Construction of Various γ34.5 Deleted Fluorescent-Expressing Oncolytic Herpes Simplex Virus Type 1 (oHSV-1) for Generation and Isolation of HSV-Based Vectors

Abdoli, Shahriyar; Roohvand, Farzin; Teimoori‐Toolabi, Ladan; Shokrgozar, Mohammad Ali; Bahrololoumi, Mina; Azadmanesh, Kayhan

doi:10.18869/acadpub.ibj.21.4.206

Cited by 10 publications

(7 citation statements)

References 43 publications

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“…This vector was used to insertion the HMGB1 coding sequence at the tk locus in the HSV-ICP34.5- Blecherry TK-GFP to generate new recombinant HSV-HMGB1. Construction of HSV-ICP34.5- Blecherry was described elsewhere [ 48 ].…”

Section: Methodsmentioning

confidence: 99%

Cell type-specific response of colon cancer tumor cell lines to oncolytic HSV-1 virotherapy in hypoxia

et al. 2022

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Background Novel strategies are required since the hypoxic tumor microenvironment is one of the important impediments for conventional cancer therapy. High mobility group box 1 (HMGB1) protein can block aerobic respiration in cancer cells. We hypothesized that HMGB1could also kill the colorectal cancer cells during hypoxia. Methods In this study, we developed oncolytic herpes simplex virus type 1 expressing HMGB1 protein (HSV-HMGB1) and investigated the cytotoxic effect of HSV-HMGB1 and its parental virus (HSV-ble) on three colorectal cancer cells (HCT116, SW480, and HT29) under normoxic (20% oxygen) and hypoxic (1% oxygen) conditions. We further identified potential autophagy- related genes in HT29 cells by retrieving mRNA expression microarray datasets from the Gene Expression Omnibus database. These genes were then detected in HT29 cells infected with HSV-HMGB1 and HSV-ble during normoxia and hypoxia by Real-Time quantitative PCR (qRT-PCR). Results The cytotoxic effect of HSV-HMGB1 was significantly higher than that of HSV-ble during normoxia; however, during hypoxia, HSV-HMGB1 enhanced the viability of HT29 cells at MOI 0.1. Analyzing the cell death pathway revealed that HSV-HMGB1 induced autophagy in HT29 cells under hypoxic conditions. Conclusion In conclusion, it appears that oncolytic virotherapy is cell context-dependent. Therefore, understanding the cancer cells’ characteristics, microenvironment, and cell signaling are essential to improve the therapeutic strategies.

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Section: Methodsmentioning

confidence: 99%

Cell type-specific response of colon cancer tumor cell lines to oncolytic HSV-1 virotherapy in hypoxia

et al. 2022

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“…This virus was derived from an ICP34.5-null mutant of HSV-1, where both copies of ICP34.5 contained a BleCherry reporter gene driven by the cytomegalovirus promoter. The Δγ34.5/HSV-1 served as a reporter virus for identifying and verifying homologous recombination (Abdoli et al 2017;Haghighi-Najafabadi et al 2021).…”

Section: Virusesmentioning

confidence: 99%

“…The construction of Δγ34.5/HSV-1 was achieved using a standard homologous recombination-based method, starting from HSV-1 obtained from Dr. Houriyeh Soleimanjahi of Tarbiat Modarres University, Tehran, Iran. The virus was available at the Virology Department of Pasteur Institute of Iran (Abdoli et al 2017;Haghighi-Najafabadi et al 2021).…”

Section: Virusesmentioning

confidence: 99%

“…The use of fluorescent reporter genes-expressing shuttle vectors such as GFP (Liu et al 2003)(green fluorescent protein), RFP (red fluorescent proteins) (Ramachandran et al 2008), mCherry (Etienne et al 2017) and YFP (yellow fluorescent protein) (Miyagawa et al 2015) has become a prevalent strategy for effectively screening recombinant HSV-1 that expresses these specific fluorescent proteins. (Abdoli et al 2017;Scanlan et al 2022). These viruses can be readily identified through fluorescent microscopy, allowing for easy detection.…”

Section: Introductionmentioning

confidence: 99%

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Exploiting non-permissive CHO cells as a rapid and efficient method for recombinant HSV-1 isolation

Kelishadi,

Shahsavarani,

Tabarraei

et al. 2024

AMB Expr

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Using herpes simplex virus type 1 (HSV-1) as a therapeutic tool has recently emerged as a promising strategy for enhancing the treatment of various cancers, particularly those associated with the nervous system, which is the virus's natural site of infection. These viruses are specifically engineered to infect and eradicate tumor cells while leaving healthy cells unharmed. To introduce targeted mutations in specific viral genes, gene-modification techniques such as shuttle vector homologous recombination are commonly employed. Plaque purification is then utilized to select and purify the recombinant virus from the parental viruses. However, plaque purification becomes problematic when the insertion of the desired gene at the target site hampers progeny virus replication, resulting in a lower titer of cell-released virus than the parental virus. This necessitates a laborious initial screening process using approximately 10–15 tissue culture dishes (10 cm), making plaque purification time-consuming and demanding. Although the recently developed CRISPR-Cas9 system significantly enhances the efficiency of homologous integration and editing precision in viral genes, the purification of recombinant variants remains a tedious task. In this study, we propose a rapid and innovative method that employs non-permissive Chinese hamster ovary (CHO) cells, representing a remarkable improvement over the aforementioned arduous process. With this approach, only 1–2 rounds of plaque purification are required. Our proposed protocol demonstrates great potential as a viable alternative to current methods for isolating and purifying recombinant HSV-1 variants expressing fluorescent reporter genes using CHO cells and plaque assays.

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“…In addition to T-VEC, other HSV-based OVs have been developed, such as G47δ, oHSV-IL12, G207, and rRp450 ( 111 , 112 ). The majority of HSV-based vectors carry deletions in ICP34.5, a neurovirulence gene that restricts virus replication to tumor cells overexpressing the Ras gene ( 113 ). The inactivation of the ICP6 gene, which encodes a viral homolog of the cellular ribonucleotide reductase (RR), is another mechanism of HSV specificity ( 114 , 115 ).…”

Section: Various Types Of Ovsmentioning

confidence: 99%

Immunovirotherapy: The role of antibody based therapeutics combination with oncolytic viruses

Jafari

Kadkhodazadeh²,

Shapourabadi³

et al. 2022

Front. Immunol.

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Despite the fact that the new drugs and targeted therapies have been approved for cancer therapy during the past 30 years, the majority of cancer types are still remain challenging to be treated. Due to the tumor heterogeneity, immune system evasion and the complex interaction between the tumor microenvironment and immune cells, the great majority of malignancies need multimodal therapy. Unfortunately, tumors frequently develop treatment resistance, so it is important to have a variety of therapeutic choices available for the treatment of neoplastic diseases. Immunotherapy has lately shown clinical responses in malignancies with unfavorable outcomes. Oncolytic virus (OV) immunotherapy is a cancer treatment strategy that employs naturally occurring or genetically-modified viruses that multiply preferentially within cancer cells. OVs have the ability to not only induce oncolysis but also activate cells of the immune system, which in turn activates innate and adaptive anticancer responses. Despite the fact that OVs were translated into clinical trials, with T-VECs receiving FDA approval for melanoma, their use in fighting cancer faced some challenges, including off-target side effects, immune system clearance, non-specific uptake, and intratumoral spread of OVs in solid tumors. Although various strategies have been used to overcome the challenges, these strategies have not provided promising outcomes in monotherapy with OVs. In this situation, it is increasingly common to use rational combinations of immunotherapies to improve patient benefit. With the development of other aspects of cancer immunotherapy strategies, combinational therapy has been proposed to improve the anti-tumor activities of OVs. In this regard, OVs were combined with other biotherapeutic platforms, including various forms of antibodies, nanobodies, chimeric antigen receptor (CAR) T cells, and dendritic cells, to reduce the side effects of OVs and enhance their efficacy. This article reviews the promising outcomes of OVs in cancer therapy, the challenges OVs face and solutions, and their combination with other biotherapeutic agents.

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Construction of Various γ34.5 Deleted Fluorescent-Expressing Oncolytic Herpes Simplex Virus Type 1 (oHSV-1) for Generation and Isolation of HSV-Based Vectors

Cited by 10 publications

References 43 publications

Cell type-specific response of colon cancer tumor cell lines to oncolytic HSV-1 virotherapy in hypoxia

Cell type-specific response of colon cancer tumor cell lines to oncolytic HSV-1 virotherapy in hypoxia

Exploiting non-permissive CHO cells as a rapid and efficient method for recombinant HSV-1 isolation

Immunovirotherapy: The role of antibody based therapeutics combination with oncolytic viruses

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