Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Micro-RNA Associated With Rheumatoid Arthritis

a technology of rheumatoid arthritis and microrna, which is applied in the field of microrna associated with rheumatoid arthritis, can solve the problems of difficult diagnosis of rheumatoid arthritis accurately, insufficient diagnosis method, and development of biological formulations (anti-tnf antibodies), and achieve the effect of suppressing inflammation and growth of ra synovial

Inactive Publication Date: 2011-05-19
KOBE UNIV
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016]The marker for rheumatoid arthritis of the present invention is non-coding RNA whose expression is specifically increased or decreased in rheumatoid arthritis. In particular, mi-RNA (miR-124a) whose expression is suppressed directly acts on mRNAs of cyclin-dependent kinase (CDK) 2, CDK6, and monocyte chemotactic protein-1 (MCP1). The fact has confirmed that the non-coding RNA serving as the marker for rheumatoid arthritis of the present invention is involved in cell cycles and the expression of an inflammation-associated protein. It is conceivable that CDK2, CDK6, and MCP1 act on RA synovial cells through the suppression of the expression of the non-coding RNA serving as the marker for rheumatoid arthritis of the present invention, which may be associated with the pathology of rheumatoid arthritis. An oligonucleotide that forms the non-coding RNA of the present invention regulates the expression of CDK2, CDK6, and MCP1, suppresses the inflammation and growth of RA synovial cells attributed to those proteins, and thus acts as an active ingredient for the treatment of rheumatoid arthritis.

Problems solved by technology

The antibody is said to be an excellent diagnosis method for rheumatoid arthritis as compared to a conventional diagnosis method, but is not a sufficiently satisfactory diagnosis method.
In particular, rheumatoid arthritis is difficult to be diagnosed accurately at the early stage of the development.
However, the cause of such hyperfunctions of synovial cells, which play important roles in forming the pathology of rheumatoid arthritis, has not been sufficiently elucidated yet, and the inhibition of the abnormal growth of synovial cells is conceivable to be extremely important in the treatment of rheumatoid arthritis.
A conventional treatment with an anti-rheumatic drug cannot completely inhibit the progression of arthritis, and there are many cases in each of which the treatment has no effect.
Further, a biological formulation (anti-TNFα antibody), which has been developed in recent years, also has many problems.
For example, there are cases in each of which the biological formulation has no effect or has a reduced effect, and high treatment costs are required.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Micro-RNA Associated With Rheumatoid Arthritis
  • Micro-RNA Associated With Rheumatoid Arthritis
  • Micro-RNA Associated With Rheumatoid Arthritis

Examples

Experimental program
Comparison scheme
Effect test

reference example 1

Expression Profile of miRNA in RA Synovial Cells

[0045]Synovial cells were obtained through the approval of an ethics committee from a patient with RA diagnosed in conformity with the standard of the American College of Rheumatology and a patient with osteoarthritis (OA) as a control who had given informed consent (RA synovial cells (RA-FLS) and OA synovial cells (OA-FLS)). Each of the cells were cultured and maintained in an RPMI medium containing 10% FCS and 4 mM L-glutamine. Small RNAs having 200 bases or less were extracted from the above-mentioned cells (mirVana™miRNA Bioarrays, manufactured by Ambion) to perform miRNA expression analysis. The small RNAs were extracted in accordance with a method known per se. The extracted 156 kinds of miRNAs were subjected to quantification by real-time RT-PCR (TaqMan® MicroRNA Assays manufactured by Applied Biosystems) and a ΔCt method.

[0046]The results confirmed that six kinds of miRNAs different by a factor of two or more in expression amou...

example 1

Expression of miRNA in RA-FLS

[0047]300 pmol of pre-miR-124a or control pre-miRNA (Ambion) were added to 1 mL of an OptiMEM medium (Invitrogen) together with 10 μL of RNAiMAX (Invitrogen). After that, the mixture was added to a plate having a size of 60 mm, and the temperature was kept at room temperature for 20 minutes. After that, 5 ml of an RA-FLS cell suspension (1×105 cells / ml) in an RPMI medium were seeded to the plate, and the mixture was cultured in a CO2 incubator for 24 to 96 hours. Quantification was performed by miR-124a-specific real-time RT-PCR (TaqMan® MicroRNA Assays, manufactured by Applied Biosystems) and a ΔCt method.

[0048]The results confirmed that the expression amount of miR-124a was high as compared to the expression amount of control miRNA in each time of culture as a result of the addition of an equal amount of pre-miRNA (FIG. 2).

experimental example 1

Influences of miR-124a on Growth of Various Cells

[0049]Influences of miR-124a on the growth of various cells were examined.

[0050]In the same manner as in Example 1, an examination was made on influences on the growth of cells in the case where the respective cells, OA-FLS (n=2), RA-FLS (n=4), E11 cells, and HeLa cells were transfected with 300 pmol of pre-miR-124a or control pre-miRNA (Ambion) and cultured in a CO2 incubator for 24 to 96 hours.

[0051]The measurement results of the degree of the growth of cells by an XTT method using a Cell Proliferation Kit II (manufactured by Roche Applied Science) confirmed that the growth of cells was clearly suppressed in RA-FLS, OA-FLS, E11, and HeLa cells, transfected with pre-miR-124a (FIG. 3).

[0052]Also in the case where the number of cells was measured by a trypan blue method, as illustrated in FIG. 4, it was confirmed that, in RA-FLS cells, the growth of cells was clearly suppressed in the cells transfected with pre-miR-124a.

[0053]Further, ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

An object of the present invention is to provide a novel marker for rheumatoid arthritis (RA), and more specifically, to provide a marker whose expression may be specifically increased or decreased in RA. Another object of the present invention is to confirm whether or not miRNA serving as the marker is involved as the etiology of RA, and to provide an inspection method for RA and a therapeutic agent for RA each using the miRNA involved. The marker includes miRNA (for example, miR124a) whose expression is specifically increased or decreased in RA synovial cells based on a small RNA expression profile in the RA synovial cells. In addition, the therapeutic agent for RA includes miRNA (for example, miR124a) as an active ingredient.

Description

TECHNICAL FIELD[0001]The present invention relates to microRNA associated with rheumatoid arthritis (RA), and also relates to an inspection method for rheumatoid arthritis using the microRNA as an indicator and a therapeutic agent for rheumatoid arthritis containing the microRNA as an active ingredient.[0002]The present application claims the priority of JP 2008-174072, which is hereby incorporated by reference.BACKGROUND ART[0003]Rheumatoid arthritis is a chronic, systemic, progressive autoimmune disease of unknown origin, which causes arthralgia (arthrosynovitis). In Japan, about 700,000 or more people suffer from the disease. Rheumatoid arthritis develops in people of any age, and more frequently develops in people who are in their thirties to fifties.[0004]Genetic predisposition and environmental factors are presumably involved in the development of rheumatoid arthritis, but the etiology of rheumatoid arthritis remains unknown. Rheumatoid arthritis is a systemic disease in which...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/68C07H21/02
CPCC12N15/113C12N2310/141C12Q2600/178C12Q1/6883C12Q2600/158C12N2330/10A61P19/02A61P29/00
Inventor KAWANO, SEIJINAKAMACHI, YUJI
Owner KOBE UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com