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Potent bystander effect and tumor tropism in suicide gene therapy using stem cells from human exfoliated deciduous teeth

Cancer Gene Therapy volume 30, pages 85–95 (2023)Cite this article

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Abstract

Herpes simplex virus thymidine kinase (HSVTK)/ganciclovir (GCV) suicide gene therapy has a long history of treating malignant gliomas. Recently, stem cells from human exfoliated deciduous teeth (SHED), which are collected from deciduous teeth and have excellent harvestability, ethical aspects, and self-renewal, have been attracting attention mainly in the field of gene therapy. In the present study, we assessed SHED as a novel cellular vehicle for suicide gene therapy in malignant gliomas, as we have previously demonstrated with various cell types. SHED was transduced with the HSVTK gene (SHEDTK). In vitro experiments showed a significant bystander effect between SHEDTK and glioma cell lines in coculture. Furthermore, apoptotic changes caused by caspase 3/7 activation were simultaneously observed in SHEDTK and glioma cells. Mice implanted with a mixture of U87 and SHEDTK and treated with intraperitoneal GCV survived for longer than 100 days. Additionally, tumors in treatment model mice were significantly reduced in size during the treatment period. SHEDTK implanted at the contralateral hemisphere migrated toward the tumor crossing the corpus callosum. These results suggested that SHEDTK-based suicide gene therapy has potent tumor tropism and a bystander-killing effect, potentially offering a new promising therapeutic modality for malignant gliomas.

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Fig. 1: In vitro bystander effect between SHEDTK and glioma cells.
Fig. 2: In vitro time-lapse imaging and apoptosis detection in the bystander effect evaluation.
Fig. 3: In vitro migration of SHEDTK toward glioma CM and representative tumor growth factors.
Fig. 4: In vivo migration ability toward a U87 glioma mass.
Fig. 5: In vivo bystander effect in the nude mouse intracranial tumor model implanted with a mixture of SHEDTK and U87-Luc glioma cells (coculture model mouse).
Fig. 6: In vivo bystander effect for U87 glioma in treatment model mouse.

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Data availability

The datasets used and analyzed during the current study are available from the last corresponding author on reasonable request.

References

  1. Anjum K, Shagufta BI, Abbas SQ, Patel S, Khan I, Shah SAA, et al. Current status and future therapeutic perspectives of glioblastoma multiforme (GBM) therapy: a review. Biomed Pharmacother. 2017;92:681–9.

    Article  CAS  Google Scholar 

  2. Hegi ME, Diserens AC, Gorlia T, Hamou MF, de Tribolet N, Weller M, et al. MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med. 2005;352:997–1003.

    Article  CAS  Google Scholar 

  3. Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, et al. Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol. 2009;10:459–66.

    Article  CAS  Google Scholar 

  4. Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987–96.

    Article  CAS  Google Scholar 

  5. Stupp R, Taillibert S, Kanner AA, Kesari S, Steinberg DM, Toms SA, et al. Maintenance therapy with tumor-treating fields plus temozolomide vs temozolomide alone for glioblastoma: a Randomized Clinical Trial. JAMA. 2015;314:2535–43.

    Article  CAS  Google Scholar 

  6. Gilbert MR, Dignam JJ, Armstrong TS, Wefel JS, Blumenthal DT, Vogelbaum MA, et al. A randomized trial of bevacizumab for newly diagnosed glioblastoma. N. Engl J Med. 2014;370:699–708.

    Article  CAS  Google Scholar 

  7. Chinot OL, de La Motte Rouge T, Moore N, Zeaiter A, Das A, Phillips H, et al. AVAglio: Phase 3 trial of bevacizumab plus temozolomide and radiotherapy in newly diagnosed glioblastoma multiforme. Adv Ther. 2011;28:334–40.

    Article  CAS  Google Scholar 

  8. Chinot OL, Wick W, Cloughesy T. Bevacizumab for newly diagnosed glioblastoma. N Engl J Med. 2014;370:2049.

    Article  Google Scholar 

  9. Chinot OL, Wick W, Mason W, Henriksson R, Saran F, Nishikawa R, et al. Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma. N Engl J Med. 2014;370:709–22.

    Article  CAS  Google Scholar 

  10. Stupp R, Taillibert S, Kanner A, Read W, Steinberg D, Lhermitte B, et al. Effect of tumor-treating fields plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma: a Randomized Clinical Trial. JAMA. 2017;318:2306–16.

    Article  CAS  Google Scholar 

  11. Westphal M, Hilt DC, Bortey E, Delavault P, Olivares R, Warnke PC, et al. A phase 3 trial of local chemotherapy with biodegradable carmustine (BCNU) wafers (Gliadel wafers) in patients with primary malignant glioma. Neuro Oncol. 2003;5:79–88.

    Article  CAS  Google Scholar 

  12. Rainov NG. A phase III clinical evaluation of herpes simplex virus type 1 thymidine kinase and ganciclovir gene therapy as an adjuvant to surgical resection and radiation in adults with previously untreated glioblastoma multiforme. Hum Gene Ther. 2000;11:2389–401.

    Article  CAS  Google Scholar 

  13. Culver KW, Ram Z, Wallbridge S, Ishii H, Oldfield EH, Blaese RM. In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors. Science. 1992;256:1550–2.

    Article  CAS  Google Scholar 

  14. Li S, Tokuyama T, Yamamoto J, Koide M, Yokota N, Namba H. Potent bystander effect in suicide gene therapy using neural stem cells transduced with herpes simplex virus thymidine kinase gene. Oncology. 2005;69:503–8.

    Article  Google Scholar 

  15. Ichikawa T, Otani Y, Kurozumi K, Date I. Phenotypic transition as a survival strategy of glioma. Neurol Med Chir. 2016;56:387–95.

    Article  Google Scholar 

  16. Aboody KS, Brown A, Rainov NG, Bower KA, Liu S, Yang W, et al. Neural stem cells display extensive tropism for pathology in adult brain: evidence from intracranial gliomas. Proc Natl Acad Sci USA. 2000;97:12846–51.

    Article  CAS  Google Scholar 

  17. Ehtesham M, Kabos P, Gutierrez MA, Chung NH, Griffith TS, Black KL, et al. Induction of glioblastoma apoptosis using neural stem cell-mediated delivery of tumor necrosis factor-related apoptosis-inducing ligand. Cancer Res. 2002;62:7170–4.

    CAS  Google Scholar 

  18. Amano S, Gu C, Koizumi S, Tokuyama T, Namba H. Tumoricidal bystander effect in the suicide gene therapy using mesenchymal stem cells does not injure normal brain tissues. Cancer Lett. 2011;306:99–105.

    Article  CAS  Google Scholar 

  19. Li S, Gao Y, Tokuyama T, Yamamoto J, Yokota N, Yamamoto S, et al. Genetically engineered neural stem cells migrate and suppress glioma cell growth at distant intracranial sites. Cancer Lett. 2007;251:220–7.

    Article  CAS  Google Scholar 

  20. Yamasaki T, Wakao S, Kawaji H, Koizumi S, Sameshima T, Dezawa M, et al. Genetically engineered multilineage-differentiating stress-enduring cells as cellular vehicles against malignant gliomas. Mol Ther Oncol. 2017;6:45–56.

    Article  CAS  Google Scholar 

  21. Yamazoe T, Koizumi S, Yamasaki T, Amano S, Tokuyama T, Namba H. Potent tumor tropism of induced pluripotent stem cells and induced pluripotent stem cell-derived neural stem cells in the mouse intracerebral glioma model. Int J Oncol. 2015;46:147–52.

    Article  CAS  Google Scholar 

  22. Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, et al. Stem cell properties of human dental pulp stem cells. J Dent Res. 2002;81:531–5.

    Article  CAS  Google Scholar 

  23. Nakamura S, Yamada Y, Katagiri W, Sugito T, Ito K, Ueda M. Stem cell proliferation pathways comparison between human exfoliated deciduous teeth and dental pulp stem cells by gene expression profile from promising dental pulp. J Endod. 2009;35:1536–42.

    Article  Google Scholar 

  24. Miura M, Gronthos S, Zhao M, Lu B, Fisher LW, Robey PG, et al. SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA. 2003;100:5807–12.

    Article  CAS  Google Scholar 

  25. Amano S, Li S, Gu C, Gao Y, Koizumi S, Yamamoto S, et al. Use of genetically engineered bone marrow-derived mesenchymal stem cells for glioma gene therapy. Int J Oncol. 2009;35:1265–70.

    CAS  Google Scholar 

  26. Kenmochi H, Yamasaki T, Koizumi S, Sameshima T, Namba H. Nicotine does not affect stem cell properties requisite for suicide gene therapy against glioma. Neurol Res. 2020;42:818–27.

    Article  CAS  Google Scholar 

  27. Serfozo P, Schlarman MS, Pierret C, Maria BL, Kirk MD. Selective migration of neuralized embryonic stem cells to stem cell factor and media conditioned by glioma cell lines. Cancer Cell Int. 2006;6:1.

    Article  Google Scholar 

  28. Hata N, Shinojima N, Gumin J, Yong R, Marini F, Andreeff M, et al. Platelet-derived growth factor BB mediates the tropism of human mesenchymal stem cells for malignant gliomas. Neurosurgery. 2010;66:144–56.

    Article  Google Scholar 

  29. Ehtesham M, Yuan X, Kabos P, Chung NH, Liu G, Akasaki Y, et al. Glioma tropic neural stem cells consist of astrocytic precursors and their migratory capacity is mediated by CXCR4. Neoplasia. 2004;6:287–93.

    Article  CAS  Google Scholar 

  30. Schichor C, Birnbaum T, Etminan N, Schnell O, Grau S, Miebach S, et al. Vascular endothelial growth factor A contributes to glioma-induced migration of human marrow stromal cells (hMSC). Exp Neurol. 2006;199:301–10.

    Article  CAS  Google Scholar 

  31. Gao Y, Gu C, Li S, Tokuyama T, Yokota N, Nakayama KI, et al. p27 modulates tropism of mesenchymal stem cells toward brain tumors. Exp Ther Med. 2010;1:695–9.

    Article  Google Scholar 

  32. Li HK, Morokoshi Y, Nagatsu K, Kamada T, Hasegawa S. Locoregional therapy with alpha-emitting trastuzumab against peritoneal metastasis of human epidermal growth factor receptor 2-positive gastric cancer in mice. Cancer Sci. 2017;108:1648–56.

    Article  CAS  Google Scholar 

  33. Li Z, Yu Y, Wang M, Xu H, Han B, Jiang P, et al. Anti-breast cancer activity of SPG-56 from Sweet potato in MCF-7 bearing mice in situ through promoting apoptosis and inhibiting metastasis. Sci Rep. 2019;9:146.

    Article  Google Scholar 

  34. Chiang Y-H, Lin C-C, Chen Y-C, Lee OKJIJOMS. Treatment of arsenite intoxication-induced peripheral vasculopathy with mesenchymal stem cells. Int J Mol Sci. 2018;19:1026.

    Article  Google Scholar 

  35. Jang EH, Kim J-H, Lee JH, Kim D-H, Youn Y-NJP. Enhanced biocompatibility of multi-layered, 3D bio-printed artificial vessels composed of autologous mesenchymal stem cells. Polymers (Basel). 2020;12:538.

  36. Thery C, Zitvogel L, Amigorena S. Exosomes: composition, biogenesis and function. Nat Rev Immunol. 2002;2:569–79.

    Article  CAS  Google Scholar 

  37. Tang Y, Zhou Y, Li HJ. Advances in mesenchymal stem cell exosomes: a review. Stem Cell Res Ther. 2021;12:71.

    Article  Google Scholar 

  38. Wei W, Ao Q, Wang X, Cao Y, Liu Y, Zheng SG, et al. Mesenchymal stem cell-derived exosomes: a promising biological tool in nanomedicine. Front Pharm. 2020;11:590470.

    Article  CAS  Google Scholar 

  39. Graepler F, Lemken ML, Wybranietz WA, Schmidt U, Smirnow I, Gross CD, et al. Bifunctional chimeric SuperCD suicide gene-YCD: YUPRT fusion is highly effective in a rat hepatoma model. World J Gastroenterol. 2005;11:6910–9.

    Article  CAS  Google Scholar 

  40. Altanerova U, Benejova K, Altanerova V, Tyciakova S, Rychly B, Szomolanyi P, et al. Dental pulp mesenchymal stem/stromal cells labeled with iron sucrose release exosomes and cells applied intra-nasally migrate to intracerebral glioblastoma. Neoplasma. 2016;63:925–33.

    Article  CAS  Google Scholar 

  41. Nakamura K, Ito Y, Kawano Y, Kurozumi K, Kobune M, Tsuda H, et al. Antitumor effect of genetically engineered mesenchymal stem cells in a rat glioma model. Gene Ther. 2004;11:1155–64.

    Article  CAS  Google Scholar 

  42. Rath P, Shi H, Maruniak JA, Litofsky NS, Maria BL, Kirk MD. Stem cells as vectors to deliver HSV/tk gene therapy for malignant gliomas. Curr Stem Cell Res Ther. 2009;4:44–49.

    Article  CAS  Google Scholar 

  43. de Melo SM, Bittencourt S, Ferrazoli EG, da Silva CS, da Cunha FF, da Silva FH, et al. The anti-tumor effects of adipose tissue mesenchymal stem cell transduced with HSV-Tk gene on U-87-driven brain tumor. PLoS ONE. 2015;10:e0128922.

    Article  Google Scholar 

  44. Shree N, Bhonde RR. Conditioned media from adipose tissue derived mesenchymal stem cells reverse insulin resistance in cellular models. J Cell Biochem. 2017;118:2037–43.

    Article  CAS  Google Scholar 

  45. Alraies A, Alaidaroos NY, Waddington RJ, Moseley R, Sloan AJ. Variation in human dental pulp stem cell ageing profiles reflect contrasting proliferative and regenerative capabilities. BMC Cell Biol. 2017;18:12.

    Article  Google Scholar 

  46. Kellner M, Steindorff MM, Strempel JF, Winkel A, Kuhnel MP, Stiesch M. Differences of isolated dental stem cells dependent on donor age and consequences for autologous tooth replacement. Arch Oral Biol. 2014;59:559–67.

    Article  Google Scholar 

  47. Kobayashi T, Torii D, Iwata T, Izumi Y, Nasu M, Tsutsui TW. Characterization of proliferation, differentiation potential, and gene expression among clonal cultures of human dental pulp cells. Hum Cell. 2020;33:490–501.

    Article  CAS  Google Scholar 

  48. Longoni A, Utomo L, van Hooijdonk IE, Bittermann GK, Vetter VC, Kruijt Spanjer EC, et al. The chondrogenic differentiation potential of dental pulp stem cells. Eur Cell Mater. 2020;39:121–35.

    Article  CAS  Google Scholar 

  49. Wu W, Zhou J, Xu CT, Zhang J, Jin YJ, Sun GL. Derivation and growth characteristics of dental pulp stem cells from patients of different ages. Mol Med Rep. 2015;12:5127–34.

    Article  CAS  Google Scholar 

  50. Yi Q, Liu O, Yan F, Lin X, Diao S, Wang L, et al. Analysis of senescence-related differentiation potentials and gene expression profiles in human dental pulp stem cells. Cells Tissues Organs. 2017;203:1–11.

    Article  CAS  Google Scholar 

  51. Iohara K, Murakami M, Nakata K, Nakashima M. Age-dependent decline in dental pulp regeneration after pulpectomy in dogs. Exp Gerontol. 2014;52:39–45.

    Article  Google Scholar 

  52. Suchanek J, Visek B, Soukup T, El-Din Mohamed SK, Ivancakova R, Mokry J, et al. Stem cells from human exfoliated deciduous teeth—isolation, long term cultivation and phenotypical analysis. Acta Med (Hradec Kralove). 2010;53:93–9.

    Article  Google Scholar 

  53. Shekar R, Ranganathan K. Phenotypic and growth characterization of human mesenchymal stem cells cultured from permanent and deciduous teeth. Indian J Dent Res. 2012;23:838–9.

    Article  Google Scholar 

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Acknowledgements

We thank all members of the laboratory for their discussions and suggestions. Part of this work was performed at the Advanced Research Facilities & Services (ARFS), Hamamatsu University School of Medicine. We also thank Mitchell Arico from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript.

Funding

This work was supported by Hamamatsu University School of Medicine (HUSM) Grant-in-Aid. This study was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers JP20K09325 (KK), JP19K09523 (TS).

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan

    Makoto Horikawa, Shinichiro Koizumi, Tomoya Oishi, Taisuke Yamamoto, Tomohiro Yamasaki, Tetsuro Sameshima & Kazuhiko Kurozumi

  2. Department of Biology, Aichi Medical University, Nagakute, Aichi, Japan

    Masashi Ikeno

  3. Department of Virology and Parasitology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan

    Masahiko Ito & Tetsuro Suzuki

  4. Department of Neurosurgery, Iwata City Hospital, Iwata, Shizuoka, Japan

    Shinji Amano

  5. Kidswell Bio Corporation, Tokyo, Japan

    Yasuyuki Mitani

  6. Department of Neurological Surgery, Okayama University Faculty of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    Yoshihiro Otani

  7. Department of Surgery, Northwestern University, Chicago, IL, USA

    Yuanqing Yan

  8. Department of Neurosurgery, Enshu Hospital, Hamamatsu, Shizuoka, Japan

    Hiroki Namba

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  1. Makoto Horikawa
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  2. Shinichiro Koizumi
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Contributions

Conception and design: MH, HN, and KK. Development of methodology: MH, MI, MI, SA, SK, TS, HN, and KK. Acquisition of data (provided animals, acquired and managed patients, provided facilities, etc.): MH. Analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): MH, TO, SK, HN, and KK. Writing, review, and/or revision of the manuscript: MH, YM, YO, YY, HN, and KK. Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): MH, MI, MI, TS, HN, and KK. Study supervision: KK.

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Correspondence to Kazuhiko Kurozumi.

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Kidswell Bio Corporation (Tokyo, Japan) generously provided SHED as experimental material. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

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Horikawa, M., Koizumi, S., Oishi, T. et al. Potent bystander effect and tumor tropism in suicide gene therapy using stem cells from human exfoliated deciduous teeth. Cancer Gene Ther 30, 85–95 (2023). https://doi.org/10.1038/s41417-022-00527-5

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