Functional & Integrative Genomics

, Volume 5, Issue 2, pp 59–69 | Cite as

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

  • Rajendrakumar S. V. Chadalavada
  • Jane Houldsworth
  • Adam B. Olshen
  • George J. Bosl
  • Lorenz Studer
  • R. S. K. Chaganti
Original Paper

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.

Keywords

Embryonal carcinoma BMP-2 Micro-array Differentiation 

Supplementary material

Figures S1–S4

s10142_2005_132_fig_s1_4.pdf (52 kb)
(PDF 52 KB)

Table S1 Gene expression data from the time course analysis in triplicate after BMP-2 treatment of NT2/D1 cells

s10142_2005_132_table_s1.xls (20 mb)
(Excel 20 MB)

Table S2 Signal log ratios of time course analysis in triplicate after BMP-2 treatment of NT2/D1 cells. Designated clusters from Fig.1 and Fig. S1 are indicated in separate and adjacent columns

s10142_2005_132_table_s2.xls (304 kb)
(PDF 1 MB)
s10142_2005_132_table_s2.xls (304 kb)
(Excel 304 KB)

Table S3 Signal log ratios of time course analysis in triplicate after ATRA treatment of NT2/D1 cells. Changes in transcript expression during ATRA-induced differentiation was analyzed using the Micro Array Suite 5.0, using the 24 hour untreated control culture as the baseline

s10142_2005_132_table_s3.xls (207 kb)
(PDF 792 KB)
s10142_2005_132_table_s3.xls (207 kb)
(Excel 207 KB)

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Rajendrakumar S. V. Chadalavada
    • 1
  • Jane Houldsworth
    • 1
    • 2
  • Adam B. Olshen
    • 3
  • George J. Bosl
    • 2
  • Lorenz Studer
    • 4
    • 5
  • R. S. K. Chaganti
    • 1
    • 2
    • 6
  1. 1.Cell Biology ProgramMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  2. 2.Department of MedicineMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  3. 3.Department of Epidemiology and BiostatisticsMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  4. 4. Developmental Biology ProgramMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  5. 5.Division of NeurosurgeryMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  6. 6.Memorial Sloan-Kettering Cancer CenterNew YorkUSA

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