# US 2016/0355809 A1 · MicroRNA-Based Methods and Assays for Osteosarcoma

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- Number: US 2016/0355809 A1
- Document ID: US20160355809A1
- Status: Published application
- Kind: A1
- Application number: 15/096,480
- Filed: 2016-04-12
- Published: 2016-12-08
- Earliest claimed: 2011-09-07
- Assignees: 3-D Matrix, Ltd.; National Cancer Center
- Applicants: 3-D Matrix, Ltd.; National Cancer Center
- Inventors: Takahiro Ochiya; Tomohiro Fujiwara
- Web page: https://patentgrep.com/patent/US20160355809A1

## CPC

- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- A61B 5/4504: {Bones (A61B5/4547 takes precedence)}
- A61B 6/032: {Transmission computed tomography [CT]}
- A61B 6/505: {for diagnosis of bone}
- A61B 10/025: {for taking bone, bone marrow or cartilage samples}
- A61P 35/00: Antineoplastic agents
- C12Q 1/6886: for cancer (immunoassay for cancer G01N33/575)
- G01N 33/5011: {for testing antineoplastic activity}
- C12N 2310/113: targeting other non-coding nucleic acids, e.g. antagomirs
- C12N 2310/141: MicroRNAs, miRNAs
- C12N 2310/3231: having an additional ring, e.g. LNA, ENA
- C12N 2320/11: for the determination of target sites, i.e. of active nucleic acids
- C12N 2320/31: Combination therapy
- C12Q 2600/178: miRNA, siRNA or ncRNA
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Provided are methods and compositions useful in the diagnosis, treatment, and monitoring of osteosarcoma. Antisense to certain microRNA (miRNA) found to be associated with cancer stem cells (CSCs) or tumor-initiating cells (TICs) of osteosarcoma are useful to suppress tumor growth and metastasis, and prolong survival. Antisense oligonucleotides to miR-133a are synergistic in combination with standard chemotherapy such as cisplatin in the treatment of osteosarcoma.

## Claims

### Claim at position 1

1-21. (canceled)

### Claim 22

A method of screening for osteosarcoma, comprising:
performing on a tissue sample from a subject an assay specifically capable of detecting at least one microRNA (miRNA) selected from the group consisting of miR-1, miR-10b, and miR-133a, wherein detection by the assay of the presence in the sample of the at least one miRNA indicates the subject is at risk of having osteosarcoma.

### Claim 23 (depends on 22)

The method of claim 22, wherein the tissue is blood.

### Claim 24 (depends on 22)

The method of claim 22, wherein the tissue is serum.

### Claim at position 5

25-27. (canceled)

### Claim 28 (depends on 22)

The method of claim 22, wherein the miRNA is miR-1.

### Claim 29 (depends on 22)

The method of claim 22, wherein the miRNA is miR-10b.

### Claim 30 (depends on 22)

The method of claim 22, wherein the miRNA is miR-133a.

### Claim 31 (depends on 22)

The method of claim 22, wherein the assay comprises RT-PCR using oligonucleotide primers suitable for reverse transcribing and amplifying at least one miRNA selected from the group consisting of miR-1, miR-10b, and miR-133a.

### Claim 32 (depends on 31)

The method of claim 31, wherein the miRNA is miR-1.

### Claim 33 (depends on 31)

The method of claim 31, wherein the miRNA is miR-10b.

### Claim 34 (depends on 31)

The method of claim 31, wherein the miRNA is miR-133a.

### Claim 35 (depends on 22)

The method of claim 22, further comprising the step of verifying the presence of osteosarcoma in the subject when the subject is determined to be at risk of having osteosarcoma.

### Claim 36 (depends on 35)

The method of claim 35, wherein verifying the presence of osteosarcoma in the subject comprises performing on the subject a diagnostic procedure selected from the group consisting of skeletal survey or specific bone imaging analysis using X-rays or other suitable bone imaging technique, magnetic resonance imaging (MRI), computed tomography (CT), biopsy, and any combination thereof.

### Claim 37 (depends on 35)

The method of claim 35, wherein the miRNA is miR-1.

### Claim 38 (depends on 35)

The method of claim 35, wherein the miRNA is miR-10b.

### Claim 39 (depends on 35)

The method of claim 35, wherein the miRNA is miR-133a.

### Claim 40 (depends on 35)

The method of claim 35, further comprising the step of treating the subject for osteosarcoma.

### Claim 41 (depends on 22)

The method of claim 22, further comprising the step of treating the subject for osteosarcoma.