Preclinical Testing of an Oncolytic Parvovirus in Ewing Sarcoma: Protoparvovirus H-1 Induces Apoptosis and Lytic Infection In Vitro but Fails to Improve Survival In Vivo
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sources and Culture Conditions for Human Cell Lines
2.2. Production of Infectious Viral Particles
2.3. Detection of Infectious H-1PV Particles
2.4. Extraction of H-1PV DNA and Qantification by Real-Time PCR
2.5. Conditions of in vitro Infection Experiments
2.6. Microscopy of H-1P-Infected Cells
2.7. Analysis of the Cell Cycle, Including Sub-G1 Apoptotic Cells, during H-1PV Infection
2.8. Quantatitation of Cell Viability and Cell Lysis
2.9. Real-Time Proliferation Measurements
2.10. Animal Experiments
2.11. Statistical Methods
3. Results
3.1. Ewing Sarcoma (EWS) Cells Are Susceptible to H-1PV Infection and Sustain the Synthesis of Viral Proteins
3.2. H-1PV Infection Interferes with the Cell Cycle in Ewing Sarcoma Cell Lines
3.3. H-1PV-Induced Cytotoxic Effects in Ewing Sarcoma Cell Lines Correlate with the Dose of Input Virus
3.4. Ewings’s Sarcoma Cells Efficently Replicate H-1PV
3.5. Intratumoral Infection with H-1PV Represses Human Ewing Sarcoma Cell Proliferation in a Xenograft Mouse Model but Fails to Improve Survival
4. Discussion
4.1. H-1PV Selectively Infects Transformed Human Cells of Mesenchymal Origin
4.2. Ewing Sarcoma Cells Are Fully Permissive to Wild-Type H-1PV Infection
4.3. Animal Data Show Antiproliferative Effects of H-1PV but Fail to Demonstrate Significant Antineoplastic Efficacy in a Ewing Sarcoma Xenograft Model
5. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Cell Line | LD50 [PFU/Cell] | Tumor of Origin | Age of Patient |
---|---|---|---|
MHH-ES-1 | 0.01 | Peritoneal metastatic EWS | 12 years |
TC-71 | 0.1 | Local recurrent EWS | 22 years |
SK-ES-1 | 5 | Primary Ewing sarcoma | 18 years |
CADO-ES1 | 5 | Pleural metastatic EWS | 19 years |
Cell Line | ED50 [PFU/Cell] | R2 | Observation Time |
---|---|---|---|
MHH-ES | 0.32 | 0.922 | 256 h |
SK-ES-1 | 0.11 | 0.972 | 215 h |
TC-71 | 1.04 | 0.978 | 120 h |
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Lacroix, J.; Kis, Z.; Josupeit, R.; Schlund, F.; Stroh-Dege, A.; Frank-Stöhr, M.; Leuchs, B.; Schlehofer, J.R.; Rommelaere, J.; Dinsart, C. Preclinical Testing of an Oncolytic Parvovirus in Ewing Sarcoma: Protoparvovirus H-1 Induces Apoptosis and Lytic Infection In Vitro but Fails to Improve Survival In Vivo. Viruses 2018, 10, 302. https://doi.org/10.3390/v10060302
Lacroix J, Kis Z, Josupeit R, Schlund F, Stroh-Dege A, Frank-Stöhr M, Leuchs B, Schlehofer JR, Rommelaere J, Dinsart C. Preclinical Testing of an Oncolytic Parvovirus in Ewing Sarcoma: Protoparvovirus H-1 Induces Apoptosis and Lytic Infection In Vitro but Fails to Improve Survival In Vivo. Viruses. 2018; 10(6):302. https://doi.org/10.3390/v10060302
Chicago/Turabian StyleLacroix, Jeannine, Zoltán Kis, Rafael Josupeit, Franziska Schlund, Alexandra Stroh-Dege, Monika Frank-Stöhr, Barbara Leuchs, Jörg R. Schlehofer, Jean Rommelaere, and Christiane Dinsart. 2018. "Preclinical Testing of an Oncolytic Parvovirus in Ewing Sarcoma: Protoparvovirus H-1 Induces Apoptosis and Lytic Infection In Vitro but Fails to Improve Survival In Vivo" Viruses 10, no. 6: 302. https://doi.org/10.3390/v10060302