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Establishment of a Parvovirus B19 NS1-Expressing Recombinant Adenoviral Vector for Killing Megakaryocytic Leukemia Cells.
Viruses 2019; 11(9)V

Abstract

Adenoviral viral vectors have been widely used for gene-based therapeutics, but commonly used serotype 5 shows poor transduction efficiency into hematopoietic cells. In this study, we aimed to generate a recombinant adenovirus serotype 5 (rAd5) vector that has a high efficiency in gene transfer to megakaryocytic leukemic cells with anticancer potential. We first modified the rAd5 backbone vector with a chimeric fiber gene of Ad5 and Ad11p (rAd5F11p) to increase the gene delivery efficiency. Then, the nonstructural protein NS1 of human parvovirus B19 (B19V), which induces cell cycle arrest at the G2/M phase and apoptosis, was cloned into the adenoviral shuttle vector. As the expression of parvoviral NS1 protein inhibited Ad replication and production, we engineered the cytomegalovirus (CMV) promoter, which governs NS1 expression, with two tetracycline operator elements (TetO2). Transfection of the rAd5F11p proviral vectors in Tet repressor-expressing T-REx-293 cells produced rAd in a large quantity. We further evaluated this chimeric rAd5F11p vector in gene delivery in human leukemic cells, UT7/Epo-S1. Strikingly, the novel rAd5F11p-B19NS1-GFP vector, exhibited a transduction efficiency much higher than the original vector, rAd5-B19NS1-GFP, in UT7/Epo-S1 cells, in particular, when they were transduced at a relatively low multiplicity of infection (100 viral genome copies/cell). After the transduction of rAd5F11p-B19NS1-GFP, over 90% of the UT7/Epo-S1 cells were arrested at the G2/M phase, and approximately 40%-50% of the cells were undergoing apoptosis, suggesting the novel rAd5F11P-B19NS1-GFP vector holds a promise in therapeutic potentials of megakaryocytic leukemia.

Authors+Show Affiliations

Hubei Engineering Research Center of Viral Vector, Wuhan University of Bioengineering, Wuhan 430415, China. xupenghuashi@163.com. Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA. xupenghuashi@163.com.Hubei Engineering Research Center of Viral Vector, Wuhan University of Bioengineering, Wuhan 430415, China. wangxiaomei_wxm@126.com. Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA. wangxiaomei_wxm@126.com.Hubei Engineering Research Center of Viral Vector, Wuhan University of Bioengineering, Wuhan 430415, China. johnli2668@hotmail.com.Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA. jqiu@kumc.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31487941

Citation

Xu, Peng, et al. "Establishment of a Parvovirus B19 NS1-Expressing Recombinant Adenoviral Vector for Killing Megakaryocytic Leukemia Cells." Viruses, vol. 11, no. 9, 2019.
Xu P, Wang X, Li Y, et al. Establishment of a Parvovirus B19 NS1-Expressing Recombinant Adenoviral Vector for Killing Megakaryocytic Leukemia Cells. Viruses. 2019;11(9).
Xu, P., Wang, X., Li, Y., & Qiu, J. (2019). Establishment of a Parvovirus B19 NS1-Expressing Recombinant Adenoviral Vector for Killing Megakaryocytic Leukemia Cells. Viruses, 11(9), doi:10.3390/v11090820.
Xu P, et al. Establishment of a Parvovirus B19 NS1-Expressing Recombinant Adenoviral Vector for Killing Megakaryocytic Leukemia Cells. Viruses. 2019 Sep 4;11(9) PubMed PMID: 31487941.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Establishment of a Parvovirus B19 NS1-Expressing Recombinant Adenoviral Vector for Killing Megakaryocytic Leukemia Cells. AU - Xu,Peng, AU - Wang,Xiaomei, AU - Li,Yi, AU - Qiu,Jianming, Y1 - 2019/09/04/ PY - 2019/07/30/received PY - 2019/08/25/revised PY - 2019/09/03/accepted PY - 2019/9/7/entrez PY - 2019/9/7/pubmed PY - 2019/9/7/medline KW - adenoviral vector KW - anti-cancer KW - apoptosis KW - cell cycle arrest KW - parvovirus B19 JF - Viruses JO - Viruses VL - 11 IS - 9 N2 - Adenoviral viral vectors have been widely used for gene-based therapeutics, but commonly used serotype 5 shows poor transduction efficiency into hematopoietic cells. In this study, we aimed to generate a recombinant adenovirus serotype 5 (rAd5) vector that has a high efficiency in gene transfer to megakaryocytic leukemic cells with anticancer potential. We first modified the rAd5 backbone vector with a chimeric fiber gene of Ad5 and Ad11p (rAd5F11p) to increase the gene delivery efficiency. Then, the nonstructural protein NS1 of human parvovirus B19 (B19V), which induces cell cycle arrest at the G2/M phase and apoptosis, was cloned into the adenoviral shuttle vector. As the expression of parvoviral NS1 protein inhibited Ad replication and production, we engineered the cytomegalovirus (CMV) promoter, which governs NS1 expression, with two tetracycline operator elements (TetO2). Transfection of the rAd5F11p proviral vectors in Tet repressor-expressing T-REx-293 cells produced rAd in a large quantity. We further evaluated this chimeric rAd5F11p vector in gene delivery in human leukemic cells, UT7/Epo-S1. Strikingly, the novel rAd5F11p-B19NS1-GFP vector, exhibited a transduction efficiency much higher than the original vector, rAd5-B19NS1-GFP, in UT7/Epo-S1 cells, in particular, when they were transduced at a relatively low multiplicity of infection (100 viral genome copies/cell). After the transduction of rAd5F11p-B19NS1-GFP, over 90% of the UT7/Epo-S1 cells were arrested at the G2/M phase, and approximately 40%-50% of the cells were undergoing apoptosis, suggesting the novel rAd5F11P-B19NS1-GFP vector holds a promise in therapeutic potentials of megakaryocytic leukemia. SN - 1999-4915 UR - https://www.unboundmedicine.com/medline/citation/31487941/Establishment_of_a_Parvovirus_B19_NS1-Expressing_Recombinant_Adenoviral_Vector_for_Killing_Megakaryocytic_Leukemia_Cells L2 - http://www.mdpi.com/resolver?pii=v11090820 DB - PRIME DP - Unbound Medicine ER -
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