Increased tumorigenesis associated with loss of the tumor suppressor gene Cadm1
- Louise van der Weyden1Email author,
- Mark J Arends†2,
- Alistair G Rust†1,
- George Poulogiannis3,
- Rebecca E McIntyre1 and
- David J Adams1
© van der Weyden et al.; licensee BioMed Central Ltd. 2012
Received: 18 February 2012
Accepted: 3 May 2012
Published: 3 May 2012
CADM1 encodes an immunoglobulin superfamily (IGSF) cell adhesion molecule. Inactivation of CADM1, either by promoter hypermethylation or loss of heterozygosity, has been reported in a wide variety of tumor types, thus it has been postulated as a tumor suppressor gene.
We show for the first time that Cadm1 homozygous null mice die significantly faster than wildtype controls due to the spontaneous development of tumors at an earlier age and an increased tumor incidence of predominantly lymphomas, but also some solid tumors. Tumorigenesis was accelerated after irradiation of Cadm1 mice, with the reduced latency in tumor formation suggesting there are genes that collaborate with loss of Cadm1 in tumorigenesis. To identify these co-operating genetic events, we performed a Sleeping Beauty transposon-mediated insertional mutagenesis screen in Cadm1 mice, and identified several common insertion sites (CIS) found specifically on a Cadm1-null background (and not wildtype background).
We confirm that Cadm1 is indeed a bona fide tumor suppressor gene and provide new insights into genetic partners that co-operate in tumorigenesis when Cadm1-expression is lost.
Cell adhesion molecule 1 (CADM1; also known as TSLC1, IGSF4, Necl-2, RA175 SgIGSF, SynCAM1) is member of the immunoglobulin superfamily of cell adhesion molecules (IGSF-CAMs) and is composed of an extracellular domain containing three immunoglobulin-like C2-type domains, a transmembrane domain and a short cytoplasmic tail. The extracellular domain of CADM1 mediates the formation of homodimers or heterodimers with other CAM members, including Necl-1, CRTAM and Nectin-3 to regulate cell adhesion. The cytoplasmic domain of CADM1 interacts with the tumour-suppressor gene DAL-1 and the group of membrane-associated guanylate kinase (MAGuK) homologues, as well as being able to modulate the activation of small Rho GTPases, thus acting as a vital bridge between extracellular adhesion and intracellular signaling cascades. In addition, CADM1 can also modulate cell cycle progression and apoptosis[2, 3].
Less than a decade since the discovery of CADM1, loss of its expression by promoter hypermethylation or loss of heterozygosity (LOH) has been reported in a wide variety of tumor types ( Additional file2: Figure S1) and frequently correlates with advanced tumor stage (poor prognosis) and metastasis. Studies in nude mice have demonstrated that re-expression of CADM1 suppresses in vivo tumorigenicity of non-small cell lung cancer and nasopharyngeal carcinoma cell lines[1, 4, 5]. In contrast, studies using Cadm1 null (Cadm1 −/− ) mice have demonstrated important roles for Cadm1 in spermatogenesis/male fertility, behavior and motor performance, lens fiber cell architecture, and epidermal adhesion and wound repair. However, there are no reports on the incidence of cancer in these mice and thus whether Cadm1 is indeed a bona fide tumor suppressor gene.
The most statistically significant CIS was in the Nr3c1 gene, which encodes the glucocorticoid receptor (GR), and the insertions would be predicted to have a loss-of-function effect on Nr3c1, consistent with the finding of promoter hypermethylation or mono-allelic deletion of NR3C1 in several cancer types including colo-rectal cancer (CRC) and leukemia, respectively. Glucocorticoids (GCs), which bind the GR and allow it to translocate to the nucleus and modulate gene expression, are effective inhibitors of proliferation and tumorigenesis and routinely used in treating T-cell acute lymphoblastic leukemia (T-ALL). Interestingly, three of the other nine CIS genes identified are known interactors/regulators of the GR. St13 encodes the Hsp70-interacting protein that is involved in the assembly process of the GR, and ST13 mRNA and protein levels are down-regulated in CRC. The Ets1 proto-oncogene is a transcription factor that can act as a “molecular switch” for auto-regulation of the GR promoter, and high ETS1 expression predicts poor prognosis in patients with ovarian cancer. The Csf3r encodes the cell-surface granulocyte colony-stimulating factor (G-CSF) receptor, and activated GR can synergize with G-CSF signals.
Cell junctions including tight junctions, adherens junctions and desmosomes, consist of multi-protein complexes that provide contact between neighboring cells or between a cell and the extracellular matrix and as such play important roles in regulation of cell proliferation and differentiation, as well as cancer. Like CADM1, two of our CIS genes are part of these multi-protein complexes. Fchsd2 encodes the FCH and double SH3 domains protein 2 (FCHSD2), which binds to epithelial junction MAGuKs, specifically MAGI-1 and CASK; CADM1 has been shown to interact with several MAGuK members, including CASK. Jup encodes junction plakoglobin (JUP), which complexes with numerous other desmosomal proteins (including cadherins, desmogleins and desmocollins)), and was recently shown to be expressed on the surface of colorectal cancer cells associated with high metastatic potential.
Finally, it is interesting to note that in addition to ‘Cadm1 null-specific’ CIS, we also identified CIS that were only found in tumors from wildtype mice (i.e., not found in Cadm1 null tumors). These CIS represent loci that are mutated in the process of tumorigenesis in the presence of an intact Cadm1 signaling pathway, and whose contribution to tumorigenesis is potentially rendered obsolete in the absence of Cadm1. Some of these genes, such as Pik3r5 and Malt1, have also been identified as CIS in leukemia/lymphomas from wildtype mice in other Sleeping Beauty transposon screens we have performed (unpublished data). Several CIS genes including Pten Notch1 and Erg are mutated in both wildtype and Cadm1 null tumors suggesting that mutation of these genes can contribute to tumorigenesis regardless of Cadm1 status.
Thus we have shown that CADM1 is a bona fide tumor suppressor gene, and loss of Cadm1 results in an increased tumor incidence. Our insertional mutagenesis screen provides new insights into Cadm1-mediated tumor suppression by identifying genes that co-operate with loss of Cadm1 in lymphomagenesis, in particular those regulating glucocorticoid signaling and cell junctions.
LvdW was supported by the Kay Kendall Leukemia Fund. GP is a Pfizer Fellow of the Life Sciences Research Foundation. MJA was supported by Cancer Research UK. DJA was supported by Cancer Research UK and the Wellcome Trust.
- Kuramochi M, Fukuhara H, Nobukuni T, Kanbe T, Maruyama T, Ghosh HP, Pletcher M, Isomura M, Onizuka M, Kitamura T, Sekiya T, Reeves RH, Murakami Y: TSLC1 is a tumor-suppressor gene in human non-small-cell lung cancer. Nat Genet. 2001, 27: 427-430. 10.1038/86934View ArticlePubMedGoogle Scholar
- Murakami Y: Involvement of a cell adhesion molecule. TSLC1/IGSF4, in human oncogenesis. Cancer Sci. 2005, 96: 543-552. 10.1111/j.1349-7006.2005.00089.xView ArticlePubMedGoogle Scholar
- Liang QL, Chen GQ, Li ZY, Wang BR: Function and histopathology of a cell adhesion molecule TSLC1 in cancer. Cancer Invest. 2011, 29: 107-112. 10.3109/07357907.2010.543211View ArticlePubMedGoogle Scholar
- Mao X, Seidlitz E, Truant R, Hitt M, Ghosh HP: Re-expression of TSLC1 in a non-small-cell lung cancer cell line induces apoptosis and inhibits tumor growth. Oncogene. 2004, 23: 5632-5642. 10.1038/sj.onc.1207756View ArticlePubMedGoogle Scholar
- Lung HL, Cheung AK, Xie D, Cheng Y, Kwong FM, Murakami Y, Guan XY, Sham JS, Chua D, Protopopov AI, Zabarovsky ER, Tsao SW, Stanbridge EJ, Lung ML: TSLC1 is a tumor suppressor gene associated with metastasis in nasopharyngeal carcinoma. Cancer Res. 2006, 66: 9385-9392. 10.1158/0008-5472.CAN-06-0590View ArticlePubMedGoogle Scholar
- van der Weyden L, Arends MJ, Chausiaux OE, Ellis PJ, Lange UC, Surani MA, Affara N, Murakami Y, Adams DJ, Bradley A: Loss of TSLC1 causes male infertility due to a defect at the spermatid stage of spermatogenesis. Mol Cell Biol. 2006, 26: 3595-3609. 10.1128/MCB.26.9.3595-3609.2006PubMed CentralView ArticlePubMedGoogle Scholar
- Takayanagi Y, Fujita E, Yu Z, Yamagata T, Momoi MY, Momoi T, Onaka T: Impairment of social and emotional behaviors in Cadm1-knockout mice. Biochem Biophys Res Commun. 2010, 396: 703-708. 10.1016/j.bbrc.2010.04.165View ArticlePubMedGoogle Scholar
- De Maria A, Shi Y, Luo X, Van Der Weyden L, Bassnett S: Cadm1 expression and function in the mouse lens. Invest Ophthalmol Vis Sci. 2011, 52: 2293-2299. 10.1167/iovs.10-6677PubMed CentralView ArticlePubMedGoogle Scholar
- Giangreco A, Jensen KB, Takai Y, Miyoshi J, Watt FM: Necl2 regulates epidermal adhesion and wound repair. Development. 2009, 136: 3505-3514. 10.1242/dev.038232PubMed CentralView ArticlePubMedGoogle Scholar
- Paulsson K, An Q, Moorman AV, Parker H, Molloy G, Davies T, Griffiths M, Ross FM, Irving J, Harrison CJ, Young BD, Strefford JC: Methylation of tumour suppressor gene promoters in the presence and absence of transcriptional silencing in high hyperdiploid acute lymphoblastic leukaemia. Br J Haematol. 2009, 144: 838-847. 10.1111/j.1365-2141.2008.07523.xView ArticlePubMedGoogle Scholar
- Fu L, Gao Z, Zhang X, Tsang YH, Goh HK, Geng H, Shimizu N, Tsuchiyama J, Srivastava G, Tao Q: Frequent concomitant epigenetic silencing of the stress-responsive tumor suppressor gene CADM1, and its interacting partner DAL-1 in nasal NK/T-cell lymphoma. Int J Cancer. 2009, 124: 1572-1578. 10.1002/ijc.24123View ArticlePubMedGoogle Scholar
- Heddle JA: A rapid in vivo test for chromosomal damage. Mutat Res. 1973, 18: 187-190. 10.1016/0027-5107(73)90035-3View ArticlePubMedGoogle Scholar
- Collier LS, Carlson CM, Ravimohan S, Dupuy AJ, Largaespada DA: Cancer gene discovery in solid tumours using transposon-based somatic mutagenesis in the mouse. Nature. 2005, 436: 272-276. 10.1038/nature03681View ArticlePubMedGoogle Scholar
- March HN, Rust AG, Wright NA, ten Hoeve J, de Ridder J, Eldridge M, van der Weyden L, Berns A, Gadiot J, Uren A, Kemp R, Arends MJ, Wessels LF, Winton DJ, Adams DJ: Insertional mutagenesis identifies multiple networks of cooperating genes driving intestinal tumorigenesis. Nat Genet. 2011, 43: 1202-1209. 10.1038/ng.990PubMed CentralView ArticlePubMedGoogle Scholar
- Lind GE, Kleivi K, Meling GI, Teixeira MR, Thiis-Evensen E, Rognum TO, Lothe RA: ADAMTS1, CRABP1, and NR3C1 identified as epigenetically deregulated genes in colorectal tumorigenesis. Cell Oncol. 2006, 28: 259-272.PubMedGoogle Scholar
- Kuster L, Grausenburger R, Fuka G, Kaindl U, Krapf G, Inthal A, Mann G, Kauer M, Rainer J, Kofler R, Hall A, Metzler M, Meyer LH, Meyer C, Harbott J, Marschalek R, Strehl S, Haas OA, Panzer-Grümayer R: ETV6/RUNX1-positive relapses evolve from an ancestral clone and frequently acquire deletions of genes implicated in glucocorticoid signaling. Blood. 2011, 117: 2658-2667. 10.1182/blood-2010-03-275347View ArticlePubMedGoogle Scholar
- Wang LB, Zheng S, Zhang SZ, Peng JP, Ye F, Fang SC, Wu JM: Expression of ST13 in colorectal cancer and adjacent normal tissues. World J Gastroenterol. 2005, 11: 336-339.PubMed CentralView ArticlePubMedGoogle Scholar
- Davidson B, Reich R, Goldberg I, Gotlieb WH, Kopolovic J, Berner A, Ben-Baruch G, Bryne M, Nesland JM: Ets-1 messenger RNA expression is a novel marker of poor survival in ovarian carcinoma. Clin Cancer Res. 2001, 7: 551-557.PubMedGoogle Scholar
- Dror Y, Ward AC, Touw IP, Freedman MH: Combined corticosteroid/granulocyte colony-stimulating factor (G-CSF) therapy in the treatment of severe congenital neutropenia unresponsive to G-CSF: Activated glucocorticoid receptors synergize with G-CSF signals. Exp Hematol. 2000, 28: 1381-1389. 10.1016/S0301-472X(00)00544-0View ArticlePubMedGoogle Scholar
- Dusek RL, Attardi LD: Desmosomes: new perpetrators in tumour suppression. Nat Rev Cancer. 2011, 11: 317-323. 10.1038/nrc3051PubMed CentralView ArticlePubMedGoogle Scholar
- Ohno H, Hirabayashi S, Kansaku A, Yao I, Tajima M, Nishimura W, Ohnishi H, Mashima H, Fujita T, Omata M, Hata Y: Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Oncogene. 2003, 22: 8422-8431. 10.1038/sj.onc.1206996View ArticlePubMedGoogle Scholar
- Biederer T, Sara Y, Mozhayeva M, Atasoy D, Liu X, Kavalali ET, Südhof TC: SynCAM, a synaptic adhesion molecule that drives synapse assembly. Science. 2002, 297: 1525-1531. 10.1126/science.1072356View ArticlePubMedGoogle Scholar
- Desai BV, Harmon RM, Green KJ: Desmosomes at a glance. J Cell Sci. 2009, 122: 4401-4407. 10.1242/jcs.037457PubMed CentralView ArticlePubMedGoogle Scholar
- Luque-García JL, Martínez-Torrecuadrada JL, Epifano C, Cañamero M, Babel I, Casal JI: Differential protein expression on the cell surface of colorectal cancer cells associated to tumor metastasis. Proteomics. 2010, 10: 940-952.PubMedGoogle Scholar
- van der Weyden L, Rust AG, McIntyre RE, Robles-Espinoza CD, Del Castillo Velasco-Herrera M, Strogantsev R, Ferguson-Smith AC, McCarthy S, Keane TM, Arends MJ, Adams DJ: Jdp2 downregulates Trp53 transcription to promote leukaemogenesis in the context of Trp53 heterozygosity. Oncogene. 2012, 10.1038/onc.2012.56. Epub ahead of printGoogle Scholar
- Dupuy AJ, Akagi K, Largaespada DA, Copeland NG, Jenkins NA: Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system. Nature. 2005, 436: 2212226-View ArticleGoogle Scholar
- Skrzypczak M, Gorycam K, Rubet T, Paziewska A, Mikula A, Jarosz D, Pachlewski J, Oledzki J, Ostrowski J: Modelling oncogenic signalling in colon tumors by multidirectional analyses of microarry data directed for maximization of analytical reliabilty. PLoS One. 2010, 5i: e13091-View ArticleGoogle Scholar
- Su H, Hu N, Yang HH, Wang C, Takikita M, Wang QH, Giffen C, Clifford R, Hewitt SM, Shou JZ, Goldstein AM, Lee MP, Taylor PR: Global gane expression profiling and validation in esophageal squamous cell carcinoma and wiht clinical phenotypes. Clin Cancer Res. 2011, 17: 2955-2966. 10.1158/1078-0432.CCR-10-2724PubMed CentralView ArticlePubMedGoogle Scholar
- Landi MT, Dracheva T, Rotunno MC, Figueroa JD, Liu H, Dasgupta A, Mnn FE, Fukuoka J, Hames M, Bergen AW, Murphy SE, Pesatori AC Yang P, Consonni D, Bertazzi PA, Wacholder S, Shih JH, Caporaso NE, Jen J: Gene expression signature of cigarette smoking and its role in long adenocarcinoma devalopment and survival. PLoS One. 2008, 3: e1651 10.1371/journal.pone.0001651PubMed CentralView ArticlePubMedGoogle Scholar
- Buchholz M, Braun M, Heidenblut A, Kertler HA, Klōppel G, Schmiegel W, Hahn SA, Luttges J, Gress TM: Transcipthelial neoplastic lesion. Oncogene. 2005, 24: 6626-6636. 10.1038/sj.onc.1208804View ArticlePubMedGoogle Scholar
- Shedden K, Taylor JM, Enkemann SA, Tsao MS, Yeatman TJ, Gerald WL, Eschrich S, Jurisica I, Goirdana TJ, Misek DE, Chang AC, Zhu CQ, Strumpt D, Hanash S, Shepherd FA, Ding K, Seymour L, Naoki , Pennell N, Wier B, Kris M, Viale A, Motoi N, Travis W, Conley B, Seshan VE, Meyerson M, Kuick R, Dubbin KK, Lively T, Jacobson JW, Beer DG, : Gene expression based survival prediction in lung adenocarcinoma: a multi-site blinded validation study. Nat Med. 2008, 14: 822-827. 10.1038/nm.1790PubMed CentralView ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.