THERAPEUTIC AGENT ASSOCIATED WITH SUPPRESSION OF PROLIFERATION AND METASTASIS OF TUMOR, WHICH COMPRISES EXOSOMES RELEASED FROM CYTOTOXIC T CELLS AND TARGETS CANCER STROMAL/MESENCHYMAL CELLS
|Posted date||Nov 11, 2016|
|International application number||2016JP058721|
|International publication number||WO 2016152786|
|Date of international filing||Mar 18, 2016|
|Date of international publication||Sep 29, 2016|
|Title||THERAPEUTIC AGENT ASSOCIATED WITH SUPPRESSION OF PROLIFERATION AND METASTASIS OF TUMOR, WHICH COMPRISES EXOSOMES RELEASED FROM CYTOTOXIC T CELLS AND TARGETS CANCER STROMAL/MESENCHYMAL CELLS|
[Problem] To provide a therapeutic agent which can suppress cell proliferation and metastasis of a tumor or the like.
[Solution] Provided is a therapeutic agent for cell-proliferative diseases, which comprises extracellular vesicles (exosomes) released from cytotoxic T cells. The therapeutic agent targets cancer stromal/mesenchymal cells and can suppress the proliferation and metastasis of a tumor including cancer.
|Outline of related art and contending technology||
Primary tumor tissue, tumor stroma, fibronectin, laminin and collagen and extracellular matrix, many other formed by the cells. These cells include, cancer-associated fibroblasts (cancer-associate fibroblasts: CAFs, fibroblast marker and platelet - derived growth factor receptor α (CD140a)+ 、α smooth muscle actin (α-SMA)+) 、(mesenchymal stem cells (MSCs) (stromal stem cells in non-patent document 1. In addition, each of the documents is, at the end are shown together.): CD140a+ Stem cell antigen Sca-1+) is observed. In addition, such as E- cadherin (non-patent document 3, 4) by tight binding to the cancer cells, the cancer cells to fill the gap between the quality of the spread around, where angiogenesis (Sca-1+ CD31+) (Non-Patent Document 2) is performed. For tumor invasion and metastasis, tumor stroma (epithelial to mesenchymal transition due to the interaction of epithelial mesenchymal transformation: EMT) including progression of cancer cells becomes a key. Epithelial mesenchymal conversion, a marker for judging the aggressiveness of tumors (non-patent document 5) capable of becoming a. As the molecules involved in epithelial mesenchymal transformation, in some reports (non-patent document 3, 6, 7).
Endosomal membrane derived microvesicles (100-200 nm diameter) is, tumor cells and tumor stromal cells released from a variety of cells, proteins and RNA contained therein by, information is transmitted between the cells are known to be (non-patent document 8, 9). Various types of tumor cells extracellular vesicles (extracellular vesicles: ECV. Exosomes or exosomes may also be referred to. In the present specification, referred to as' exosomes' or 'ECV'.) Release, this is self-perpetuating, immune tolerance, involved in the tumor environment such as adjustment of a report (non-patent document 8,10-13). On the other hand, exosomes may be released by the tumor, epithelial mesenchymal transformation, and tumor growth and progression has been a report that promoted (non-patent document 9, 14, 15). For this reason, the studies about exosomes, in order to evaluate the degree of progression of the tumor, and are therefore important. Studies have shown that human and mouse model, CD8 tumor infiltrating activated+ Tcells, infiltrate the tumor or tumor stroma (non-patent document 16) is. In addition, to tumor-associated antigens in immunotherapy with monoclonal antibodies, cytotoxic T lymphocytes (CTL) CD8 including+ Tcells, integrated on a tumor tissue or tumor in the growth of known (non-patent document 17, 18). Is CTL, tumor-specific but, through the basement membrane remodeling, and permeates from the blood vessel to the tumor (non-patent document 19) since, CD8+ Tcells, tumor development and progression to achieve various estimated to be involved.
And it is an object of the present invention to offer a means to solve this
In the above-described situation, cytotoxic T cells release exosomes, regarding the aggressiveness of tumors, in any conditions, the user has what effect will be, has not been little is known. Therefore, the inventors of the present invention, cytotoxic T cells release exosomes, an influence exerted on tumor progression were examined in detail. As a result, among the various types of cytotoxic T cells, in particular CD8+ Tcell-derived exosomes, instead of the tumor tissue of the cancer, to kill the surrounding mesenchymal cells, the growth of cancer, inhibiting the progress of cancer metastasis including that in fact, essentially resulted in the completion of the present invention. 1 Of the first aspect of the present invention, cytotoxic T extracellular vesicles released from the cells (exosomes) including a therapeutic agent for diseases related to cell proliferation. 2 Aspect of the present invention, said cytotoxic T cells among human CD4+ 、CD8+ 、CD9+ 、CD63+、 TCR+ Tcells released from the at least one of 1 or 2 or more extracellular vesicles (exosomes) 1 comprising a first aspect of the therapeutic agent for diseases related to cell proliferation. 3 Aspect of the present invention, said cytotoxic T cells among CD8+ Textracellular vesicles released from the cells in an extracellular vesicles (exosomes) (exosomes) 2 comprising a first aspect of the therapeutic agent for diseases related to cell proliferation. 4 Of the first aspect of the present invention, the extracellular vesicles (exosomes) including miRNA in cell growth effective in suppressing 2 and 3 of the first aspect of the embodiment or the second therapeutic agent for diseases related to cell proliferation.
5 Aspect of the present invention, in an effective anti-cell proliferation The miRNA 4 characterized in that it comprises a first aspect of the therapeutic agent for diseases related to cell proliferation. 6 Aspect of the present invention, the therapeutic agent for cell proliferative disease, a bactericide, mucosal removal agent, isotonic agent, pH adjusting agents, stabilizers, thickeners, preservatives, pressure-sensitive adhesive, or immune-enhancing agent selected from the group consisting of 1 or more of the first embodiment 2 further comprising a therapeutic agent for diseases related to cell proliferation. 7 Of the first aspect of the present invention, the therapeutic agent in the tumor tissue, mesenchymal cells into the tumor tissue, vein or subcutaneously administered to the fourth aspect of the embodiment 2 of a therapeutic agent for diseases related to cell proliferation. 8 Aspect of the present invention, cytotoxic T cells were collected and emitted from the exosomes, in an effective anti-cell proliferation that miRNA from the exosomes specific for a cell proliferative disease treatment method for extracting miRNA. 9 Aspect of the present invention, in an effective anti-cell proliferation comprises miRNA and a therapeutic agent for diseases related to cell proliferation.
10 Aspect of the present invention, cytotoxic T cells included in the light emitted from the exosome miRNA miRNA having a base sequence the same as the cultured human mesenchymal stem cells (MSC) and added to the culture, by examining the cytotoxic activity with respect to the MSC, MSC MSC miRNA to evaluate the cytotoxicity of cytotoxic miRNA identification method. Cytotoxic MSC after miRNA have been identified, including its miRNA or exosomes, having the same sequence as the synthesized miRNA miRNA, miRNA or as it is synthesized by reconstructing as exosomes, cells used as a therapeutic agent for proliferative diseases. Thus, 11 of the first aspect of the present invention, including cytotoxic MSC miRNA for a therapeutic agent for cell proliferative disorder. In addition, in yet another aspect, method for treating a cell proliferative disease, to a patient, each of the above aspects of a method of administering a therapeutic agent. At this time, method of administration, the tumor tissue, mesenchymal cells into the tumor tissue, any vein or subcutaneously is preferred. And exosomes, secreted to the outside from the various cells in the lipid bilayer membrane vesicles formed means that, in the degree of a diameter of about 40 nm - 200 nm. Is a living body, saliva, blood, urine, amniotic fluid, body fluids such as observed among malignant ascites. In addition, secreted into the culture medium from the cultured cells. The exosome, various proteins, are included in the RNA, information is transmitted between cells may be responsible for pointed out. In addition, according to the present invention, cytotoxic T cells by administering an acquired from the exosomes, the exosomes mesenchymal cell death around the cancer cells (cancer stromal collapse) and, as a result, the growth of cancer cells/metastasis can be suppressed. CD8 Cells among said cytotoxic T+ T/particularly exosomes derived from a cell growth inhibiting effect on metastasis. As the mechanism of action, the exosomes, the cancer cells and be captured in both the mesenchymal cells, mesenchymal cells only were the death (apoptosis) and, as a result, cancer cells or stromal cells necessary for transitions lose, isolated, growth/metastasis. In addition, the exosome specific miRNA similarly included in the growth of cancer cells/transition suppression effect can be obtained.
|Scope of claims||
前記細胞傷害性T細胞は、ヒトCD4 ＋、CD8 ＋、CD9 ＋、CD63 ＋、 TCR +T細胞のうち少なくとも1または2以上から放出された細胞外小胞（エキソソーム）を含む請求項1記載の細胞増殖性疾病用の治療薬。
|IPC(International Patent Classification)||
National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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