Top

European Journal of Nuclear Medicine and Molecular Imaging

Published in:

01-01-2008 | Original Article

Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter

Authors: Do Won Hwang, Joo Hyun Kang, Jae Min Jeong, June-Key Chung, Myung Chul Lee, Soonhag Kim, Dong Soo Lee

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 1/2008

Abstract

Purpose

We imaged neuronal differentiation in vivo using dual reporters (sodium iodide symporter [NIS] and luciferase) coupled to a neuron-specific enolase (NSE) promoter.

Methods

PC12 (NSE positive) and F11 cells were transfected with a bicistronic (NIS and luciferase; pNSE-NF) or a luciferase (pNSE-Fluc) reporter coupled to the NSE promoter. Weak NSE promoter activity was overcome by a two-step transcriptional amplification (TSTA) system (pNSE-TSTA-Fluc). In vivo, NIS and luciferase expression were examined using a ^99mTc-pertechnetate gamma camera and bioluminescence imaging, respectively.

Results

pNSE-NF-transfected PC12 cells showed 3-fold higher radioiodine uptakes and >100-fold higher luciferase activity than parental cells. NIS or luciferase activity was not detected in pNSE-NF-transfected HeLa cells. When F11 cells were differentiated into neurons by db-cAMP, NIS and luciferase activities increased 4-fold compared to those without treatment, which was confirmed by Western blot and RT-PCR of NSE. In vivo in pNSE-NF-transfected F11 cells, db-cAMP treatment increased the luciferase activity but not the scintigraphic activity. In vitro, pNSE-TSTA-Fluc produced 130-fold higher luciferase activity than pNSE-Fluc and neuronal differentiation showed 4-fold higher activity from both pNSE-TSTA-Fluc and pNSE-Fluc than before differentiation. In vivo, in pNSE-TSTA-Fluc-transfected F11 cells, luciferase activity increased after neuronal differentiation. In vivo luciferase activity persisted up to 2 days after db-cAMP-induced neuronal differentiation.

Conclusion

NSE promoter-driven dual reporter transgenes revealed the possibility of in vivo imaging of neuronal differentiation, which was further enabled by high amplification using a TSTA system. We propose that this strategy be used to follow the transplanted stem cells during differentiation in live animals.

Pluchino S, Zanotti L, Deleidi M, Martino G. Neural stem cells and their use as therapeutic tool in neurological disorders. Brain Res Brain Res Rev 2005;48:211–9.PubMedCrossRef

Baizabal JM, Furlan-Magaril M, Santa-Olalla J, Covarrubias L. Neural stem cells in development and regenerative medicine. Arch Med Res 2003;34:572–88.PubMedCrossRef

Deng J, Petersen BE, Steindler DA, Jorgensen ML, Laywell ED. Mesenchymal stem cells spontaneously express neural proteins in culture and are neurogenic after transplantation. Stem Cells 2006;24:1054–64.PubMedCrossRef

Mokry J, Karbanova J, Filip S. Differentiation potential of murine neural stem cells in vitro and after transplantation. Transplant Proc 2005;37:268–72.PubMedCrossRef

Dziewczapolski G, Lie DC, Ray J, Gage FH, Shults CW. Survival and differentiation of adult rat-derived neural progenitor cells transplanted to the striatum of hemiparkinsonian rats. Exp Neurol 2003;183:653–64.PubMedCrossRef

Gage FH, Coates PW, Palmer TD, Kuhn HG, Fisher LJ, Suhonen JO, et al. Survival and differentiation of adult neuronal progenitor cells transplanted to the adult brain. Proc Natl Acad Sci USA 1995;92:11879–83.PubMedCrossRef

Chu K, Kim M, Park KI, Jeong SW, Park HK, Jung KH, et al. Human neural stem cells improve sensorimotor deficits in the adult rat brain with experimental focal ischemia. Brain Res 2004;1016:145–53.PubMedCrossRef

Navarro-Galve B, Villa A, Bueno C, Thompson L, Johansen J, Martinez-Serrano A. Gene marking of human neural stem/precursor cells using green fluorescent proteins. J Gene Med 2005;7:18–29.PubMedCrossRef

Wernig M, Benninger F, Schmandt T, Rade M, Tucker KL, Bussow H, et al. Functional integration of embryonic stem cell-derived neurons in vivo. J Neurosci 2004;24:5258–68.PubMedCrossRef

10.

Herschman HR, MacLaren DC, Iyer M, Namavari M, Bobinski K, Green LA, et al. Seeing is believing: non-invasive, quantitative and repetitive imaging of reporter gene expression in living animals, using positron emission tomography. J Neurosci Res 2000;59:699–705.PubMedCrossRef

11.

Wang X, Rosol M, Ge S, Peterson D, McNamara G, Pollack H, et al. Dynamic tracking of human hematopoietic stem cell engraftment using in vivo bioluminescence imaging. Blood 2003;102:3478–82.PubMedCrossRef

12.

Okada S, Ishii K, Yamane J, Iwanami A, Ikegami T, Katoh H, et al. In vivo imaging of engrafted neural stem cells: its application in evaluating the optimal timing of transplantation for spinal cord injury. Faseb J 2005;19:1839–41.PubMed

13.

Min JJ, Ahn Y, Moon S, Kim YS, Park JE, Kim SM, et al. In vivo bioluminescence imaging of cord blood derived mesenchymal stem cell transplantation into rat myocardium. Ann Nucl Med 2006;20:165–70.PubMedCrossRef

14.

Shah K, Weissleder R. Molecular optical imaging: applications leading to the development of present day therapeutics. NeuroRx 2005;2:215–25.PubMedCrossRef

15.

Andersen JK, Frim DM, Isacson O, Breakefield XO. Herpesvirus-mediated gene delivery into the rat brain: specificity and efficiency of the neuron-specific enolase promoter. Cell Mol Neurobiol 1993;13:503–15.PubMedCrossRef

16.

Morelli AE, Larregina AT, Smith-Arica J, Dewey RA, Southgate TD, Ambar B, et al. Neuronal and glial cell type-specific promoters within adenovirus recombinants restrict the expression of the apoptosis-inducing molecule Fas ligand to predetermined brain cell types, and abolish peripheral liver toxicity. J Gen Virol 1999;80:571–83.PubMed

17.

Sakimura K, Kushiya E, Ogura A, Kudo Y, Katagiri T, Takahashi Y. Upstream and intron regulatory regions for expression of the rat neuron-specific enolase gene. Brain Res Mol Brain Res 1995;28:19–28.PubMedCrossRef

18.

Marangos PJ, Schmechel DE, Parma AM, Goodwin FK. Developmental profile of neuron-specific (NSE) and non-neuronal (NNE) enolase. Brain Res 1980;190:185–93.PubMedCrossRef

19.

Niu G, Gaut AW, Ponto LL, Hichwa RD, Madsen MT, Graham MM, et al. Multimodality noninvasive imaging of gene transfer using the human sodium iodide symporter. J Nucl Med 2004;45:445–9.PubMed

20.

Chung JK. Sodium iodide symporter: its role in nuclear medicine. J Nucl Med 2002;43:1188–200.PubMed

21.

Dai G, Levy O, Carrasco N. Cloning and characterization of the thyroid iodide transporter. Nature 1996;379:458–60.PubMedCrossRef

22.

Groot-Wassink T, Aboagye EO, Wang Y, Lemoine NR, Reader AJ, Vassaux G. Quantitative imaging of Na/I symporter transgene expression using positron emission tomography in the living animal. Mol Ther 2004;9:436–42.PubMedCrossRef

23.

Iyer M, Wu L, Carey M, Wang Y, Smallwood A, Gambhir SS. Two-step transcriptional amplification as a method for imaging reporter gene expression using weak promoters. Proc Natl Acad Sci USA 2001;98:14595–600.PubMedCrossRef

24.

Zhang L, Adams JY, Billick E, Ilagan R, Iyer M, Le K, et al. Molecular engineering of a two-step transcription amplification (TSTA) system for transgene delivery in prostate cancer. Mol Ther 2002;5:223–32.PubMedCrossRef

25.

Crocker SJ, Liston P, Anisman H, Lee CJ, Smith PD, Earl N, et al. Attenuation of MPTP-induced neurotoxicity and behavioural impairment in NSE-XIAP transgenic mice. Neurobiol Dis 2003;12:150–61.PubMedCrossRef

26.

So MK, Kang JH, Chung JK, Lee YJ, Shin JH, Kim KI, et al. In vivo imaging of retinoic acid receptor activity using a sodium/iodide symporter and luciferase dual imaging reporter gene. Mol Imaging 2004;3:163–71.PubMedCrossRef

27.

Kosugi S, Sasaki N, Hai N, Sugawa H, Aoki N, Shigemasa C, et al. Establishment and characterization of a Chinese hamster ovary cell line, CHO-4J, stably expressing a number of Na+/I- symporters. Biochem Biophys Res Commun 1996;227:94–101.PubMedCrossRef

28.

Min JJ, Chung JK, Lee YJ, Shin JH, Yeo JS, Jeong JM, et al. In vitro and in vivo characteristics of a human colon cancer cell line, SNU-C5N, expressing sodium-iodide symporter. Nucl Med Biol 2002;29:537–45.PubMedCrossRef

29.

Ghil SH, Kim BJ, Lee YD, Suh-Kim H. Neurite outgrowth induced by cyclic AMP can be modulated by the alpha subunit of Go. J Neurochem 2000;74:151–8.PubMedCrossRef

30.

Acton PD, Zhou R. Imaging reporter genes for cell tracking with PET and SPECT. Q J Nucl Med Mol Imaging 2005;49:349–60.PubMed

31.

Kirik D, Breysse N, Bjorklund T, Besret L, Hantraye P. Imaging in cell-based therapy for neurodegenerative diseases. Eur J Nucl Med Mol Imaging 2005;32:417–34.CrossRef

32.

Roy NS, Benraiss A, Wang S, Fraser RA, Goodman R, Couldwell WT, et al. Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone. J Neurosci Res 2000;59:321–31.PubMedCrossRef

33.

Tang F, Shang K, Wang X, Gu J. Differentiation of embryonic stem cell to astrocytes visualized by green fluorescent protein. Cell Mol Neurobiol 2002;22:95–101.PubMedCrossRef

34.

Kang JH, Lee DS, Paeng JC, Lee JS, Kim YH, Lee YJ, et al. Development of a sodium/iodide symporter (NIS)-transgenic mouse for imaging of cardiomyocyte-specific reporter gene expression. J Nucl Med 2005;46:479–83.PubMed

35.

Tsuchiya R, Yoshiki F, Kudo Y, Morita M. Cell type-selective expression of green fluorescent protein and the calcium indicating protein, yellow cameleon, in rat cortical primary cultures. Brain Res 2002;956:221–9.PubMedCrossRef

36.

Jakobsson J, Ericson C, Jansson M, Bjork E, Lundberg C. Targeted transgene expression in rat brain using lentiviral vectors. J Neurosci Res 2003;73:876–85.PubMedCrossRef

37.

Klein RL, Meyer EM, Peel AL, Zolotukhin S, Meyers C, Muzyczka N, et al. Neuron-specific transduction in the rat septohippocampal or nigrostriatal pathway by recombinant adeno-associated virus vectors. Exp Neurol 1998;150:183–94.PubMedCrossRef

38.

Brecht WJ, Harris FM, Chang S, Tesseur I, Yu GQ, Xu Q, et al. Neuron-specific apolipoprotein e4 proteolysis is associated with increased tau phosphorylation in brains of transgenic mice. J Neurosci 2004;24:2527–34.PubMedCrossRef

39.

Martinou JC, Dubois-Dauphin M, Staple JK, Rodriguez I, Frankowski H, Missotten M, et al. Overexpression of BCL-2 in transgenic mice protects neurons from naturally occurring cell death and experimental ischemia. Neuron 1994;13:1017–30.PubMedCrossRef

40.

Forss-Petter S, Danielson PE, Catsicas S, Battenberg E, Price J, Nerenberg M, et al. Transgenic mice expressing beta-galactosidase in mature neurons under neuron-specific enolase promoter control. Neuron 1990;5:187–97.PubMedCrossRef

41.

Navarro V, Millecamps S, Geoffroy MC, Robert JJ, Valin A, Mallet J, et al. Efficient gene transfer and long-term expression in neurons using a recombinant adenovirus with a neuron-specific promoter. Gene Ther 1999;6:1884–92.PubMedCrossRef

42.

Hannas-Djebbara Z, Didier-Bazs M, Sacchettoni S, Prod’hon C, Jouvet M, Belin MF, et al. Transgene expression of plasmid DNAs directed by viral or neural promoters in the rat brain. Brain Res Mol Brain Res 1997;46:91–9.PubMedCrossRef

43.

Bhattacharjee AK, Ueyama T, Kondoh T, Hayashi S, Abouelfetouh A, Sakai N, et al. In vivo transgene expression using an adenoviral tetracycline-regulated system with neuron-specific enolase promoter. Biochem Biophys Res Commun 2004;317:1144–8.PubMedCrossRef

44.

Shin JH, Chung JK, Kang JH, Lee YJ, Kim KI, So Y, et al. Noninvasive imaging for monitoring of viable cancer cells using a dual-imaging reporter gene. J Nucl Med 2004;45:2109–15.PubMed

45.

Lang KJ, Rathjen J, Vassilieva S, Rathjen PD. Differentiation of embryonic stem cells to a neural fate: a route to re-building the nervous system. J Neurosci Res 2004;76:184–92.PubMedCrossRef

Title: Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter
Authors: Do Won Hwang
Joo Hyun Kang
Jae Min Jeong
June-Key Chung
Myung Chul Lee
Soonhag Kim
Dong Soo Lee
Publication date: 01-01-2008
Publisher: Springer-Verlag
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 1/2008
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-007-0561-8

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2008

Tumor hypoxia imaging in orthotopic liver tumors and peritoneal metastasis: a comparative study featuring dynamic 18F-MISO and 124I-IAZG PET in the same study cohort

Intestinal accumulation of 99mTc-MDP in a patient with protein-losing enteropathy

Activity-based costing evaluation of [18F]-fludeoxyglucose production

Comparative analysis of MR imaging, Ictal SPECT and EEG in temporal lobe epilepsy: a prospective IAEA multi-center study

Staging with PET and the “Will Rogers” effect: redefining prognosis and survival in patients with cancer

Could choline PET play a role in malignancies other than prostate cancer?