Coxsackievirus B3 used as a gene therapy vector to express functional FGF2

Abstract

Current gene therapies are predominantly based on a handful of viral vectors. The limited choice of delivery vectors has been one of the stumbling blocks to the advancement of gene therapy. Therefore, the development of novel recombinant vectors should facilitate the application of gene therapies. In this study, we examined coxsackievirus B3 (CVB3) as a novel recombinant vector for the delivery and expression of a foreign gene in vitro and in vivo. A recombinant CVB3 complementary DNA was constructed by inserting a gene encoding human fibroblast growth factor 2 (FGF2). The recombinant virus (CVB3 --FGF2) efficiently expressed FGF2 in HeLa cells and human cardiomyocytes in vitro and in mouse hindlimbs in vivo. The injection of the recombinant virus into mice with ischemic hindlimbs protected the hindlimbs from ischemic necrosis. CVB3 --FGF2 injection significantly improved the blood flow in the ischemic limbs for over 3 weeks compared with that in the phosphate-buffered saline-or CVB3-injected controls, suggesting that FGF2 expressed from CVB3 --FGF2 is functional and therapeutically effective. The virulence of CVB3 was also drastically attenuated in the recombinant virus. Thus, CVB3 can be modified to express a functional foreign protein, supporting its use as a novel viral vector for gene therapy.

Recent progress in understanding coxsackievirus replication, dissemination, and pathogenesis

“…Many of these recombinant viral constructs have proven to be quite stable, although the loss of the foreign insert can appear in the viral population within five passages, depending upon the size of the insert, nucleotide sequence, nature of the gene, and passage conditions. The utilization of these rCVBs in tissue culture and in vivo have clarified questions regarding viral tropism (Feuer, Pagarigan et al, 2005) (Puccini, Ruller et al, 2014), activation of the adaptive immune response against infection (Kemball, Harkins et al, 2009), mechanisms of virulence and disease (Zeng, Chen et al, 2013), and possible uses of CVB as therapeutic/vaccine vectors (Kim, Kim et al, 2012) (Miller, Geng et al, 2009). …”

Global landscape of coxsackieviruses in human health

“…Within this context, both CVB3 and CVA21 have proven useful for gene therapy and vaccine development ( Lundstrom, 2023 ; Bradley et al, 2014 ). In particular, recombinant CVB3 expressing the human fibroblast growth factor 2 (FGF2) demonstrated protection against ischemic necrosis after administration into mice with ischemic hindlimbs ( Kim et al, 2012 ). Additionally, it has been shown that recombinant CVB3 expressing IFN- γ is an important approach for preventing CVB3-induced myocarditis and for providing long-term immune response ( Henke et al, 2008 ).…”

Viral Vectors in Gene Therapy

“…Coxsackieviruses have been used for several gene therapy applications [ 23 ]. For instance, the coxsackievirus B3 (CVB3) expressing the human fibroblast growth factor 2 (FGF2) was injected into ischemic hindlimbs of mice showing protection from ischemic necrosis [ 76 ]. The treatment improved the blood flow in ischemic limbs for more than 3 weeks.…”