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Methods for diagnosing, prognosing and therapeutically monitoring neurological, neurodegenerative and inflammatory based diseases comprising microRNA-based microglia microvesicles

一种神经胶质细胞、微泡的技术，应用在生物化学设备和方法、DNA / RNA片段、重组DNA技术等方向，能够解决不清楚分子释放、小神经胶质细胞和神经元通讯机制未明确阐明等问题

Active Publication Date: 2022-05-10
BRAINDTECH SRL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Despite evidence that chronic inflammation may influence the pathogenesis of degenerative diseases, the mechanisms of communication between microglia and neurons have not been clearly elucidated, and it remains unclear which molecules are released by these cells and how the damage occurs. Occurs at the neuron level

Method used

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  • Methods for diagnosing, prognosing and therapeutically monitoring neurological, neurodegenerative and inflammatory based diseases comprising microRNA-based microglia microvesicles
  • Methods for diagnosing, prognosing and therapeutically monitoring neurological, neurodegenerative and inflammatory based diseases comprising microRNA-based microglia microvesicles
  • Methods for diagnosing, prognosing and therapeutically monitoring neurological, neurodegenerative and inflammatory based diseases comprising microRNA-based microglia microvesicles

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Experimental program
Comparison scheme
Effect test

Embodiment

[0096] Materials and methods

[0097] Microglial cell culture

[0098] Microglial cells were obtained from mixed glial cell cultures of P2 postnatal CD1 mice (Harlan Laboratories). In the absence of Ca ++ and Mg ++ Cortices were isolated in ice-cold balanced salt solution (HBSS). Brains were collected and meninges were manually removed under a dissecting microscope under a sterile hood. The tissue was then minced with a scalpel. All these manipulations were performed at 4°C.

[0099] Place the tissue fragments in a HBSS solution with 2.5 mg / mL trypsin, 0.2 mg / mL EDTA, 1 mg / mL glucose and 0.1 mg / mL bovine pancreatic DNase I at 37 °C under constant gentle shaking in a water bath incubator Incubate for 20-30 minutes.

[0100] After culturing, a fresh medium (DMEM / F12 (3:1)) containing 20% ​​heat-inactivated fetal calf serum, 100 U / mL penicillin-streptomycin was added, and the suspension was centrifuged at 1500 rpm for 5 minute. Finally, a single cell suspension was o...

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Abstract

The present invention describes a method for in vitro diagnosis, prognosis and / or therapeutic monitoring based on neurodegenerative, neurological and inflammatory diseases, wherein said method comprises the steps of: a) isolating from a biological fluid obtained from an individual microglial microvesicles (MVs); b) collecting the microRNAs (miRNAs) contained in the MVs; c) determining the expression profile of a predetermined set of miRNAs; d) comparing the expression profile to one or more Comparison of said determined expression profile with said one or more reference expression profiles allows diagnosis, prognosis and / or therapeutic monitoring of said disease.

Description

technical field [0001] The present invention describes a method for in vitro diagnosis, prognosis and / or therapeutic monitoring of neurodegenerative, neurological and inflammation-based diseases, wherein said method comprises the following steps: [0002] a) isolating microglial microvesicles (MVs) from a biological fluid obtained from an individual; [0003] b) collecting said microRNA (miRNA) contained in said MV; [0004] c) determining an expression profile of a predetermined set of miRNAs; [0005] d) comparing said expression profile to one or more reference expression profiles, [0006] Wherein the comparison of said determined expression profile with said one or more reference expression profiles allows for diagnosis, prognosis and / or treatment monitoring of said disease. Background technique [0007] Emerging evidence suggests that inflammation underlies many CNS diseases, including chronic neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12N15/113C12Q2600/178C12Q2600/158C12N2310/141C12Q1/6886
Inventor 法比奥·比安科诺埃米·汤纳
Owner BRAINDTECH SRL
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