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Medical uses of exosomes

A technology of exosomes and adult stem cells, which is applied in the field of treatment of diseases or conditions that exhibit or are caused by inflammatory processes, and can solve problems such as increasing serious side effects

Inactive Publication Date: 2019-07-30
VBC HLDG LLC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Anti-inflammatory drugs and pain relievers sometimes have limited long-term effectiveness, and short- and long-term use of these drugs increases the risk of potentially serious side effects

Method used

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  • Medical uses of exosomes
  • Medical uses of exosomes
  • Medical uses of exosomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0166] Isolation of exosomes from synovial fluid by polymer precipitation

[0167] By polymer precipitation (ExoQuick TM , from SBI, Palo Alto, CA) to isolate exosomes. The technology captures and collects exosomes in a "polymer mesh" that is recovered by low-speed centrifugation. Once the exosome pellet was obtained, the supernatant containing excess polymer was removed, and then the pelleted exosomes were resuspended in phosphate-buffered saline (PBS) solution to dissolve the polymer network and release intact exosomes.

Embodiment 2

[0169] Validation of isolated exosomes

[0170] To confirm the functional exosomes isolated by the polymer precipitation method described in Example 1, the identity of the exosomes was verified by measuring the presence of appropriate biomarkers, as described below.

[0171] Exosomes were isolated from the synovial fluid of patients diagnosed with osteoarthritis (n=6) by the method described above, and the exosomes were verified by Western blot using a specific exosomal marker: tetraspanin ( tetraspanins), CD9, CD63, CD81 (located on the surface of exosomes), and proteins expressed by tumor susceptibility gene 101 ("TSG101") present in exosomes.

[0172] The isolated exosomes were added to RIPA buffer supplemented with protease inhibitors (Sigma-Aldrich) (150mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 50mM Lysis was carried out in Tris (Tris, pH 8.0) on ice for 30 minutes. Lysates were quantified for Bradford assay (Bio-Rad) and 25 μg of protein was mix...

Embodiment 3

[0179] Effects of exosomes isolated from synovial fluid on M1 macrophages

[0180] To verify the immunomodulatory properties of synovial fluid-derived exosomes, exosomes were isolated by polymer precipitation as described in Example 1. CD14+ monocytes were purified from healthy donor peripheral blood mononuclear cells (“PBMCs”) using the Human CD14+ Cell Enrichment Kit (StemCell Technologies) by adding 100 ng / ml granulocyte-macrophage colony-stimulating factor (GM-CSF, Peprotech) for 10 days to differentiate into M1 macrophages, then treated with 150 UI / ml interferon (“IFN”) gamma (“γ”) (Peprotech) and 10 ng / ml lipopolysaccharide (LPS) (Sigma-Aldrich ) stimulated for 24 hours in the presence of exosomes, as Figure 4A shown.

[0181] M1 macrophages are pro-inflammatory cells with potent antimicrobial activity that promote T helper cell responses. by using supernatants derived from M1-macrophages Assay (Bio-Rad Laboratories) measures cytokine production in M1 macrophages. ...

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Abstract

The invention provides anti-inflammatory exosomes that, when administered locally or systemically, downregulate an inflammatory process in a subject in need of such treatment, as well as methods of making and using the anti-inflammatory exosomes.

Description

technical field [0001] This document relates generally to anti-inflammatory exosomes, methods of obtaining or producing anti-inflammatory exosomes, and treating subjects exhibiting or caused by an inflammatory process by administering anti-inflammatory exosomes to a subject in need of such treatment The method of causing a disease or condition. Background technique [0002] The Background Art includes information useful for understanding the present invention. It is not an admission that the information presented herein is prior art or relevant to the present invention, or that publications cited directly or indirectly are prior art. [0003] Inflammation is the normal response of the immune system to various types of injury, infection and / or other damage to living tissue. Typically, inflammation results from an acute injury or disease process, and symptoms of inflammation (eg, pain, heat, redness, swelling, and loss of function) are limited in scope and duration. The inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K31/7088C12N5/077C12N5/0775C12N15/113A61K35/28C12N5/0786
CPCA61K31/7088C12N5/0645C12N5/0662C12N5/0669C12N15/113C12N2501/22C12N2501/24C12N2501/25C12N2502/1317C12N2502/1382A61K35/24A61P29/00C12N2310/141A61K45/06A61K2300/00A61K9/0073C12N5/0663C12N5/0667C12N5/0668
Inventor K·尼亚齐F·柯西奥
Owner VBC HLDG LLC
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