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| Status |
Public on Jun 05, 2015 |
| Title |
Outer ECM-attached vs. inner cells of MCF10A acini at day 6 of morphogenesis |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Basal-like carcinoma is a subtype of breast cancer that is characterized by poor prognosis and high intratumor heterogeneity. Using a basal-like breast epithelial line, we have identified two anti-correlated gene-expression programs that arise among single extracellular matrix (ECM)-attached cells during organotypic 3D culture. The first program contains TGFBR3, a high-affinity receptor for transforming growth factor β (TGFβ) and other related ligands. The second program contains the JUND transcription factor together with the basal-like marker, KRT5. By disrupting the TGFBR3 and JUND programs individually, we reveal an important circuit for 3D morphogenesis that is wired together by four negative-feedback loops. Computational modeling of this circuit showed that it could exhibit damped, antiphase oscillations when excited with small impulses of TGFβ-like ligand. We directly visualize the circuit's spontaneous dynamics in organotypic cultures by using live-cell imaging with engineered pathway reporters. Importantly, we show that the essence of the JUND-TGFBR3 expression circuit holds true in early basal-like tumors that heterogeneously express KRT5. Correlated JUND-KRT5 expression depends critically on contact with stromal ECM and local expression of tenascin C. This work illustrates how complex tumor heterogeneities can be deconstructed into intrinsic single-cell expression circuits that are modulated by the microenvironment.
Gene expression analysis of outer ECM-attached vs. inner cells of MCF10A-5E clones grown in organotypic 3D culture at day 6.
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| Overall design |
Total RNA was isolated from ~50 outer ECM-attached cells and ~50 inner cells, each separately microdissected from 8 micron sections of MCF10A structures at day 6 of morphogenesis. Total RNA was amplified in quadruplicate, and hybridized to HumanRef-8 v2.0 Expression BeadChips (Illumina).
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| Contributor(s) |
Janes KA |
| Citation(s) |
24658685 |
| Submission date |
Oct 11, 2012 |
| Last update date |
Jun 05, 2015 |
| Contact name |
Leen Jamal |
| E-mail(s) |
lj2p@virginia.edu, ljamal@ucsd.edu
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| Organization name |
University of Virginia
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| Department |
Biomedical Engineering
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| Lab |
The Cell-signaling Bioanalysis Laboratory
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| Street address |
415 Lane Road, MR5-Rm. 2225
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| City |
Charlottesville |
| State/province |
VA |
| ZIP/Postal code |
22908 |
| Country |
USA |
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| Platforms (1) |
| GPL6255 |
Illumina humanRef-8 v2.0 expression beadchip |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA177354 |
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