# US 10,308,980 B2 · Methods and biomarkers for analysis of colorectal cancer

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- Number: US 10,308,980 B2
- Document ID: US10308980B2
- Status: Granted
- Kind: B2
- Application number: 14/354,310
- Filed: 2012-11-05
- Published: 2014-10-09
- Granted: 2019-06-04
- Earliest claimed: 2011-11-04
- Assignees: Oslo Universitetssykehus Hf
- Applicants: Ragnhild A. Lothe; Trude Holmeide Agesen; Anita Sveen; Guro Elisabeth Lind; Arild Nesbakken; Rolf Inge Skotheim
- Inventors: Ragnhild A. Lothe; Trude Holmeide Agesen; Anita Sveen; Guro Elisabeth Lind; Arild Nesbakken; Rolf Inge Skotheim
- Web page: https://patentgrep.com/patent/US10308980B2

## CPC

- C12Q 1/6813: Hybridisation assays
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 35/00: Antineoplastic agents
- C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575)
- C12Q 2563/159: Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components
- C12Q 2600/118: Prognosis of disease development
- 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
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present invention relates to methods and biomarkers (e.g., gene expression biomarkers) for detection of colorectal cancer in biological samples (e.g., tissue samples, biopsy samples, stool samples, blood samples, plasma samples, serum samples). In some embodiments, methods and biomarkers of the present invention find use in detection of colon cancer and in providing a prognosis to colorectal cancer patients.

## Claims

### Claim 1

A method for treating a human subject that has stage II colorectal cancer, comprising:
a) extracting and reverse transcribing mRNA from a tumor sample from a human subject that has stage II colorectal cancer to provide complementary DNA;
b) detecting the expression level of, SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to, SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said mRNA transcripts;
c) determining that said human subject has altered expression of, SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts,
administering adjuvant chemotherapy to said human subject determined to have altered levels of expression of, SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, as compared to reference levels of expression, wherein said reference level is selected from the group consisting of the level in stage I colorectal cancer, the level in non-cancerous colorectal tissue, and the level in pre-cancerous colorectal tissue.

### Claim 2 (depends on 1)

The method of claim 1, wherein said reference level is determined from multiple samples.

### Claim 3 (depends on 1)

The method of claim 1, further comprising detecting the expression level of PIGR mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said PIGR mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said PIGR mRNA transcripts.

### Claim 4 (depends on 1)

The method of claim 1, further comprising detecting the expression level of CXCL13 mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said CXCL13 mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said CXCL13 mRNA transcripts.

### Claim 5 (depends on 1)

The method of claim 1, further comprising detecting the expression level of MMP3 mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said MMP3 mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said MMP3 mRNA transcripts.

### Claim 6 (depends on 1)

The method of claim 1, further comprising detecting the expression level of TUBA1B mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said TUBA1B mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said TUBA1B mRNA transcripts.

### Claim 7 (depends on 1)

The method of claim 1, further comprising detecting the expression level of DSC3 mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said DSC3 mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said DSC3 mRNA transcripts.

### Claim 8 (depends on 1)

The method of claim 1, further comprising detecting the expression level of CXCL10 mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said CXCL10 mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said CXCL10 mRNA transcripts.

### Claim 9 (depends on 1)

The method of claim 1, further comprising detecting the expression level of ENPP3 mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said ENPP3 mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said ENPP3 mRNA transcripts.

### Claim 10 (depends on 1)

The method of claim 1, further comprising detecting the expression level of BNIP3 mRNA transcripts in addition to said SESN1, AZGP1, KLK6, EPHA7, and SEMA3A, mRNA transcripts, wherein said detection comprises contacting said complementary DNA with primers specific for complementary DNAs corresponding to said BNIP3mRNA transcripts, amplifying said complementary DNAs; and quantifying the expression levels of said BNIP3 mRNA transcripts.
