Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors

Irina Kiprianova, Alexandra Just, Barbara Leuchs, Jean Rommelaere, Assia L. Angelova

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Fluorescence in situ hybridization (FISH) is a specific, sensitive, accurate, and reliable technique widely applied in both research and clinic. Here we describe the detailed protocol of a FISH method established by us to serve the scientific purposes of the first oncolytic parvovirus clinical trial (ParvOryx01). This trial was launched in Germany in 2011. After trial completion in 2015, results were published in Molecular Therapy in 2017. The primary purpose of the trial was to evaluate the safety of an oncolytic parvovirus, H-1PV (ParvOryx), in recurrent glioblastoma patients. In addition, the efficiency of H-1PV tumor targeting after intratumoral or systemic virus administration was assessed by FISH detection of viral nucleic acids (genomic single-stranded DNA, mRNA and parvovirus double-stranded replicative forms) in formalin-fixed paraffin-embedded glioblastoma tissues resected at day 10 after ParvOryx treatment. The FISH method allowed the detection—for the first time in humans—of H-1PV replication markers in brain tumors of parvovirus-treated patients. A protocol combining mRNA FISH with simultaneous immunofluorescent staining for tumor and tumor microenvironment markers was also developed and is described here, in order to better characterize H-1PV cellular targets and H-1PV treatment-associated tumor microenvironment changes.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages295-306
Number of pages12
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2058
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

H-1 parvovirus
Tumor Microenvironment
Fluorescence In Situ Hybridization
Parvovirus
Brain Neoplasms
Nucleic Acids
Glioblastoma
Messenger RNA
Single-Stranded DNA
Tumor Biomarkers
Paraffin
Formaldehyde
Germany
Neoplasms
Therapeutics
Clinical Trials
Staining and Labeling
Viruses
Safety
Research

Keywords

  • Combined FISH/immunofluorescence (IF)
  • First oncolytic parvovirus clinical trial
  • Fluorescence in situ hybridization (FISH)
  • Oncolytic H-1 parvovirus (H-1PV)
  • Oncolytic parvovirus replication markers
  • Paraffin sections
  • Recurrent glioblastoma
  • Tumor microenvironment

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Kiprianova, I., Just, A., Leuchs, B., Rommelaere, J., & Angelova, A. L. (2020). Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors. In Methods in Molecular Biology (pp. 295-306). (Methods in Molecular Biology; Vol. 2058). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9794-7_20

Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors. / Kiprianova, Irina; Just, Alexandra; Leuchs, Barbara; Rommelaere, Jean; Angelova, Assia L.

Methods in Molecular Biology. Humana Press Inc., 2020. p. 295-306 (Methods in Molecular Biology; Vol. 2058).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kiprianova, I, Just, A, Leuchs, B, Rommelaere, J & Angelova, AL 2020, Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2058, Humana Press Inc., pp. 295-306. https://doi.org/10.1007/978-1-4939-9794-7_20
Kiprianova I, Just A, Leuchs B, Rommelaere J, Angelova AL. Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors. In Methods in Molecular Biology. Humana Press Inc. 2020. p. 295-306. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9794-7_20
Kiprianova, Irina ; Just, Alexandra ; Leuchs, Barbara ; Rommelaere, Jean ; Angelova, Assia L. / Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors. Methods in Molecular Biology. Humana Press Inc., 2020. pp. 295-306 (Methods in Molecular Biology).
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