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Loss of adenomatous polyposis coli gene function disrupts thymic development

Nature Immunology volume 6, pages 800–809 (2005)Cite this article

Abstract

Loss of the adenomatous polyposis coli (APC) protein is a common initiating event in colon cancer. Here we show that thymocyte-specific loss of APC deregulated β-catenin signaling and suppressed Notch-dependent transcription. These events promoted the proliferation of cells of the double-negative 3 and 4 stages and reduced rearrangements between the variable, diversity and joining regions of the gene encoding T cell receptor (TCR) β, encouraging developmental progression of aberrant thymocytes lacking pre-TCR and αβ TCR. Simultaneously, the loss of APC prolonged the mitotic metaphase-to-anaphase checkpoint and impaired chromosome segregation, blocking development beyond the double-negative 4 stage. The result was extensive thymic atrophy and increased frequencies of thymocytes with chromosomal abnormalities. Thus, loss of APC in immature thymocytes has consequences distinct from those of deregulation of β-catenin signaling and is essential for T cell differentiation.

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Figure 1: Conditional Apc ablation in mouse thymocytes.
Figure 2: Mice with conditional Apc deficiency have reduced thymic cellularity and a blockade at the DN4 stage of thymocyte development.
Figure 3: Survival and proliferation of APC deficient thymocytes.
Figure 4: Suppression of Tcrb VDJ rearrangements after ablation of APC or stabilization of β-catenin.
Figure 5: Suppression of Notch activity by APC truncation and stabilization of β-catenin.
Figure 6: Expression of activated Notch1 partially restores intracellular TCRβ expression in LckCre-Apclox/lox468 and LckCre-Ctnnb1+/lox(ex3) thymocytes differentiating on OP9-DL1 stroma cells.
Figure 7: LckCre-Apclox/lox468 DN4 thymocytes divide and differentiate more slowly than LckCre or LckCre-Ctnnb1+/lox(ex3) DN4 thymocytes on OP9-DL1 stroma cells.
Figure 8: Ablation of APC results in inhibition of chromosome segregation after chromatide separation and the accumulation of cells with aberrant chromosome numbers.

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Acknowledgements

The authors thank J.C. Zúñiga-Pflücker (University of Toronto, Toronto, Canada) for advice, support and the OP9-DL1 cells; I. Khan for help with the OP9-DL1 cultures; K. Georgopoulos (Massachusetts General Hospital), P. Sicinski and R. Van Etten for critical comments on the manuscript; I. Aifantis (University of Chicago), X. Li and A. Campese for the NotchIC retrovirus; and H. von Boehmer and R.M. Meyer for their interest and support. Supported by the National Institutes of Health (R01 AI059676-01 to F.G. and R01 CA104547-01A1 to K.K.), the Medical Foundation (Smith Family New Investigator Award to F.G.), the Dana Farber Cancer Institute (National Colorectal Cancer Research Alliance Award to K.K.) and the Claudia Adams Barr Program (K.K.).

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Authors and Affiliations

  1. Molecular Oncology Research Institute, Tufts–New England Medical Center, Boston, 02111, Massachusetts, USA

    Fotini Gounari, Rui Chang, Zhuyan Guo & Marei Dose

  2. Department of Pediatrics, Tufts–New England Medical Center, Boston, 02111, Massachusetts, USA

    Janet Cowan

  3. Dana Farber Cancer Institute, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Elias Gounaris & Khashayarsha Khazaie

  4. Center for Molecular Imaging Research, Massachusetts General Hospital and Harvard Medical School, Charlestown, 02129, Massachusetts, USA

    Elias Gounaris & Khashayarsha Khazaie

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Supplementary Fig. 1

Detection of mitotic abnormalities in DN LckCre-APClox/loxD468 thymocytes. (PDF 570 kb)

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Gounari, F., Chang, R., Cowan, J. et al. Loss of adenomatous polyposis coli gene function disrupts thymic development. Nat Immunol 6, 800–809 (2005). https://doi.org/10.1038/ni1228

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