardiac fibroblasts were infected with doxycycline inducible, 11 cardiac factors. Addition of doxycycline resulted in appearance of proliferative cells that had high nuclear-to-cytoplasmic ratio. We called these cells iCPCs. iCPC were stably reporgrammed and could be maintined in proliferative state without doxycycline. Here we compared the transcriptome of iCPC with their parental cells; adult cardiac fibroblasts. We found that iCPCs have upregulated genes involved in cardiovascular development and downregulated fibroblasts specific genes. iCPCs did not expressed endoderm, ectoderm or pluripotency associated genes indicating that they were cardiac mesoderm restricted progenitors.

ORGANISM(S): Mus musculus

PROVIDER: GSE61486 | GEO | 2015/09/16

SECONDARY ACCESSION(S): PRJNA261286

REPOSITORIES: GEO

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Project description:The germ layer concept has been one of the foremost organizing principles in developmental biology, classification, systematics and evolution for 150 years. Of the three germ layers, the mesoderm is found in bilaterian animals but is absent in species in the phyla Cnidaria and Ctenophora, which has been taken as evidence that the mesoderm was the final germ layer to evolve. The origin of the ectoderm and endoderm layers, however, remains unclear with models supporting the antecedence of each as well as a simultaneous origin. We hypothesized that global analysis of gene expression in each layer throughout development may resolve the early events of animal evolution. Here, we determine the temporal and spatial components of gene expression spanning embryonic development for all C. elegans genes and use it to determine the evolutionary age of the germ layers. The gene expression programs of the mesoderm is generally induced after the ectoderm and endoderm germ layers, thus making it the last germ layer to both evolve and develop. Strikingly, the C. elegans endoderm and ectoderm expression programs do not co-induce; rather the endoderm activates earlier. We also observed early expression of endoderm orthologs during the embryology of Xenopus tropicalis, Nematostella vectensis, and the sponge Amphimedon queenslandica. Querying for the phylogenetic ages of specifically expressed genes revealed that the endoderm is comprised of older genes, supporting its antecedence among the germ layers. Taken together, we propose that the endoderm program has retained the feeding functions of the last common ancestor with the choanoflagellates, thus allowing for the specialization of an ectoderm germ layer. Our work reveals that the evolutionary appearance of the germ layers continues to constrain regulatory networks in metazoans. Two temporal assays of C. elegans embryonic development, starting at the zygote: (a) Embryos collected at fixed (~10 minute) time intervals. (b) Embryo segregates, up to five lines of blastomeres, isolated in reference to mitotic events. There were 184 samples in total, representing 100 distnict data points.

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Lineage Reprogramming of mouse fibroblasts into induced cardiac progenitor cells by defined factors

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