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MARCH1 gene serving as diagnosis and treatment marker of rheumatoid arthritis and osteoarthritis

A technology for rheumatoid and osteoarthritis, applied in the field of diagnosis with the detection of MARCH1 abnormality as a means, can solve the problems of difficult diagnosis and difficult identification of clinical features.

Active Publication Date: 2018-09-21
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In primary care, the diagnosis of early rheumatoid arthritis is as difficult as other musculoskeletal problems: clinical features are poorly discernible, inflammatory markers such as erythrocyte sedimentation rate and C-reactive protein are not characteristic, and specific markers such as rheumatoid factor ( Seronegative in 31% of patients) and anti-cyclic citrullinated peptide antibodies (anti-CCP, seronegative in 33% of patients) were also commonly seronegative
There are no molecular markers for the differential diagnosis of rheumatoid arthritis and osteoarthritis, and the applicant carried out this study under such a background

Method used

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  • MARCH1 gene serving as diagnosis and treatment marker of rheumatoid arthritis and osteoarthritis
  • MARCH1 gene serving as diagnosis and treatment marker of rheumatoid arthritis and osteoarthritis
  • MARCH1 gene serving as diagnosis and treatment marker of rheumatoid arthritis and osteoarthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Screening of differentially expressed genes in osteoarthritis synovial tissue, rheumatoid arthritis synovial tissue and normal synovial tissue

[0075] The synovial tissue of 5 patients with rheumatoid arthritis came from patients with rheumatoid arthritis who underwent knee replacement or synovectomy in the orthopedics department of the hospital. The diagnosis of all cases met the classification criteria of rheumatoid arthritis revised by the American Rheumatology Association in 1987 , including 3 females and 2 males, with an average age of (55±8) years and an average disease duration of (12±7) years. The synovial tissues of 5 patients with osteoarthritis were obtained from osteoarthritis patients undergoing knee joint replacement or synovectomy in the Department of Orthopedics of the hospital. Age (63±5) years old, mean disease duration (12±8) years. 6 cases of normal synovial tissues came from joint synovial tissues of patients undergoing trauma surgery. ...

Embodiment 2

[0122] Example 2 Large sample verification of differentially expressed genes

[0123] Based on the previous high-throughput sequencing results and according to the size of the P value, we selected the MARCH1 gene for verification. The up-regulation was even greater in patients with osteoarthritis.

[0124] 1. Detection of differential expression of the MARCH1 gene at the transcriptional level

[0125] 1. According to the method of Example 1, 100 cases of rheumatoid arthritis synovial tissue, 100 cases of osteoarthritis synovial tissue, and 100 cases of normal synovial tissue were collected.

[0126] 2. Carry out the extraction and quality detection of RNA according to the method of Example 1

[0127] 3. Reverse transcription

[0128] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC wate...

Embodiment 3

[0149] Example 3 MARCH1 gene expression plasmid construction

[0150] 1. Design and synthesis of interference RNA

[0151] According to the MARCH1 gene sequence, siRNA was designed and synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd. Shanghai Gemma Pharmaceutical Technology Co., Ltd. also provides a negative control siRNA (siRNA-NC) that has no sequence homology with the MARCH1 gene.

[0152] siRNA-MARCH1:

[0153] The sense strand is 5'-UGUUGUUUGGAAUUCUGUGAG-3' (SEQ ID NO.5);

[0154] The antisense strand is 5'-CACAGAAUUCCAAACAACACG-3'(SEQ ID NO.6),

[0155] 2. In vitro culture of synovial tissue cells

[0156] After the aseptically obtained synovial tissues of rheumatoid arthritis and osteoarthritis were washed with PBS, they were repeatedly cut into tissue pieces of about 1mm x 1mm x 1mm with sterile surgical scissors, and collagenase II (0.5mg / ml) After digestion at 37°C for 2 hours, filter through a 200-mesh gauze, centrifuge to remove the supernata...

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Abstract

The invention discloses an MARCH1 gene capable of being used as a molecular marker for diagnosing rheumatoid arthritis and / or osteoarthritis. The experiment of the invention proves that compared witha normal person, the expression of the MARCH1 in a rheumatoid arthritis and / or osteoarthritis patient is high. The invention also discloses application of MARCH1 gene to preparation of a drug for treating the rheumatoid arthritis and / or osteoarthritis. A research achievement of the invention provides a novel method for clinically diagnosing the rheumatoid arthritis and / or osteoarthritis, and alsoprovides a novel drug target for treating the rheumatoid arthritis and / or osteoarthritis.

Description

technical field [0001] The present invention relates to the fields of molecular diagnosis and treatment, more specifically, the present invention relates to a diagnostic method by means of detecting MARCH1 abnormality; and a therapeutic agent for inhibiting MARCH1 gene or protein. Background technique [0002] Rheumatoid arthritis is an autoimmune disease characterized by polyarticular involvement that can progress to joint destruction and deformity, often involving peripheral joints. The etiology is unclear, but extraarticular organ involvement such as pulmonary interstitial disease and Sjogren's syndrome are also common. Appropriate early treatment can improve joint symptoms and function, reduce mortality, and reduce complications. [0003] A 2010 systematic review found that the prevalence of rheumatoid arthritis in North America and Northern Europe was estimated to range from 0.5% to 1.1%. The incidence in developing countries is relatively lower (0.1% to 0.5%). Rheum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68A61K45/00A61P19/02A61P19/08A61P29/00
CPCA61K45/00A61P19/02A61P19/08A61P29/00C12Q1/6883C12Q2600/158G01N33/6893
Inventor 肖枫李曙光孙耀兰常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD