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Transcriptional program of bone morphogenetic protein-2-induced epithelial and smooth muscle differentiation of pluripotent human embryonal carcinoma cells

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Abstract

Pluripotent human embryonal carcinoma NTera2/cloneD1 (NT2/D1) cells respond to multiple vertebrate patterning factors and offer a unique model system to investigate the signaling events associated with lineage determination and cell differentiation. Here, we define the temporal changes in global gene expression patterns in NT2/D1 cells upon treatment with bone morphogenetic protein-2 (BMP-2). Exposure to BMP-2 rapidly induced the expression of several transcription factors involved in establishing non-neural ectodermal fate followed by the appearance of epithelial-specific markers. Subsequent loss of stem cell markers was coupled to gene expression changes associated with decreased proliferative activity. Temporal clustering of gene expression patterns revealed a concurrent down-regulation of multiple transcripts involved in neurogenesis, neurite outgrowth, and axonal guidance, suggesting that the BMP-mediated differentiation process involves pro-epithelial as well as anti-neurogenic mechanisms. In addition, increased expression of smooth muscle markers both by gene expression and immunohistochemistry was detected. Several neural crest markers were induced preceding such a differentiation, compatible with a neural crest origin of NT2/D1-derived smooth muscle cells. Comparison of changes in transcript expression between BMP-2-induced epithelial versus all-trans-retinoic acid (ATRA)-induced neural differentiation revealed potential candidates for regulation of BMP-2 signaling and suppression of neural fate by BMP-2. This study suggests that BMP-2-induced differentiation of NT2/D1 cells provides a powerful assay to study early human epithelial and smooth muscle development.

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Acknowledgements

We thank the MSKCC Genomics Core Facility for hybridization, washing, and scanning of the arrays. This work was supported by grants from the NIH and the Byrne Research Fund.

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Authors and Affiliations

  1. Cell Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 391, New York, NY, 10021, USA

    Rajendrakumar S. V. Chadalavada, Jane Houldsworth & R. S. K. Chaganti

  2. Department of Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 391, New York, NY, 10021, USA

    Jane Houldsworth, George J. Bosl & R. S. K. Chaganti

  3. Department of Epidemiology and Biostatistics, Memorial Sloan-Kettering Cancer Center, 307 East 63rd Street, Suite 349, New York, NY, 10021, USA

    Adam B. Olshen

  4. Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 256, New York, NY, 10021, USA

    Lorenz Studer

  5. Division of Neurosurgery, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 256, New York, NY, 10021, USA

    Lorenz Studer

  6. Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 30, New York, NY, 10021, USA

    R. S. K. Chaganti

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  1. Rajendrakumar S. V. Chadalavada
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  2. Jane Houldsworth
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  6. R. S. K. Chaganti
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Correspondence to R. S. K. Chaganti.

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Chadalavada, R.S.V., Houldsworth, J., Olshen, A.B. et al. Transcriptional program of bone morphogenetic protein-2-induced epithelial and smooth muscle differentiation of pluripotent human embryonal carcinoma cells. Funct Integr Genomics 5, 59–69 (2005). https://doi.org/10.1007/s10142-005-0132-7

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