# US 9,434,995 B2 · Breast cancer biomarker signatures for invasiveness and prognosis > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,434,995 B2 - Document ID: US9434995B2 - Status: Granted - Kind: B2 - Application number: 14/483,232 - Filed: 2014-09-11 - Published: 2014-12-25 - Granted: 2016-09-06 - Earliest claimed: 2012-01-20 - Assignees: The Ohio State University - Applicants: The Ohio State University - Inventors: Carlo M. Croce; Stefano Volinia - Web page: https://patentgrep.com/patent/US9434995B2 ## CPC - C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575) - G06F 16/2468: {Fuzzy queries} - C12Q 2600/112: Disease subtyping, staging or classification - C12Q 2600/118: Prognosis of disease development - C12Q 2600/158: Expression markers - C12Q 2600/178: miRNA, siRNA or ncRNA - 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 - G06F: ELECTRIC DIGITAL DATA PROCESSING (computer systems based on specific computational models G06N) ## Abstract MicroRNA profiles transition from normal breast to ductal carcinoma in situ and transition to invasive ductal carcinoma (IDC) and methods of use thereof are described. Methods of diagnosis and prognosis using microRNA signatures to differentiate invasive from in situ carcinoma are described. Also described is the use of microRNA expression for predicting overall survival and time to metastasis. ## Claims ### Claim 1 A method of diagnosing whether a subject has breast invasive ductal carcinoma (IDC) as compared to ductal carcinoma in situ (DCIS), comprising: a) reverse transcribing RNA from a test sample obtained from the subject to provide a set of target oligodeoxynucleotides wherein the subject has breast IDC; b) hybridizing the target oligodeoxynucleotides to a microarray comprising miR-210, let-7d and miR-221 specific probe oligonucleotides to provide a hybridization profile for the test sample; and c) comparing the test sample hybridization profile to a hybridization profile generated from a control sample, wherein an increase in the expression of miR-210, let-7d and miR-221 is indicative of the subject having IDC as compared to ductal carcinoma in situ (DCIS). ### Claim 2 (depends on 1) The method of claim 1, wherein step c) comprises comparing the test sample hybridization profile to a database, statistics, or table of miR levels associated with non-cancerous samples. ### Claim 3 (depends on 1) The method of claim 1, wherein a level of expression of miR-210, let-7d and miR-221 is assessed by detecting the presence of a transcribed polynucleotide or portion thereof, wherein the transcribed polynucleotide comprises a coding region of miR-210, let-7d and miR-221. ### Claim 4 (depends on 1) The method of claim 1, wherein the sample comprises cells obtained from the subject taken over time. ### Claim 5 A method of determining disease progression of breast invasive ductal carcinoma (IDC) in a subject as compared to ductal carcinoma in situ (DCIS), comprising: a) identifying the relative miR-210 expression compared to a control by: i) extracting a test sample of tissue from a human subject, wherein the extracting is by hypodermic needle, microdissection, or laser capture; and ii) measuring by microarray analysis the level of a miRNA/mRNA signature in the test sample of tissue from the human subject, the miRNA/mRNA signature consisting of miR-210, let-7d and miR-221; and, b) determining the disease progression as IDC in the subject if the subject has increased miR-210, let-7d and miR-221 expression compared to the control; or, determining no disease progression if the subject does not have increased miR-210, let-7d and miR-221 expression compared to the control. ### Claim 6 (depends on 5) A method of claim 5, which further comprises designing a treatment plan based on the diagnosis. ### Claim 7 (depends on 5) A method of claim 5, which further comprises administration of a treatment based on the diagnosis. ### Claim 8 (depends on 5) A method of claim 5, which further comprises determining prognosis based on the diagnosis. ### Claim 9 A method for determining the likelihood of breast cancer progression in a human subject having breast cancer, comprising: a) determining the expression level of a signature of hsa-miR-210, hsa-let-7d and hsa-miR-221 in a sample containing breast cancer cells from the subject with breast cancer, by assaying by microarray analysis a nucleic acid sample obtained from the breast cancer cells to determine the expression level of the miRNA signature in the nucleic acid sample; b) comparing the expression level from step a) to a standard miRNA expression level in a control sample, and determining that the subject has a poor survival outcome, if there is an increase in the expression levels of miRNA signature in the nucleic acid sample, as compared to a control sample. ### Claim 10 (depends on 9) The method of claim 9, wherein the control sample comprises tissue from a representative individual or pool of individuals with breast cancer wherein the breast cancer has not progressed. ### Claim 11 (depends on 9) The method of claim 9, wherein the control sample comprises tissue from the subject taken at an earlier point in time, as compared to the time of determining the expression level of step a). ### Claim 12 (depends on 9) The method of claim 9, wherein the standard miRNA expression level is from the representative pool of individuals and is a mean, median or other statistically manipulated or otherwise summarized or aggregated representative miRNA expression level for the miRNA level in the control tissues in the subject. ### Claim 13 (depends on 9) The method of claim 9, wherein the expression level of one or more of: miR-10b, miR-126, miR-143, miR-218 and miR-335-5p, is also measured relative to the expression level in the control sample, and wherein a decreased expression level of one or more of: miR-10b, miR-126, miR-143, miR-218 and miR-335-5p correlates with a higher risk of progression.