COMPOSITION FOR THE DIAGNOSIS, PREVENTION OR TREATMENT OF DISEASES RELATED TO CELLS EXPRESSING IL-8 OR GRO-ALPHA, COMPRISING UCB-MSCs

Abstract

Provided is a gene therapy composition for transferring one of a therapeutical gene, a maker gene, or a mixture thereof to a cell that expresses interleukin-8(IL-8) or GRO-α and induces tropism of mesenchymal stem cells isolated from umbilical cord blood and / or the mesenchymal stem cells expanded from said mesenchymal stem cells (UCB-MSCs), wherein the cell-treating composition includes UCB-MSCs. Provided is a composition for treating disease related to a cell expressing IL-8 or GRO-α, that is, a brain tumor in gene therapy, by using UCB-MSCs. Provided is a composition or kit for diagnosing brain tumors, preventing brain tumors, treating brain tumors, or monitoring brain tumor treatment progression by using UCB-MSCs.

Description

TECHNICAL FIELD[0001]The present invention relates to a gene therapy composition for transferring a therapeutical gene, a marker gene, or a product thereof to a cell that expresses interleukin-8(IL-8) or GRO-α and induces tropism of umbilical cord blood-derived mesenchymal stem cells or mesenchymal stem cells isolated from umbilical cord blood and / or the mesenchymal stem cells expanded from said mesenchymal stem cells (UCB-MSCs), wherein the gene therapy composition includes UCB-MSCs.[0002]The present invention also relates to preventing or treating disease related to a cell expressing IL-8 or GRO-α, or brain tumors in gene therapy, using the composition includes UCB-MSCs.[0003]The present invention also relates to a composition or kit for diagnosing brain tumors, preventing brain tumors, treating brain tumors, or monitoring brain tumor treatment progression, using UCB-MSCs.BACKGROUND ART[0004]It is known that stem cells migrate toward sites of pathology. Recently, it was found that...

AI Technical Summary

Benefits of technology

[0016]Umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) in the composition according to the present invention has a selective tropism for cells that express IL-8 or GRO-α and thus induce the tropism of UCB-MSCs or brain tumor cells. The tropism capability of the UCB-MSCs is better than that of other stem cells and thus, therapeutical genes or a product thereof can be more effectively delivered than when other conventional stem cells are used. Accordingly, a pharmaceutical composition or a kit comprising UCB-MSCs according to the present invention can be used to diagnose, prevent, and treat diseases related to cells expressing interleukin (IL)-8 or GRO-α or brain tumors.MODE OF INVENTIONS

[0017]The inventors of the present invention have studied stem cells having an effective tropism for tumors, and surprisingly found that umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) have a strong tropism for brain tumors, specifically a stronger tropism for brain tumors than bone marrow-derived mesenchymal stem cells (BM-MSCs), which has never been known before. Also, the inventors found that at least one selected from the group consisting of interleukin (IL)-8 and GRO-α relates to the tropism of UCB-MSCs.

[0018]The inventors co-cultured UCB-MSCs and representative tumor cell lines to identify characteristics of the tropism of UCB-MSCs and cytokine related to those tumor cell lines. Specifically, UCB-MSCs, and one selected from human brain tumor cell lines, such as U-87 MG, LN18, U138, or U251 cells; human rectal cancer cell line such as LS-174T; human B lymphocyte such as NC37; mouse's fibroblast (NIH3T3); a gastric cancer cell line such as KATO III; a lung cancer cell line such as A549, and a liver cancer cell line such as PLC / PRF5 were co-cultured in a transwell chamber to measure mobility of UCB-MSCs. As a result, it was found that UCB-MSCs have a strong tropism for U-87 MG, LN18, U138, and U251 cells which are brain tumor cells (see FIGS. 3 and 4). UCB-MSCs also had tropism for a conditioned media that did not include U-87 MG cells and was obtained by culturing U-87 MG (see FIG. 4).

[0019]Also, the tropism for a brain tumor cell line of UCB-MSCs was compared with that of BM-MSCs that is currently used as a source of stem cells. As a result, it was found that UCB-MSCs have a stronger tropism for a brain tumor cell line than BM-MSCs (FIG. 5). Also, the chemotactic index of UCB-MSCs was largest with respect to the brain tumor cell line among various cancer cell lines (FIG. 6). Such a high chemotactic index with respect to the brain tumor cell is an additional advantage of UCB-MSCs, in addition to the fact that UCB-MSCs can be obtained more easily and have more immunological stability than BM-MSCs, proving that UCB-MSCs are a very suitable medium for gene therapy of a brain tumor because a therapeutical gene can be efficiently transferred to the inside or neighboring portions of the brain tumor.

[0020]The tropism of UCB-MSCs in the transwell chamber may be derived by cytokines that are derived to be secreted in the co-culture of two cells. Thus, two cells were co-cultured in a transwell chamber to prepare a medium and then, the medium was analyzed using the cytokine array. As a result, it was identified that high levels of cytokines, such as IL-8 or GRO-α, were secreted in a medium in which UCB-MSCs and U87 MG had been co-cultured (see FIG. 7). Accordingly, it is highly likely that these cytokines may derive a tropism of the UCB-MSCs.

[0021]The inventors of the present invention cultured UCB-MSCs alone, cultured U-87 MG alone, and co-cultured UCB-MSCs and U-87 MG, and then analyzed IL-8 mRNA levels of these cells using RT-PCR. As a result, UCB-MSCs did not express IL-8 in either the presence or absence of U-87 MG cells. However, U-87 MG expressed IL-8 constitutively in both the presence and absence of UCB-MSCs (see FIG. 8). Treatment of UCB-MSCs with IL-8 significantly enhanced its migration when compared to untreated cells (see (A) of FIG. 9). However, when UCB-MSCs were pre-incubated with anti-CXCR1 antibodies that are antibodies with respect to an IL-8 receptor and recombinant IL-8 was applied to UCB-MSCs, IL-8 mediated migration of UCB-MSCs were reduced in a dose-dependent manner by anti-CXCR1 treatment ((B) of FIG. 9). Anti-CXCR2 treatment also showed the same effect. Similarly, GRO-α treatment also enhanced UCB-MSCs migration when compared to untreated UCB-MSCs ((C) of FIG. 9). In contrast, there were no significant differences in UCB-MSCs migration in cultures treated with MCP-1 ((D) of FIG. 9). This data indicates that IL-8 and GRO-α participate in UCB-MSCs migration toward U-87 MG cells.

Problems solved by technology

However, when this regulation collapses, cells are continuously divided and tumors are formed.

Treating brain tumors are difficult due to the sites of the tumors.

For physical surgery, when tumor sites are completely removed, complications are likely to occur.

For chemotherapy, a high-concentration anticancer drug needs to be injected due to a brain-blood barrier, and thus, it seriously damages other organs.

However, there is a limit to migrate a sufficient amount of virus vectors to the target cancer site.

However, mechanisms regulating stem cells trafficking to tumors are unclear.

However, using neural stem cell in clinical experiments causes ethical problems related to how neural stem cells are taken, and immunological rejection caused by allogenic transplantation.

In this case, however, since BM-MSCs are taken through a plurality of complex processes, subjects from which the BM-MSCs are taken suffer from mental and physical stress for a long period of time.

However, such attempts for identifying availability of UCB-MSCs have not yet been made.

Also, all the information disclosed in the present specification is used only to help understanding of the background of the present inventive concept and cannot be prior art.

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