Recruitment of Insulin Receptor Substrate-1 and Activation of NF-Jb Essential for Midkine Growth Signaling Through Anaplastic Lymphoma Kinase
Total Page：16
File Type：pdf, Size：1020Kb
Oncogene (2007) 26, 859–869 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc ORIGINAL ARTICLE Recruitment of insulin receptor substrate-1 and activation of NF-jB essential for midkine growth signaling through anaplastic lymphoma kinase AH Kuo, GE Stoica, AT Riegel and A Wellstein Department of Oncology, Lombardi Cancer Center, Georgetown University, Washington, DC, USA Anaplastic lymphoma kinase (ALK) is a transmembrane oncogenic fusion tyrosine kinase, nucleophosmin receptor tyrosine kinase in the insulin receptor super- (NPM)-ALK, involved in anaplastic large-cell lympho- family. We recently demonstrated that the growth factors ma (Morris et al., 1994). This NPM-ALK fusion protein pleiotrophin (PTN) and midkine (MK) are ligands for (residing in the cytoplasm and nucleus) is a constitu- ALK and that upon ALK activation, insulin receptor tively activated tyrosine kinase, owing to the dimeriza- substrate-1 (IRS-1) and other substrates are phosphory- tion driven by the N-terminal NPM fusion partner and lated. Here, the role of IRS-1 in ligand-mediated ALK acts as a transforming oncogene by virtue of its kinase signaling is investigated in interleukin-3 (IL-3)-dependent activity (Morris et al., 1994; Kuefer et al., 1997). 32D murine myeloid cells. These cells do not express ALK Subsequent studies identified several other ALK kinase and IRS family members, and do not respond to fusion proteins that support oncogenesis (Hernandez exogenously added PTN or MK. We show that expression et al., 1999; Colleoni et al., 2000; Lawrence et al., 2000; of ALK plus IRS-1 renders these cells independent of IL-3 Ma et al., 2000; Touriol et al., 2000; Trinei et al., 2000). owing to the activation of ALK by endogenous MK. Full-length ALK was initially described as an orphan Mutational analysis reveals that this transformed pheno- receptor tyrosine kinase that shows restricted tissue type of 32D cells requires kinase-active ALK as well as distribution and is regulated during organ development the interaction of ALK with IRS-1. Furthermore, 32D/ (Iwahara et al., 1997; Morris et al., 1997). Several IRS-1/ALK cells display an enhanced activation of studies show expression of full-length ALK protein in mitogen-activated protein kinase and PI3-kinase path- cultured fibroblasts and endothelial cells as well as cell ways, and a selective transcriptional activation of nuclear lines derived from epithelial cancers such as pancreatic factor (NF)-jB. Small interfering RNA-mediated knock- and breast carcinoma (Stoica et al., 2001, 2002) and the down of the endogenous MK or p65/NF-jB revealed that neuroectoderm, that is, melanoma (Dirks et al., 2002), both these are rate limiting for the transformed phenotype neuroblastoma (Lamant et al., 2000; Stoica et al., 2002) induced by ALK plus IRS-1. We conclude that the glioblastoma (Powers et al., 2002) and non-Hodgkin’s recruitment of IRS-1 to activated ALK and the activation lymphoma (Delsol et al., 1997), reviewed by Pulford of NF-jB are essential for the autocrine growth and et al. (2004). survival signaling of MK. We identified ALK as a receptor for the growth factor Oncogene (2007) 26, 859–869. doi:10.1038/sj.onc.1209840; pleiotrophin (PTN) using phage display of a human published online 31 July 2006 brain cDNA library with immobilized PTN as a bait and characterized the ligand-receptor interaction in cell-free Keywords: ALK; MK; IRS-1; NF-kB; autocrine and cell-based assays (Stoica et al., 2001). The phage display had revealed a fragment of the ALK extra- cellular domain (ECD) as the ligand-binding domain (LBD) and we found that PTN binds the recombinant Introduction ALK ECD protein as well as the receptor expressed in intact cells with dissociation constants (Kd) of approxi- Anaplastic lymphoma kinase (ALK), a 220 kDa trans- mately 30 pM ( ¼ 0.5 ng/ml). Also, antibodies raised to membrane receptor tyrosine kinase, and its smaller the ALK LBD as well as to PTN inhibited receptor relative, the 110 kDa leukocyte tyrosine kinase (LTK), binding of the ligand and signal transduction via ALK belong to the insulin receptor superfamily based on the as well as biologic effects were observed at EC50 values amino-acid sequence homology of their intracellular close to the Kd of PTN binding to the ALK receptor kinase domains. ALK was first described as part of an (Stoica et al., 2001; Bowden et al., 2002; Powers et al., 2002). In support of this, recent work from the laboratory of Paul Mischel (Lu et al., 2005) using Correspondence: Dr A Wellstein, Department of Oncology, Lombardi different glioblastoma cell line models also reported that Cancer Center, Georgetown University, Research Bldg. E311, 3970 ALK phosphorylation and signaling are induced by Reservoir Road, Washington, DC 20007, USA. E-mail: [email protected] PTN. This corroborates our earlier work in U87 Received 5 March 2006; revised 19 June 2006; accepted 19 June 2006; glioblastoma cells in which depletion of their endo- published online 31 July 2006 genous ALK by ribozymes specifically blocked growth ALK and IRS-1 in MK-mediated signaling AH Kuo et al 860 factor signaling by PTN and tumor growth (Powers reporters responsive to growth factor signaling showed a et al., 2002). In extension of the studies with PTN, its selective activation of the nuclear factor (NF)-kB homolog midkine (MK) was also shown to bind to the pathway by the ALK/IRS-1 autocrine activation. ALK receptor, albeit with a slightly lower affinity (Kd of Finally, small interfering RNA (siRNA)-mediated approximately 100 pM), to induce phosphorylation of targeting of the endogenous MK or of the p65 subunit the ALK substrate insulin receptor substrate (IRS)-1, of NF-kB revealed that both of these factors are rate- activate mitogen-activated protein kinase (MAPK) and limiting for the ALK-mediated effects through IRS-1. PI3-kinase and induce colony formation in soft agar (Stoica et al., 2002). Blockade of the effect of exogenously added MK by monoclonal antibodies raised against the LBD of ALK revealed the contribu- Results tion of ALK (Stoica et al., 2002). Overall, PTN and MK induce a variety of effects in normal and transformed Analysis of the endogenous expression of ALK, LTK and cells or tissues including neurite outgrowth, prolifera- MK in 32D cells tion, angiogenesis, antiapoptosis and tumor metastasis The 32D murine myeloid progenitor cells are IL-3- (see e.g. Fang et al., 1992; Czubayko et al., 1996; Schulte dependent and have been used extensively to evaluate et al., 1996; reviewed by Schulte and Wellstein, 1997; the role of IRS-1 in IGF-1 receptor and insulin receptor Zhang and Deuel, 1999; Muramatsu, 2002). signaling (Wang et al., 1993). These cells do not express Interestingly, in Drosophila, the 560 amino-acid any known IRS family member (Wang et al., 1993) and protein jelly belly (Jeb) was shown by mutation and we reasoned that they thus offer a good model system to phenotypic analysis to interact with the fly ALK analyse the contribution of IRS-1 to ALK signaling. homolog dALK (Englund et al., 2003; Lee et al., 2003) Reverse transcription–polymerase chain reaction (RT– and to induce MAPK signaling. No known homolog of PCR) for murine and human ALK mRNA expression Jeb has yet been described for vertebrates and our recent showed an amplified product of the expected size in the searches did not reveal a Jeb-related vertebrate protein positive controls, that is, NIH3T3 fibroblasts and SW13, or gene in the databases of either National Center for but no detectable product in 32D cells (Figure 1a). The Biotechnology Information or EMBL. It was proposed ALK homolog LTK is expressed in Ba/F3 cells that that the activation of dALK by Jeb occurs by have been used to evaluate LTK kinase signaling heterodimerization of dALK with the membrane-bound through a chimeric epidermal growth factor (EGF)/ Jeb (Freeman, 2003), and this suggests that other LTK receptor (Ueno et al., 1996) whereas 32D cells do proteins in addition to the PTN/MK growth factors not express this receptor (Figure 1b). As we reported can activate the receptor at least in invertebrates. earlier that 32D cells do not express PTN or respond to Here, we report on the contribution of IRS-1 to PTN stimulation (Stoica et al., 2001), we decided to signaling through the ALK receptor. The IRS family evaluate whether these cells express detectable levels of has four known members, and of the four, IRS-1 has the second member of this growth factor family, MK. been most extensively studied (White, 1998). The role of As shown in Figure 1c, SW13 cells were negative for IRS-1 in insulin signaling is well established owing to the MK expression whereas both NIH3T3 and 32D cells fact that IRS-1 is regarded as a critical player in expressed MK mRNA (see below, Figure 3a, for MK maintaining basic cellular function such as metabolism, protein expression). These results indicated that 32D cell growth and survival (Chang et al., 2002). Aberrant expression or activation of IRS-1 has been associated with breast, pancreatic and prostate cancers as well as a b several rare tumor types (White, 1998, 2002; Burks and White, 2001; Chang et al., 2002). In addition to already NIH 3T3 SW13 32D known interactions, IRS-1 is also thought to be a central ALK 32D Ba /F3 element of ERa/insulin-like growth factor (IGF)-1 LTK crosstalk (Surmacz, 2000; Sachdev and Yee, 2001; GAPDH GAPDH Surmacz and Bartucci, 2004). In the present paper, we analyze the contribution of c IRS-1 to ALK signaling using the interleukin-3 (IL-3)- dependent, 32D murine hematopoietic cells that have no SW13 NIH 3T3 32D detectable ALK and IRS protein expression (Wang MK et al., 1993; Stoica et al., 2001; White, 2002).