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The RABiT-II DCA in the Rhesus Macaque Model
Ekaterina Royba1, Mikhail Repin1, Adayabalam S Balajee2
1Center for Radiological Research, Columbia University Irving Medical Center, New York, New York 10032.
Radiation Research
|October 6, 2020
Summary
The Rapid Automated Biodosimetry Tool II (RABiT-II) validates the dicentric chromosome assay (DCA) for radiation dose assessment. Rhesus macaques show similar radiation dose responses to humans, validating this high-throughput biodosimetry method.
Area of Science:
- Cytogenetics
- Radiation Biology
- Biodosimetry
Background:
- The dicentric chromosome assay (DCA) is a standard method for biological dosimetry.
- High-throughput methods are needed for population screening after radiological events.
- Validation in an in vivo model is recommended by the FDA for automated assays.
Purpose of the Study:
- To validate the accuracy of the high-throughput Rapid Automated Biodosimetry Tool II (RABiT-II) using an in vivo model.
- To assess the suitability of rhesus macaques as a model for human radiation biodosimetry.
- To develop and optimize tools for automated chromosome analysis in non-human primates.
Main Methods:
- Development of custom 17/22-mer peptide nucleic acid (PNA) probes for rhesus macaque centromere binding.
- Optimization of low-temperature telomere PNA FISH staining in multiwell plates.
- Utilizing the FluorQuantDic V4 software with added telomere detection for automated chromosome analysis.
- Ex vivo irradiation of rhesus macaque and human blood samples for comparative analysis.
Main Results:
- Custom PNA probes demonstrated high specificity for rhesus macaque centromere detection.
- Optimized FISH staining and software improved automated chromosome analysis accuracy.
- Rhesus macaques exhibited dose responses to gamma radiation similar to humans in lymphocyte systems.
- Statistically significant, though small, differences were observed between the two model systems.
Conclusions:
- The RABiT-II platform, utilizing optimized DCA with PNA probes and advanced software, is validated in an in vivo model.
- Rhesus macaques serve as a suitable in vivo model for validating human radiation biodosimetry assays.
- The study supports the use of high-throughput automated biodosimetry for large-scale radiological event response.
