Apparatus and Method for Testing Relationships Between Gene Expression and Physical Appearance of Skin
First Claim
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1. A method of testing to identify genes associated with one or more physical attributes of skin aging comprising:
- exposing a first sample of human skin tissue to an agent;
determining a first set of expression levels of a plurality of genes in the first sample of human skin;
comparing the first set of expression levels to a second set of expression levels, the second set of expression levels corresponding to expression levels of human skin tissue not exposed to the agent, to identify a first subset of genes having a fold change difference in expression level between the exposed and unexposed samples that meets a first, selected biological relevance level;
selecting from the first subset of genes a second subset of genes, each gene being associated with a biochemical pathway associated with physical appearance of skin aging;
selecting from the second subset of genes, at least one skin attribute subset of genes, each gene in the skin attribute subset being associated with a biochemical pathway relating to the skin attribute that is shown in the comparing step to have been regulated in a more youthful direction for that biochemical pathway and skin attribute;
exposing a second sample of human skin tissue to the agent;
determining the levels of expression for the at least one skin attribute subset of genes in the second sample of human skin tissue using a method for determining expression levels that is different than that used for the first sample of human skin tissue; and
selecting a third subset of genes from the at least one skin attribute subset of genes whose expression levels in the second sample of human skin tissue meet a second, selected biological relevance level and whose direction of regulation conforms to the more youthful direction used in selecting the at least one skin attribute subset of genes.
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Abstract
The disclosure is directed to apparatus and methods for testing relationships between gene expression and physical appearance of skin and methods of assessing the efficacy of skin anti-aging agents.
17 Citations
22 Claims
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1. A method of testing to identify genes associated with one or more physical attributes of skin aging comprising:
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exposing a first sample of human skin tissue to an agent; determining a first set of expression levels of a plurality of genes in the first sample of human skin; comparing the first set of expression levels to a second set of expression levels, the second set of expression levels corresponding to expression levels of human skin tissue not exposed to the agent, to identify a first subset of genes having a fold change difference in expression level between the exposed and unexposed samples that meets a first, selected biological relevance level; selecting from the first subset of genes a second subset of genes, each gene being associated with a biochemical pathway associated with physical appearance of skin aging; selecting from the second subset of genes, at least one skin attribute subset of genes, each gene in the skin attribute subset being associated with a biochemical pathway relating to the skin attribute that is shown in the comparing step to have been regulated in a more youthful direction for that biochemical pathway and skin attribute; exposing a second sample of human skin tissue to the agent; determining the levels of expression for the at least one skin attribute subset of genes in the second sample of human skin tissue using a method for determining expression levels that is different than that used for the first sample of human skin tissue; and selecting a third subset of genes from the at least one skin attribute subset of genes whose expression levels in the second sample of human skin tissue meet a second, selected biological relevance level and whose direction of regulation conforms to the more youthful direction used in selecting the at least one skin attribute subset of genes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A computer based system of testing to identify genes associated with one or more physical attributes of skin aging comprising:
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a first instrument for exposing a first sample of human skin tissue to an agent and determining a first set of expression levels of a plurality of genes in the first sample of human skin; a computer module for comparing the first set of expression levels to a second set of expression levels, the second set of expression levels corresponding to expression levels of human skin tissue not exposed to the agent to identify a first subset of genes having a fold change difference in expression level between the exposed and unexposed samples that meet a first, selected biological relevance level; a computer module for accessing a stored data set identifying genes, each gene being associated with a biochemical pathway associated with physical appearance of skin aging and for selecting from the first subset a second subset comprising those genes also in the second subset; a computer module for selecting from the second subset of genes, at least one skin attribute subset of genes, each gene in the skin attribute subset being associated with a biochemical pathway relating to the skin attribute that is shown in the comparing step to have been regulated in a more youthful direction for that biochemical pathway and skin attribute; a second instrument for exposing a second sample of human skin tissue to the agent and for determining the levels of expression for the at least one skin attribute subset of genes in the second sample of human skin tissue using a method for determining expression levels that is different than that used for the first sample of human tissue; and a computer module for selecting a third subset of genes from the at least one skin attribute subset of genes whose expression levels in the second sample of human skin tissue meet a second, selected biological relevance level and whose direction of regulation conforms to the more youthful direction used in selecting the at least one skin attribute subset of genes. - View Dependent Claims (12, 13, 14, 15, 16)
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17. A method of assessing the efficacy of a skin anti-aging agent comprising:
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exposing a first sample of human skin tissue to an agent; determining a first set of expression levels of a plurality of genes in the first sample of human skin; comparing the first set of expression levels to a second set of expression levels, the second set of expression levels corresponding to expression levels of human skin tissue not exposed to the agent, to identify a first subset of genes having a fold change difference in expression level between the exposed and unexposed samples that meets a first, selected biological relevance level; selecting from the first subset of genes a second subset of genes, each gene being associated with a biochemical pathway associated with physical appearance of skin aging; selecting from the second subset of genes, at least one skin attribute subset of genes, each gene in the skin attribute subset being associated with a biochemical pathway relating to the skin attribute that is shown in the comparing step to have been regulated in a more youthful direction for that biochemical pathway and skin attribute; exposing a second sample of human skin tissue to the agent; determining the levels of expression for the at least one skin attribute subset of genes in the second sample of human skin tissue using a method for determining expression levels that is different than that used for the first sample of human skin tissue; selecting a third subset of genes from the at least one skin attribute subset of genes whose expression levels in the second sample of human skin tissue meet a second, selected biological relevance level and whose direction of regulation conforms to the more youthful direction used in selecting the at least one skin attribute subset of genes; and comparing the third subset of genes to a previously determined third subset of genes for a second agent, thereby showing the efficacy of the skin anti-aging agent. - View Dependent Claims (18, 19)
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20. A method of formulating a composition with a plurality of skin anti-aging agents, comprising:
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assessing the efficacy of each of two or more skin anti-aging agents by a method comprising; exposing a first sample of human skin tissue to an agent; determining a first set of expression levels of a plurality of genes in the first sample of human skin; comparing the first set of expression levels to a second set of expression levels, the second set of expression levels corresponding to expression levels of human skin tissue not exposed to the agent, to identify a first subset of genes having a fold change difference in expression level between the exposed and unexposed samples that meets a first, selected biological relevance level; selecting from the first subset of genes a second subset of genes, each gene being associated with a biochemical pathway associated with physical appearance of skin aging; selecting from the second subset of genes, at least one skin attribute subset of genes, each gene in the skin attribute subset being associated with a biochemical pathway relating to the skin attribute that is shown in the comparing step to have been regulated in a more youthful direction for that biochemical pathway and skin attribute; exposing a second sample of human skin tissue to the agent; determining the levels of expression for the at least one skin attribute subset of genes in the second sample of human skin tissue using a method for determining expression levels that is different than that used for the first sample of human skin tissue; selecting a third subset of genes from the at least one skin attribute subset of genes whose expression levels in the second sample of human skin tissue meet a second, selected biological relevance level and whose direction of regulation conforms to the more youthful direction used in selecting the at least one skin attribute subset of genes; and comparing the third subset of genes to a previously determined third subset of genes for a second agent, thereby showing the efficacy of the skin anti-aging agent; selecting from the two or more skin anti-aging agents assessed two or more agents found to have efficacy for at least one skin attribute; and formulating a composition with such two or more agents found to have efficacy as active ingredients and a pharmaceutically acceptable carrier. - View Dependent Claims (21, 22)
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Specification