Protein tyrosine phosphorylation is thought to be important for regulation of the proliferation, differentiation, and rapid turnover of intestinal epithelial cells (IECs). The role of protein tyrosine phosphatases in such homeostatic regulation of IECs has remained largely unknown, however. Src homology 2–containing protein tyrosine phosphatase (Shp2) is a ubiquitously expressed cytoplasmic protein tyrosine phosphatase that functions as a positive regulator of the Ras–mitogen-activated protein kinase (MAPK) signaling pathway operative downstream of the receptors for various growth factors and cytokines, and it is thereby thought to contribute to the regulation of cell proliferation and differentiation. We now show that mice lacking Shp2 specifically in IECs (Shp2 CKO mice) develop severe colitis and die as early as 3 to 4 weeks after birth. The number of goblet cells in both the small intestine and colon of Shp2 CKO mice was markedly reduced compared with that for control mice. Furthermore, Shp2 CKO mice showed marked impairment of both IEC migration along the crypt-villus axis in the small intestine and the development of intestinal organoids from isolated crypts. The colitis as well as the reduction in the number of goblet cells apparent in Shp2 CKO mice were normalized by expression of an activated form of K-Ras in IECs. Our results thus suggest that Shp2 regulates IEC homeostasis through activation of Ras and thereby protects against the development of colitis.
References
[1]
Blanpain C, Horsley V, Fuchs E (2007) Epithelial stem cells: turning over new leaves. Cell 128: 445–458. doi: 10.1016/j.cell.2007.01.014
[2]
van der Flier LG, Clevers H (2009) Stem cells, self-renewal, and differentiation in the intestinal epithelium. Annu Rev Physiol 71: 241–260. doi: 10.1146/annurev.physiol.010908.163145
[3]
Hall PA, Coates PJ, Ansari B, Hopwood D (1994) Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. J Cell Sci 107 (Pt 12): 3569–3577.
[4]
Eisenhoffer GT, Loftus PD, Yoshigi M, Otsuna H, Chien CB, et al. (2012) Crowding induces live cell extrusion to maintain homeostatic cell numbers in epithelia. Nature 484: 546–549. doi: 10.1038/nature10999
[5]
Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, et al. (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459: 262–265. doi: 10.1038/nature07935
[6]
Holmberg J, Genander M, Halford MM, Anneren C, Sondell M, et al. (2006) EphB receptors coordinate migration and proliferation in the intestinal stem cell niche. Cell 125: 1151–1163. doi: 10.1016/j.cell.2006.04.030
[7]
Batlle E, Henderson JT, Beghtel H, van den Born MM, Sancho E, et al. (2002) Beta-catenin and TCF mediate cell positioning in the intestinal epithelium by controlling the expression of EphB/ephrinB. Cell 111: 251–263. doi: 10.1016/s0092-8674(02)01015-2
[8]
Sadakata H, Okazawa H, Sato T, Supriatna Y, Ohnishi H, et al. (2009) SAP-1 is a microvillus-specific protein tyrosine phosphatase that modulates intestinal tumorigenesis. Genes Cells 14: 295–308. doi: 10.1111/j.1365-2443.2008.01270.x
[9]
Noguchi T, Tsuda M, Takeda H, Takada T, Inagaki K, et al. (2001) Inhibition of cell growth and spreading by stomach cancer-associated protein-tyrosine phosphatase-1 (SAP-1) through dephosphorylation of p130cas. J Biol Chem 276: 15216–15224. doi: 10.1074/jbc.m007208200
[10]
Matozaki T, Murata Y, Saito Y, Okazawa H, Ohnishi H (2009) Protein tyrosine phosphatase SHP-2: a proto-oncogene product that promotes Ras activation. Cancer Sci 100: 1786–1793. doi: 10.1111/j.1349-7006.2009.01257.x
[11]
Neel BG, Gu H, Pao L (2003) The 'Shp'ing news: SH2 domain-containing tyrosine phosphatases in cell signaling. Trends Biochem Sci 28: 284–293. doi: 10.1016/s0968-0004(03)00091-4
[12]
Kodama A, Matozaki T, Fukuhara A, Kikyo M, Ichihashi M, et al. (2000) Involvement of an SHP-2-Rho small G protein pathway in hepatocyte growth factor/scatter factor-induced cell scattering. Mol Biol Cell 11: 2565–2575. doi: 10.1091/mbc.11.8.2565
[13]
Schoenwaelder SM, Petch LA, Williamson D, Shen R, Feng GS, et al. (2000) The protein tyrosine phosphatase Shp-2 regulates RhoA activity. Curr Biol 10: 1523–1526. doi: 10.1016/s0960-9822(00)00831-9
[14]
Saxton TM, Henkemeyer M, Gasca S, Shen R, Rossi DJ, et al. (1997) Abnormal mesoderm patterning in mouse embryos mutant for the SH2 tyrosine phosphatase Shp-2. EMBO J 16: 2352–2364. doi: 10.1093/emboj/16.9.2352
[15]
Fornaro M, Burch PM, Yang W, Zhang L, Hamilton CE, et al. (2006) SHP-2 activates signaling of the nuclear factor of activated T cells to promote skeletal muscle growth. J Cell Biol 175: 87–97. doi: 10.1083/jcb.200602029
[16]
Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21: 70–71.
[17]
Jackson EL, Willis N, Mercer K, Bronson RT, Crowley D, et al. (2001) Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras. Genes Dev 15: 3243–3248. doi: 10.1101/gad.943001
[18]
Madison BB, Dunbar L, Qiao XT, Braunstein K, Braunstein E, et al. (2002) Cis elements of the villin gene control expression in restricted domains of the vertical (crypt) and horizontal (duodenum, cecum) axes of the intestine. J Biol Chem 277: 33275–33283. doi: 10.1074/jbc.m204935200
[19]
Murata Y, Mori M, Kotani T, Supriatna Y, Okazawa H, et al. (2010) Tyrosine phosphorylation of R3 subtype receptor-type protein tyrosine phosphatases and their complex formations with Grb2 or Fyn. Genes Cells 15: 513–524. doi: 10.1111/j.1365-2443.2010.01398.x
[20]
Siegmund B, Sennello JA, Lehr HA, Batra A, Fedke I, et al. (2004) Development of intestinal inflammation in double IL-10- and leptin-deficient mice. J Leukoc Biol 76: 782–786. doi: 10.1189/jlb.0404239
[21]
Kanazawa Y, Saito Y, Supriatna Y, Tezuka H, Kotani T, et al. (2010) Role of SIRPα in regulation of mucosal immunity in the intestine. Genes Cells 15: 1189–1200. doi: 10.1111/j.1365-2443.2010.01453.x
[22]
Barker N, Clevers H (2010) Lineage tracing in the intestinal epithelium. Curr Protoc Stem Cell Biol Chapter 5 : Unit5A 4.
[23]
van Es JH, Sato T, van de Wetering M, Lyubimova A, Nee AN, et al. (2012) Dll1+ secretory progenitor cells revert to stem cells upon crypt damage. Nat Cell Biol 14: 1099–1104. doi: 10.1038/ncb2581
[24]
van der Flier LG, Haegebarth A, Stange DE, van de Wetering M, Clevers H (2009) OLFM4 is a robust marker for stem cells in human intestine and marks a subset of colorectal cancer cells. Gastroenterology 137: 15–17. doi: 10.1053/j.gastro.2009.05.035
[25]
Noguchi T, Matozaki T, Horita K, Fujioka Y, Kasuga M (1994) Role of SH-PTP2, a protein-tyrosine phosphatase with Src homology 2 domains, in insulin-stimulated Ras activation. Mol Cell Biol 14: 6674–6682.
[26]
Takahashi A, Tsutsumi R, Kikuchi I, Obuse C, Saito Y, et al. (2011) SHP2 tyrosine phosphatase converts parafibromin/Cdc73 from a tumor suppressor to an oncogenic driver. Mol Cell 43: 45–56. doi: 10.1016/j.molcel.2011.05.014
[27]
Grossmann KS, Wende H, Paul FE, Cheret C, Garratt AN, et al. (2009) The tyrosine phosphatase Shp2 (PTPN11) directs Neuregulin-1/ErbB signaling throughout Schwann cell development. Proc Natl Acad Sci U S A 106: 16704–16709. doi: 10.1073/pnas.0904336106
[28]
Korinek V, Barker N, Willert K, Molenaar M, Roose J, et al. (1998) Two members of the Tcf family implicated in Wnt/beta-catenin signaling during embryogenesis in the mouse. Mol Cell Biol 18: 1248–1256.
[29]
Haigis KM, Kendall KR, Wang Y, Cheung A, Haigis MC, et al. (2008) Differential effects of oncogenic K-Ras and N-Ras on proliferation, differentiation and tumor progression in the colon. Nat Genet 40: 600–608. doi: 10.1038/ng.115
[30]
Feng Y, Bommer GT, Zhao J, Green M, Sands E, et al.. (2011) Mutant KRAS promotes hyperplasia and alters differentiation in the colon epithelium but does not expand the presumptive stem cell pool. Gastroenterology 141: : 1003–1013 e1001–1010.
Van der Sluis M, De Koning BA, De Bruijn AC, Velcich A, Meijerink JP, et al. (2006) Muc2-deficient mice spontaneously develop colitis, indicating that MUC2 is critical for colonic protection. Gastroenterology 131: 117–129. doi: 10.1053/j.gastro.2006.04.020
[33]
Heazlewood CK, Cook MC, Eri R, Price GR, Tauro SB, et al. (2008) Aberrant mucin assembly in mice causes endoplasmic reticulum stress and spontaneous inflammation resembling ulcerative colitis. PLoS Med 5: e54. doi: 10.1371/journal.pmed.0050054
[34]
Coulombe G, Leblanc C, Cagnol S, Maloum F, Lemieux E, et al. (2013) Epithelial tyrosine phosphatase SHP-2 protects against intestinal inflammation in mice. Mol Cell Biol 33: 2275–2284. doi: 10.1128/mcb.00043-13
[35]
Ouellette AJ (2005) Paneth cell alpha-defensins: peptide mediators of innate immunity in the small intestine. Springer Semin Immunopathol 27: 133–146. doi: 10.1007/s00281-005-0202-x
[36]
Shi J (2007) Defensins and Paneth cells in inflammatory bowel disease. Inflamm Bowel Dis 13: 1284–1292. doi: 10.1002/ibd.20197
[37]
Gunther C, Martini E, Wittkopf N, Amann K, Weigmann B, et al. (2011) Caspase-8 regulates TNF-α-induced epithelial necroptosis and terminal ileitis. Nature 477: 335–339. doi: 10.1038/nature10400
