Nonclinical pharmacokinetics and biodistribution of VSV-GP using methods to decouple input drug disposition and viral replication

Dambra, Richard; Matter, Andrea; Graca, Kaitlynn; Akhand, Saeed Salehin; Mehta, Saurin; Bell-Cohn, Ashlee; Swenson, Joyce M.; Abid, Sadia; Xin, Dongyue; Lewis, Cedric; Coyle, Luke; Wang, Min; Bunosso, Kathleen; Maugiri, Michelle; Ruiz, Richard; Cirillo, Corey M.; Fogal, Birgit; Grimaldi, Christine; Vigil, Adam; Wood, Charles E.; Ashour, Joseph

doi:10.1016/j.omtm.2022.12.013

Cited by 4 publications

(5 citation statements)

References 85 publications

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“…In addition, this result is consistent with other studies reporting short values of halflife (Garcia-Carbonero et al, 2017;Béguin et al, 2021;Parra-Guillen et al, 2021). Development of an immunogenic reaction against biological therapies is a known fact that it is reflected by an increase in CL with time (Béguin et al, 2021;Parra-Guillen et al, 2021;Dambra et al, 2023), as has been the case in the current investigation. As a representative instance, the study by Dambra et al divided the viral kinetics into early and late time points, with the clearance rate estimated to be 21.8 and 282.8 mL/h, respectively.…”

Section: Discussionsupporting

confidence: 93%

“…However, the data reveal that a maximum of 1.44 × 10 3 copies/µg RNA is reached during the final sampling period. This accumulation of V937 in the spleen may be attributed to the delayed activation of the antiviral response in this particular tissue, as suggested by the previous literature ( Dambra et al, 2023 ).…”

Section: Discussionsupporting

confidence: 67%

“…Although the current study was performed in non-tumor-bearing organs and replication was not identified, the complete absence of replication cannot be ruled out. A recent preclinical experiment using methods to discriminate between viral distribution and viral replication showed that the latter process occurred in healthy tissues but was restricted to those with resident macrophages ( Dambra et al, 2023 ). This could suggest that macrophages play a role in shaping viral dynamics.…”

Section: Discussionmentioning

confidence: 99%

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A physiologically based pharmacokinetic model for V937 oncolytic virus in mice

Peribañez-Dominguez,

Parra-Guillen,

Freshwater

et al. 2023

Front. Pharmacol.

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Introduction: Oncolytic viruses (OVs) represent a novel therapeutic strategy in oncology due to their capability to selectively infect and replicate in cancer cells, triggering a direct and/or immune-induced tumor lysis. However, the mechanisms governing OV pharmacokinetics are still poorly understood. This work aims to develop a physiologically based pharmacokinetic model of the novel OV, V937, in non-tumor-bearing mice to get a quantitative understanding of its elimination and tissue uptake processes.Materials and methods: Model development was performed using data obtained from 60 mice. Viral levels were quantified from eight tissues after a single intravenous V937 dose. An external dataset was used for model validation. This test set included multiple-dose experiments with different routes of administration. V937 distribution in each organ was described using a physiological structure based on mouse-specific organ blood flows and volumes. Analyses were performed using the non-linear mixed-effects approach with NONMEM 7.4.Results: Viral levels showed a drop from 108 to 105 copies/µg RNA at day 1 in blood, reflected in a high estimate of total clearance (18.2 mL/h). A well-stirred model provided an adequate description for all organs except the muscle and heart, where a saturable uptake process improved data description. The highest numbers of viral copies were observed in the brain, lymph node, kidney, liver, lung, and spleen on the first day after injection. On the other hand, the maximum amount of viral copies in the heart, muscle, and pancreas occurred 3 days after administration.Conclusion: To the best of our knowledge, this is the first physiologically based pharmacokinetic model developed to characterize OV biodistribution, representing a relevant source of quantitative knowledge regarding the in vivo behavior of OVs. This model can be further expanded by adding a tumor compartment, where OVs could replicate.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 67%

Section: Discussionmentioning

confidence: 99%

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A physiologically based pharmacokinetic model for V937 oncolytic virus in mice

Peribañez-Dominguez,

Parra-Guillen,

Freshwater

et al. 2023

Front. Pharmacol.

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“…VSV-GP virus batches were produced in HEK-293F cells in either shaker flask or bioreactor formats (depending on batch ID). After infecting cells at an MOI of 0.0005, virus was purified from clarified harvest via centrifugation and/or chromatographic methods similar to previous reports (58, 59). The downstream processes employed were slightly unique for each batch (for example the use of different purification columns or formulations).…”

Section: Methodsmentioning

confidence: 99%

“…The cell pellets were frozen at -80 o C until further processing. Supernatants were collected at the same time points as the cells and were analyzed by tissue culture infectious dose 50% (TCID 50 ) assay as reported previously (58).…”

Section: Time Point Cell-infection Samples With Vsv-gpmentioning

confidence: 99%

Absolute Quantification of Viral Proteins from Pseudotyped Vesicular Stomatitis Virus (VSV-GP) using Ultra High-Performance Liquid Chromatography- Multiple Reaction Monitoring (UPLC-MRM)

Basu,

Dambra,

Jiang

et al. 2023

Preprint

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The rapidly developing field of oncolytic virus (OV) therapy necessitates development of new and improved analytical approaches for characterization of the virus during production and development. Accurate monitoring and absolute quantification of viral proteins is crucial for OV product characterization and can facilitate the understanding of infection, immunogenicity, and development stages of viral replication. Targeted mass spectrometry methods, like multiple reaction monitoring (MRM), offers a robust way to directly detect and quantify specific targeted proteins represented by surrogate peptides. We have leveraged the power of MRM by combining ultra-high performance liquid chromatography (UPLC) with a Sciex 6500 triple stage quadrupole mass spectrometer to develop an assay that accurately and absolutely quantifies the structural proteins of a pseudotyped vesicular stomatitis virus intended for use as a new biotherapeutic (designated hereafter as VSV-GP) to differentiate it from native VSV. The new UPLC-MRM method provides absolute quantification with the use of heavy labeled reference standard surrogate peptides. When added in known exact amounts to standards and samples, the reference standards normalize and account for any small perturbations during sample preparation and/or instrument performance, resulting in accurate and precise quantification. Because of the multiplexed nature of MRM all targeted proteins are quantified at the same time. The optimized assay has been enhanced to quantify the ratios of the processed GP1 and GP2 proteins while simultaneously measuring any remaining or unprocessed form of the envelope protein GPC (full-length GPC).IMPORTANCEDevelopment of oncolytic viral therapy has gained considerable momentum in the recent years. VSV-GP is a new biotherapeutic emerging in the oncolytic viral therapy platform. Novel analytical assays that can accurately and precisely quantify the viral proteins are a necessity for the successful development of viral vector as a biotherapeutic. We developed a UPLC-MRM based assay to quantify the absolute concentrations of the different structural proteins of VSV-GP. The complete processing of GPC is a pre-requisite for infectivity of the virus. The assay extends the potential for quantifying full-length GPC, which provides an understanding of the processing of GPC (along with the quantification of GP1 and GP2 separately). We used this assay in tracking GPC processing in HEK-293-F production cell lines infected with VSV-GP.

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Absolute quantification of viral proteins from pseudotyped VSV-GP using UPLC-MRM

Basu,

Dambra,

Jiang

et al. 2024

Microbiol Spectr

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The rapidly developing field of oncolytic virus (OV) therapy necessitates the development of new and improved analytical approaches for the characterization of the virus during production and development. Accurate monitoring and absolute quantification of viral proteins are crucial for OV product characterization and can facilitate the understanding of infection, immunogenicity, and development stages of viral replication. Targeted mass spectrometry methods like multiple reaction monitoring (MRM) offer a robust way to directly detect and quantify specific targeted proteins represented by surrogate peptides. We have leveraged the power of MRM by combining ultra-high performance liquid chromatography (UPLC) with a Sciex 6500 triple-stage quadrupole mass spectrometer to develop an assay that accurately and absolutely quantifies the structural proteins of a pseudotyped vesicular stomatitis virus (VSV) intended for use as a new biotherapeutic (designated hereafter as VSV-GP to differentiate it from native VSV). The new UPLC-MRM method provides absolute quantification with the use of heavy-labeled reference standard surrogate peptides. When added in known exact amounts to standards and samples, the reference standards normalize and account for any small perturbations during sample preparation and/or instrument performance, resulting in accurate and precise quantification. Because of the multiplexed nature of MRM, all targeted proteins are quantified at the same time. The optimized assay has been enhanced to quantify the ratios of the processed GP1 and GP2 proteins while simultaneously measuring any remaining or unprocessed form of the envelope protein GP complex (GPC; full-length GPC). IMPORTANCE The development of oncolytic viral therapy has gained considerable momentum in recent years. Vesicular stomatitis virus glycoprotein (VSV-GP) is a new biotherapeutic emerging in the oncolytic viral therapy platform. Novel analytical assays that can accurately and precisely quantify the viral proteins are a necessity for the successful development of viral vector as a biotherapeutic. We developed an ultra-high performance liquid chromatography multiple reaction monitoring-based assay to quantify the absolute concentrations of the different structural proteins of VSV-GP. The complete processing of GP complex (GPC) is a prerequisite for the infectivity of the virus. The assay extends the potential for quantifying full-length GPC, which provides an understanding of the processing of GPC (along with the quantification of GP1 and GP2 separately). We used this assay in tracking GPC processing in HEK-293-F production cell lines infected with VSV-GP.

show abstract

Nonclinical pharmacokinetics and biodistribution of VSV-GP using methods to decouple input drug disposition and viral replication

Cited by 4 publications

References 85 publications

A physiologically based pharmacokinetic model for V937 oncolytic virus in mice

A physiologically based pharmacokinetic model for V937 oncolytic virus in mice

Absolute Quantification of Viral Proteins from Pseudotyped Vesicular Stomatitis Virus (VSV-GP) using Ultra High-Performance Liquid Chromatography- Multiple Reaction Monitoring (UPLC-MRM)

Absolute quantification of viral proteins from pseudotyped VSV-GP using UPLC-MRM

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