Discovery and structure-activity relationship of BMP Inhibitors, in-vivo SAR, zebrafish embryo 24-well plate
Because the heart has negligible intrinsic capacity to regenerate new tissues to replace those lost to injury, there is currently no definitive heart failure treatment, other than organ transplantation. Recent studies have introduced the prospect of replacing damaged heart tissues with healthy cardiomyocytes derived from pluripotent stem cells. However, realizing the full therapeutic potential of more ..
BioActive Compounds: 5
Assay Provider: Charles Hong
Assay Provider Affiliation: Vanderbilt University
Because the heart has negligible intrinsic capacity to regenerate new tissues to replace those lost to injury, there is currently no definitive heart failure treatment, other than organ transplantation. Recent studies have introduced the prospect of replacing damaged heart tissues with healthy cardiomyocytes derived from pluripotent stem cells. However, realizing the full therapeutic potential of stem cells faces numerous hurdles, including the potential for tumor formation, a low rate of cardiomyocyte formation, and an inadequate mechanistic understanding of cardiomyogenesis. Additionally, translational efforts are hampered by a lack of pharmaceutical agents to boost therapeutic effects of stem cells. Dorsomorphin, the first known small molecule inhibitor of the bone morphogenetic protein (BMP) signaling, is one of the most potent chemical inducers of cardiomyogenesis in mouse embryonic stem (ES) cells. Dorsomorphin treatment during the initial 24 to 48 hours of ES cell differentiation was sufficient for robust cardiomyocyte induction. Strikingly, the massive cardiac induction occurs apparently in the absence of mesoderm induction and at the expense of other mesoderm-derived lineages, including endothelial, smooth muscle and hematopoietic lineages. From these results, we hypothesize that atimely BMP signal inhibition commits the primitive multipotent progenitor cells toward the cardiomyocyte development. The aim is to develop potent and selective BMP inhibitors with excellent pharmaceutical properties (no cellular toxicity, high solubility, limited off-target activity) for use in directed differentiation of pluripotent stem cell toward cardiac development. Here, the Assessment of specific perturbations in embryonic dorsoventral patterning without additional toxicity or nonspecific defects is performed.
3-hour old zebrafish embryos will be arrayed in 24-well culture plates, and compounds delivered at various concentrations from 0.1mM to 100mM. 20 embryos will be tested for each condition. At 24-48 hours, embryos will be examined for specific perturbations in embryonic dorsoventral pattern, or nonspecific lethality or developmental delay. Compounds that dorsalize the embryonic pattern at lowest tested concentrations are deemed most potent, while compounds that only dorsalize and do not cause additional defects or nonspecific toxicities at the highest tested concentrations will be deemed most selective.
The in vivo assay is very robust with negligible false positive rate (<0.1%). Positive criterion is dorsalization of 2 or more embryos out of 20 tested at concentrations tested (at EC50, 10 embryos out of 20 tested will be dorsalized). Negative criterion is 1 or fewer embryos out of 20 tested. Compounds with EC50< 2.5mM on in vivo testing will be pursued further.
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