PgmNr M279: A Cross-Species Novel Genetic Cell Ablation Technology Involving hCD59 and Intermedilysin.

Authors:
E. C. Bryda 1 ; Marina Hanson 1 ; Suman Gurung 1 ; Fengming Liu 2 ; Shen Dai 2 ; Alison Kearnes 2 ; Dechun Feng 3 ; Bin Gao 3 ; Anand Chandrasekhar 1 ; Xuebin Qin 2


Institutes
1) University of Missouri, Columbia, MO; 2) Temple University School of Medicine, Philadelphia, PA; 3) National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD.


Abstract:

The ability to selectively ablate cell types in model organisms is a powerful tool for understanding cell lineage, cell function, developmental processes and disease mechanisms. Current methodologies have limitations including lack of cell-type specificity, off–target effects, narrow pharmacological windows, and relatively slow onset of cell death. Our novel system utilizes human CD59 (hCD59), a membrane receptor, and intermedilysin (ILY), a bacterial toxin that binds hCD59 selectively.  We have generated multiple rodent models that express hCD59 in a cell-specific manner. We demonstrated 1) ablation of circulating cells in a rat anemia model which expresses hCD59 on erythrocytes for the study of the pathogenesis of hemolysis-associated complications, 2) ablation of cells in a solid organ by generating a mouse model in which epithelial cells in the liver were ablated for the study of liver damage and regeneration, and 3) ablation of neuronal cells in an inducible rat model carrying a transgene with constitutive expression of ZsGreen and hCD59 in the presence of Cre recombinase.  The Cre-inducible model can be bred with any conditional Cre recombinase-expressing rat to generate animals with expression of hCD59 and ZsGreen in the cell type/tissue of choice.  These models have shown that cell ablation is dose-dependent, rapid, and has a large pharmacological window.  Extending these experiments to zebrafish, we performed dose-response studies with hCD59 RNA-injected and control zebrafish embryos treated with ILY and found that ILY rapidly induced extensive tissue lysis in the embryos expressing hCD59. Current efforts are focused on further characterization of the effectiveness of hCD59-ILY-mediated cell ablation in zebrafish. Together, these studies demonstrate that the hCD59-ILY cell ablation technology has wide utility and can be applied to study virtually any tissue or cell type in the animal species of choice.