Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders
Abstract Background B-cell proliferative disorders, such as post-transplant lymphoproliferative disease (PTLD), are increased among persons afflicted by T-cell compromise. Most are Epstein–Barr virus (EBV) + and can first present with a focal lesion. Direct introduction of oncolytic viruses into loc...
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| Format: | Article |
| Language: | English |
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BMC
2021-11-01
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| Series: | Virology Journal |
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| Online Access: | https://doi.org/10.1186/s12985-021-01695-w |
| _version_ | 1818842373576196096 |
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| author | Elham Ahmadi Mehrdad Ravanshad Jun Xie Rajesh Panigrahi Sandeep S. Jubbal Santosh Kumar Guru Gao Guangping Mazyar Ziyaeyan Joyce Fingeroth |
| author_facet | Elham Ahmadi Mehrdad Ravanshad Jun Xie Rajesh Panigrahi Sandeep S. Jubbal Santosh Kumar Guru Gao Guangping Mazyar Ziyaeyan Joyce Fingeroth |
| author_sort | Elham Ahmadi |
| collection | DOAJ |
| description | Abstract Background B-cell proliferative disorders, such as post-transplant lymphoproliferative disease (PTLD), are increased among persons afflicted by T-cell compromise. Most are Epstein–Barr virus (EBV) + and can first present with a focal lesion. Direct introduction of oncolytic viruses into localized tumors provides theoretical advantages over chemotherapy, immunotherapy and radiation therapy by reducing systemic toxicity. Despite extensive study as a vehicle for gene therapy, adeno-associated viruses (AAV) have rarely been applied to human cancer research due to technical and theoretical obstacles. Moreover, human B-cells have historically been described as resistant to AAV infection. Nonetheless, advances using different recombinant (r)AAV serotypes with unique tropisms to deliver cytotoxic therapy suggested a localized anti-tumor approach was feasible. Methods As a prelude to the development of a therapeutic vehicle, the ability of fifteen distinct EGFP-bearing rAAV serotypes to transduce human B-cells, including primary, immortalized, and B-cell tumor lines ± EBV was assessed by confocal microscopy, flow cytometry and subsequently cell viability assay. Results Rank order analysis revealed augmented transduction by rAAV6.2 and closely related virions. EBV infection of EBV-negative B-cell tumor lines and EBV immortalization of primary B-cells increased susceptibility to rAAV6.2 transduction. As a proof of concept, transduction by rAAV6.2 encoding herpes simplex virus type 1 (HSV1)-thymidine kinase (TK) eliminated TK-negative rhabdomyosarcoma cells and diminished viability of transduced B-cell lines upon incubation with ganciclovir. Conclusions rAAV serotypes differentially transduce human B-cell lines reversing the dogma that human B-cells are refractory to AAV infection. EBV + B-cells display increased susceptibility to rAAV6.2 infection, uncovering a new method for improved nucleic acid transfer into transfection-resistant B-cell lines. The introduction of a functional suicide gene into the rAAV6.2 genome identifies a candidate vector for the development of rAAV-based oncolytic therapy targeting focal EBV-bearing B-lymphoproliferative disorders. |
| first_indexed | 2024-12-19T04:40:57Z |
| format | Article |
| id | doaj.art-fee1d2b15166479ea75c08b4d27cf077 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-19T04:40:57Z |
| publishDate | 2021-11-01 |
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| series | Virology Journal |
| spelling | doaj.art-fee1d2b15166479ea75c08b4d27cf0772022-12-21T20:35:38ZengBMCVirology Journal1743-422X2021-11-0118111210.1186/s12985-021-01695-wSerotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disordersElham Ahmadi0Mehrdad Ravanshad1Jun Xie2Rajesh Panigrahi3Sandeep S. Jubbal4Santosh Kumar Guru5Gao Guangping6Mazyar Ziyaeyan7Joyce Fingeroth8Department of Virology, Faculty of Medical Sciences, Tarbiat Modares UniversityDepartment of Virology, Faculty of Medical Sciences, Tarbiat Modares UniversityHorae Gene Therapy Center, University of Massachusetts Medical SchoolDivision of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical SchoolDivision of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical SchoolDepartment of Molecular, Cell and Cancer Biology, University of Massachusetts Medical SchoolHorae Gene Therapy Center, University of Massachusetts Medical SchoolAlborzi Clinical Microbiology Research Center, Namazi Hospital, Shiraz University of Medical SciencesDivision of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical SchoolAbstract Background B-cell proliferative disorders, such as post-transplant lymphoproliferative disease (PTLD), are increased among persons afflicted by T-cell compromise. Most are Epstein–Barr virus (EBV) + and can first present with a focal lesion. Direct introduction of oncolytic viruses into localized tumors provides theoretical advantages over chemotherapy, immunotherapy and radiation therapy by reducing systemic toxicity. Despite extensive study as a vehicle for gene therapy, adeno-associated viruses (AAV) have rarely been applied to human cancer research due to technical and theoretical obstacles. Moreover, human B-cells have historically been described as resistant to AAV infection. Nonetheless, advances using different recombinant (r)AAV serotypes with unique tropisms to deliver cytotoxic therapy suggested a localized anti-tumor approach was feasible. Methods As a prelude to the development of a therapeutic vehicle, the ability of fifteen distinct EGFP-bearing rAAV serotypes to transduce human B-cells, including primary, immortalized, and B-cell tumor lines ± EBV was assessed by confocal microscopy, flow cytometry and subsequently cell viability assay. Results Rank order analysis revealed augmented transduction by rAAV6.2 and closely related virions. EBV infection of EBV-negative B-cell tumor lines and EBV immortalization of primary B-cells increased susceptibility to rAAV6.2 transduction. As a proof of concept, transduction by rAAV6.2 encoding herpes simplex virus type 1 (HSV1)-thymidine kinase (TK) eliminated TK-negative rhabdomyosarcoma cells and diminished viability of transduced B-cell lines upon incubation with ganciclovir. Conclusions rAAV serotypes differentially transduce human B-cell lines reversing the dogma that human B-cells are refractory to AAV infection. EBV + B-cells display increased susceptibility to rAAV6.2 infection, uncovering a new method for improved nucleic acid transfer into transfection-resistant B-cell lines. The introduction of a functional suicide gene into the rAAV6.2 genome identifies a candidate vector for the development of rAAV-based oncolytic therapy targeting focal EBV-bearing B-lymphoproliferative disorders.https://doi.org/10.1186/s12985-021-01695-wrAAVEBVB-cellsFocal tumorsPTLDMononucleosis |
| spellingShingle | Elham Ahmadi Mehrdad Ravanshad Jun Xie Rajesh Panigrahi Sandeep S. Jubbal Santosh Kumar Guru Gao Guangping Mazyar Ziyaeyan Joyce Fingeroth Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders Virology Journal rAAV EBV B-cells Focal tumors PTLD Mononucleosis |
| title | Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders |
| title_full | Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders |
| title_fullStr | Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders |
| title_full_unstemmed | Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders |
| title_short | Serotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders |
| title_sort | serotype dependent recombinant adeno associated vector aav infection of epstein barr virus positive b cells towards recombinant aav based therapy of focal ebv lymphoproliferative disorders |
| topic | rAAV EBV B-cells Focal tumors PTLD Mononucleosis |
| url | https://doi.org/10.1186/s12985-021-01695-w |
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