Skip to main content

Advertisement

SpringerLink
Log in

Can Carrier-Mediated Delivery System Promote the Development of Antisense Imaging?

  • Research Article
  • Published:
Molecular Imaging and Biology Aims and scope Submit manuscript

Abstract

Purpose

We aimed to explore the feasibility of transfection methods for antisense imaging.

Procedures

Antisense oligonucleotides (ASON) targeted to the mRNA of hTERT gene were synthesized and labeled with Technetium-99m and fluorescein isothiocyanate (FITC), respectively. Then, ASON was combined with transfection reagent Lipofectamine 2000 and XfectTM, named Lipo-ASON and Xfect-ASON, respectively. After transfection, the labeled ASON was characterized in hNPCs-G3 and hRPE cells. Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were performed to assay the hTERT mRNA and protein levels after hNPCs-G3 cells were incubated with Lipo-ASON, Xfect-ASON, and naked ASON. In addition, Lipo-ASON, Xfect-ASON, and naked ASON were injected into tumor-bearing mice, and the biodistribution in vivo was performed.

Results

The presence of two transfection reagents significantly increased intracellular uptake of radiolabeled ASON in both cell lines compared with naked ASON (p < 0.05). However, there was no significant difference in cellular uptake rates of Lipo-ASON and Xfect-ASON between hNPCs-G3 and hRPE cells. In comparison with naked ASON, the fluorescence intensity was strongly enhanced after binding to transfection reagents. Furthermore, the levels of hTERT mRNA and protein were significantly reduced in cells treated with Lipo-ASON and Xfect-ASON (p < 0.05), but naked ASON had no significant effect on hTERT expression level. The biodistribution study indicated that tumor radioactivity uptake of radiolabeled ASON for naked ASON, Lipo-ASON, and Xfect-ASON group was low and shown no significant difference in vivo.

Conclusions

Lipofectamine transfection and XfectTM transfection were not effective delivery methods of ASON for antisense imaging.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.

Similar content being viewed by others

References

  1. Massoud TF, Gambhir SS (2003) Molecular imaging in living subjects: seeing fundamental biological processes in a new light. Genes Dev 17:545–580

    Article  CAS  PubMed  Google Scholar 

  2. Dewanjee MK, Ghafouripour AK, Kapadvanjwala M et al (1994) Noninvasive imaging of c-myc oncogene messenger RNA with Indium-1l-antisense probes in a mammary tumor-bearing mouse model. J Nucl Med 35:1054–1063

    CAS  PubMed  Google Scholar 

  3. Hnatowich D, Winnard P Jr, Virzi F et al (1995) Labeling deoxyribonucleic acid oligonucleotides with 99mTc. J Nucl Med 36:2306–2314

    CAS  PubMed  Google Scholar 

  4. Shi N, Boado RJ, Pardridge WM (2000) Antisense imaging of gene expression in the brain in vivo. Proc Nat Acad Sci 97:14709–14714

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Opalinska JB, Gewirtz AM (2002) Nucleic acid therapeutics: basic principles and recent applications. Nat Rev Drug Discov 1:503–514

    Article  CAS  PubMed  Google Scholar 

  6. Kole R, Krainer AR, Altman S (2012) RNA therapeutics: beyond RNA interference and antisense oligonucleotides. Nat Rev Drug Discov 11:125–140

    CAS  PubMed  Google Scholar 

  7. Visser ME, Witztum JL, Stroes ES, Kastelein JJ (2012) Antisense oligonucleotides for the treatment of dyslipidaemia. Eur Heart J ehs084

  8. Southwell AL, Skotte NH, Bennett CF, Hayden MR (2012) Antisense oligonucleotide therapeutics for inherited neurodegenerative diseases. Trends Mol Med 18:634–643

    Article  CAS  PubMed  Google Scholar 

  9. Gutschner T, Hammerle M, Eissmann M et al (2013) The long non-coding RNA MALAT1 is an essential gene regulator for lung cancer metastasis in a novel human knockout model. Cancer Res 73:1121

    Article  Google Scholar 

  10. Kamimura K, Suda T, Zhang G, Liu D (2011) Advances in gene delivery systems. Pharm Med 25:293–306

    Article  Google Scholar 

  11. Zobel H-P, Junghans M, Maienschein V et al (2000) Enhanced antisense efficacy of oligonucleotides adsorbed to monomethylaminoethylmethacrylate methylmethacrylate copolymer nanoparticles. Eur J Pharm Biopharm 49:203–210

    Article  CAS  PubMed  Google Scholar 

  12. Vinogradov S, Batrakova E, Kabanov A (1999) Poly (ethylene glycol)–polyethyleneimine NanoGel™ particles: novel drug delivery systems for antisense oligonucleotides. Colloids Surf B Biointerfaces 16:291–304

    Article  CAS  Google Scholar 

  13. Zelphati O, Szoka FC Jr (1996) Liposomes as a carrier for intracellular delivery of antisense oligonucleotides: a real or magic bullet? J Control Release 41:99–119

    Article  CAS  Google Scholar 

  14. Pakunlu RI, Wang Y, Saad M, Khandare JJ, Starovoytov V, Minko T (2006) In vitro and in vivo intracellular liposomal delivery of antisense oligonucleotides and anticancer drug. J Control Release 114:153–162

    Article  CAS  PubMed  Google Scholar 

  15. Fu P, Shen B, Zhao C, Tian G (2010) Molecular imaging of MDM2 messenger RNA with 99mTc-labeled antisense oligonucleotides in experimental human breast cancer xenografts. J Nucl Med 51:1805–1812

    Article  CAS  PubMed  Google Scholar 

  16. Liu M, Wang RF, Yan P, Zhang CL, Cui YG (2014) Molecular imaging and pharmacokinetics of 99mTc‐hTERT antisense oligonucleotide as a potential tumor imaging probe. J Label Compd Radiopharm 57:97–101

    Article  CAS  Google Scholar 

  17. Koo OM, Rubinstein I, Onyuksel H (2005) Role of nanotechnology in targeted drug delivery and imaging: a concise review. Nanomedicine: Nanotechnol Biol Med 1:193–212

    Article  CAS  Google Scholar 

  18. Lambert G, Fattal E, Couvreur P (2001) Nanoparticulate systems for the delivery of antisense oligonucleotides. Adv Drug Deliv Rev 47:99–112

    Article  CAS  PubMed  Google Scholar 

  19. Peddada LY, Garbuzenko OB, Devore DI, Minko T, Roth CM (2014) Delivery of antisense oligonucleotides using poly (alkylene oxide)–poly (propylacrylic acid) graft copolymers in conjunction with cationic liposomes. J Control Release 194:103–112

    Article  CAS  PubMed  Google Scholar 

  20. Brown PK, Qureshi AT, Hayes DJ, Monroe WT (2011) Targeted gene silencing with light and a silver nanoparticle antisense delivery system [abstract]. 573-574P

  21. Rudin CM, Marshall JL, Huang CH et al (2004) Delivery of a liposomal c-raf-1 antisense oligonucleotide by weekly bolus dosing in patients with advanced solid tumors: a phase I study. Clin Cancer Res 10:7244–7251

    Article  CAS  PubMed  Google Scholar 

  22. Koslowsky I, Shahhosseini S, Mirzayans R et al (2011) Evaluation of an 18F-labeled oligonucleotide probe targeting p21(WAF1) transcriptional changes in human tumor cells. Oncol Res Featuring Preclin Clin Cancer Ther 19:265–274

    CAS  Google Scholar 

  23. Zhang Y-M, Rusckowski M, Liu N et al (2001) Cationic liposomes enhance cellular/nuclear localization of 99mTc-antisense oligonucleotides in target tumor cells. Cancer Biother Radiopharm 16:411–419

    Article  CAS  PubMed  Google Scholar 

  24. Kraemer K, Fuessel S, Schmidt U et al (2003) Antisense-mediated hTERT inhibition specifically reduces the growth of human bladder cancer cells. Clin Cancer Res 9:3794–3800

    CAS  PubMed  Google Scholar 

  25. Beltinger C, Saragovi H, Smith R et al (1995) Binding, uptake, and intracellular trafficking of phosphorothioate-modified oligodeoxynucleotides. J Clin Invest 95:1814

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Zelphati O, Szoka FC (1996) Mechanism of oligonucleotide release from cationic liposomes. Proc Nat Acad Sci 93:11493–11498

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Crooke RM, Graham MJ, Cooke ME, Crooke ST (1995) In vitro pharmacokinetics of phosphorothioate antisense oligonucleotides. J Pharmacol Exp Ther 275:462–473

    CAS  PubMed  Google Scholar 

  28. Hnatowich DJ, Nakamura K (2004) Antisense targeting in cell culture with radiolabeled DNAs—a brief review of recent progress. Ann Nucl Med 18:363–368

    Article  CAS  PubMed  Google Scholar 

  29. Hnatowich DJ (1999) Antisense and nuclear medicine. J Nucl Med 40:693–703

    CAS  PubMed  Google Scholar 

  30. Liu M, Wang RF, Zhang CL et al (2007) Noninvasive imaging of human telomerase reverse transcriptase (hTERT) messenger RNA with 99mTc-radiolabeled antisense probes in malignant tumors. J Nucl Med 48:2028–2036

    Article  CAS  PubMed  Google Scholar 

  31. Zheng J-G, Tan T-Z (2004) Antisense imaging of colon cancer-bearing nude mice with liposome-entrapped 99m-technetium-labeled antisense oligonucleotides of c-myc mRNA. World J Gastrolenterol 10:2563–2566

    CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by the Natural Science Foundation of Hainan Province (814328). We wish to express our warm thanks to Fenghe (Shanghai) Information Technology Co., Ltd. Their ideas and help gave a valuable added dimension to our research.

Conflict of Interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Nuclear Medicine, The Chinese PLA General Hospital, 28 Fuxing Road, Beijing, 100853, People’s Republic of China

    Chang-bin Liu, Bai-xuan Xu, Jin-ming Zhang, Ying-mao Chen, Rui-min Wang & Jia-he Tian

  2. Department Of Radiology, Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, Shaanxi, People’s Republic of China

    Jun-qing Xu

Authors
  1. Chang-bin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun-qing Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bai-xuan Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jin-ming Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ying-mao Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rui-min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jia-he Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jia-he Tian.

Additional information

Chang-bin Liu and Jun-qing Xu are co-first authors.

Highlights

Carrier-mediated delivery systems enhanced cellular uptake of ASON in vitro.

Carrier-mediated delivery systems resulted in low target/non-target ratios.

Transfection reagents were useless for the cellular uptake of ASON in vivo.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, Cb., Xu, Jq., Xu, Bx. et al. Can Carrier-Mediated Delivery System Promote the Development of Antisense Imaging?. Mol Imaging Biol 17, 625–632 (2015). https://doi.org/10.1007/s11307-015-0827-7

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11307-015-0827-7

Key words

Search

Navigation