Journal of Neuro-Oncology

, Volume 72, Issue 1, pp 35–46

Analysis of target genes induced by IL-13 cytotoxin in human glioblastoma cells

Laboratory Investigation


IL-13 cytotoxin comprised of IL-13 and a mutated form of Pseudomonas exotoxin (fusion protein termed IL-13-PE38QQR) has been shown to inhibit protein synthesis leading to necrotic and apoptotic cell death in glioblastoma cells that express high levels of interleukin-13 receptors (IL-13R). To identify target genes of cell death and other cellular genes with IL-13 receptors in glioblastoma cells, we utilized the cDNA microarrays to analyze global gene expression profiles after IL-13 cytotoxin and IL-13 treatment. IL-13 cytotoxin mediated cytotoxicity to U251 cells in a dose-dependant manner. Hierarchical cluster analysis of differentially expressed genes in U251 glioma cells at different time points after IL-13 cytotoxin treatment showed three major groups, each representing a specific expression pattern. Randomly selected differentially expressed genes from each group were confirmed by RT-PCR analysis. Most down-regulated genes belong to cell adhesion, motility, angiogenesis, DNA repair, and metabolic pathways. While up-regulated genes belong to cell cycle arrest, apoptosis, signaling and various metabolic pathways. Unexpectedly, at early time points, both IL-13 and IL-13 cytotoxin induced several genes belonging to different pathways most notably IL-8, DIO2, END1, and ALDH1A3 indicating that these genes are early response genes and their products may be associated with IL-13R. In addition, IL-13 cytotoxin induced IL-13Rα2 mRNA expression during the treatment in glioma cells. Our results indicate that novel cellular genes are involved with IL-13 receptors and that IL-13 cytotoxin induced cell death involves various target genes in human glioblastoma cells. On going studies will determine the role of associated genes and their products in the IL-13R functions in glioma cells.


cDNA microarray gene expression profiles IL-13 cytotoxin IL-13Rα2 receptor 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ries LAG, Eisner MP, Kosary CL, Hankey BF, Miller BA, Clegg LX, Edwards BK (eds) SEERS Cancer Statistics Review, 1973–1997, National cancer Institute. NIH Pub. No. 00-2789. Bethesda, MD 2000.Google Scholar
  2. Puri, RK, Leland, P, Kreitman, RJ, Pastan, I 1994Human neurological cancer cells express interleukin-4 (IL-4) receptors which are targets for the toxic effects of IL-4 Pseudomonas exotoxin chimeric proteinInt J Cancer58574581Google Scholar
  3. Laske, DW, Youle, RJ, Oldfield, EH 1997Tumor regression with regional distribution of the targeted toxin TF-CRM107 in patients with malignant brain tumorsNature Med313621368Google Scholar
  4. Hao, CH, Parney, IF, Roa, WH,  et al. 2002Cytokine and cytokine receptor mRNA expression in human glioblastoma: evidence of Th1, Th2 and Th3 cytokine dysregulationActa Neuropathol103171178Google Scholar
  5. Obiri, NI, Debinski, W, Leonard, WJ, Puri, RK 1995Receptor for interleukin 13: interaction with interleukin 4 by a mechanism that dose not involve the common γ chain shared by receptors for interleukin 2, 4, 7, 9, and 15J Biol Chem27087978804Google Scholar
  6. Debinski, W, Obiri, NI, Powers, SK, Pastan, I, Puri, RK 1995Human glioma cells overexpress receptors for interleukin 13 and are extremely sensitive to a novel chimeric protein composed of interleukin 13 and Pseudomonas exotoxinClin Cancer Res112531258Google Scholar
  7. Husain, SR, Obiri, NI, Gill, P, Zhang, T, Pastan, I, Debinski, W, Puri, RK 1997Receptor for interleukin 13 on AIDS-associated Kaposi’s sarcoma cells serves as a new target for poten Pseudomonas exotoxin-based chimeric toxin proteinClin Cancer Res3151156Google Scholar
  8. Joshi, BH, Plautz, GE, Puri, RK 2000Interleukin-13 receptor a chain: a novel tumor-associated transmembrane protein in primary explants of human malignant gliomasCancer Res6011681172Google Scholar
  9. Husain, SR, Joshi, BH, Puri, RK 2001Interleukin-13 receptor as a unique target for anti-glioblastoma therapyInt J Cancer92168175Google Scholar
  10. Hilton, DJ, Zhang, JG, Metcalf, D, Alexander, WS, Nicola, NA, Willson, TA 1996Cloning and characterization of a binding subunit of the interleukin 13 receptor that is also a component of the interleukin 4 receptorProc Natl Acad Sci USA93497501Google Scholar
  11. Aman, MJ, Tayebi, N, Obiri, NI, Puri, RK, Modi, WS, Leonard, WJ 1996cDNA cloning and characterization of the human interleukin 13 receptor alpha chainJ Biol Chem2712926529270Google Scholar
  12. Murata, T, Obiri, NI, Puri, RK 1998Structure of and signal transduction through interleukin-4 and interleukin-13 receptorsInt J Mol Med1551557Google Scholar
  13. Miloux, B, Laurent, P, Bonnin, O,  et al. 1997Cloning of the human IL-13Rα1chain and reconstitution with the IL-4Rα of a functional IL-4/Il-13 receptor complexFEBS Lett401163166Google Scholar
  14. Orchansky, PL, Kwan, R, Lee, F, Schrader, JW 1999Characterization of the cytoplasmic domain of interleukin-13 receptor-αJ Biol Chem2742081820825Google Scholar
  15. Caput, D, Laurent, P, Kaghad, M,  et al. 1996Cloning and characterization of a specific interleukin (IL)-13 binding protein structurally related to the IL-5 receptor α. chain.J Biol Chem2711692116926Google Scholar
  16. Kawakami, K, Taguchi, J, Murata, T, Puri, RK 2001The interleukin-13 receptor α2 chain: an essential component for binding and internalization but not for interleukin-13-induced signal transduction through STAT6 pathwayBlood9726732679Google Scholar
  17. Kawakami, K, Takeshita, F, Puri, RK 2001Identification of distinct roles for a dileucine and a tyrosine internalization motif in the interleukin (IL)-13 binding component IL-13 receptor alpha 2 chainJ Biol Chem2762511425120Google Scholar
  18. Debinski, W, Obiri, NI, Pastan, I, Puri, RK 1995A novel chimeric protein composed of interleukin 13 and Pseudomonas exotoxin is highly cytotoxic to human carcinoma cells expressing receptors for interleukin 13 and interleukin 4J Biol Chem2701677516780Google Scholar
  19. Joshi, BH, Kawakami, K, Leland, P, Puri, RK 2002Heterogeneity in interleukin-13 receptor expression and subunit structure in squamous cell carcinoma of head and neck: differential sensitivity to chimeric fusion proteins comprised of interleukin-13 and a mutated form of Pseudomonas exotoxinClin Cancer Res819481956Google Scholar
  20. Puri, RK, Leland, P, Obiri, NI, Husain, SR, Kreitman, RJ, Haas, GP, Pastan, I, Debinski, W 1996Targeting of IL-13 receptor on human renal cell carcinoma cells by a recombinant chimeric protein composed of interleukin-13 and a truncated from of Pseudomonas exotoxin A (PE38QQR)Blood8743334339Google Scholar
  21. Husain, SR, Puri, RK 2000Interleukin-13 fusion cytotoxin as a potent targeted agent for AIDS-Kaposi’s sarcoma xenograftBlood9535063513Google Scholar
  22. Kawakami, K, Husain, SR, Kawakami, M, Puri, RK 2002Improved anti-tumor activity and safety of interleukin-13 receptor targeted cytotoxin by systemic continuous administration in head and neck cancer xenograft modelMol Med8487494Google Scholar
  23. Kawakami, K, Kawakami, M, Puri, RK 2002Cytokine receptor as a sensitizer for targeted cancer therapyAnti-Cancer Drugs13693699Google Scholar
  24. Kawakami, M, Kawakami, K, Puri, RK 2002Intratumor administration of interleukin 13 receptor-targeted cytotoxin induces apoptotic cell death in human malignant glioma tumor xenograftsMol Cancer Therap19991007Google Scholar
  25. Kawakami, K, Joshi, BH, Puri, RK 2000Sensitization of cancer cells to interleukin 13-Pseudomonas exotoxin-induced cell death by gene transfer of interleukin 13 receptor a chainHuman Gene Therapy1118291835Google Scholar
  26. Weingart J, Grossman SA, Bohan E, Fisher JD, Strauss L, Puri RK: Phase I/II study of interstitial infusion of IL13-PE38QQR cytotoxin in recurrent malignant glioma. First Quadrennial meeting – World Federation of Neuro-Oncology, November 15–17, 2001, Washington, DC.Google Scholar
  27. Weingart, J, Strauss, LC, Grossman, SA, Markett, J, Tatter, S, Fisher, JD, Fleming, CK, Puri, RK 2002Phase I/II study: intra-tumoral infusion of IL13-PE38QQR cytotoxin for recurrent supratentorial malignant glioma.Neoro-oncol4379Google Scholar
  28. Prados MD, Lang FF, Strauss L, Fleming C, Alalpe K, Kunwar S, Yung WKA, Chang S, Husain SR, Gutin PH, Raizer J, Piepmeier JM, Berger M, McDermott M, Puri RK: Intratumoral and intracerebral microinfusion of IL13-PE38QQR cytotoxin: phase I/II study of pre- and post-resection infusions in recurrent resectable malignant glioma. Proc. Am. Soc. Clin. Oncol. 21, 69 (Abstract 2087), 2002.Google Scholar
  29. Lang F, Kunwar S, Strauss L, Gutin P, Piepmeier J, McDermott M, Fleming C, Sherman J, Raizer J, Alalpe K, Yung WKA, Husain SR, Chang S, Berger M, Prados M, Puri RK: A clinical study of convection-enhanced delivery of IL13-PE38QQR cytotoxin Pre- and post- resection of recurrent GBM. American Society of Neuro-Oncologists Chicago, April 2002.Google Scholar
  30. Prados MD, Lang FF, Sherman JW, Strauss LC, Fleming CK, Alalpe K, Kunwar S, Yung WKA, Chang SM, Husain SR, Gutin PH, Raizer J, Piepmeier JM, Berger M, McDermott M, Puri RK: Convection-enhanced delivery (CED) by positive pressure infusion for intra- tumoral and peri-tumoral administration of IL13-PE38QQR a recombinant tumor-targeted cytotoxin in recurrent malignant glioma. Neuro-Oncol 4: S78, 2002(B).Google Scholar
  31. Schena, M, Shalon, D, Davis, RW, Brown, PO 1995Quantitative monitoring of gene expression patterns with a complementary DNA microarrayScience270467470PubMedGoogle Scholar
  32. Bowtell, DDL 1999Options available- from start to finish-for obtaining expression data by microarrayNat Genet212532Google Scholar
  33. Pollack, JR, Perou, CM, Alizadeh, AA, Eisen, MB, Pergamenschikov, A, Williams, CF, Jeffrey, SS, Botstein, D, Brown, PO 1999Genome-wide analysis of DNA copy-number changes using cDNA microarraysNat Genet234146Google Scholar
  34. King, HC, Sinha, AA 2001Gene expression profile analysis by DNA microarrayJAMA28622802288Google Scholar
  35. Shoemaker, DD, Schadt, EE, Armour, CD, He, YD, Garrett-Engele, P, McDonagh, PD, Loerch, PM, Leonardson, A, Lum, PY, Cavet, G, Wu, LF, Altschuler, SJ, Edwards, S, King, J, Tsang, JS, Schimmack, G, Schelter, JM, Koch, J, Ziman, M, Marton, MJ, Li, B, Cundiff, P, Ward, T, Castlw, J, Krolewski, M, Meyer, MR, Mao, M, Burchard, J, Kidd, MJ, Dal, H, Pillips, JW, Linsley, PS, Stoughton, R, Scherer, S, Bouguski, MS 2001Experimental annotation of the human genome using microarray technologyNature409922927Google Scholar
  36. Veer, LJ, Dai, H, Vijver, MJ, He, YD, Hart, AAM, Mao, M, Peterse, HL, Kooy, KV, Marton, MJ, Witteveen, AT, Schreiber, GJ, kerkhoven, RM, Roberts, C, Linsley, PS, Bernards, R, Friend, SH 2002Gene expression profiling predicts clinical outcome of breast cancerNature415530536CrossRefPubMedGoogle Scholar
  37. Brazma, A, Higamp, P, Quackenbush, J, Sherlock, G, Spellman, P, Stoeckert, C, Aach, J, Anssorge, W, Ball, CA, Causton, HC, Gaasterland, T, Glenisson, P, Holstege, FCP, Kim, IF, Markowitz, V, Matese, JC, Parkinson, H, Robinson, A, Sarkans, U, Schulze-Kremer, S, Stewart, J, Taylor, R, Vilo, J, Vingron, M 2001Minimum information about a microarray experiment (MIAME)-toward standards for microarray dataNat Genet29365371CrossRefGoogle Scholar
  38. Calvo, A, Xiao, N, Kang, J, Best, CJ, Leiva, I, Emmert-Buck, MR, Jorcyk, C, Green, JE 2002Alterations in gene expression profiles during prostate cancer progression: functional correlations to tumorigenicity and down-regulation of selenoprotein-P in mouse and human tumorsCancer Res6253255335Google Scholar
  39. Risinger, JI, Maxwell, GL, Chandramouli, GV, Jazaeri, A, Aprelikova, O, Patterson, T, Berchuck, A, Barrett, JC 2003Microarray analysis reveals distinct gene expression profiles among different histologic types of endometrial cancerCancer Res63611Google Scholar
  40. Irvine, AD, Christiano, AM 2001Hair on a gene string: recent advances in understanding the molecular genetics of hair lossClin Exp Dermatol265971Google Scholar
  41. Sargiannidou, I, Zhou, J, Tuszynski, GP 2001The role of thrombospondin-1 in tumor progressionExp Biol Med226726733Google Scholar
  42. Montfort, WR, Weichsel, A 1997Thymidylate synthase: structure, inhibition, and strained conformations during catalysisPharmacol Ther762943Google Scholar
  43. Babic, AM, Kireeva, ML, Kolesnikova, TV, Lau, LF 1998CYR61, a product of a growth factor-inducible immediate early gene, promotes angiogenesis and tumor growthPNAS USA9563556360Google Scholar
  44. Wagner, M, Klussmann, JP, Fangmann, R, Linder, R, Elewa, ME, Eidt, S, Rose, VM, Jungehulsing, M, Schulze, HJ 2001Cyclin-dependent kinase-inhibitor 1 (CDKN1A) in the squamous epithelium of the oropharynx: possible implications of molecular biology and compartmentationAnticancer Res21333334Google Scholar
  45. Zhou, R, Diehl, D, Hoeflich, A, Lahm, H 2003IGF-binding protein-4: biochemical characteristics and functional consequencesJ Endocrinol178177193PubMedGoogle Scholar
  46. Heinrich, PC, Behrmann, I, Haan, S, Hermanns, HM, Muller-Newen, G, Schaper, F 2003Principles of interleukin (IL)-6-type cytokine signalling and its regulationBiochem J374120Google Scholar
  47. Zhang, W, Bae, I, Krishnaraju, K, Azam, N, Fan, W, Smith, K, Hoffman, B, Leibermann, DA 1999CR6: A third member in the MyD118 and Gadd 45 family which functions in negative growth controlOncogene18749757Google Scholar
  48. Smith, ML, Ford, JM, Hollander, MC, Bortnick, RA, Amundson, SA, Seo, YR, Deng, CX, Hanawalt, PC, Fornace, AJ 2000p53-mediated DNA repair responses to UV radiation: studies of mouse cells lacking p53, p21, and /or gadd45 genesMol Cell Biol2037053714Google Scholar
  49. Harkin, DP, Bean, JM, Miklos, D, Song, YH, Truong, VB, Englert, C, Christians, FC, Ellison, LW, Maheswaran, S, Oliner, JD, Haber, DA 1999Induction of GADD45 and JNK/SARK-dependent apoptosis following inducible expression of BRC1Cell97575586Google Scholar
  50. Liebermann, DA, Hoffman, B 2002Myeloid differentiation (MyD)/growth arrest DNA damage (GADD) genes in tumor suppression, immunity and inflammationLeukemia16527541Google Scholar
  51. Aschele, C, Lonardi, S, Monfardini, S 2002Thymidylate synthase expression as a predictor of clinical response to fluoropyrimidine-base chemotherapy in advanced colorectal cancerCancer Treat Rev282747CrossRefPubMedGoogle Scholar
  52. Chen, H, Herndon, ME, Lawler, J 2000The cell biology of thrombospondin-1Matrix Biol19597614Google Scholar
  53. Blease, K, Jakubzick, C, Schuh, JM, Joshi, BH, Puri, RK, Hogaboam, CM 2001IL-13 fusion cytotoxin ameliorates chronic fungal-induced allergic airway disease in miceJ Immunol16765836592Google Scholar
  54. Murata, T, Noguchi, PD, Puri, RK 1996Il-13 induces phosphorylation and activation of JAK2 janus kinase in human colon carcinoma cell lineJ Immunol15629722978Google Scholar
  55. Krebs, DL, Hilton, DJ 2000SOCS: physiological suppressors of cytokine signaling.J Cell Sci11328132819PubMedGoogle Scholar
  56. Weaver, M, Laske, DW 2003Transferrin receptor ligand-targeted toxin conjugate (Tf-CRM107) for therapy of malignant gliomasJ Neuro Oncol65313Google Scholar
  57. Muldoon, LL, Neuwelt, EA 2003BR96-DOX immunoconjugate targeting of chemotherapy in brain tumor modelsJ Neuro Oncol654962Google Scholar
  58. Kawakami, M, Kawakami, K, Puri, RK 2003Interleukin-4 Pseudomonas exotoxin chimeric fusion protein for malignant glioma therapyJ Neuro Oncol651525Google Scholar
  59. Guo, N, Zabrenetzky, VS, Chandrasekaran, L, Sipes, JM, Lawler, J, Krutzsch, HC, Roberts, DD 1998Differential roles of protein kinase C and pertussis toxin-sensitive G-binding proteins in modulation of melanoma cell proliferation and motility by thrombospondin-1Cancer Res5831543162Google Scholar
  60. Volpert, OV, Lawler, J, Bouck, NP 1998A human fibrosacoma inhibits systemic angiogenesis and the growth of experimental metastases via thrombospondin-1Proc Natl Acad Sci USA9563436348Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  1. 1.Laboratory of Molecular Tumor Biology, CBER/NCI Genomics Program, Division of Cellular and Gene Therapies, Center for Biologics Evaluation and ResearchFood and Drug Administration, CBER/FDABethesdaUSA
  2. 2.Bioinformatics & Molecular Analysis Section, Center for Information TechnologyNational Institutes of HealthBethesdaUSA

Personalised recommendations