Methods and compositions for the modulation of acid-sphingomyelinase-related apoptosis
AssigneeSloan-Kettering Institute for Cancer Research
InventorsRichard Kolesnick, Edward H. Schuchman
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Abstract
The present invention relates, first, to methods and compositions for the modulation of acid sphingomyelinase (ASM)-related processes, including apoptosis. Such apoptosis can include, but is not limited to, environmental stress-induced apoptosis such as, for example, ionizing radiation and/or chemotherapeutic agent-induced apoptosis. Apoptosis can be characterized by a cellular morphology comprising cellular condensation, nuclear condensation or zeiosis. The present invention further relates to methods for the identification of compounds which modulate (i.e., either increase or decrease) sensitivity to ASM-related processes, including apoptosis.
Claims9
A method for identifying a compound which increases a cell's sensitivity to acid sphingomyelinase-related apoptosis, comprising:
- (a) contacting an acid sphingomyelinase-deficient cell with a test compound;
- (b) exposing the cell to a stress stimulus for a time sufficient to induce apoptosis in a cell exhibiting normal acid sphingomyelinase activity;
- (c) exposing an acid sphingomyelinase-deficient cell, in the absence of the test compound, to the stress stimulus for a time sufficient to induce apoptosis in a cell exhibiting normal acid sphingomyelinase activity;
- and
- (d) monitoring the exposed cells of steps (b) and (c) for the presence of an apoptotic morphology,
- such that if the cell from step (b) exhibits a more severe apoptotic morphology, than that of the cell from step (c) the test compound represents a compound which increases a cell's sensitivity to acid sphingomyelinase-related apoptosis.
A method for identifying a compound which increases a cell's sensitivity to acid sphingomyelinase-related apoptosis, comprising:
- (a) contacting an acid sphingomyelinase-deficient cell with a test compound;
- (b) exposing the cell to a stress stimulus;
- (c) exposing an acid sphingomyelinase-deficient cell, in the absence of the test compound, to the stress stimulus;
- and
- (d) comparing the levels of sphingomyelin and ceramide present in the exposed cell of step (b) to the levels present in the exposed cell of step (c),
- such that if the level of sphingomyelin in the cell of step (b) is less than that of the cell of step (c), or the level of ceramide in the cell of step (b) is greater than that of the cell in step (c), the test compound represents a compound which increases a cell's sensitivity to acid sphingomyelinase-related apoptosis.
A method for identifying a compound which decreases a cell's sensitivity to acid sphingomyelinase-related apoptosis, comprising:
- (a) contacting a cell exhibiting acid sphingomyelinase activity with a test compound;
- (b) exposing the cell to a stress stimulus;
- (c) exposing a cell which exhibits acid sphingomyelinase activity to the stress stimulus, in the absence of the test compound;
- and
- (d) monitoring the exposed cells of steps (b) and (c) for the presence of an apoptotic morphology,
- such that if the cell from step (b) exhibits a less severe apoptotic morphology, than the cell from step (c) the test compound represents a compound which decreases a cell's sensitivity to acid sphingomyelinase-related apoptosis.
A method for identifying a compound which decreases a cell's sensitivity to acid sphingomyelinase-related apoptosis, comprising:
- (a) contacting a cell exhibiting acid sphingomyelinase activity with a test compound;
- (b) exposing the cell to a stress stimulus;
- (c) exposing a cell exhibiting acid sphingomyelinase activity to the stress stimulus, in the absence of the test compound;
- and
- (d) comparing the levels of sphingomyelin and ceramide present in the exposed cell of step (b) to the levels present in the exposed cell of step (c),
- such that if the level of sphingomyelin in the cell of step (b) is greater than that of the cell of step (c), or the level of ceramide in the cell of step (b) is less than that of the cell in step (c), the test compound represents a compound which decreases a cell's sensitivity to acid sphingomyelinase-related apoptosis.
Dependencies are inferred from the claim text.