Recurrent spontaneous abortion biomarker and applications thereof

A technology of recurrent miscarriage and biomarkers, applied in the field of biomedicine, can solve problems such as no reports

Pending Publication Date: 2020-08-28
THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

What causes the abnormal expression of circRNA in RSA, and how the abnormally expressed circRNA participates in the occurrence and development of RSA has not been reported so far

Method used

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  • Recurrent spontaneous abortion biomarker and applications thereof
  • Recurrent spontaneous abortion biomarker and applications thereof
  • Recurrent spontaneous abortion biomarker and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Expression and regulation of hsa_circRNA_100737 in normal early pregnancy villi and recurrent abortion villi

[0032] 1. Experimental method

[0033] 1.1 Extraction of RNA from HTR8 / SVneo and villus tissue

[0034] 1) Pour the collected villous tissue into a petri dish filled with sterile PBS, use ophthalmic forceps to remove blood clots, chorion and thick villi as much as possible, and collect villus columns. Weigh the collected villous tissue, add 1ml Triozol for every 25mg, fully homogenize in a tissue homogenizer for 2min, centrifuge at 4°C, 4000rpm×10min, take the supernatant into a clean 1.5ml EP tube, and proceed to step 3 .

[0035] 2) Aspirate the supernatant of the treated HTR8 / SVneo in the 6-well plate / 12-well plate, wash gently with PBS 3 times, and remove excess liquid. Add 1ml Triozol, pipette gently for more than 10 minutes to fully lyse the cells, then transfer the liquid to a 1.5ml EP tube, and proceed to step 3.

[0036] 3) Add 200 ul of ...

Embodiment 2

[0122] Example 2 Hsa_circRNA_100737 negatively regulates the biological behavior of trophoblast cells through the miR-28-5p-IL-34 axis

[0123] 1. Experimental method

[0124] The hsa_circRNA_100737-miR-28-5p-IL-34 interaction network in villi was predicted by database and bioinformatics analysis. Normal early pregnancy villi and RSA villi were collected, and the expression levels of miR-28-5p, IL-34, and IL-34 receptor (CSF1R) were detected by RT-qPCR and Western Blot. Dual luciferase experiments verified the interaction between hsa_circRNA_100737 and miR-28-5p, miR-28-5p and IL-34. FISH experiments detected the co-localization of hsa_circRNA_100737 and miR-28-5p on HTR8 / SVneo and villus tissues. Interference or overexpression of hsa_circRNA_100737-miR-28-5p-IL-34 axis layer by layer in HTR8 / SVneo cells, flow cytometry to detect the expression of MMP9 and ki67, Transwell invasion assay to detect the invasion ability of HTR8, EdU cell proliferation assay The proliferative a...

Embodiment 3

[0135] Example 3 Hsa_circRNA_100737-miR-28-5p-IL-34 regulates trophoblast function and affects maternal-fetal immune tolerance

[0136] 1. Experimental method

[0137] Direct treatment of decidua CD4 with exogenous IL-34 + T cells, CD8 + T cells, macrophages, or HTR8 / SVneo supernatants treated with IL-34 and decidua CD4 + T cells, CD8 +T cells and macrophages were co-cultured; the levels of transcription factors, cytokines and functional molecules in each group of cells were detected by flow cytometry.

[0138] Up-regulate or down-regulate the hsa_circRNA_100737-miR-28-5p-IL-34 axis in HTR8 / SVneo, and then take its supernatant and decidua CD4 + T cells, CD8 + T cells and macrophages were co-cultured, and the levels of transcription factors, cytokines and functional molecules of immune cells were detected by flow cytometry.

[0139] 2. Results

[0140] IL-34 directly processes decidua CD4 + T cells, CD8 + T cells, macrophages, and T cell phenotype and function did not ...

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Abstract

The invention discloses a recurrent spontaneous abortion biomarker and applications thereof, and belongs to the technical field of biomedicine. The circular RNA used for diagnosing recurrent spontaneous abortion is hsa_circRNA_100737, can be highly expressed in human RSA villi tissue in early pregnancy, can regulate the expression of IL-34 through miR-28-5p and can influence the proliferation, invasion and tube-forming functions of trophoblast through a miR-28-5p-IL-34 axis. The differential expression of the hsa_circRNA_100737 in normal early pregnancy and RSA villi tissue is taken as a breakthrough point to discuss the influences of the hsa_circRNA_100737 on the biological behavior of the trophoblast and mechanisms and to research whether the function of the hsa_circRNA_100737 of regulating the trophoblast can influence maternal-fetal immune tolerance, so that the pathogenesis of RSA can be hopefully analyzed, and new ideas can be provided for the prevention and treatment of diseasessuch as the RSA.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a recurrent abortion biomarker and its application. Background technique [0002] Recurrent spontaneous abortion (RSA) refers to three or more fetal losses before 28 weeks of pregnancy. The causes of RSA are complex and diverse, and the risk of RSA increases with the increase in the number of miscarriages, which has an impact that cannot be underestimated on the health of mothers and infants, family and social harmony. The etiology of RSA is complex, including genetic factors, anatomical abnormalities, immune function abnormalities, infection factors, prethrombotic state, and social and psychological factors. Even with the rapid development of assisted reproductive technology today, there is still a lack of effective prevention and treatment strategies. The prevention and treatment of RSA has become an urgent problem in the field of reproductive medicine, and the analysis of the mole...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113A61K45/00A61P15/06
CPCC12Q1/6883A61K45/00A61P15/06C12Q2600/158C12Q2600/178
Inventor 杜美蓉王松存李梦蝶
Owner THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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