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PTEN-deficient intestinal stem cells initiate intestinal polyposis

Nature Genetics volume 39, pages 189–198 (2007)Cite this article

Abstract

Intestinal polyposis, a precancerous neoplasia, results primarily from an abnormal increase in the number of crypts, which contain intestinal stem cells (ISCs). In mice, widespread deletion of the tumor suppressor Phosphatase and tensin homolog (PTEN) generates hamartomatous intestinal polyps with epithelial and stromal involvement. Using this model, we have established the relationship between stem cells and polyp and tumor formation. PTEN helps govern the proliferation rate and number of ISCs and loss of PTEN results in an excess of ISCs. In PTEN-deficient mice, excess ISCs initiate de novo crypt formation and crypt fission, recapitulating crypt production in fetal and neonatal intestine. The PTEN-Akt pathway probably governs stem cell activation by helping control nuclear localization of the Wnt pathway effector β-catenin. Akt phosphorylates β-catenin at Ser552, resulting in a nuclear-localized form in ISCs. Our observations show that intestinal polyposis is initiated by PTEN-deficient ISCs that undergo excessive proliferation driven by Akt activation and nuclear localization of β-catenin.

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Figure 1: Inactivation of PTEN leads to intestinal polyposis.
Figure 2: PTEN expression in the intestine and the impact of PTEN inactivation on the activity of the PI3K-Akt pathway and on the expression of cell cycle regulators.
Figure 3: PTEN-deficient intestinal stem cells were found at the initiation of crypt budding and fission.
Figure 4: PI3K-Akt pathway and its downstream targets operate in ISCs when PTEN is inactivated.
Figure 5: Identification of the Akt phosphorylation site at the C terminus of β-catenin.
Figure 6: Akt activity coincides with nuclear p-β-cat-Ser552 and β-catenin–dependent transcriptional activity in ISCs.
Figure 7: Cells with nuclear p-β-cat-Ser552 initiate crypt fission and budding.

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Acknowledgements

We are grateful to B. Neaves and R. Krumlauf for scientific support. We thank H. Okano for providing anti-Musashi1 (14-H1) and A. Ouellette for anti-cryptdin; S. Peck and G. Yang for comments on the manuscript; D. di Natale for assistance on manuscript editing; P. Kulesa and D. Stark for imaging assistance; C. Seidel, K. Zueckert-Gaudenz and M. Coleman for assistance in microarray analysis; H. Marshall for technology support and J. Chen for assistance in statistical analysis. L. Li is supported in part by research grant 5-FY05-31 from the March of Dimes Birth Defects Foundation, by grant R01 DK070001 from the National Institute of Diabetes and Digestive and Kidney Diseases and by the Stowers Institute for Medical Research.

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Authors and Affiliations

  1. Stowers Institute for Medical Research, Kansas City, 64110, Missouri, USA

    Xi C He, Tong Yin, Justin C Grindley, Toshiro Sato, Kimberly S Porter-Westpfahl, Mark Hembree, Teri Johnson, Leanne M Wiedemann & Linheng Li

  2. Institute for Systems Biology, Seattle, 98103, Washington, USA

    Qiang Tian & Leroy Hood

  3. Department of Biochemistry, Purdue University, West Lafayette, 47907, Indiana, USA

    W Andy Tao

  4. Department of Medicine and Microbiology/Immunology, Division of Gastroenterology, Northwestern University Medical School, Chicago, 60611, Illinois, USA

    Raminarao Dirisina & Terrence A Barrett

  5. Department of Pathology and Laboratory Medicine, Kansas University Medical Center, Kansas City, 66160, Kansas, USA

    Leanne M Wiedemann & Linheng Li

  6. Department of Molecular and Medical Pharmacology, University of California, Los Angeles, School of Medicine, Los Angeles, 90095, California, USA

    Hong Wu

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Contributions

X.C.H., J.C.G. and L.L. designed the research; X.C.H., T.Y., T.S. and J.C.G. performed the research; Q.T., W.A.T. and L.H. performed the mass spectrometry; R.D., K.S.P-W., M.H., T.J. and T.A.B. provided technical support; H.W. contributed critical reagents; J.C.G., L.M.W. and L.L. wrote the paper.

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Correspondence to Linheng Li.

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Supplementary information

Supplementary Fig. 1

Comparison of expression patterns of p-PTEN, p-Akt, 14-3-3ζ, pGSK3β and Musashi1. (PDF 1831 kb)

Supplementary Fig. 2

Detection of Musashi1 expression using an antibody to Musashi1 (clone 14H1). (PDF 380 kb)

Supplementary Fig. 3

Cluster of Musashi1+ cells detected at the initiation site of crypt budding or crypt fission in PTEN mutants. (PDF 367 kb)

Supplementary Fig. 4

Comparison of expression patterns of pβ-cat-Ser552, Top-Gal, p27kip1 and cyclin D1. (PDF 555 kb)

Supplementary Fig. 5

Distribution of cells with pβ-cat-Ser552. (PDF 414 kb)

Supplementary Video 1

PTEN-deficient stem cells initiate crypt fission. Crypts were dissociated from the polyp region of PTEN mutant intestine. Whole-mount staining was performed using two antibodies: anti-β-cat-S552 (green) to recognize stem cells (normal or abnormal) and anti-cryptdin (red) to recognize Paneth cells. One β-cat-S552+ cell was located just above the Paneth cells in the proposed stem cell position; the other dividing β-cat-S552+ cell was found at the tip of the ridge formed between two dividing crypts. (AVI 2092 kb)

Supplementary Video 2

PTEN-deficient stem cells initiate crypt budding. Crypts were dissociated from the polyp region of PTEN mutant intestine. Whole-mount staining was performed using two antibodies: anti-β-cat-S552 (green) to recognize stem cells (normal or abnormal) and anti-cryptdin (red) to recognize Paneth cells. One β-cat-S552+ cell was located just above the Paneth cells in the proposed stem cell position; a cluster of β-cat-S552+ cells was found at the junction between a budding crypt and the edge of the original crypt. (AVI 2168 kb)

Supplementary Methods (PDF 105 kb)

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He, X., Yin, T., Grindley, J. et al. PTEN-deficient intestinal stem cells initiate intestinal polyposis. Nat Genet 39, 189–198 (2007). https://doi.org/10.1038/ng1928

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