Methods to identify compounds affecting mitochondria
AssigneeThe Johns Hopkins University
InventorsEduardo Marban, Brian O'Rourke
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Abstract
The present invention relates to methods for identifying a compound capable of modulating mitochondrial function, comprising contacting a eukaryotic cell with one or more candidate compounds, and detecting a change in the mitochondrial redox state of the cell. The methods further relates to such methods wherein endogenous fluorescence of the cell mitochondria is indicative of a change of redox state.
Claims31
A method for identifying a compound capable of increasing or decreasing the redox state of mitochondria, comprising
- (a) contacting a eukaryotic cell with one or more candidate compounds;
- and
- (b) measuring fluorescence of an endogenous mitochondrial component;
- (c) detecting a change in fluorescence upon contact with a candidate compound, wherein a change in fluorescence is indicative of an increase or decrease in the mitochondrial redox state of the cell;
- and
- (d) selecting a compound or compounds which causes a change in fluorescence.
A method of identifying a compound capable of increasing or decreasing the activity of a mitochondrial ion channel or mitochondrial transporter, comprising
- (a) contacting a eukaryotic cell with one or more candidate compounds, and
- (b) monitoring an fluorescence of an endogenous mitochondrial component;
- (c) measuring an activity of an ion channel current or a mitochondrial transporter by detecting a change in fluorescence upon contact with a candidate compound wherein the change in fluorescence is indicative of an increase or decrease in the activity of a mitochondrial transporter or mitochondrial ion channel;
- and
- (d) selecting a compound or compounds which causes a change in fluorescence.
A method for identifying a compound capable of increasing or decreasing mitochondrial redox potential, comprising:
- a) providing a population of eukaryotic cells;
- b) contacting a first portion of the cells with one or more candidate compounds;
- c) contacting a second portion of the cells with a known mitochondrial oxidizing or reducing agent;
- d) monitoring fluorescence of an endogenous mitochondrial component;
- e) measuring a difference between mitochondrial fluorescence produced in steps a) and b) and a) and c), wherein a change in fluorescence is indicative of an increase or decrease in the mitochondrial redox state of the cell;
- f) correlating a detected change in steps a) and b) to a change in steps a) and c)
- g) selecting a compound or compounds which causes a change in fluorescence.
Dependencies are inferred from the claim text.