Opportunity:
- Perform toxicological assessments with cerebral spinal fluid (CSF) collected at termination
Challenges:
- Ensure that RNA from CSF and tissue samples are of high quality and free of hemolysis, and that tissues have an RNA integrity number (RIN) greater than 8.5.
- Weigh and preserve eight tissue samples in 10% NBF and liquid nitrogen within three minutes of humane euthanasia.
- Establish and validate methods for measuring hemoglobin and optimizing blood collection methodology to ensure low hemolysis in CSF, plasma and serum samples, and prevent interference with downstream miRNA profiling.
- Organize and catalog for transfer the large number of samples generated, and make sure that sample quality is not compromised during shipping.
- Complete 5 range finders and 5 definitive studies within 6 months.
Outcome:
- Implemented team-based necropsy approach to excise, weigh and appropriately fix tissues within the three-minute time window.
- Developed methodology to accurately measure hemoglobin concentration in plasma and serum.
- After initial testing showed hemoglobin levels higher than the acceptable threshold (< 20 mg/dL), optimized procedures in necropsy to reliably obtain high-quality serum and plasma samples within the acceptable hemoglobin limit..
- Conducted dose range finders to ensure any toxicity observed at top dose levels was a true test article-related effect and not due to dosing error.
- Two ILS staff members were included as co-authors on the published manuscript describing the studies: Toxicol Sci. 2016 Jan;149(1):67-88. doi: 10.1093/toxsci/kfv214. Epub 2015 Sep 22.
