Metabolism-based drug screening platform in bioengineered zebrafish
The present application provides a novel drug screening platform designed to identify drugs that favorably modulate cellular bioenergetics in the human brain of a zebrafish which has been modified by altering a teleost gene that corresponds to a human gene associated with a brain dysfunction disorder (such as epilepsy). The drug screening platform are also useful for determining a mutation in a human gene associated with a brain dysfunction disorder that is associated with a human individual responsive to the treatment of a compound. Also provided are modified teleosts useful for the methods described herein.
1. A method of identifying a compound as a candidate compound for modulating mitochondrial function in the human brain, comprising:
a) contacting a compound with a teleost exhibiting decreased expression of a teleost gene that results in altered mitochondrial function, wherein the teleost is selected from the group consisting of: a morphant teleost comprising a morpholino that decreases expression of the teleost gene, and a knockout teleost comprising a knockout of the teleost gene; and
b) assaying for mitochondrial function in the contacted morphant or knockout teleost,
wherein restoration of normal mitochondrial function in the contacted morphant or knockout teleost compared to a control morphant or knockout teleost not contacted with the compound is indicative that the compound is a candidate compound for modulating mitochondrial function in the human brain.
2. The method of claim 1 , wherein the teleost is a zebrafish embryo, a zebrafish larva or a zebrafish adult.
3. The method of claim 1 , wherein said contacting comprises homogeneously distributing the compound in media containing the teleost or injecting said compound into the teleost.
4. The method of claim 1 , wherein the compound is selected from the group consisting: of a small molecule, a nucleic acid, a peptide, a protein, a carbohydrate, and a lipid.
5. The method of claim 1 , wherein the mitochondrial function is assayed by measuring one of more mitochondria outputs selected from the group consisting of ATP level, a level of an ATP metabolite, mitochondria respiration rate, reactive oxygenated species, reactive nitrogen species, mitochondrial uncoupling protein, mitochondrial permeability transition threshold, mitochondrial calcium homeostasis, mitochondrial appearance change, transcriptional changes in mitochondrial teleost genes, and mitochondrial membrane potential.
6. The method of claim 1 , wherein the teleost exhibits neuronal hyperactivity or loss of neuronal function.
7. The method of claim 1 , wherein the teleost gene is selected from the group consisting of: scn1a, kcna1, idh1, smarcb1a, and shank3.
8. The method of claim 1 , wherein the teleost gene is scn1a.
9. The method of claim 1 , wherein the teleost gene is kcna1.