[en] Strategies for non-invasive and quantitative imaging of gene expression in vivo have been developed over the past decade. Non-invasive assessment of the dynamics of gene regulation is of interest for the detection of endogenous disease-specific biological alterations (e.g., signal transduction) and for monitoring the induction and regulation of therapeutic genes (e.g., gene therapy). To demonstrate that non-invasive imaging of regulated expression of any type of gene after in vivo transduction by versatile vectors is feasible, we generated regulatable herpes simplex virus type 1 (HSV-1) amplicon vectors carrying hormone (mifepristone) or antibiotic (tetracycline) regulated promoters driving the proportional co-expression of two marker genes. Regulated gene expression was monitored by fluorescence microscopy in culture and by positron emission tomography (PET) or bioluminescence (BLI) in vivo. The induction levels evaluated in glioma models varied depending on the dose of inductor. With fluorescence microscopy and BLI being the tools for assessing gene expression in culture and animal models, and with PET being the technology for possible application in humans, the generated vectors may serve to non-invasively monitor the dynamics of any gene of interest which is proportionally co-expressed with the respective imaging marker gene in research applications aiming towards translation into clinical application.
Disciplines :
Oncology
Author, co-author :
Winkeler, Alexandra; Laboratory for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Department of Neurology, University of Cologne, Cologne, Germany
Sena-Esteves, Miguel; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Paulis, Leonie E M; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Li, Hongfeng; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Waerzeggers, Yannic; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Rückriem, Benedikt; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Himmelreich, Uwe; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Klein, Markus; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Monfared, Parisa; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Rueger, Maria A; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
HENEKA, Michael ; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Vollmar, Stefan; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Hoehn, Mathias; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Fraefel, Cornel; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Graf, Rudolf; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Wienhard, Klaus; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Heiss, Wolf D; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Jacobs, Andreas H; Lab. for Gene Therapy and Molecular Imaging at the Max Planck-Institute for Neurological Research, Center for Molecular Medicine (CMMC) and Departments of Neurology and Radiology, The University of Cologne, Cologne, Germany
Gambhir SS, Barrio JR, Herschman HR, Phelps ME, (1999) Assays for noninvasive imaging of reporter gene expression. Nucl Med Biol 26: 481-490.
Jacobs AH, Dittmar C, Winkeler A, Garlip G, Heiss WD, (2002) Molecular imaging of gliomas. Mol Imaging 1: 309-335.
Blasberg RG, (2003) In vivo molecular-genetic imaging: multi-modality nuclear and optical combinations. Nucl Med Biol 30: 879-888.
Contag CH, Bachmann MH, (2002) Advances in in vivo bioluminescence imaging of gene expression. Annu Rev Biomed Eng 4: 235-260.
Hoehn M, Kustermann E, Blunk J, Wiedermann D, Trapp T, et al.(2002) Monitoring of implanted stem cell migration in vivo: a highly resolved in vivo magnetic resonance imaging investigation of experimental stroke in rat. Proc Natl Acad Sci U S A 99: 16267-16272.
Jacobs AH, Rueger MA, Winkeler A, Li H, Vollmar S, et al.(2007) Imaging-guided gene therapy of experimental gliomas. Cancer Res 67: 1706-1715.
Jacobs A, Dubrovin M, Hewett J, Sena-Esteves M, Tan CW, et al.(1999) Functional coexpression of HSV-1 thymidine kinase and green fluorescent protein: implications for noninvasive imaging of transgene expression. Neoplasia 1: 154-161.
Tjuvajev JG, Joshi A, Callegari J, Lindsley L, Joshi R, et al.(1999) A general approach to the non-invasive imaging of transgenes using cis-linked herpes simplex virus thymidine kinase. Neoplasia 1: 315-320.
Yu Y, Annala AJ, Barrio JR, Toyokuni T, Satyamurthy N, et al.(2000) Quantification of target gene expression by imaging reporter gene expression in living animals. Nat Med 6: 933-937.
Yaghoubi SS, Wu L, Liang Q, Toyokuni T, Barrio JR, et al.(2001) Direct correlation between positron emission tomographic images of two reporter genes delivered by two distinct adenoviral vectors. Gene Ther 8: 1072-1080.
Saito Y, Price RW, Rottenberg DA, Fox JJ, Su TL, et al.(1982) Quantitative autoradiographic mapping of herpes simplex virus encephalitis with a radiolabeled antiviral drug. Science 217: 1151-1153.
Tjuvajev JG, Stockhammer G, Desai R, Uehara H, Watanabe K, et al.(1995) Imaging the expression of transfected genes in vivo. Cancer Res 55: 6126-6132.
Iyer M, Salazar FB, Lewis X, Zhang L, Carey M, et al.(2004) Noninvasive imaging of enhanced prostate-specific gene expression using a two-step transcriptional amplification-based lentivirus vector. Mol Ther 10: 545-552.
Doubrovin M, Ponomarev V, Beresten T, Balatoni J, Bornmann W, et al.(2001) Imaging transcriptional regulation of p53-dependent genes with positron emission tomography in vivo. Proc Natl Acad Sci U S A 98: 9300-9305.
Green LA, Yap CS, Nguyen K, Barrio JR, Namavari M, et al.(2002) Indirect monitoring of endogenous gene expression by positron emission tomography (PET) imaging of reporter gene expression in transgenic mice. Mol Imaging Biol 4: 71-81.
Ponomarev V, Doubrovin M, Lyddane C, Beresten T, Balatoni J, et al.(2001) Imaging TCR-dependent NFAT-mediated T-cell activation with positron emission tomography in vivo. Neoplasia 3: 480-488.
Gossen M, Bujard H, (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A 89: 5547-5551.
Hynes NE, Kennedy N, Rahmsdorf U, Groner B, (1981) Hormone-responsive expression of an endogenous proviral gene of mouse mammary tumor virus after molecular cloning and gene transfer into cultured cells. Proc Natl Acad Sci U S A 78: 2038-2042.
Wurm FM, Gwinn KA, Kingston RE, (1986) Inducible overproduction of the mouse c-myc protein in mammalian cells. Proc Natl Acad Sci U S A 83: 5414-5418.
Christopherson KS, Mark MR, Bajaj V, Godowski PJ, (1992) Ecdysteroid-dependent regulation of genes in mammalian cells by a Drosophila ecdysone receptor and chimeric transactivators. Proc Natl Acad Sci U S A 89: 6314-6318.
Mayo KE, Warren R, Palmiter RD, (1982) The mouse metallothionein-I gene is transcriptionally regulated by cadmium following transfection into human or mouse cells. Cell 29: 99-108.
Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, et al.(1995) Transcriptional activation by tetracyclines in mammalian cells. Science 268: 1766-1769.
Baron U, Freundlieb S, Gossen M, Bujard H, (1995) Co-regulation of two gene activities by tetracycline via a bidirectional promoter. Nucleic Acids Res 23: 3605-3606.
Wang Y, O'Malley BW Jr, Tsai SY, O'Malley BW, (1994) A regulatory system for use in gene transfer. Proc Natl Acad Sci U S A 91: 8180-8184.
Burcin MM, Schiedner G, Kochanek S, Tsai SY, O'Malley BW, (1999) Adenovirus-mediated regulable target gene expression in vivo. Proc Natl Acad Sci U S A 96: 355-360.
Furth PA, St Onge L, Boger H, Gruss P, Gossen M, et al.(1994) Temporal control of gene expression in transgenic mice by a tetracycline-responsive promoter. Proc Natl Acad Sci U S A 91: 9302-9306.
Shockett PE, Schatz DG, (1996) Diverse strategies for tetracycline-regulated inducible gene expression. Proc Natl Acad Sci U S A 93: 5173-5176.
Kistner A, Gossen M, Zimmermann F, Jerecic J, Ullmer C, et al.(1996) Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. Proc Natl Acad Sci U S A 93: 10933-10938.
Wang Y, DeMayo FJ, Tsai SY, O'Malley BW, (1997) Ligand-inducible and liver-specific target gene expression in transgenic mice. Nat Biotechnol 15: 239-243.
Sun X, Annala AJ, Yaghoubi SS, Barrio JR, Nguyen KN, et al.(2001) Quantitative imaging of gene induction in living animals. Gene Ther 8: 1572-1579.
Fraefel C, Song S, Lim F, Lang P, Yu L, et al.(1996) Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells. J Virol 70: 7190-7197.
Black ME, Newcomb TG, Wilson HM, Loeb LA, (1996) Creation of drug-specific herpes simplex virus type 1 thymidine kinase mutants for gene therapy. Proc Natl Acad Sci U S A 93: 3525-3529.
Jacobs AH, Winkeler A, Hartung M, Slack M, Dittmar C, et al.(2003) Improved herpes simplex virus type 1 amplicon vectors for proportional coexpression of positron emission tomography marker and therapeutic genes. Hum Gene Ther 14: 277-297.
Freundlieb S, Schirra-Muller C, Bujard H, (1999) A tetracycline controlled activation/repression system with increased potential for gene transfer into mammalian cells. J Gene Med 1: 4-12.
Lipshutz GS, Gruber CA, Cao Y, Hardy J, Contag CH, et al.(2001) In utero delivery of adeno-associated viral vectors: intraperitoneal gene transfer produces long-term expression. Mol Ther 3: 284-292.
Bhaumik S, Gambhir SS, (2002) Optical imaging of Renilla luciferase reporter gene expression in living mice. Proc Natl Acad Sci U S A 99: 377-382.
Wu JC, Sundaresan G, Iyer M, Gambhir SS, (2001) Noninvasive optical imaging of firefly luciferase reporter gene expression in skeletal muscles of living mice. Mol Ther 4: 297-306.
Wang Y, Yu YA, Shabahang S, Wang G, Szalay AA, (2002) Renilla luciferase-Aequorea GFP (Ruc-GFP) fusion protein, a novel dual reporter for real-time imaging of gene expression in cell cultures and in live animals. Mol Genet Genomics 268: 160-168.
Tannous BA, Kim DE, Fernandez JL, Weissleder R, Breakefield XO, (2005) Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Mol Ther 11: 435-443.
Shah K, Tang Y, Breakefield X, Weissleder R, (2003) Real-time imaging of TRAIL-induced apoptosis of glioma tumors in vivo. Oncogene 22: 6865-6872.
Yu YA, Szalay AA, (2002) A Renilla luciferase-Aequorea GFP (ruc-gfp) fusion gene construct permits real-time detection of promoter activation by exogenously administered mifepristone in vivo. Mol Genet Genomics 268: 169-178.
Dumortier J, Schonig K, Oberwinkler H, Low R, Giese T, et al.(2005) Liver-specific expression of interferon gamma following adenoviral gene transfer controls hepatitis B virus replication in mice. Gene Ther.
Doubrovin M, Serganova I, Mayer-Kuckuk P, Ponomarev V, Blasberg RG, (2004) Multimodality in vivo molecular-genetic imaging. Bioconjug Chem 15: 1376-1388.
Weissleder R, (2002) Scaling down imaging: molecular mapping of cancer in mice. Nat Rev Cancer 2: 11-18.
Weissleder R, (2006) Molecular imaging in cancer. Science 312: 1168-1171.
Heiss W-D, Habedank B, Klein JC, Herholz K, Wienhard K, et al.(2004) Metabolic Rates in Small Brain Nuclei Determined by High-Resolution PET. J Nucl Med 45: 1811-1815.
