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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Autores principales:	Elham Ahmadi, Mehrdad Ravanshad, Jun Xie, Rajesh Panigrahi, Sandeep S. Jubbal, Santosh Kumar Guru, Gao Guangping, Mazyar Ziyaeyan, Joyce Fingeroth
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	rAAV EBV B-cells Focal tumors PTLD Mononucleosis Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/fee1d2b15166479ea75c08b4d27cf077
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spelling	oai:doaj.org-article:fee1d2b15166479ea75c08b4d27cf0772021-11-21T12:02:05ZSerotype-dependent recombinant adeno-associated vector (AAV) infection of Epstein–Barr virus-positive B-cells, towards recombinant AAV-based therapy of focal EBV + lymphoproliferative disorders10.1186/s12985-021-01695-w1743-422Xhttps://doaj.org/article/fee1d2b15166479ea75c08b4d27cf0772021-11-01T00:00:00Zhttps://doi.org/10.1186/s12985-021-01695-whttps://doaj.org/toc/1743-422XAbstract 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.Elham AhmadiMehrdad RavanshadJun XieRajesh PanigrahiSandeep S. JubbalSantosh Kumar GuruGao GuangpingMazyar ZiyaeyanJoyce FingerothBMCarticlerAAVEBVB-cellsFocal tumorsPTLDMononucleosisInfectious and parasitic diseasesRC109-216ENVirology Journal, Vol 18, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	rAAV EBV B-cells Focal tumors PTLD Mononucleosis Infectious and parasitic diseases RC109-216
spellingShingle	rAAV EBV B-cells Focal tumors PTLD Mononucleosis Infectious and parasitic diseases RC109-216 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
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.
format	article
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
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_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_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_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
publisher	BMC
publishDate	2021
url	https://doaj.org/article/fee1d2b15166479ea75c08b4d27cf077
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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

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