23 - High Throughput Drug Screening in Zebrafish for Identifying Novel Compounds That Influence Engraftment Efficiency

Poster Board Number: 23
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Co-authors:

Sarah Jacobson - Research Technician, Pediatrics, Albert Einstein College of Medicine; Kyle Lorentzen - Developmental and Molecular Biology - Albert Einstein College of Medicine; Teresa Bowman - Principal Investigator, Developmental and Molecular Biology, Albert Einstein College of Medicine; Ellen Fraint - Pediatric Hematology, Oncology, and Cellular Therapy, Children's Hospital at Montefiore - Albert Einstein College of Medicine

Abstract Body : As hematopoietic cell transplantation (HCT) outcomes have improved over time due to better optimized regimens and supportive care, the population of patients deemed eligible candidates for HCT is expanding. But despite the significant advances in the field, HCT brings a risk of morbidity and mortality even for the most fit patients. Slow or failed engraftment is a potentially modifiable cause of non-relapse mortality. Research that aims to hasten or strengthen engraftment may decrease morbidity from organ and tissue damage as well as shorten the period of profound immune compromise. Our stem cell transplant assay in zebrafish larvae is a cost-effective high throughput way to investigate novel compounds with a potential to influence engraftment. By taking advantage of the small size, translucent nature of the casper zebrafish strain, we used this HCT assay to screen 121 drugs for more efficient or more robust hematopoietic engraftment. We transplanted fluorescent donor marrow cells into recipient larvae, arranged them in multi-well plates, then treated them with drugs from a library of pharmacologically active compounds. Engraftment was assessed at two time points to determine early (4 days post-transplant) and sustained (8 weeks after transplantation) engraftment. Quantification of fluorescent donor cells at early stages was performed via fluorescent microscopy and at later stages by flow cytometry of recipient marrow. Our results identified 10 potentials hits out of 121 candidate compounds. One candidate passed secondary validation. This compound hastened engraftment in the early stages but did not sustain long term engraftment. Using live imaging, we will evaluate if donor cell homing, fitness, cell division kinetics, and/or if endogenous hematopoietic cells or their niche are affected by treatment with the validated candidate compound. Our ultimate goal is to identify ways to improve early engraftment after hematopoietic cell transplantations.