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FIGURE 3 from The Oncolytic Activity of Zika Viral Therapy in Human Neuroblastoma In Vivo Models Confers a Major Survival Advantage in a CD24-dependent Manner

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posted on 2024-01-09, 14:20 authored by Joseph Mazar, Jeanne K. Brooks, Matthew Peloquin, Rosa Rosario, Emma Sutton, Matthew Longo, Dennis Drehner, Tamarah J. Westmoreland

Time course of the In vivo modeling of the Zika viral treatment of neuroblastoma tumors. ZIKV was introduced once at a concentration of 2 × 10⁶ pfu for all tumors in NCr nude mice. A, Time course assessment of the application of ZIKV to IMR-32 tumors. Tumor size was measured at day 0, 2, 5, and 7 posttreatment compared with vehicle control treatment. Each timepoint within the study utilized an n = 3 for both vehicle- and Zika-treated cohorts. Error bars represent SD. *, P < 0.05 from Vehicle, unpaired t test, days 2, 5, and 7. B, Viral copy number was also measured in vehicle control treated mice for each timepoint in the IMR-32 tumor model, measuring the tumor, kidneys, spleen, liver, heart, and brain by absolute quantification PCR. C, Time course assessment of the application of ZIKV to SK-N-AS tumors. Tumor size was measured at day 0, 4, 7, and 10 posttreatment compared with vehicle control treatment. Each timepoint within the study utilized an n = 3 for both vehicle- and Zika-treated cohorts. Error bars represent SD. *, P < 0.05 from vehicle, unpaired t test, days 4, 7, and 10. D, Viral copy number was also measured in vehicle control treated mice for each timepoint in SK-N-AS tumor models, measuring the tumor, kidneys, spleen, liver, heart, and brain by absolute quantification PCR. E, Relative expression for CD24 was assessed using qRT-PCR at each timepoint for IMR-32 tumors. F, Relative expression for CD24 was assessed using qRT-PCR at each timepoint for SK-N-AS tumors. Both qRT-PCR studies compare cells with tumors for each neuroblastoma. Expression was normalized to GAPDH. All qPCR data shown are the composite of triplicate wells acquired from triplicate experiments. Error bars represent SD. *, P < 0.05 from In vitro cells, one-way ANOVA for tumor day 0, 4, 7, and10.

Funding

Nemours Children's Hospital

History

ARTICLE ABSTRACT

Neuroblastoma is the most common extracranial tumor, accounting for 15% of all childhood cancer-related deaths. The long-term survival of patients with high-risk tumors is less than 40%, and MYCN amplification is one of the most common indicators of poor outcomes. Zika virus (ZIKV) is a mosquito-borne flavivirus associated with mild constitutional symptoms outside the fetal period. Our published data showed that high-risk and recurrent neuroblastoma cells are permissive to ZIKV infection, resulting in cell type–specific lysis. In this study, we assessed the efficacy of ZIKV as an oncolytic treatment for high-risk neuroblastoma using in vivo tumor models. Utilizing both MYCN-amplified and non-amplified models, we demonstrated that the application of ZIKV had a rapid tumoricidal effect. This led to a nearly total loss of the tumor mass without evidence of recurrence, offering a robust survival advantage to the host. Detection of the viral NS1 protein within the tumors confirmed that a permissive infection preceded tissue necrosis. Despite robust titers within the tumor, viral shedding to the host was poor and diminished rapidly, correlating with no detectable side effects to the murine host. Assessments from both primary pretreatment and recurrent posttreatment isolates confirmed that permissive sensitivity to ZIKV killing was dependent on the expression of CD24, which was highly expressed in neuroblastomas and conferred a proliferative advantage to tumor growth. Exploiting this viral sensitivity to CD24 offers the possibility of its use as a prognostic target for a broad population of expressing cancers, many of which have shown resistance to current clinical therapies. Sensitivity to the tumoricidal effect of ZIKV on high-risk neuroblastoma tumors is dependent on CD24 expression, offering a prognostic marker for this oncolytic therapy in an extensive array of CD24-expressing cancers.