Abstract
Glioblastoma multiforme is the most aggressive form of human brain tumor, which has no effective cure. Previously, we have demonstrated that overexpression of the C-terminal fragment of the human telomerase reverse transcriptase (hTERTC27) inhibits the growth and tumorigenicity of human cervical cancer HeLa cells. In this study, the therapeutic effect and molecular mechanisms of hTERTC27-mediated cancer gene therapy were further explored in vivo in established human glioblastoma xenografts in nude mice. We showed that intratumoral injection of adeno-associated virus carrying hTERTC27 (rAAV-hTERTC27) is highly effective in reducing the growth of the subcutaneously transplanted glioblastoma tumors. Histological analyses showed that rAAV-hTERTC27 treatment leads to profound necrosis, apoptosis, infiltration of polymorphonuclear neutrophils and reduced microvessel density in the tumor samples. To study the molecular mechanism of rAAV-hTERTC27-mediated antitumor effects, we analyzed the global gene expression profiles of the rAAV-hTERTC27-treated tumor tissues and cell line as compared with that of the control rAAV-green fluorescent protein-treated samples by DNA microarray. Our results suggest that hTERTC27 exerts its effect through complex mechanisms, which involve genes regulating apoptosis, cell adhesion, cell cycle, immune responses, metabolism, signal transduction, transport, transcription and telomere maintenance.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
Accessions
GenBank/EMBL/DDBJ
References
Lantos PL, VandenBerg SR, Kleihues P . Tumors of the nervous system. In: Graham DI, Lantos PL (eds). GreenField's Neuropathology. Oxford University Press: London, 1996 pp 583–879.
Kanzawa T, Ito H, Kondo Y, Kondo S . Current and future gene therapy for malignant gliomas. J Biomed Biotechnol 2003; 2003: 25–34.
Preston-Martin S . Epidemiology. In: Berger MS, Wilson CB (eds). The Gliomas. WB Saunders: Philadelphia, 1999, pp 1–11.
Mahaley Jr MS, Mettlin C, Natarajan N, Laws Jr ER, Peace BB . National survey of patterns of care for brain-tumor patients. J Neurosurg 1989; 71: 826–836.
Deen DF, Chiarodo A, Grimm EA, Fike JR, Israel MA, Kun LE et al. Brain Tumor Working Group Report on the 9th International Conference on Brain Tumor Research and Therapy. Organ System Program, National Cancer Institute. J Neurooncol 1993; 16: 243–272.
Hiraga S, Ohnishi T, Izumoto S, Miyahara E, Kanemura Y, Matsumura H et al. Telomerase activity and alterations in telomere length in human brain tumors. Cancer Res 1998; 58: 2117–2125.
Blackburn EH . Structure and function of telomeres. Nature 1991; 350: 569–573.
Blackburn EH . Telomeres and telomerase: their mechanisms of action and the effects of altering their functions. FEBS Lett 2005; 579: 859–862.
Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL et al. Specific association of human telomerase activity with immortal cells and cancer. Science 1994; 266: 2011–2015.
Broccoli D, Young JW, de LT . Telomerase activity in normal and malignant hematopoietic cells. Proc Natl Acad Sci USA 1995; 92: 9082–9086.
Langford LA, Piatyszek MA, Xu R, Schold Jr SC, Shay JW . Telomerase activity in human brain tumours. Lancet 1995; 346: 1267–1268.
Huang JJ, Lin MC, Bai YX, Jing DD, Wong BC, Han SW et al. Ectopic expression of a COOH-terminal fragment of the human telomerase reverse transcriptase leads to telomere dysfunction and reduction of growth and tumorigenicity in HeLa cells. Cancer Res 2002; 62: 3226–3232.
Newbold RF . Telomerase as an anti-cancer drug target: will it fulfil its early promise? Anticancer Drug Des 1999; 14: 349–354.
Shay JW, Zou Y, Hiyama E, Wright WE . Telomerase and cancer. Hum Mol Genet 2001; 10: 677–685.
Huo LF, Tang JW, Huang JJ, Huang PT, Huang CF, Kung HF et al. Cancer immunotherapy targeting the telomerase reverse transcriptase. Cell Mol Immunol 2006; 3: 1–11.
Chen Y, Luk KD, Cheung KM, Xu R, Lin MC, Lu WW et al. Gene therapy for new bone formation using adeno-associated viral bone morphogenetic protein-2 vectors. Gene Therapy 2003; 10: 1345–1353.
Tu SP, Cui JT, Liston P, Huajiang X, Xu R, Lin MC et al. Gene therapy for colon cancer by adeno-associated viral vector-mediated transfer of survivin Cys84Ala mutant. Gastroenterology 2005; 128: 361–375.
Qing K, Mah C, Hansen J, Zhou S, Dwarki V, Srivastava A . Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. Nat Med 1999; 5: 71–77.
Mitchell JR, Wood E, Collins K . A telomerase component is defective in the human disease dyskeratosis congentia. Nature 1999; 402: 551–555.
McConnell BB, Vertino PM . Activation of a caspase-9-mediated apoptotic pathway by subcellular redistribution of the novel caspase recruitment domain protein TMS1. Cancer Res 2000; 60: 6243–6247.
Durand B, Sperisen P, Emery P, Barras E, Zufferey M, Mach B et al. RFXAP, a novel subunit of the RFX DNA binding complex is mutated in MHC class II deficiency. EMBO J 1997; 16: 1045–1055.
Edelman GM, Crossin KL . Cell adhesion molecules: implications for a molecular histology. Annu Rev Biochem 1991; 60: 155–190.
Song XT, Aldrich M, Chen SY . Suppressor of cytokine signaling 1 inhibition strategy to enhance anti-HIV vaccination. Expert Rev Vaccines 2006; 5: 495–503.
Sanjo H, Takeda K, Tsujimura T, Ninomiya-Tsuji J, Matsumoto K, Akira S . TAB2 is essential for prevention of apoptosis in fetal liver but not for interleukin-1 signaling. Mol Cell Biol 2003; 23: 1231–1238.
Peinado H, Del Carmen Iglesias-de la Cruz M, Olmeda D, Csiszar K, Fong KS, Vega S et al. A molecular role for lysyl oxidase-like 2 enzyme in snail regulation and tumor progression. EMBO J 2005; 24: 3446–3458.
Prasad NK, Papoff G, Zeuner A, Bonnin E, Kazatchkine MD, Ruberti G et al. Therapeutic preparations of normal polyspecific IgG (IVIg) induce apoptosis in human lymphocytes and monocytes: a novel mechanism of action of IVIg involving the Fas apoptotic pathway. J Immunol 1998; 161: 3781–3790.
Chen T, Richard S . Structure-function analysis of Qk1: a lethal point mutation in mouse quaking prevents homodimerization. Mol Cell Biol 1998; 18: 4863–4871.
Croy BA, Linder KE, Yager JA . Primer for non-immunologists on immune-deficient mice and their applications in research. Comp Med 2001; 51: 300–313.
Kelland LR . Of mice and men: values and liabilities of the athymic nude mouse model in anticancer drug development. Eur J Cancer 2004; 40: 827–836.
Carter PJ, Samulski RJ . Adeno-associated viral vectors as gene delivery vehicles. Int J Mol Med 2000; 6: 17–27.
Ma HI, Guo P, Li J, Lin SZ, Chiang YH, Xiao X et al. Suppression of intracranial human glioma growth after intramuscular administration of an adeno-associated viral vector expressing angiostatin. Cancer Res 2002; 62: 756–763.
Ma HI, Lin SZ, Chiang YH, Li J, Chen SL, Tsao YP et al. Intratumoral gene therapy of malignant brain tumor in a rat model with angiostatin delivered by adeno-associated viral (AAV) vector. Gene Therapy 2002; 9: 2–11.
Cunningham J, Oiwa Y, Nagy D, Podsakoff G, Colosi P, Bankiewicz KS . Distribution of AAV-TK following intracranial convection-enhanced delivery into rats. Cell Transplant 2000; 9: 585–594.
Mizuno M, Yoshida J, Colosi P, Kurtzman G . Adeno-associated virus vector containing the herpes simplex virus thymidine kinase gene causes complete regression of intracerebrally implanted human gliomas in mice, in conjunction with ganciclovir administration. Jpn J Cancer Res 1998; 89: 76–80.
Okada H, Miyamura K, Itoh T, Hagiwara M, Wakabayashi T, Mizuno M et al. Gene therapy against an experimental glioma using adeno-associated virus vectors. Gene Therapy 1996; 3: 957–964.
Masumoto J, Taniguchi S, Ayukawa K, Sarvotham H, Kishino T, Niikawa N et al. ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells. J Biol Chem 1999; 274: 33835–33838.
Conway KE, McConnell BB, Bowring CE, Donald CD, Warren ST, Vertino PM . TMS1, a novel proapoptotic caspase recruitment domain protein, is a target of methylation-induced gene silencing in human breast cancers. Cancer Res 2000; 60: 6236–6242.
Heiss NS, Knight SW, Vulliamy TJ, Klauck SM, Wiemann S, Mason PJ et al. X-linked dyskeratosis congenita is caused by mutations in a highly conserved gene with putative nucleolar functions. Nat Genet 1998; 19: 32–38.
Knight SW, Heiss NS, Vulliamy TJ, Greschner S, Stavrides G, Pai GS et al. X-linked dyskeratosis congenita is predominantly caused by missense mutations in the DKC1 gene. Am J Hum Genet 1999; 65: 50–58.
Cayuela ML, Flores JM, Blasco MA . The telomerase RNA component Terc is required for the tumour-promoting effects of Tert overexpression. EMBO R 2005; 6: 268–274.
Briggs RC, Kao WY, Dworkin LL, Briggs JA, Dessypris EN, Clark J . Regulation and specificity of MNDA expression in monocytes, macrophages, and leukemia/B lymphoma cell lines. J Cell Biochem 1994; 56: 559–567.
Kehlen A, Thiele K, Riemann D, Rainov N, Langner J . Interleukin-17 stimulates the expression of IkappaB alpha mRNA and the secretion of IL-6 and IL-8 in glioblastoma cell lines. J Neuroimmunol 1999; 101: 1–6.
Goswami S, Gupta A, Sharma SK . Interleukin-6-mediated autocrine growth promotion in human glioblastoma multiforme cell line U87MG. J Neurochem 1998; 71: 1837–1845.
Hirose K, Hakozaki M, Nyunoya Y, Kobayashi Y, Matsushita K, Takenouchi T et al. Chemokine gene transfection into tumour cells reduced tumorigenicity in nude mice in association with neutrophilic infiltration. Br J Cancer 1995; 72: 708–714.
Rossi D, Zlotnik A . The biology of chemokines and their receptors. Annu Rev Immunol 2000; 18: 217–242.
Di CE, Forni G, Lollini P, Colombo MP, Modesti A, Musiani P . The intriguing role of polymorphonuclear neutrophils in antitumor reactions. Blood 2001; 97: 339–345.
Midorikawa Y, Yamashita T, Sendo F . Modulation of the immune response to transplanted tumors in rats by selective depletion of neutrophils in vivo using a monoclonal antibody: abrogation of specific transplantation resistance to chemical carcinogen-induced syngeneic tumors by selective depletion of neutrophils in vivo. Cancer Res 1990; 50: 6243–6247.
Matsumoto Y, Saiki I, Murata J, Okuyama H, Tamura M, Azuma I . Recombinant human granulocyte colony-stimulating factor inhibits the metastasis of hematogenous and non-hematogenous tumors in mice. Int J Cancer 1991; 49: 444–449.
Colombo MP, Ferrari G, Stoppacciaro A, Parenza M, Rodolfo M, Mavilio F et al. Granulocyte colony-stimulating factor gene transfer suppresses tumorigenicity of a murine adenocarcinoma in vivo. J Exp Med 1991; 173: 889–897.
Musiani P, Allione A, Modica A, Lollini PL, Giovarelli M, Cavallo F et al. Role of neutrophils and lymphocytes in inhibition of a mouse mammary adenocarcinoma engineered to release IL-2, IL-4, IL-7, IL-10, IFN-alpha, IFN-gamma, and TNF-alpha. Lab Invest 1996; 74: 146–147.
Acknowledgements
We thank the Genome Research Center (HKU) for conducting the microarray analysis. The work was supported by Innovation and Technology Fund (ITS/105/02) to MCL and grants from the Hong Kong Research Grant Council (CUHK 7422/03M to HFK; HKU 7243/02M to MCL), Li Ka Shing Institute of Health Sciences (to HFK), Shanghai Metropolitan Fund for Research and Development (04JC14096) and Guangzhou Metropolitan Fund (2005Z1-E0131).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Ng, S., Gao, Y., Chau, D. et al. A novel glioblastoma cancer gene therapy using AAV-mediated long-term expression of human TERT C-terminal polypeptide. Cancer Gene Ther 14, 561–572 (2007). https://doi.org/10.1038/sj.cgt.7701038
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.cgt.7701038
Keywords
This article is cited by
-
Telomerase gene therapy: a remission toward cancer
Medical Oncology (2022)
-
Gene Delivery in Neuro-Oncology
Current Oncology Reports (2017)
-
Emerging Role of Combination of All-trans Retinoic Acid and Interferon-gamma as Chemoimmunotherapy in the Management of Human Glioblastoma
Neurochemical Research (2007)