Meta-Analysis and Optimization of the <i>In Vitro</i> Immortalization Assay for Safety Assessment of Retroviral Vectors in Gene Therapy
The underlying risk of retroviral vector-induced insertional oncogenesis in gene therapies requires a reliable preclinical safety assessment. Dysregulation of genes neighboring the vector's integration sites has triggered hematopoietic malignancies in patients treated with different vector genera and designs. With ca. 18 years in practical use, the <i>in vitro</i> immortalization (IVIM) assay can quantify this mutagenic potential and is actively requested by regulatory authorities during preclinical stages. Here, we present a thorough meta-analysis of IVIM data alongside a step-by-step cell culture protocol. On this basis, we propose clonal outgrowth as the single indicator of mutagenicity, simplifying the IVIM assay cost- and time-wise.
Authors
- Gabi Paul
- Philipp John-Neek
- Violetta Dziadek
- Bettina Weigel (ORCID: https://orcid.org/0000-0003-1684-9087)
- Axel Schambach (ORCID: https://orcid.org/0000-0003-2743-0070)
- Antonella Lucía Bastone (ORCID: https://orcid.org/0000-0003-0356-2909)
- Maike Hagedorn
- Jenni Fleischauer (ORCID: https://orcid.org/0009-0000-1850-9006)
- Friederike Mansel (ORCID: https://orcid.org/0009-0005-1484-1286)
- Michael Rothe (ORCID: https://orcid.org/0000-0002-6813-4705)
Institutions
- Boston Children's Hospital (US)
- Harvard University (US)
- Medizinische Hochschule Hannover (DE)
Publication Details
- Journal
- Human Gene Therapy
- Published
- 2025-04-09
- DOI
- https://doi.org/10.1089/hum.2024.221
- Citations
- 3
- Primary Topic
- Virus-based gene therapy research
- Type
- article
- Field-Weighted Citation Impact
- 5.45