# US 12,180,547 B2 · Detection of expression of markers useful for predicting risk of catastrophic injuries in athletic animals > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,180,547 B2 - Document ID: US12180547B2 - Status: Granted - Kind: B2 - Application number: 17/198,897 - Filed: 2021-03-11 - Published: 2021-09-16 - Granted: 2024-12-31 - Earliest claimed: 2020-03-11 - Assignees: University of Kentucky Research Foundation - Applicants: University of Kentucky Research Foundation - Inventors: Allen E. Page; David W. Horohov - Web page: https://patentgrep.com/patent/US12180547B2 ## CPC - C12Q 1/6883: for diseases caused by alterations of genetic material - C12Q 2600/124: Animal traits, i.e. production traits, including athletic performance or the like - C12Q 2600/156: Polymorphic or mutational markers - C12Q 2600/158: Expression markers - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract The present disclosure relates to methods of detecting gene expression in a biological sample from an animal. More specifically, this disclosure relates to methods of detecting risk for a catastrophic injury in an animal, such as a non-human athletic animal, based on the expression of select genes within a biological sample from the animal, which can be determined using mRNA expression analysis. ## Claims ### Claim 1 A method of detecting biomarker expression indicative of increased risk for catastrophic injury in a non-human athletic animal, comprising: (a) obtaining a blood sample from a subject horse; (b) detecting in the blood sample from the subject horse the mRNA expression level of at least one of interleukin 1 receptor antagonist (IL1RN), insulin-like growth factor (IGF-1), and matrix metallopeptidase 2 (MMP2); and (c) detecting at least one of (i) a decreased mRNA expression level of IL1RN in the blood sample obtained from the subject horse relative to a control expression level of IL1RN from one or more control blood samples obtained from one or more control horses without catastrophic injury, (ii) an increased mRNA expression level of IGF-1 in the blood sample obtained from the subject horse relative to a control expression level of IGF-1 from the one or more control blood samples obtained from the one or more control horses without catastrophic injury, and (iii) an increased mRNA expression level of MMP2 in the blood sample obtained from the subject horse relative to a control expression level of MMP2 from the one or more control blood samples obtained from the one or more control horses without catastrophic injury; (d) identifying the subject horse as being at risk for catastrophic injury based on the detection of the at least one of the decreased mRNA expression level of IL1RN in the blood sample obtained from the subject horse, the increased mRNA expression level of IGF-1 in the blood sample obtained from the subject horse, and the increased mRNA expression level of MMP2 in the blood sample obtained from the subject horse; and (e) treating the subject horse subsequent to being identified as at risk for catastrophic injury; wherein treating the subject horse includes implementing advanced diagnostics to localize potential injury locations. ### Claim 2 A method for detecting biomarker expression indicative of increased risk for catastrophic injury in a non-human athletic animal, comprising: (a) obtaining a first blood sample from a subject horse; (b) detecting in the first blood sample from the subject horse the mRNA expression level of at least one of interleukin 1 receptor antagonist (IL1RN), insulin-like growth factor (IGF-1), and matrix metallopeptidase 2 (MMP2); (c) obtaining a second blood sample from the subject horse at a time subsequent to when the first blood sample was obtained; and (d) detecting in the second blood sample the mRNA expression level of at least one of IL1RN, IGF-1, and MMP2; and (e) detecting at least one of (i) a decreased level of IL1RN in the second blood sample obtained from the subject horse relative to the expression level of IL1RN in the first blood sample, (ii) an increased level of IGF-1 in the second blood sample obtained from the subject horse relative to the expression level of IGF-1 in the first blood sample, and (iii) an increased level of MMP2 in the second blood sample obtained from the subject horse relative to the expression level of MMP2 in the first blood sample; (f) identifying the subject horse as being at risk for catastrophic injury based on the detection of the at least one of the decreased mRNA expression level of IL1RN in the second blood sample obtained from the subject horse, the increased mRNA expression level of IGF-1 in the second blood sample obtained from the subject horse, and the increased mRNA expression level of MMP2 in the second blood sample obtained from the subject horse; (g) treating the subject horse subsequent to being identified as at risk for catastrophic injury; wherein treating the subject horse includes implementing advanced diagnostics to localize potential injury locations. ### Claim 3 (depends on 1) The method of claim 1, wherein the blood sample from the subject horse is from whole peripheral blood. ### Claim 4 (depends on 3) The method of claim 3, wherein the blood sample from the subject horse is plasma or serum from the whole peripheral blood. ### Claim 5 (depends on 3) The method of claim 3, wherein the blood sample from the subject horse is a buffy coat fraction of the whole peripheral blood. ### Claim 6 (depends on 1) The method of claim 1, and further comprising extracting mRNA from the blood sample from the subject horse. ### Claim 7 (depends on 6) The method of claim 6, and further comprising measuring in the extracted mRNA the levels of mRNA corresponding to the at least one of IL1RN, IGF-1, and MMP2. ### Claim 8 (depends on 7) The method of claim 7, wherein quantitative polymerase chain reaction (qPCR) is used to measure the mRNA by measuring cDNA of the mRNA. ### Claim 9 (depends on 1) The method of claim 1, wherein an increased expression level of at least one of IGF-1 and MMP2 in the blood sample from the subject animal is detected relative to the control expression level of a corresponding gene of the one or more control blood samples obtained from the one or more control animals without catastrophic injury. ### Claim 10 (depends on 2) The method of claim 2, wherein an increased mRNA expression level of at least one of IGF-1 and MMP2 in the first blood sample is detected relative to the mRNA expression level of an mRNA expression level of a corresponding gene in the second blood sample.