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Application of micro RNA group related to Th17 differentiation to preparation of drugs for treatment and effect judgment

1. The technology of microrna4426 and arthritis, which is applied in the field of diagnostic reagents for judging the clinical curative effect of hydroxychloroquine, achieves a broad prospect of promotion

Inactive Publication Date: 2016-08-24
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, regarding the micro RNA related to Th17 differentiation of the present invention, and its application as a target in the diagnosis and / or treatment of autoimmune diseases mediated by IL-17, and as a diagnostic reagent for judging the clinical efficacy of hydroxychloroquine, There is no relevant report yet

Method used

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  • Application of micro RNA group related to Th17 differentiation to preparation of drugs for treatment and effect judgment
  • Application of micro RNA group related to Th17 differentiation to preparation of drugs for treatment and effect judgment
  • Application of micro RNA group related to Th17 differentiation to preparation of drugs for treatment and effect judgment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Effect of Hydroxychloroquine on Th17 Differentiation of Naive T Cells

[0034] Sorting peripheral blood mononuclear cells from healthy people's peripheral blood, and further sorting CD4 by flow cytometry + Naive T cells were cultured in vitro. On day 0, the sorted naive CD4 + T cells were resuspended in RPMI 1640 medium containing 10% fetal bovine serum, and anti-CD3 / CD28 activated magnetic beads (5 μl / ml), IL-1β (50ng / ml), TGF-β (1ng / ml), IL-6(30ng / ml), IL-23(30ng / ml), anti-IL-4(5ng / ml), anti-IL-12(5ng / ml) and anti-IL-2(20U / ml) Differentiation was induced; on the 6th and 9th days, the medium was changed in half, and the corresponding cytokines were supplemented; on the 12th day, the cells were harvested. In the hydroxychloroquine intervention group, 20 μM hydroxychloroquine was added on the 0th day of culture, and hydroxychloroquine was supplemented when half the medium was changed. The results suggest that naive T cells can be effectively induced to diff...

Embodiment 2

[0035] Example 2 Hydroxychloroquine Time-Dependent Inhibition of Th17 Differentiation of Naive T Cells

[0036] On day 0, the sorted naive CD4 + T cells were resuspended in RPMI1640 medium containing 10% fetal bovine serum, and anti-CD3 / CD28 activated magnetic beads (5 μl / ml), IL-1β (50ng / ml), TGF-β (1ng / ml), IL -6 (30ng / ml), IL-23 (30ng / ml), anti-IL-4 (5ng / ml), anti-IL-12 (5ng / ml) and anti-IL-2 (20U / ml) Differentiation was induced; on the 6th and 12th day, the cells were harvested respectively, and half of the medium was changed on the 6th day in the 12-day group, and the corresponding cytokines and hydroxychloroquine were supplemented. The results suggest that cytokines can induce Th17 differentiation in a time-dependent manner, and hydroxychloroquine can inhibit Th17 differentiation in a time-dependent manner. The result is as figure 2 shown. This figure illustrates that hydroxychloroquine can time-dependently inhibit the conversion of naive T cells to Th17.

Embodiment 3

[0037] Example 3 Differentially expressed genes of naive T cells induced to differentiate into Th17

[0038] Sorting peripheral blood mononuclear cells from healthy people's peripheral blood, and further sorting CD4 by flow cytometry + Naive T cells were cultured in vitro. On day 0, the sorted naive CD4 + T cells were resuspended in RPMI 1640 medium containing 10% fetal bovine serum, and anti-CD3 / CD28 activated magnetic beads (5 μl / ml), IL-1β (50ng / ml), TGF-β (1ng / ml), IL-6(30ng / ml), IL-23(30ng / ml), anti-IL-4(5ng / ml), anti-IL-12(5ng / ml) and anti-IL-2(20U / ml) Differentiation was induced; on the 6th and 9th days, the medium was changed in half, and the corresponding cytokines were supplemented; on the 12th day, the cells were harvested. BGI performed RNA-Seq to understand gene expression. The specific steps are to extract the total RNA of the sample and digest the DNA with DNaseI, enrich the mRNA with magnetic beads containing Oligo(dT), add an appropriate amount of fragment...

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Abstract

The invention provides a micro RNA group related to Th17 differentiation. The micro RNA group comprises micro RNA 4426, micro RNA 3615, micro RNA 106b, micro RNA 590 and micro RNA 573. The micro RNA changes remarkably in the Th17 differentiation process and can be used as Th17 control targets for preparing drugs for treating or preventing Th17-related diseases including lupus erythematosus, dermatomyositis, vasculitis, sicca syndromes, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, scleroderma, multiple sclerosis, malaria, solar dermatitis, eczema, leucoderma, psoriasis, atopic dermatitis, lichen planus, diabetes, coronary heart diseases, hyperlipemia and the like. The micro RNA group is sensitive to hydroxychloroquine intervention and can serve as a sensitive index for judgment of the clinical effect of hydroxychloroquine, and the micro RNA is convenient to test clinically and can be rapidly used for judging the clinical effect of hydroxychloroquine.

Description

technical field [0001] The present invention relates to the field of biotechnology, specifically, a group of microRNAs related to Th17 differentiation are used as targets in the diagnosis and / or treatment of autoimmune diseases and chronic inflammatory diseases mediated by IL-17, and as hydroxychloroquine clinical Application in diagnostic reagents for judging curative effect. Background technique [0002] CD4 secreting IL-17 + T cells (Th17) are a group of CD4 + T cell subsets, which mainly secrete interleukin-17A (IL-17A), interleukin-17F (IL-17F), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) and other cytokines mediate the infiltration and damage of inflammatory cells to tissues; at the same time, Th17 cells can also secrete cytotoxic substances to directly damage tissues. A large number of studies have shown that Th17 cells play a role in some autoimmune diseases such as: lupus erythematosus, dermatomyositis, vasculitis, Sjogren's syndrome, scleroderma, r...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/7088A61P37/02A61P29/00A61P17/00A61P21/00A61P9/00A61P19/02A61P33/06A61P3/10A61P3/06A61P9/10C12Q1/68
CPCA61K31/7088A61K45/00C12Q1/6883C12Q2600/118C12Q2600/178Y02A50/30
Inventor 杨骥李明
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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