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Melanoma hücrelerinin Sendai viral vektörleri ile verimli transdüksüyonu

Yıl 2017, Cilt: 74 Sayı: 2, 113 - 120, 01.06.2017

Açelya Yılmazer Aktuna Hadiseh Taheri Alp Can

Öz

Amaç: Yaşayan hücrelere gen salımı yapmak üzere pek çok viral vektör geliştirilmiştir. Sendai viral SeV vektörleri geçici gen ifadesi, geniş konak özgüllüğü, düşük patojenite ve yüksek immünojenite gibi özellikleri sayesinde gen aktarımı için önemli vektörlerdir. SeV vektörleri gen tedavisi, aşı teknolojileri ve rejeneratif amaçlı moleküler tıpta sıklıkla kullanılır.Yöntem: Bu çalışmada, farklı melanoma hücre dizilerinde SeV vektörlerinin gen aktarım verimlilikleri floresan mikroskop ve konfokal lazer taramalı mikroskop görüntüleme teknikleri ile değerlendirilmiştir. A375, MDA- MB- 435, G361 ve WM115 hücreleri yeşil flüoresan proteini GFP ifade eden SeV vektörleri tarafından farklı virüs derişimlerinde enfeksiyon çarpanı MOI : 1, 3 ve 9 transdükte edilmiştir. GFP ifadesi virüs inkübasyonundan 24 ve 48 saat sonrasında kontrol edilmiştir. Konfokal lazer taramalı mikroskop görüntüleme ile gen salım verimliliği hesaplanmıştır. Bulgular: Floresan mikroskop görüntüleme ile düşük virüs derişimlerinde dahi enfeksiyon çarpanı: 1 , A375, MDA -MB- 435, G361 ve WM115 hücrelerinin SeV tarafından verimli şekilde transdükte edildiği gösterilmiştir. Viral transdüksiyonu takiben, GFP kontrol gen aktivitesi 24 saat içerisinde gözlemlenmeye başlanmış ve 48 saatte artış göstermiştir. Transdüksiyondan 24 saat sonrasında hücrelerde hafif toksisite gözlemlenmiş olsa da 48 saat sonrasında hücreler toksisite etkisinden kurtularak çoğalmış ve verimli şekilde gen ifadesi göstermişlerdir. Konfokal lazer taramalı mikroskop görüntüleme sonucuna göre 48 saat sonunda tüm hücre dizilerinde hücrelerin %80’inden fazlası başarılı bir şekilde GFP genini ifade etmiştir. Sonuç: Sonuç olarak, SeV vektörleri melanoma hücrelerini yüksek verimlilikle transdükte edip gen ifadesini sağlamıştır. Bu çalışma SeV vektörlerinin melanoma orijinli hücrelerdeki kullanımını açığa çıkarmış ve SeV vektörlerinin kullanımını içeren kanser tedavi ve hücre programlama alanındaki gelecek çalışmalarına destek sağlamıştır

Anahtar Kelimeler

Melanoma Sendai virüsleri GFP transdüksüyon gen aktarımı

Kaynakça

  • 1. Lamb RA, Kolakofsky D. Paramyxoviridae: The Viruses and Their Replication In: D. M. Knipe, P. M. Howley, D. E. Griffin, R. A. Lamb, M. A. Martin, B. Roizman, and S. E. Straus (eds.), Fields virology, 4th ed., Philadelphia, Lippincott Williams & Wilkins, 2001:1305-40
  • 2. Li H-O, Zhu Y-F, Asakawa M, Kuma H, Hirata T, Ueda Y, et al. A Cytoplasmic RNA Vector Derived from Nontransmissible Sendai Virus with Efficient Gene Transfer and Expression. J Virology, 2000;74(14):6564-9.
  • 3. Eguchi A, Kondoh T, Kosaka H, Suzuki T, Momota H, Masago A, et al. Identification and Characterization of Cell Lines with a Defect in a Post-adsorption Stage of Sendai Virus-mediated Membrane Fusion. J Biol Chem, 2000;275(23):17549-55.
  • 4. Bitzer M, Armeanu S, Lauer UM, Neubert WJ. Sendai virus vectors as an emerging negative-strand RNA viral vector system. J Gene Med, 2003;5(7):543-53.
  • 5. Yonemitsu Y, Kitson C, Ferrari S, Farley R, Griesenbach U, Judd D, et al. Efficient gene transfer to airway epithelium using recombinant Sendai virus. Nat Biotech. 2000;18(9):970-3.
  • 6. Masaki I, Yonemitsu Y, Komori K, Ueno H, Nakashima Y, Nakagawa K, et al. Recombinant Sendai virusmediated gene transfer to vasculature: a new class of efficient gene transfer vector to the vascular system. FASEB J, 2001;15(7)1294-6.
  • 7. Murakami Y, Ikeda Y, Yonemitsu Y, Tanaka S, Kondo H, Okano S, et al. Newly-developed Sendai virus vector for retinal gene transfer: reduction of innate immune response via deletion of all enveloperelated genes. J Gene Med, 2008;10(2):165-76.
  • 8. Fujita S, Eguchi A, Okabe J, Harada A, Sasaki K, Ogiwara N, et al. Sendai Virus-Mediated Gene Delivery into Hepatocytes via Isolated Hepatic Perfusion. Biol Pharm Bull, 2006;29(8):1728-34.
  • 9. Goto T, Morishita M, Nishimura K, Nakanishi M, Kato A, Ehara J, et al. Novel Mucosal Insulin Delivery Systems Based on Fusogenic Liposomes. Pharmaceut Res, 2006;23(2):384-91.
  • 10. Shibata S, Okano S, Yonemitsu Y, Onimaru M, Sata S, Nagata-Takeshita H, et al. Induction of Efficient Antitumor Immunity Using Dendritic Cells Activated by Recombinant Sendai Virus and Its Modulation by Exogenous IFN-β Gene. J Immunol, 2006;177(6):3564-76.
  • 11. Nishimura K, Sano M, Ohtaka M, Furuta B, Umemura Y, Nakajima Y, et al. Development of Defective and Persistent Sendai Virus Vector: A unique gene delivery/expression system ideal for cell reprogramming. J Biol Chem, 2011;286(6):4760-71.
  • 12. Mochiduki Y, Okita K. Methods for iPS cell generation for basic research and clinical applications. Biotechnol J, 2012;7(6):789-97.
  • 13. Saga K, Kaneda Y. Virosome Presents Multimodel Cancer Therapy without Viral Replication. Biomed Res Int, 2013;2013:764706.
  • 14. dUra T, Okuda K, Shimada M. Developments in Viral Vector-Based Vaccines. Vaccines, 2014;2(3):624.
  • 15. Mahito N, Makoto O. Development of Sendai Virus Vectors and their Potential Applications in Gene Therapy and Regenerative Medicine. Curr Gene Ther, 2012;12(5):410-6.
  • 16. Oishi K, Noguchi H, Yukawa H, Inoue M, Takagi S, Iwata H, et al. Recombinant Sendai Virus-Mediated Gene Transfer to Mouse Pancreatic Stem Cells. Cell Transplant, 2009;18(5):573-80
  • 17. Takahashi K, Yamanaka S. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell, 2006;126(4):663-76.
  • 18. de Lázaro I, Yilmazer A, Kostarelos K. Induced pluripotent stem (iPS) cells: A new source for cell-based therapeutics? J Control Release, 2014;185:37-44.
  • 19. Choi J, Lee S, Mallard W, Clement K, Tagliazucchi GM, Lim H, et al. A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs. Nat Biotech, 2015;33(11):1173-81.
  • 20. Zhang X-B. Cellular Reprogramming of Human Peripheral Blood Cells. Genomics Proteomics Bioinformatics, 2013;11(5):264-74
  • 21. Bueno C, Sardina JL, Di Stefano B, Romero-Moya D, Munoz-Lopez A, Ariza L, et al. Reprogramming human B cells into induced pluripotent stem cells and its enhancement by C/EBP[alpha]. Leukemia, 2015;30(3):674-82.
  • 22. Miere C, Devito L, Ilic D. Sendai Virus-Based Reprogramming of Mesenchymal Stromal/Stem Cells from Umbilical Cord Wharton’s Jelly into Induced Pluripotent Stem Cells. In: Turksen K, Nagy A, editors. Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology. 1357: Springer New York; 2016:33- 44
  • 23. Yilmazer A, de Lázaro I, Taheri H. Reprogramming cancer cells: A novel approach for cancer therapy or a tool for disease-modeling? Cancer Lett, 2015;369(1):1-8.

Efficient transduction of melanoma cells with Sendai viral vectors

Yıl 2017, Cilt: 74 Sayı: 2, 113 - 120, 01.06.2017

Açelya Yılmazer Aktuna Hadiseh Taheri Alp Can

Öz

Objective: Various viral vectors have been developed in order to delivery genes to living cells. Sendai virus SeV vectors are important viral vectors due to their properties suitable for gene delivery including transient gene expression, wide host cell specificity, low pathogenicity and strong immunogenicity. SeVs vectorss are highly used in molecular medicine in gene therapy, vaccine technology and regenerative.Methods: It was evaluated the gene delivery efficiency of SeV particles in various melanoma cell lines by using fluorescence microscope and confocal laser scanning microscope imaging techniques. A375, MDAMB-435, G361 and WM115 cells have been transduced with SeV vectors expressing green fluorescent protein GFP at different multiplicity of infections MOI : 1, 3, and 9. GFP expression was checked at 24 and 48 hours later following transduction. Confocal laser scanning microscopy imaging was calculated to gene delivery efficiency.Results: It was showed that A375, MDA-MB-435, G361 and WM115 cells are efficiently tranduced by seV even at low virus concentration with fluorescence microscopy imaging. GFP reporter gene activity started to be observed in 24 hours and peaked in 48 hours following viral transduction. Slight toxicity was observed hücrelerde hafif toksisite gözlemlenmiş olsa da 48 saat sonrasında hücreler toksisite etkisinden kurtularak çoğalmış ve verimli şekilde gen ifadesi göstermişlerdir. Konfokal lazer taramalı mikroskop görüntüleme sonucuna göre 48 saat sonunda tüm hücre dizilerinde hücrelerin %80’inden fazlası başarılı bir şekilde GFP genini ifade etmiştir. Sonuç: Sonuç olarak, SeV vektörleri melanoma hücrelerini yüksek verimlilikle transdükte edip gen ifadesini sağlamıştır. Bu çalışma SeV vektörlerinin melanoma orijinli hücrelerdeki kullanımını açığa çıkarmış ve SeV vektörlerinin kullanımını içeren kanser tedavi ve hücre programlama alanındaki gelecek çalışmalarına destek sağlamıştır

Anahtar Kelimeler

Melanoma Sendai virus particles GFP transduction gene delivery

Kaynakça

  • 1. Lamb RA, Kolakofsky D. Paramyxoviridae: The Viruses and Their Replication In: D. M. Knipe, P. M. Howley, D. E. Griffin, R. A. Lamb, M. A. Martin, B. Roizman, and S. E. Straus (eds.), Fields virology, 4th ed., Philadelphia, Lippincott Williams & Wilkins, 2001:1305-40
  • 2. Li H-O, Zhu Y-F, Asakawa M, Kuma H, Hirata T, Ueda Y, et al. A Cytoplasmic RNA Vector Derived from Nontransmissible Sendai Virus with Efficient Gene Transfer and Expression. J Virology, 2000;74(14):6564-9.
  • 3. Eguchi A, Kondoh T, Kosaka H, Suzuki T, Momota H, Masago A, et al. Identification and Characterization of Cell Lines with a Defect in a Post-adsorption Stage of Sendai Virus-mediated Membrane Fusion. J Biol Chem, 2000;275(23):17549-55.
  • 4. Bitzer M, Armeanu S, Lauer UM, Neubert WJ. Sendai virus vectors as an emerging negative-strand RNA viral vector system. J Gene Med, 2003;5(7):543-53.
  • 5. Yonemitsu Y, Kitson C, Ferrari S, Farley R, Griesenbach U, Judd D, et al. Efficient gene transfer to airway epithelium using recombinant Sendai virus. Nat Biotech. 2000;18(9):970-3.
  • 6. Masaki I, Yonemitsu Y, Komori K, Ueno H, Nakashima Y, Nakagawa K, et al. Recombinant Sendai virusmediated gene transfer to vasculature: a new class of efficient gene transfer vector to the vascular system. FASEB J, 2001;15(7)1294-6.
  • 7. Murakami Y, Ikeda Y, Yonemitsu Y, Tanaka S, Kondo H, Okano S, et al. Newly-developed Sendai virus vector for retinal gene transfer: reduction of innate immune response via deletion of all enveloperelated genes. J Gene Med, 2008;10(2):165-76.
  • 8. Fujita S, Eguchi A, Okabe J, Harada A, Sasaki K, Ogiwara N, et al. Sendai Virus-Mediated Gene Delivery into Hepatocytes via Isolated Hepatic Perfusion. Biol Pharm Bull, 2006;29(8):1728-34.
  • 9. Goto T, Morishita M, Nishimura K, Nakanishi M, Kato A, Ehara J, et al. Novel Mucosal Insulin Delivery Systems Based on Fusogenic Liposomes. Pharmaceut Res, 2006;23(2):384-91.
  • 10. Shibata S, Okano S, Yonemitsu Y, Onimaru M, Sata S, Nagata-Takeshita H, et al. Induction of Efficient Antitumor Immunity Using Dendritic Cells Activated by Recombinant Sendai Virus and Its Modulation by Exogenous IFN-β Gene. J Immunol, 2006;177(6):3564-76.
  • 11. Nishimura K, Sano M, Ohtaka M, Furuta B, Umemura Y, Nakajima Y, et al. Development of Defective and Persistent Sendai Virus Vector: A unique gene delivery/expression system ideal for cell reprogramming. J Biol Chem, 2011;286(6):4760-71.
  • 12. Mochiduki Y, Okita K. Methods for iPS cell generation for basic research and clinical applications. Biotechnol J, 2012;7(6):789-97.
  • 13. Saga K, Kaneda Y. Virosome Presents Multimodel Cancer Therapy without Viral Replication. Biomed Res Int, 2013;2013:764706.
  • 14. dUra T, Okuda K, Shimada M. Developments in Viral Vector-Based Vaccines. Vaccines, 2014;2(3):624.
  • 15. Mahito N, Makoto O. Development of Sendai Virus Vectors and their Potential Applications in Gene Therapy and Regenerative Medicine. Curr Gene Ther, 2012;12(5):410-6.
  • 16. Oishi K, Noguchi H, Yukawa H, Inoue M, Takagi S, Iwata H, et al. Recombinant Sendai Virus-Mediated Gene Transfer to Mouse Pancreatic Stem Cells. Cell Transplant, 2009;18(5):573-80
  • 17. Takahashi K, Yamanaka S. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell, 2006;126(4):663-76.
  • 18. de Lázaro I, Yilmazer A, Kostarelos K. Induced pluripotent stem (iPS) cells: A new source for cell-based therapeutics? J Control Release, 2014;185:37-44.
  • 19. Choi J, Lee S, Mallard W, Clement K, Tagliazucchi GM, Lim H, et al. A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs. Nat Biotech, 2015;33(11):1173-81.
  • 20. Zhang X-B. Cellular Reprogramming of Human Peripheral Blood Cells. Genomics Proteomics Bioinformatics, 2013;11(5):264-74
  • 21. Bueno C, Sardina JL, Di Stefano B, Romero-Moya D, Munoz-Lopez A, Ariza L, et al. Reprogramming human B cells into induced pluripotent stem cells and its enhancement by C/EBP[alpha]. Leukemia, 2015;30(3):674-82.
  • 22. Miere C, Devito L, Ilic D. Sendai Virus-Based Reprogramming of Mesenchymal Stromal/Stem Cells from Umbilical Cord Wharton’s Jelly into Induced Pluripotent Stem Cells. In: Turksen K, Nagy A, editors. Induced Pluripotent Stem (iPS) Cells. Methods in Molecular Biology. 1357: Springer New York; 2016:33- 44
  • 23. Yilmazer A, de Lázaro I, Taheri H. Reprogramming cancer cells: A novel approach for cancer therapy or a tool for disease-modeling? Cancer Lett, 2015;369(1):1-8.
Toplam 23 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Bölüm Araştırma Makalesi
Yazarlar

Açelya Yılmazer Aktuna Bu kişi benim

Hadiseh Taheri Bu kişi benim

Alp Can Bu kişi benim

Yayımlanma Tarihi 1 Haziran 2017
Yayımlandığı Sayı Yıl 2017 Cilt: 74 Sayı: 2

Kaynak Göster

APA Aktuna, A. Y., Taheri, H., & Can, A. (2017). Efficient transduction of melanoma cells with Sendai viral vectors. Türk Hijyen Ve Deneysel Biyoloji Dergisi, 74(2), 113-120.
AMA Aktuna AY, Taheri H, Can A. Efficient transduction of melanoma cells with Sendai viral vectors. Turk Hij Den Biyol Derg. Haziran 2017;74(2):113-120.
Chicago Aktuna, Açelya Yılmazer, Hadiseh Taheri, ve Alp Can. “Efficient Transduction of Melanoma Cells With Sendai Viral Vectors”. Türk Hijyen Ve Deneysel Biyoloji Dergisi 74, sy. 2 (Haziran 2017): 113-20.
EndNote Aktuna AY, Taheri H, Can A (01 Haziran 2017) Efficient transduction of melanoma cells with Sendai viral vectors. Türk Hijyen ve Deneysel Biyoloji Dergisi 74 2 113–120.
IEEE A. Y. Aktuna, H. Taheri, ve A. Can, “Efficient transduction of melanoma cells with Sendai viral vectors”, Turk Hij Den Biyol Derg, c. 74, sy. 2, ss. 113–120, 2017.
ISNAD Aktuna, Açelya Yılmazer vd. “Efficient Transduction of Melanoma Cells With Sendai Viral Vectors”. Türk Hijyen ve Deneysel Biyoloji Dergisi 74/2 (Haziran 2017), 113-120.
JAMA Aktuna AY, Taheri H, Can A. Efficient transduction of melanoma cells with Sendai viral vectors. Turk Hij Den Biyol Derg. 2017;74:113–120.
MLA Aktuna, Açelya Yılmazer vd. “Efficient Transduction of Melanoma Cells With Sendai Viral Vectors”. Türk Hijyen Ve Deneysel Biyoloji Dergisi, c. 74, sy. 2, 2017, ss. 113-20.
Vancouver Aktuna AY, Taheri H, Can A. Efficient transduction of melanoma cells with Sendai viral vectors. Turk Hij Den Biyol Derg. 2017;74(2):113-20.