A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin

Vapniarsky, Natalia; Lamé, Michael W.; McDonnel, Samantha J.; Murphy, Brian G

doi:10.1371/journal.pone.0045099

Cited by 3 publications

(3 citation statements)

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“…This study builds on previous results demonstrating the effectiveness of a replication-defective lentiviral gene therapy vector as a method of efficiently delivering the feEPO gene in vitro. 29 The Vector D lentivirus features the constitutively active SV40 promoter, which is functionally attenuated relative to the CMV promoter. Prior studies 35 have shown more stable, long-term transgene expression with the SV40 promoter relative to with the CMV promoter.…”

Section: Discussionmentioning

confidence: 99%

“…feEPO mRNA was previously PCR amplified from feline renal tissue-derived RNA, and the resulting reverse-transcribed cDNA was incorporated into an initial lentiviral vector system, referred to here as Vector A. 29 The feEPO gene (cDNA) was subsequently utilized in the design and construction of a novel third-generation, replication-defective lentiviral vector, referred to here as Vector D. A UC Davis Biological Use Authorization (BUA; #895) was approved for the use of this vector. The plasmidencoded feEPO cDNA was confirmed by sequencing (GenBank JQ413414).…”

Section: Design and Packaging Of Lentiviral Vectorsmentioning

confidence: 99%

“…Importantly, the reported sequence of the feEPO transgene utilized in both of these studies had a single nucleotide misincorporation resulting in an amino acid substitution at the 44th codon of the feEPO gene (glycine for glutamic acid). 29 Whether this amino acid misincorporation had any effect in the study outcome is unknown.…”

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confidence: 99%

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A lentivirus-vectored feline erythropoietin gene therapy strategy in tissue culture and rodent models for the potential treatment of chronic renal disease-associated anemia

Cook,

Castillo,

Wolf

et al. 2024

ajvr

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OBJECTIVE The aim of this study was to assess the efficacy and safety of a third-generation lentivirus-based vector encoding the feline erythropoietin (EPO) (feEPO) gene in vitro and in rodent models in vivo. This vector incorporates a genetic mechanism to facilitate the termination of the therapeutic effect in the event of supraphysiologic polycythemia, the herpes simplex virus thymidine kinase (HSV-TK) “suicide gene.” ANIMALS CFRK cells and replication-defective lentiviral vectors encoding feEPO were used for in vitro experiments. Eight Fischer rats were enrolled in the pilot in vivo study, 24 EPO-deficient mice were used in the initial mouse study, and 15 EPO-deficient mice were enrolled in the final mouse study. METHODS Efficacy of a third-generation lentivirus encoding feEPO was determined in vitro using western blot assays. Subsequently, in a series of rodent experiments, animals were administered the viral vector in progressively increasing inoculation doses with serial measurements of blood packed cell volume (PCV) over time. RESULTS We documented production of feEPO protein in transduced CRFK cells with subsequent cessation of production when treated with the HSV-TK substrate ganciclovir. In vivo, we demonstrated variably persistent elevated PCV values in treated rats and mice with eventual return to baseline values over time. CLINICAL RELEVANCE These results provide justification for a lentiviral gene therapy approach to the treatment of nonregenerative anemia associated with chronic renal disease in cats.

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Section: Discussionmentioning

confidence: 99%

Section: Design and Packaging Of Lentiviral Vectorsmentioning

confidence: 99%