Ravi Kane, Ph.D., (School of Chemical & Biomolecular Engineering, Georgia Institute of Technology)
- Andrés García, Ph.D. (School of Mechanical Engineering, Georgia Institute of Technology)
- Manu Platt, Ph.D. (Department of Biomedical Engineering, Georgia Institute of Technology)
- Todd Sulchek, Ph.D. (School of Mechanical Engineering, Georgia Institute of Technology)
- Ronghu Wu, Ph.D. (School of Chemistry & Biochemistry, Georgia Institute of Technology)
"Engineering Tools to Promote and Characterize Wnt-Mediated Stem Cell Differentiation"
The Wnt signaling pathway plays an important role in the development of many tissues in the body from the very earliest stage of the process. However, the precise mechanisms of the pathway and the specific roles it has in development in the context of different tissue types remain poorly understood. This is in part due to the complexity of embryonic development and in part due to the hydrophobicity of Wnt ligands which renders them expensive and difficult to purify in a usable form.
To overcome issues associated with the use of natural Wnt ligands, we have developed a heterodimer of Fabs which bind to the Wnt co-receptors LRP6 and Frizzled. We have demonstrated that this dimer can activate Wnt signaling with an efficacy comparable to that of the natural ligand. We will explore the ability to improve upon this tool by linking these dimer units to create a multivalent construct with higher avidity for Wnt receptors and will test its ability to activate signaling even more potently.
To better characterize the downstream effects of Wnt signaling during the manufacturing of therapeutic cells, we are also generating CRISPR/Cas9 edited reporter iPSC lines which we hope will be able to detect the expression of Wnt-regulated marker genes with high sensitivity and specificity. Luminescent signals generated by these cell lines during directed differentiation into cardiomyocytes may be able to guide the optimization of the manufacturing process to produce cardiomyocytes with a more mature phenotype. These cells will also be equipped with an inducible suicide mechanism to enable their easy and selective removal for cell manufacturing applications involving co-culture with unedited cells.