Method for detecting radiation exposure
First Claim
1. A method of identifying cells that have been exposed to radiation induced biological stress, comprising:
- exposing a probe set to a labeled nucleic acid composition from a test cell which specifically hybridizes to members of the probe set when the cell has been exposed to an amount of ionizing radiation sufficient to cause differential expression of stress genes in the cell; and
determining whether the nucleic acid composition hybridizes to the nucleic acid molecules representing genes that are differentially expressed.
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Abstract
A method is disclosed for detecting exposure of organisms to biologically significant or hazardous amounts of ionizing radiation. The method uses nucleic acid microarray hybridization to evaluate biological effects, such as patterns of expression of genes after radiation exposure. Numerous genes are provided which have been found to be responsive to radiation exposure in a variety of cell lines. These genes are incorporated into probe sets, which are exposed to a labeled nucleic acid composition from a test cell, such as cDNA reverse transcribed from mRNA in the test cell, which specifically hybridizes to members of the probe set when the cell has been exposed to a biologically significant amount of ionizing radiation. Whether the nucleic acid composition hybridizes to the nucleic acid molecules representing genes that are differentially expressed is determined. The invention also includes methods for determining a dose response relationship between radiation exposure and differential expression of one or more genes, for example to determine a probable radiation dose in cells that have actually or potentially been exposed to the ionizing radiation. The invention also includes probe sets and microarrays used in this method.
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Citations
74 Claims
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1. A method of identifying cells that have been exposed to radiation induced biological stress, comprising:
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exposing a probe set to a labeled nucleic acid composition from a test cell which specifically hybridizes to members of the probe set when the cell has been exposed to an amount of ionizing radiation sufficient to cause differential expression of stress genes in the cell; and
determining whether the nucleic acid composition hybridizes to the nucleic acid molecules representing genes that are differentially expressed. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 33, 34, 35, 36, 37, 38, 73, 74)
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14-20. -20. (canceled)
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21. A probe set, comprising at least 10% of the probes identified in any one of Tables 9, 10, 11, 12, 13 or 14.
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22-23. -23. (canceled)
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24. A method of identifying cells that have been exposed to radiation induced biological stress, comprising:
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providing a plurality of probe elements bound to a surface, each probe element comprising a nucleic acid that is differentially expressed by a cell following ionizing radiation induced biological stress;
contacting the plurality of probe elements with a plurality of test nucleic acid sequences obtained from a test cell, under conditions that allow the test nucleic acid sequences to specifically hybridize to one or more of the probe elements, and provide a signal which indicates differential expression of one or more genes if the cell has been exposed to an amount of ionizing radiation sufficient to cause differential expression of stress genes in the cell; and
detecting the presence or absence of the signal. - View Dependent Claims (25, 26, 27, 28, 29, 30, 39, 40, 41, 43, 44, 45, 46, 47)
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31-32. -32. (canceled)
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42. (canceled)
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48. (canceled)
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49. A method of making a microarray for identifying cells that have actually or potentially been exposed to an amount of ionizing radiation sufficient to cause differential expression of stress genes in the cell, comprising:
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identifying genes that are differentially expressed by a cell following exposure to the ionizing radiation; and
providing a plurality of probe elements bound to a substrate surface, the probe elements comprising probe nucleic acids from genes that are identified as differentially expressed by a cell following ionizing radiation induced biological stress, and wherein the probe nucleic acid is capable of hybridizing to a nucleic acid differentially expressed by the cell following exposure of the cell to the ionizing radiation. - View Dependent Claims (50, 51, 52, 58)
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53. A method of diagnosing ionizing radiation exposure in a subject, comprising:
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obtaining a biological sample from the subject;
synthesizing cDNA from mRNA expressed in one or more cells of the biological sample, and labeling the cDNA with a detectable label;
exposing the labeled cDNA to a probe set which represents genes that are differentially expressed in the biological sample following exposure to ionizing radiation, and determining if the labeled cDNA selectively hybridizes to one or more probes of the probe set that are associated with the radiation exposure. - View Dependent Claims (54, 62)
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55-57. -57. (canceled)
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59-61. -61. (canceled)
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63. (canceled)
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64. A method for measuring a biological response to in a subject comprising:
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obtaining a biological sample from the subject;
synthesizing cDNA from mRNA expressed in one or more cells of the biological sample, labeling the cDNA with a detectable label; and
exposing the labeled cDNA to a probe set; and
determining if the labeled cDNA selectively hybridizes to one or more probes of the probe set differentially expressed following exposure to ionizing radiation. - View Dependent Claims (65, 67, 70, 71, 72)
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66. (canceled)
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68-69. -69. (canceled)
Specification