Identification of ALK Fusion Proteins in Non-Small Cell Lung Cancer
ALK fusion proteins have been identified in research studies in a range of human cancers.
Summary of ALK Fusion Partners and Associated Cancers
|RTKs||Inhibitors||Fusion Partners||Associated Cancers|
|ALK||Crizotinib5||EML42, TFG1, KIF5B6, NPM7||NSCLC1,2, ALCL7|
Table 1. Summary of ALK and ROS1 fusion partners and associated cancers.
Abbreviations: ALCL=Anaplastic large cell lymphoma EML4=Echinoderm microtubule-associated protein-like 4 NPM=Nucleophosmin NSCLC=Non-small cell lung cancer TFG=TRK-fused gene
CST's Research to Identify ALK and ROS1 Fusion Proteins in NSCLC
CST performed an unbiased, large-scale survey of tyrosine kinase activity in lung cancer using PTMScan® Technology. This proprietary technology, developed at Cell Signaling Technology(11), uses a CST™ Motif Antibody for immunoaffinity purification of peptides from digested cell extracts combined with LC tandem mass spectrometry to identify and quantify changes in post-translational modifications such as phosphorylation, acetylation, or ubiquitination. For this study, we used a phospho-tyrosine motif antibody to analyze changes in phosphorylation across the proteome in NSCLC cell lines and tissues.
Using PTMScan® Technology, we surveyed the phosphotyrosine status of receptor tyrosine kinases (RTK) and non-receptor tyrosine kinases in 41 NSCLC cell lines and over 150 NSCLC tumors. Over 50 tyrosine kinases and more than 2,500 downstream substrates that play roles in NSCLC growth and progression were identified. Two very exciting findings from this study were the identification of novel anaplastic lymphoma kinase (ALK) and c-ros oncogene 1 (ROS1) C-terminal fusion proteins in some NSCLC cell lines and tumors.
IHC analysis of paraffin-embedded human lung carcinoma using ROS1 (D4D6) Rabbit mAb #3287. Note: Staining is of FIG-ROS1 fusion protein(4).
Rabbit monoclonal antibodies specific to ALK and ROS1 [ALK (D5F3) XP® Rabbit mAb #3633; ROS1 (D4D6) Rabbit mAb #3287] were developed to detect both full-length and C-terminal fusion proteins. These antibodies have been validated for IHC and can detect ALK and ROS1 fusion protein expression in NSCLC sample(12, 4).
IHC analysis of paraffin-embedded human lung carcinoma with high (upper) and low levels (lower) of ALK expression using ALK (D5F3) XP® Rabbit mAb #3633.
A partnership with Pfizer, Inc., creator of the ALK inhibitor crizotinib, and Ventana Medical Systems, Inc., a leader in companion diagnostic testing, was formed to develop the use of ALK (D5F3) XP® Rabbit mAb in an automated diagnostic IHC screening assay to detect ALK fusion proteins in NSCLC patient samples.
Patient samples that stain positive for ALK expression may be candidates for crizotinib, which was approved for use in the U.S. in August 2011.
CST scientists are highlighted in bold.
- Katayama R, Shaw AT, Khan TM, Mino-Kenudson M, Solomon BJ, Halmos B, Jessop NA, Wain JC, Yeo AT, Benes C, Drew L, Saeh JC, Crosby K, Sequist LV, Iafrate AJ, Engelman JA (2012) Mechanisms of acquired crizotinib resistance in ALK-rearranged lung Cancers. Sci Transl Med 4(120), 120ra17.
- Ren H, Tan ZP, Zhu X, Crosby K, Haack H, Ren JM, Beausoleil S, Moritz A, Innocenti G, Rush J, Zhang Y, Zhou XM, Gu TL, Yang YF, Comb MJ (2012) Identification of anaplastic lymphoma kinase as a potential therapeutic target in ovarian cancer. Cancer Res. 72(13), 3312–23.
- Rimkunas VM, Crosby KE, Li D, Hu Y, Kelly ME, Gu TL, Mack JS, Silver MR, Zhou X, Haack H (2012) Analysis of Receptor Tyrosine Kinase ROS1-Positive Tumors in Non-Small Cell Lung Cancer: Identification of a FIG-ROS1 Fusion. Clin. Cancer Res. 18(16), 4449–57.
- Gu TL, Deng X, Huang F, Tucker M, Crosby K, Rimkunas V, Wang Y, Deng G, Zhu L, Tan Z, Hu Y, Wu C,Nardone J, MacNeill J, Ren J, Reeves C, Innocenti G, Norris B, Yuan J, Yu J, Haack H, Shen B, Peng C, Li H, Zhou X, Liu X, Rush J, Comb MJ (2011) Survey of tyrosine kinase signaling reveals ROS kinase fusions in human cholangiocarcinoma.
- Carretero J, Shimamura T, Rikova K, Jackson AL, Wilkerson MD, Borgman CL, Buttarazzi MS, Sanofsky BA, McNamara KL, Brandstetter KA, Walton ZE, Gu TL, Silva JC, Crosby K, Shapiro GI, Maira SM, Ji H, Castrillon DH, Kim CF, García-Echeverría C, Bardeesy N, Sharpless NE, Hayes ND, Kim WY, Engelman JA, Wong KK (2010) Integrative genomic and proteomic analyses identify targets for Lkb1-deficient metastatic lung tumors. Cancer Cell 17(6), 547-59.
- Boccalatte FE, Voena C, Riganti C, Bosia A, D'Amico L, Riera L, Cheng M, Ruggeri B, Jensen ON, Goss VL, Lee K,Nardone J, Rush J, Polakiewicz RD, Comb MJ, Chiarle R, Inghirami G (2009) The enzymatic activity of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase is enhanced by NPM-ALK: new insights in ALK-mediated pathogenesis and the treatment of ALCL. Blood 113(12), 2776-90.
- Yu J, Kane S, Wu J, Benedettini E, Li D, Reeves C, Innocenti G, Wetzel R, Crosby K, Becker A, Ferrante M,Cheung WC, Hong X, Chirieac LR, Sholl LM, Haack H, Smith BL, Polakiewicz RD, Tan Y, Gu TL, Loda M, Zhou X, Comb MJ (2009) Mutation-specific antibodies for the detection of EGFR mutations in non-small-cell lung cancer. Clin. Cancer Res. 15(9), 3023-8.
- Guo A, Villén J, Kornhauser J, Lee KA, Stokes MP, Rikova K, Possemato A, Nardone J, Innocenti G,Wetzel R, Wang Y, MacNeill J, Mitchell J, Gygi SP, Rush J, Polakiewicz RD, Comb MJ (2008) Signaling networks assembled by oncogenic EGFR and c-Met. Proc. Natl. Acad. Sci. U.S.A. 105(2), 692-7.
- Rikova K, Guo A, Zeng Q, Possemato A, Yu J, Haack H, Nardone J, Lee K, Reeves C, Li Y, Hu Y, Tan Z,Stokes M, Sullivan L, Mitchell J, Wetzel R, Macneill J, Ren JM, Yuan J, Bakalarski CE, Villen J, Kornhauser JM,Smith B, Li D, Zhou X, Gygi SP, Gu TL, Polakiewicz RD, Rush J, Comb MJ (2007) Global survey of phosphotyrosine signaling identifies oncogenic kinases in lung cancer. Cell 131(6), 1190-203..
- Rush J, Moritz A, Lee KA, Guo A, Goss VL, Spek EJ, Zhang H, Zha XM, Polakiewicz RD, Comb MJ (2005) Immunoaffinity profiling of tyrosine phosphorylation in cancer cells. Nat. Biotechnol. 23(1), 94-101.
- Pan S, Zhang H, Rush J, Eng J, Zhang N, Patterson D, Comb MJ, Aebersold R (2005) High throughput proteome screening for biomarker detection. Mol. Cell Proteomics 4(2), 182-90. PMID:15637048]
- Hornbeck PV, Chabra I, Kornhauser JM, Skrzypek E, Zhang B (2004) PhosphoSite: A bioinformatics resource dedicated to physiological protein phosphorylation. Proteomics 4(6), 1551-61.
- Zhang H, Zha X, Tan Y, Hornbeck PV, Mastrangelo AJ, Alessi DR, Polakiewicz RD, Comb MJ (2002)Phosphoprotein analysis using antibodies broadly reactive against phosphorylated motifs. J. Biol. Chem. 277(42), 39379-87.
- Cell Signaling Technology, Inc. Announces License in Personalized Cancer Diagnostics | July 29, 2013
- Cell Signaling Technology Awarded Patents Critical to Lung Cancer Therapy | July 9, 2013
- Cell Signaling Technology Granted Patent for PCR Methods for Detection of a Subset of Treatable Non-Small Cell Lung Cancers | January 7, 2013
- Ventana to collaborate with Pfizer and Cell Signaling Technology, Inc. on companion diagnostic to identify lung cancer patients with ALK gene rearrangements | January 9, 2012
Order a copy of the newly updated Pathways in Human Cancer poster. The recently published textbook The Biology of Cancer by Robert A. Weinberg and published by Garland Science includes a copy of this poster.
ROS1 is an RTK of the insulin receptor family that stimulates cell proliferation and survival. Like ALK, ROS1 has been shown to undergo a number of gene rearrangements that result in an oncogenic fusion protein, such as FIG-ROS1 in glioblastoma, and SLC34A2-ROS1 and CD74-ROS1 in NSCLC(9). In a recent immunohistochemistry (IHC) screening assay of >500 NSCLC samples, researchers found 1.6% of the tumors contained oncogenic ROS1 rearrangements, with the CD74-ROS1 fusion being the most prevalent(4). ROS1 activity can be inhibited with crizotinib, as shown in preclinical and early clinical studies(13).
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