Diabetes treatment using stem cell migration agents

Abstract

The present disclosure provides a therapy for diabetes that targets abnormal stem cells in combination with stem cell migration. In one embodiment, the present disclosure provides a therapy for diabetes and / or diabetes-related diseases and disorders and / or symptoms that targets abnormal stem cells in combination with stem cell migration. In one embodiment, the present disclosure provides diagnosis of diabetes and / or diabetes-related diseases and disorders and / or symptoms, or the risk thereof, using abnormal stem cell migration and / or residence as an indicator.

Description

[Technical Field]

[0001] The present disclosure relates to a treatment for diabetes and / or its associated diseases that combines stem cell migration and inhibition of abnormal stem cells, and to a diagnosis of diabetes and / or its associated diseases using stem cell migration and / or retention as indicators. More particularly, the present disclosure relates to a treatment for diabetes and / or its associated diseases by migrating and inhibiting abnormal hematopoietic stem cells, and to a diagnosis of diabetes and / or its associated diseases by detecting the migration of abnormal hematopoietic stem cells from and / or retention in specific niches. [Background technology]

[0002] Diabetes is generally classified as type 1 or type 2 (Non-Patent Document 1). However, because the details of the onset of both types of diabetes are unknown, the classification of type 1 and type 2 as a pathogenesis-based entity is unreasonable. Currently, various drugs are being developed for the treatment of diabetes, but most are aimed at blood glucose control, and while they may be effective in preventing the progression of diabetes, they may be insufficient to cure diabetes. Furthermore, diabetic complications such as neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders commonly occur in both type 1 and type 2 diabetes, but once they develop, they can be difficult to cure. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Diabetes Treatment Guidelines 2016, Japan Diabetes Society, Nankodo Summary of the Invention [Means for solving the problem]

[0004] The present inventors have found that the therapeutic strategy for diabetes and / or its related diseases by suppressing abnormal stem cells is more effective when combined with stem cell migration. Based on this new finding, the present disclosure provides a means for the treatment and diagnosis of diabetes and / or its related diseases.

[0005] Thus, the present disclosure provides: (Item 1) A composition for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising an inhibitor of abnormal hematopoietic stem cells (HSCs), wherein the inhibitor is administered in combination with a stem cell migration agent. (Item 2) The composition of any of the preceding items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and functional equivalents thereof is not expressed and / or does not function at normal levels. (Item 3) The composition of any of the preceding items, wherein the expression at less than normal levels is overexpression. (Item 4) The composition of any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item 5) The composition of any of the preceding items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α) and proinsulin not being expressed at normal levels. (Item 6) The composition of any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof. (Item 7) The composition of any of the preceding items, wherein the disease, disorder and / or condition comprises a diabetic complication. (Item 8) The composition of any of the preceding items, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item 9) The composition of any of the preceding items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item 10) The composition of any of the above items, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item 11) The composition of any of the preceding items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item 12) A pharmaceutical for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising a combination of an inhibitor of abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent. (Item 13) A composition for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising a stem cell migration agent, wherein the stem cell migration agent is administered in combination with an inhibitor of abnormal hematopoietic stem cells (HSCs). (Item 14) A composition for selecting a treatment for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or conditions, comprising an agent that detects migration and / or persistence of abnormal hematopoietic stem cells (HSCs). (Item 15) The composition of any of the preceding items, wherein said migration and / or retention is migration from and / or retention in the bone marrow niche. (Item 16) The composition of any of the preceding items, wherein the migration detection agent comprises a detection agent for CD106 or a functional equivalent. (Item 17) A method for treating and / or preventing diabetes or diabetes-related diseases, disorders and / or symptoms in a subject, comprising administering to the subject effective amounts of an agent that reduces or eliminates abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent. (Item 18) A method for using migration and / or persistence of abnormal hematopoietic stem cells (HSCs) as an indicator of treatment for the treatment and / or prevention of diabetes or diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, the method comprising detecting migration and / or persistence of abnormal hematopoietic stem cells (HSCs) in the subject. (Item 19) The composition of any of the preceding items, wherein the composition is a pharmaceutical composition. (Item 20) The composition of any of the preceding items, further comprising a pharmaceutically acceptable excipient. (Item A1) A method for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, comprising administering to the subject an effective amount of an inhibitor of abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent. (Item A2) The method of any of the preceding items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and functional equivalents thereof is not expressed and / or does not function at normal levels. (Item A3) The method according to item A2, wherein the expression at an abnormal level is overexpression. (Item A4) The method according to any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item A5) The method of any of the preceding items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α) and proinsulin not being expressed at normal levels. (Item A6) The method according to any one of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof. (Item A7) The method of any of the preceding items, wherein the disease, disorder and / or condition comprises a diabetic complication. (Item A8) The method of any of the preceding items, wherein the disease, disorder and / or condition is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item A9) The method of any of the preceding items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item A10) Any of the methods described above, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item A11) The method of any of the preceding items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item A12) A method for selecting a treatment for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, comprising administering to the subject an effective amount of an agent that detects migration and / or retention of abnormal hematopoietic stem cells (HSCs). (Item A13) The method of any of the preceding items, wherein said migration and / or persistence is migration from and / or persistence in the bone marrow niche. (Item A14) The method of any of the preceding items, wherein the migration detection agent comprises a detection agent for CD106 or a functional equivalent. (Item A15) A method for diagnosing diabetes or diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, comprising detecting migration and / or persistence of abnormal hematopoietic stem cells (HSCs) in the subject. (Item B1) A composition for the suppression of abnormal hematopoietic stem cells (HSCs) for use in combination with a stem cell migration agent for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item B2) The composition of any of the preceding items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and functional equivalents thereof is not expressed and / or does not function at normal levels. (Item B3) The composition of any of the preceding items, wherein the expression at less than normal levels is overexpression. (Item B4) The inhibitor according to any of the preceding items, comprising at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item B5) The composition of any of the preceding items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α) and proinsulin not being expressed at normal levels. (Item B6) The composition of any of the preceding items, comprising at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof. (Item B7) The composition of any of the preceding items, wherein the disease, disorder and / or condition comprises a diabetic complication. (Item B8) The composition of any of the preceding items, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item B9) The composition of any of the preceding items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item B10) The composition of any of the above items, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item B11) The composition of any of the preceding items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item B13) A composition for stem cell migration for use in combination with an inhibitor of abnormal hematopoietic stem cells (HSCs) for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item B14) A composition for detecting migration and / or persistence of abnormal hematopoietic stem cells (HSCs) for selection of treatments for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item B15) The composition of any of the preceding items, wherein said migration and / or retention is migration from and / or retention in the bone marrow niche. (Item B16) The composition of any of the preceding items, comprising a detection agent for CD106 or a functional equivalent. (Item B17) The composition of any of the preceding items, wherein the composition is a pharmaceutical composition. (Item B18) The composition of any of the preceding items, further comprising a pharmaceutically acceptable excipient. (Item C1) Use of an inhibitor of abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent in the manufacture of a medicament for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item C2) The use of any of the above items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and its functional equivalents is not expressed and / or does not function at normal levels. (Item C3) The use of any of the preceding items, wherein said non-normal expression is overexpression. (Item C4) The use of any of the above items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item C5) The use of any of the preceding items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α) and proinsulin not being expressed at normal levels. (Item C6) The use of any of the above items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof. (Item C7) The use of any of the above items, wherein said disease, disorder and / or condition comprises diabetic complications. (Item C8) The use of any of the preceding items, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item C9) The use of any of the above items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item C10) Use of any of the above items, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item C11) Use of any of the above items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item C14) 1. Use of an agent that detects migration and / or persistence of abnormal hematopoietic stem cells (HSCs) in the manufacture of a medicament for selecting a treatment for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item C15) The use of any of the preceding items, wherein said migration and / or retention is migration from and / or retention in the bone marrow niche. (Item C16) The use of any of the preceding items, wherein the migration detection agent comprises a detection agent for CD106 or a functional equivalent. (Item D1) A composition for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising an inhibitor of abnormal hematopoietic stem cells (HSCs), wherein the inhibitor is administered in combination with a stem cell migration agent. (Item D2) The composition of any of the preceding items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and functional equivalents thereof is not expressed and / or does not function at normal levels. (Item D3) The composition of any of the preceding items, wherein the expression at less than normal levels is overexpression. (Item D4) The composition of any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item D5) The composition of any of the preceding items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α), histone deacetylase (HDAC), and proinsulin that is not expressed at normal levels. (Item D6) The composition of any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof and an HDAC inhibitor (e.g., trichostatin A). (Item D7) The composition of any of the preceding items, wherein the disease, disorder and / or condition comprises a diabetic complication. (Item D8) The composition of any of the preceding items, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item D9) The composition of any of the preceding items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item D10) The composition of any of the above items, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item D11) The composition of any of the preceding items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item D12) A pharmaceutical for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising a combination of an inhibitor of abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent. (Item D13) A composition for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising a stem cell migration agent, wherein the stem cell migration agent is administered in combination with an inhibitor of abnormal hematopoietic stem cells (HSCs). (Item D14) A composition for selecting a treatment for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or conditions, comprising an agent that detects migration and / or persistence of abnormal hematopoietic stem cells (HSCs). (Item D15) The composition of any of the preceding items, wherein said migration and / or retention is migration from and / or retention in the bone marrow niche. (Item D16) The composition of any of the preceding items, wherein the migration detection agent comprises a detection agent for CD106 or a functional equivalent. (Item D17) A method for treating and / or preventing diabetes or diabetes-related diseases, disorders and / or symptoms in a subject, comprising administering to the subject effective amounts of an agent that reduces or eliminates abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent. (Item D18) A method for using migration and / or persistence of abnormal hematopoietic stem cells (HSCs) as an indicator of treatment for the treatment and / or prevention of diabetes or diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, the method comprising detecting migration and / or persistence of abnormal hematopoietic stem cells (HSCs) in the subject. (Item D19) The composition of any of the preceding items, wherein the composition is a pharmaceutical composition. (Item D20) The composition of any of the preceding items, further comprising a pharmaceutically acceptable excipient. (Item E1) A method for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, comprising administering to the subject an effective amount of an inhibitor of abnormal hematopoietic stem cells (HSCs) and a stem cell migration agent. (Item E2) The method of any of the preceding items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and functional equivalents thereof is not expressed and / or does not function at normal levels. (Item E3) The method of any of the preceding items, wherein the expression at a level other than normal is overexpression. (Item E4) The method according to any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item E5) The method of any of the preceding items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α), histone deacetylase (HDAC), and proinsulin not being expressed at normal levels. (Item E6) The method of any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof and an HDAC inhibitor (e.g., trichostatin A). (Item E7) The method of any of the preceding items, wherein the disease, disorder and / or condition comprises a diabetic complication. (Item E8) The method of any of the preceding items, wherein the disease, disorder and / or condition is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item E9) The method of any of the preceding items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item E10) Any of the methods described above, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item E11) The method of any of the preceding items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item E12) A method for selecting a treatment for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, comprising administering to the subject an effective amount of an agent that detects migration and / or retention of abnormal hematopoietic stem cells (HSCs). (Item E13) The method of any of the preceding items, wherein said migration and / or persistence is migration from and / or persistence in the bone marrow niche. (Item E14) The method of any of the preceding items, wherein the migration detection agent comprises a detection agent for CD106 or a functional equivalent. (Item E15) A method for diagnosing diabetes or diabetes and / or diabetes-related diseases, disorders and / or symptoms in a subject, comprising detecting migration and / or persistence of abnormal hematopoietic stem cells (HSCs) in the subject. (Item F1) A stem cell migration agent or an abnormal hematopoietic stem cell (HSC) inhibitor for use in combination with a stem cell migration agent and an abnormal HSC inhibitor for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item F2) The stem cell migration agent or abnormal HSC suppressor according to any of the preceding items, wherein the abnormal HSC is one in which a gene or protein selected from the group consisting of CD106 and functional equivalents thereof is not expressed and / or does not function at normal levels. (Item F3) The stem cell migration agent or abnormal HSC suppressor of any of the preceding items, wherein the expression that is not at a normal level is overexpression. (Item F4) The inhibitor according to any of the preceding items, comprising at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item F5) The stem cell migration agent or abnormal HSC suppressor according to any of the above items, wherein the abnormal HSC further lacks normal levels of expression of a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α), histone deacetylase (HDAC), and proinsulin. (Item F6) The stem cell migration agent or abnormal HSC suppressor according to any of the preceding items, comprising at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof and an HDAC inhibitor (e.g., trichostatin A). (Item F7) The stem cell migration agent or abnormal HSC inhibitor of any of the preceding items, wherein the disease, disorder and / or condition comprises diabetic complications. (Item F8) The stem cell migration agent or abnormal HSC inhibitor according to any of the preceding items, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorder, delayed fracture healing, eating disorder, and skin disorder. (Item F9) The stem cell migration agent or abnormal HSC suppressor according to any of the preceding items, wherein the stem cell migration agent has the ability to cause the abnormal HSC to migrate from a niche. (Item F10) The stem cell migration agent or abnormal HSC suppressor according to any of the above items, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item F11) The stem cell migration agent or abnormal HSC inhibitor according to any of the preceding items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item F13) A stem cell migration agent or an abnormal hematopoietic stem cell (HSC) inhibitor for stem cell migration, for use in combination with an inhibitor of abnormal HSC, for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item F14) Agents that detect migration and / or persistence of abnormal hematopoietic stem cells (HSCs) for the selection of treatments for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or conditions. (Item F15) The agent of any of the preceding items, wherein said migration and / or retention is migration from and / or retention in the bone marrow niche. (Item F16) The agent of any of the above items, comprising an agent for detecting CD106 or a functional equivalent. (Item G1) Use of at least one of an abnormal hematopoietic stem cell (HSC) inhibitor or a stem cell migration agent in the manufacture of a medicament for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms using a combination of an abnormal HSC inhibitor and a stem cell migration agent. (Item G2) The use of any of the above items, wherein the abnormal HCS is one in which a gene or protein selected from the group consisting of CD106 and its functional equivalents is not expressed and / or does not function at normal levels. (Item G3) The use of any of the preceding items, wherein said non-normal expression is overexpression. (Item G4) The use of any of the above items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof. (Item G5) The use of any of the above items, wherein the abnormal HCS further comprises a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α), histone deacetylase (HDAC), and proinsulin that is not expressed at normal levels. (Item G6) The use of any of the preceding items, wherein the inhibitor comprises at least one selected from the group consisting of an anti-TNF-α antibody or a functional variant thereof and an HDAC inhibitor (e.g., trichostatin A). (Item G7) The use of any of the above items, wherein said disease, disorder and / or condition comprises diabetic complications. (Item G8) The use of any of the preceding items, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorders, delayed fracture healing, eating disorders, and skin disorders. (Item G9) The use of any of the above items, wherein the stem cell migration agent has the ability to migrate the abnormal HSCs from the niche. (Item G10) Use of any of the above items, wherein the stem cell migration agent comprises at least one agent selected from the group consisting of a CXCR4 antagonist, a CXCR2 stimulator, an epidermal growth factor receptor (EGFR) inhibitor, and a granulocyte colony-stimulating factor (G-CSF) agent. (Item G11) Use of any of the above items, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, GROβ2 (MIP2), gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, and pegfilgrastim. (Item G14) 1. Use of an agent that detects migration and / or persistence of abnormal hematopoietic stem cells (HSCs) in the manufacture of a medicament for selecting a treatment for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms. (Item G15) The use of any of the preceding items, wherein said migration and / or retention is migration from and / or retention in the bone marrow niche. (Item G16) The use of any of the preceding items, wherein the migration detection agent comprises a detection agent for CD106 or a functional equivalent.

[0006] It is contemplated that one or more of the above features may be provided in combinations other than those explicitly stated. Still further embodiments and advantages of the present disclosure will be recognized by those skilled in the art upon reading and understanding the following detailed description, if necessary. [Effects of the Invention]

[0007] The present disclosure provides new treatments and diagnoses for diabetes and / or related diseases. This eliminates the need for donors, which has been required for pancreas transplantation and pancreatic islet transplantation, traditional methods for restoring insulin secretion in diabetic patients, and overcomes supply capacity limitations relative to the number of recipients. It also avoids the burden on recipients, such as surgery. Thus, the treatment of the present disclosure can be achieved by drug administration, making it much simpler and less burdensome. Furthermore, because it allows for radical treatment, it is expected to result in a better prognosis. [Brief explanation of the drawings]

[0008] [Figure 1]Fluorescence-activated cell sorting (FACS) analysis of proinsulin-positive cells in the c-kit-positive, Sca-1-positive, lineage marker-negative (KSL) cell fraction in mononuclear cells from non-diabetic (non-DM) and streptozotocin-induced diabetic (STZ-DM) patients. The top two left panels (left: non-DM, right: STZ-DM) show KSL cells (boxed) among the lineage-negative cells. The vertical axis represents c-kit fluorescence intensity, and the horizontal axis represents Sca-1 fluorescence intensity. The bottom two left panels (left: non-DM, right: STZ-DM) show the distribution of proinsulin-positive cells (boxed). The vertical axis represents the number of KSL cells, and the horizontal axis represents proinsulin fluorescence intensity. The right graph (left: non-DM, right: STZ-DM) shows the percentage of proinsulin-positive cells among KSL cells (n = 3 per group). **: P < 0.01. [Figure 2] FACS analysis of tumor necrosis factor alpha (TNF-α)-positive cells among KSL cells in mononuclear cells from non-DM and STZ-DM mice. The top two left panels (left: non-DM, right: STZ-DM) show KSL cells (boxed) among lineage-negative cells; the vertical axis represents c-kit fluorescence intensity, and the horizontal axis represents Sca-1 fluorescence intensity. The bottom two left panels (left: non-DM, right: STZ-DM) show the distribution of TNF-α-positive cells (boxed); the vertical axis represents the number of KSL cells, and the horizontal axis represents TNF-α fluorescence intensity. The graph on the right shows the percentage of TNF-α-positive cells among KSL cells (n = 5 per group). Data are shown as mean ± standard error. **: P < 0.01. [Figure 3] FACS analysis of CD106 expression of KSL cells in mononuclear cells from non-DM and STZ-DM mice. The left two panels (top: non-DM, bottom: STZ-DM) show KSL cells (boxed) among lineage-negative cells; the vertical axis represents c-kit fluorescence intensity, and the horizontal axis represents Sca-1 fluorescence intensity. The right two panels (top: non-DM, bottom: STZ-DM) show the distribution of CD106-positive cells (boxed); the vertical axis represents side scatter (SSC) intensity, and the horizontal axis represents CD106 fluorescence intensity. The right graph (left: non-DM, right: STZ-DM) shows the percentage of CD106-positive cells among KSL cells (1 scale represents 1%) (n = 4 per group). **: P < 0.01. [Figure 4] FACS analysis of mononuclear cells from ICR and NOD mice. Two panels (left: ICR, right: NOD) show KSL cells (boxed) among lineage-negative cells. The vertical axis shows the c-kit fluorescence intensity, and the horizontal axis shows the Sca-1 fluorescence intensity. [Figure 5] FACS analysis of tumor necrosis factor alpha (TNF-α)- and CD106-positive cells in KSL cells in mononuclear cells from ICR and NOD mice. The top two panels (left: ICR, right: NOD) show the distribution of TNF-α-positive cells (boxed), with the vertical axis representing forward scatter (FSC) intensity and the horizontal axis representing TNF-α fluorescence intensity. The bottom two panels (left: ICR, right: NOD) show the distribution of CD106-positive cells (boxed), with the vertical axis representing forward scatter (FSC) intensity and the horizontal axis representing CD106 fluorescence intensity. [Figure 6] FACS analysis of the side population (SP) and non-SP fractions in non-DM and STZ-DM KSL cells. The two left panels (top: non-DM, bottom: STZ-DM) show the SP fraction (circled in the lower left) and non-SP fraction (circled in the upper right) of KSL cells. The vertical axis represents Hoechst Blue fluorescence intensity, and the horizontal axis represents Hoechst Red fluorescence intensity. The right graph (left: SP, right: non-SP) shows the percentage of the SP and non-SP fractions in non-DM (left bar) and STZ-DM (right bar) KSL cells (n = 3). Data are shown as mean ± standard error. **: P < 0.01. [Figure 7]a) FACS analysis of CD106-positive cells in KSL-non-SP cells from non-DM and STZ-DM mice. Two panels (top: non-DM, bottom: STZ-DM) show the distribution of CD106-positive cells (boxed). The vertical axis represents side scatter (SSC) intensity, and the horizontal axis represents CD106 fluorescence intensity. Graphs (left: non-DM, right: STZ-DM) show the percentage of CD106-positive cells in KSL-non-SP cells (n = 3 per group). b) FACS analysis of CD106-positive cells in KSL-SP cells from non-DM and STZ-DM mice. Two panels (top: non-DM, bottom: STZ-DM) show the distribution of CD106-positive cells (boxed; 0%). The vertical axis represents side scatter (SSC) intensity, and the horizontal axis represents CD106 fluorescence intensity. The graphs (left: non-DM, right: STZ-DM) show the percentage of CD106-positive cells in KSL-SP cells (n = 3 per group). c) FACS analysis of TNF-α-positive cells in KSL-non-SP cells with non-DM and STZ-DM. The two panels (top: non-DM, bottom: STZ-DM) show the distribution of TNF-α-positive cells (boxed), with the vertical axis representing side-scattered light intensity (SSC) and the horizontal axis representing TNF-α fluorescence intensity. The graphs (left: non-DM, right: STZ-DM) show the percentage of TNF-α-positive cells in KSL-non-SP cells (n = 3 per group). d) FACS analysis of TNF-α-positive cells in KSL-SP cells with non-DM and STZ-DM. The two panels (top: non-DM, bottom: STZ-DM) show the distribution of TNF-α-positive cells (boxed, 0%). The vertical axis represents side-scattered light (SSC) intensity, and the horizontal axis represents TNF-α fluorescence intensity. The graphs (left: non-DM, right: STZ-DM) show the percentage of TNF-α-positive cells among KSL-SP cells (n = 3 per group). Data are shown as mean ± standard error. *: P < 0.05. [Figure 8]This figure shows a comparison of histone deacetylase gene expression (HDACs) in KSL cells from non-DM and STZ-DM mice. From left to right, the expression levels of Hdac3, Hdac4, Hdac8, and Hdac9 are shown. In each bar graph, the left column shows the results for non-DM mice, and the right column shows the results for STZ-DM mice. The vertical axis shows the relative mRNA expression levels in both mice, with the mRNA expression level in non-DM mice set to 1. *: P<0.05. [Figure 9] This diagram shows the experimental setup for transplantation of KSL cells derived from non-DM or STZ-DM mice into normoglycemic mice. Transgenic mice carrying green fluorescent protein (GFP-Tg) were treated with STZ to induce diabetes (STZ-DM GFP), and non-DM mice were treated with intravenous citrate buffer injection (non-DM GFP). Three months later, KSL cells from the non-DM and STZ-DM mice were transplanted into normoglycemic wild-type mice that had been lethally irradiated with 9 Gy (non-DM-derived KSL-T and STZ-DM-derived KSL-T). [Figure 10] Blood glucose concentrations (mg / gL) (left) in non-DM-derived KSL-T (n = 9) and STZ-DM-derived KSL-T (n = 10) are shown, and the relative ratio of sensory nerve conduction velocity (SNCV) in the sciatic nerve (right) is shown, with the measured value in non-DM-derived KSL-T set to 1. NS indicates no significant difference, and ** indicates P < 0.01. [Figure 11] Immunofluorescent staining images of MAP2, proinsulin, and TNF-α in dorsal root ganglia (DRG). The four columns from the left show the results of STZ-DM-derived KSL-T. From left, they show nuclei (blue), GFP (green), target molecules (top row: MAP2, middle row: proinsulin, bottom row: TNF-α) (red), and merged images. The right column shows the results of non-DM-derived KSL-T. They show nuclei (blue), GFP (green), and target molecules (top row: MAP2, middle row: proinsulin, bottom row: TNF-α) (red) merged images. Arrowheads indicate fused cells. Scale bar = 10 μm. [Figure 12]FACS analysis of proinsulin-positive cells in the KSL fraction of mononuclear cells from non-DM-derived KSL-T and STZ-DM-derived KSL-T mice. The top two left panels (left: non-DM-derived KSL-T; right: STZ-DM-derived KSL-T) show KSL cells (boxed) among lineage-negative cells. The vertical axis shows c-kit fluorescence intensity, and the horizontal axis shows Sca-1 fluorescence intensity. The bottom two left panels (left: non-DM-derived KSL-T; right: STZ-DM-derived KSL-T) show the distribution of proinsulin-positive cells (boxed). The vertical axis shows the number of KSL cells, and the horizontal axis shows proinsulin fluorescence intensity. The right graph (left: non-DM-derived KSL-T; right: STZ-DM-derived KSL-T) shows the percentage of proinsulin-positive cells among KSL cells (n = 3 per group). *: P < 0.05. [Figure 13] FACS analysis of TNF-α-positive cells in the KSL fraction of mononuclear cells from non-DM-derived KSL-T and STZ-DM-derived KSL-T mice. The top two left panels (left: non-DM-derived KSL-T; right: STZ-DM-derived KSL-T) show KSL cells (boxed) among lineage-negative cells. The vertical axis shows c-kit fluorescence intensity, and the horizontal axis shows Sca-1 fluorescence intensity. The bottom two left panels (left: non-DM-derived KSL-T; right: STZ-DM-derived KSL-T) show the distribution of TNF-α-positive cells (boxed). The vertical axis shows the number of KSL cells, and the horizontal axis shows TNF-α fluorescence intensity. The right graph (left: non-DM-derived KSL-T; right: STZ-DM-derived KSL-T) shows the percentage of TNF-α-positive cells among KSL cells (n = 3 per group). **: P < 0.01. [Figure 14] Comparison of the expression levels of each gene in blood samples from human diabetic (DM) and healthy (non-DM) subjects. The panels show insulin (upper left), CD34 (upper right), TNF-α (lower left), and CD106 (lower right), respectively. In each panel, the left bar shows the results for healthy subjects, and the right bar shows the results for human diabetic patients. The vertical axis shows the relative mRNA expression level, with the average result for healthy subjects set at 1. [Figure 15]This figure shows changes in blood glucose levels and body weight in streptozotocin-induced diabetic mice treated with a stem cell migration agent and anti-CD106 antibody (250 μg / mouse, weekly administration via tail vein). The upper graph shows blood glucose levels (vertical axis, mg / dL) on days 0 and 18. The lower graph shows body weights (vertical axis, g) on ​​days 0 and 18. The blue line shows the results for the control treatment, the orange line shows the results for the stem cell migration agent and anti-CD106 antibody, and the gray line shows the results for the anti-CD106 antibody treatment. [Figure 16] These are images of insulin and nuclei stained in mouse cryosections. The upper left shows the results for a non-DM mouse, the upper right shows the results for an STZ-DM mouse, and the bottom two show the results for two STZ-DM mice treated with a stem cell migration agent and an anti-CD106 antibody. [Figure 17] Approximately 8-10 islets were randomly selected from each mouse, and the ratio of insulin-positive area to the size of the islets was calculated using Image J software. From left to right, the results are for a non-DM mouse, an STZ-DM mouse, and two STZ-DM mice (#1 and #2) treated with a stem cell migration agent and anti-CD106 antibody. **P<0.01. [Figure 18] This figure shows changes in blood glucose levels and body weight when NOD mice were treated with a stem cell chemotactic agent and anti-CD106 antibody (250 μg / mouse, weekly administration via tail vein). The upper graph shows blood glucose levels (vertical axis, mg / dL) on days 7 and 14. The lower graph shows body weights (vertical axis, g) on ​​days 7 and 14. The blue line (#670) and orange line (#671) show the results of two mice treated with the chemotactic agent and anti-CD106 antibody twice weekly, while the gray line (#673) shows the results of an mouse treated with the chemotactic agent and anti-CD106 antibody only once. [Figure 19] These are images of insulin and nuclei stained in mouse frozen sections. From the left, the results are for a non-DM mouse, a NOD mouse before the onset of diabetes, and a NOD mouse after the onset of diabetes. The upper row is a 20x magnification image, and the lower row is a 40x magnification image. The arrowheads indicate pancreatic islets. [Figure 20]Proinsulin staining of bone marrow from patients without DM (DM(-)) and with DM (DM(+)). Bone marrow cells stained with proinsulin antibodies (arrows) are observed in DM(+) cases but not in DM(-) cases. The scale bar indicates 50 μm. DETAILED DESCRIPTION OF THE INVENTION

[0009] The present disclosure will now be described, illustrating the best mode thereof. Throughout this specification, singular expressions should be understood to include the plural concept unless otherwise specified. Therefore, singular articles (e.g., "a," "an," "the," etc. in English) should be understood to include the plural concept unless otherwise specified. Furthermore, terms used in this specification should be understood to have the meaning commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the event of conflict, the present specification (including definitions) will prevail.

[0010] The following provides definitions of terms particularly used in this specification and / or explains basic technical content as appropriate.

[0011] (definition) As used herein, "diabetes" is used in the usual sense in the art. Excluding cases associated with pregnancy or specific genetic abnormalities, diabetes is generally classified into type 1 diabetes, which occurs when pancreatic beta cells are destroyed, resulting in insulin depletion, and type 2 diabetes, which occurs when obesity or other factors reduce insulin secretion from the beta cells in the pancreatic islets, resulting in impaired glucose uptake into muscle and adipose tissue. The results of this study suggest that both type 1 and type 2 diabetes are caused by a common cellular etiology mediated by immune abnormalities, distinct from previously thought causes. Diabetes can be diagnosed, for example, by two or more tests showing fasting blood glucose levels of ≥ 126 mg / dL, HbA1c levels of ≥ 6.5%, and a 2-hour oral glucose tolerance test (75g OGTT) of ≥ 200 mg / dL. As used herein, "diabetes" also includes early-onset maturity-onset diabetes and borderline diabetes.

[0012] As used herein, "diabetes-related disease" or "diabetes-related disease, disorder and / or symptom" may encompass any disease, disorder and symptom associated with diabetes, examples of which include diabetic neuropathy, diabetic retinopathy, diabetic nephropathy, secondary diabetes, diabetic coma, impaired consciousness, abdominal pain, cramps, neuropathy, hyperosmolar hyperglycemic syndrome, albuminuria, edema, renal failure, blindness, Alzheimer's dementia, myocardial infarction, arteriosclerosis obliterans, and cerebral infarction. These include blockage, fatty liver, skin symptoms (such as diabetic lipoid necrosis), decreased wound healing ability, susceptibility to infection (such as sepsis), cancer (liver cancer, kidney cancer, pancreatic cancer, colon cancer, stomach cancer, liver cancer, ovarian cancer, colorectal cancer, colon cancer, etc.), diabetic ketoacidosis, heart disease, cerebrovascular disease, steroid diabetes, constipation, dizziness when standing up (orthostatic hypotension), erectile dysfunction, myocardial infarction, chest pain, severe appendicitis, peritoneal irritation symptoms, low-temperature burns, gestational diabetes, dry mouth, polydipsia, and polyuria.

[0013] Unless otherwise specified, the term "non-diabetic subject" as used herein means a subject who does not have diabetes or its related diseases.

[0014] As used herein, "treatment" refers to preventing, preferably maintaining the current state, more preferably alleviating, and even more preferably eliminating, the worsening of a certain disease or disorder when that condition has developed, and includes exerting a symptom-improving or preventive effect on the patient's disease or one or more symptoms associated with the disease. Preliminary diagnosis followed by appropriate treatment is called "companion treatment," and diagnostic agents used for this purpose are sometimes called "companion diagnostic agents."

[0015] As used herein, the term "prevention" refers to preventing a certain disease or disorder from becoming a certain state before that state occurs. Diagnosis can be performed using the agent of the present disclosure, and if necessary, the agent of the present disclosure can be used to prevent, for example, diabetes, or other conditions, or preventive measures can be taken.

[0016] As used herein, "diagnosis" refers to identifying various parameters related to a condition (e.g., a disease, a disorder) in a subject and determining the current or future state of such a condition. Using the methods, compositions, and systems disclosed herein, internal conditions can be examined, and such information can be used to select various parameters, such as the condition in the subject, and the treatment or prophylactic formulation or method to be administered. In the narrow sense, "diagnosis" herein refers to assessing the current condition, but in the broad sense, it also includes "early diagnosis," "predictive diagnosis," "pre-diagnosis," and the like. The diagnostic methods disclosed herein are industrially useful because, in principle, they can utilize substances excreted from the body and can be performed independently of medical professionals such as physicians. To clarify that the methods can be performed independently of medical professionals such as physicians, the term "assistance in predictive diagnosis, pre-diagnosis, or diagnosis" is sometimes used. The technology disclosed herein is applicable to such diagnostic techniques.

[0017] As used herein, "inhibition" of target cells means reducing the proliferation rate of target cells, damaging target cells, reducing the number of target cells, and / or killing target cells. Inhibition of target cells can be achieved by direct and / or indirect effects. For example, a direct effect of inhibiting target cells is achieved by targeting any characteristic of the cells (e.g., expressed molecules) to damage, reduce, and / or kill target cells, and examples of mechanisms that can be used include, but are not limited to, irradiation, induction of apoptosis, and induction of attack by immune cells. An indirect effect of inhibiting target cells can include, but is not limited to, teaching the immune system to recognize and inhibit the target cells.

[0018] As used herein, an "inhibitor" of a target cell refers to a drug that inhibits a target cell by any means. In the inhibitors described herein, the means for inhibiting a target cell may be any, including, but not limited to, attack by immune cells triggered by binding of the target cell to a targeting molecule (e.g., an antibody), the use of a radioactive molecule, and the like.

[0019] As used herein, the term "migration" of cells refers to the movement of cells into peripheral blood and / or circulating blood. Typically, migration refers to the movement of cells from bone marrow (or a specific site therein) into peripheral blood and / or circulating blood. Such a specific site in the bone marrow is sometimes referred to as a niche or a specific niche, and a specific niche refers to a microenvironment required for stem cells to maintain their properties in a living organism. Therefore, in certain embodiments, the term "migration" as used herein refers to, but is not limited to, the migration of stem cells from a niche.

[0020] As used herein, a "cell migration agent" refers to an agent that induces migration of target cells by any means.

[0021] As used herein, "hematopoietic stem cells" or "HSCs" refer to stem cells that can differentiate into blood cells. In human adults, they are primarily present in the bone marrow and give rise to white blood cells (neutrophils, eosinophils, basophils, lymphocytes, monocytes, macrophages), red blood cells, platelets, mast cells, and dendritic cells. Human hematopoietic stem cells can be characterized, for example, by CD34 positivity, Thy-1 positivity, and also by Lineage negativity, CD34 positivity, CD38 negativity, CD90 positivity, and CD45RA negativity (i.e., Lin - CD34 + CD38 - CD90 + CD45RA - ) (see, for example, Proc Natl Acad Sci USA. 2011 Dec 13;108(50):20012-20017). Mouse hematopoietic stem cells can be characterized as c-kit positive, Sca-1 positive, lineage marker negative (KSL) cells. Hematopoietic stem cells can also be characterized by bone marrow cells stained with Hoechest 33342 dye being negative when excited with ultraviolet light (350 nm) and developed using two optical filters, Hoechst blue and Hoechst red. Hematopoietic stem cells can be further classified into long-term hematopoietic stem cells (LT-HSCs), short-term hematopoietic stem cells (ST-HSCs), etc. For the characteristics of hematopoietic stem cells, see, for example, Cytometry Research 19(2):25-32, 2009.

[0022] As used herein, "long-term hematopoietic stem cells (LT-HSCs)" refer to hematopoietic stem cells with long-term bone marrow cell repopulation potential. LT-HSCs are typically identified by surface antigen markers, and (for example, in mice) are CD34-negative, CD150-positive, CD48-negative, Lin-negative, sca1-positive, and c-kit-positive (or Lineage I-positive). - c-Kit + Sca-1 + CD34 - / low CD150 + LT-HSCs can also be expressed as CD34-negative, CD38-negative (or CD34-negative in humans, for example). - CD38 -These may be expressed as inflammatory cells (e.g., Nat Immunol. 2010 Jul;11(7):585-93; Blood 108, 2446-2454, 2006).

[0023] As used herein, "short-term hematopoietic stem cells (ST-HSCs)" refer to hematopoietic stem cells with short-term bone marrow repopulation capacity. ST-HSCs are usually identified by surface antigen markers and can be described as CD34-positive, CD150-positive, CD48-negative, Lin-negative, sca-1-positive, and c-kit-positive (e.g., in mice). ST-HSCs can also be described as CD34-positive, CD38-negative (or CD34-negative) (e.g., in humans). + CD38 - These may be expressed as inflammatory cells (e.g., Nat Immunol. 2010 Jul;11(7):585-93; Blood 108, 2446-2454, 2006).

[0024] As used herein, "abnormal hematopoietic stem cells" or "abnormal HSCs" refer to hematopoietic stem cells that express abnormal functions and / or lack at least some of their normal functions. Typically, abnormal HSCs refer to cells in which the gene or protein of CD106 or its functional equivalent is not expressed and / or does not function at normal levels.

[0025] As used herein, the phrase "not expressed at normal levels" for a gene or protein refers to the expression of the gene or protein at an amount or level that is not found in cells with normal function. For example, in the case of CD106, abnormality can be determined when CD106 is expressed outside of normal levels (normal values ​​found in normal cells), and preferably when CD106 is expressed at levels higher than normal. For example, KSL cells can be sorted by FACS from non-diabetic and diabetic mice, RNA can be extracted, and then comparative analysis of gene expression levels can be performed using QT-PCR to test for abnormal gene expression (e.g., overexpression). For example, FACS analysis can be performed to detect surface antigens on abnormal hematopoietic stem cells, and abnormal protein expression can be tested by comparing diabetic and non-diabetic subjects.

[0026] As used herein, the phrase "a gene or protein is not functioning at a normal level" refers to the absence of a level of function observed in cells with normal function. For example, in the case of CD106, a level of CD106 that is outside of the normal functioning level (the normal level or value observed in normal cells) can be determined to be abnormal, and preferably, a level of CD106 function observed that is higher than normal can be determined to be abnormal. For example, FACS analysis can be performed to detect surface antigens on abnormal hematopoietic stem cells, and protein activation can be observed in diabetic subjects compared to non-diabetic subjects to test whether a protein is not functioning at a normal level.

[0027] As used herein, "CXCR4" refers to a seven-transmembrane G protein-coupled receptor (GPCR), also known as CD184 or Fusin. The physiological ligand for CXCR4 is stromal cell-derived factor-1 (SDF-1), a CXC chemokine that potently induces monocyte and lymphocyte migration. Inhibition of CXCR4 can induce hematopoietic stem cell migration (Future Oncol. 2007 Feb;3(1):19-27).

[0028] As used herein, "CD106" refers to a surface antigen, also known as the adhesion molecule VCAM-1 (vascular cell adhesion molecule-1) or INCAM-100. It is a type I membrane protein of the Ig superfamily, a cell surface sialoglycoprotein expressed by cytokine-activated endothelium, and is known to mediate leukocyte-endothelial cell adhesion and signal transduction. Representative examples in humans include the VCAM-1 isoform a precursor (nucleic acid sequence: NM_001078.4, amino acid sequence: NP_001069.1), VCAM-1 isoform b precursor (nucleic acid sequence: NM_080682.2, amino acid sequence: NP_542413.1), and VCAM-1 isoform c precursor (nucleic acid sequence: NM_001199834.1, amino acid sequence: NP_001186763.1).

[0029] As used herein, "CD34" refers to a surface protein that is thought to be involved in the adhesion of stem cells to bone marrow extracellular matrix or stromal cells, and is known to be highly glycosylated and phosphorylated by protein kinase C. In humans, representative variants are CD34 transcript variant 1 (nucleic acid sequence: NM_001025109.2, amino acid sequence: NP_001020280.1) and CD34 transcript variant 2 (nucleic acid sequence: NM_001773.3, amino acid sequence: NP_001764.1).

[0030] As used herein, "tumor necrosis factor alpha" or its abbreviation "TNF-α" refers to a multifunctional proinflammatory cytokine belonging to the tumor necrosis factor (TNF) superfamily, also known as TNF, DIF, TNFA, TNFSF2, and TNLG1F, and primarily secreted by macrophages. TNF-α functions via receptors TNFRSF1A / TNFR1 and TNFRSF1B / TNFBR and is involved in the regulation of a wide range of biological processes, including cell proliferation, differentiation, apoptosis, lipid metabolism, and hemocoagulation. Representative human TNF-α sequences are NM_000594.4 and NP_000585.2, respectively.

[0031] As used herein, "proinsulin" refers to the precursor protein of insulin. Proinsulin is processed in the endoplasmic reticulum of pancreatic β cells, and insulin composed of A and B chains is produced in immature secretory granules by removing the C-peptide region. In humans, representative variants include proinsulin transcript variant 1 (nucleic acid sequence: NM_000207.3, amino acid sequence: NP_000198.1), proinsulin transcript variant 2 (nucleic acid sequence: NM_001185097.2, amino acid sequence: NP_001172026.1), proinsulin transcript variant 3 (nucleic acid sequence: NM_001185098.1, amino acid sequence: NP_001172027.1), and proinsulin transcript variant 4 (nucleic acid sequence: NM_001291897.2, amino acid sequence: NP_001278826.1).

[0032] As used herein, "c-Kit" refers to the type 3 transmembrane receptor for MGF (also known as mast cell growth factor or stem cell factor), also known as PBT, SCFR, KIT, CD117, or MASTC. In humans, the most representative are Kit isoform 1 precursor (nucleic acid sequence: NM_000222.2, amino acid sequence: NP_000213.1) and Kit isoform 2 precursor (nucleic acid sequence: NM_001093772.1, amino acid sequence: NP_001087241.1).

[0033] As used herein, "CD20" refers to a B lymphocyte surface molecule that is a member of the transmembrane 4A gene family, also known as MS4A1, B1, S7, Bp35, CD20, CVID5, MS4A2, or LEU-16, and plays a role in the development and differentiation of B cells into plasma cells. In humans, CD20 transcript variant 1 (nucleic acid sequence: NM_152866.2, amino acid sequence: NP_690605.1) and CD20 transcript variant 3 (nucleic acid sequence: NM_021950.3, amino acid sequence: NP_068769.2) are representative.

[0034] Unless otherwise specified, it is understood that reference to each protein in this specification contemplates not only the protein having the amino acid sequence set forth in the particular accession number (or the nucleic acid encoding it), but also its functional equivalents.

[0035] As used herein, "functional equivalents" of a molecule are understood to include mutants or variants of the molecule (e.g., amino acid sequence variants, etc.) that have the function of a characteristic (e.g., a marker) described herein, that exhibit a similar biological function to the molecule (although not necessarily to the same extent), and that can be altered into the molecule itself at the time of acting.

[0036] As used herein, a "functional variant" of a molecule includes a substance obtained by modifying the molecule while maintaining the function of the molecule (although the degree of modification may be changed). For example, functional variants of binding molecules such as antibodies include those conjugated with other moieties (e.g., labels, other functional molecules (e.g., proteins)) or fragmented so as to maintain the binding function to the target molecule. Thus, as used herein, a functional variant is one that maintains the basic function prior to modification.

[0037] The functional equivalents of the present disclosure can be those in which one or more amino acids have been inserted, substituted, or deleted in the amino acid sequence, or added to one or both termini thereof. As used herein, "one or more amino acids have been inserted, substituted, or deleted in the amino acid sequence, or added to one or both termini thereof" means that the amino acid has been modified by a well-known technical method such as site-directed mutagenesis, or by natural mutation, through substitution of a number of amino acids to the extent that would occur naturally.

[0038] As used herein, the term "biological function," when referring to a gene or its associated nucleic acid molecule or polypeptide, refers to a specific function that the gene, nucleic acid molecule, or polypeptide may have in a living organism, including, but not limited to, the production of a specific antibody, enzymatic activity, and the conferring of resistance. For such biological activities, reference can be made to the references cited in the accession numbers, Entrez numbers, and other references in the above-mentioned tables, which are also incorporated herein by reference. As used herein, biological function can be exerted through "biological activity." As used herein, "biological activity" refers to the activity that a certain factor (e.g., polynucleotide, protein, etc.) may have in a living organism, including the ability to exert various functions (e.g., transcription-promoting activity), including the activation or inactivation of another molecule through interaction with another molecule. When two factors interact, their biological activity is determined by the binding between the two molecules and the resulting biological change. For example, if one molecule is co-precipitated when the other molecule is co-precipitated with an antibody, the two molecules are considered to be bound. Therefore, observing such co-precipitation is one method of assessment. For example, if a factor is an enzyme, its biological activity includes its enzymatic activity. In another example, if a factor is a ligand, it includes the binding of the ligand to a corresponding receptor. Such biological activities can be measured by techniques well known in the art.

[0039] As used herein, "derivatives," "analogs," or "variants" of proteins or nucleic acids include, but are not limited to, molecules containing regions of substantial homology to the protein or nucleic acid, which in various embodiments are at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical across the same size amino acid or nucleic acid sequence, or when compared to sequences aligned by computer homology programs known in the art, or such nucleic acid "derivatives," "analogs," or "variants" are capable of hybridizing to the original nucleic acid under stringent, moderately stringent, or non-stringent conditions. Typically, a "derivative," "analog," or "variant" of a protein refers to a product of modifications, such as amino acid substitutions, deletions, and additions, of a naturally occurring protein, which still exhibit the biological function of the naturally occurring protein, although not necessarily to the same degree. For example, the biological function of such proteins can be examined by suitable and available in vitro assays described herein or known in the art. While this disclosure primarily discusses humans, it is understood that it also applies to other species, such as other species within the primate family, or other genera of animals, and that these mammals are also within the scope of this disclosure.

[0040] The term "activity" as used herein refers to the function of a molecule in the broadest sense. Activity generally includes, but is not limited to, the biological, biochemical, physical, or chemical functions of a molecule. Activity includes, for example, enzymatic activity, the ability to interact with other molecules, and the ability to activate, promote, stabilize, inhibit, suppress, or destabilize the function of other molecules, stability, and the ability to localize to a particular subcellular location. Where applicable, the term also relates to the function of protein complexes in the broadest sense.

[0041] As used herein, a "functionally active" protein, polypeptide, fragment or derivative possesses a structural, regulatory, or biochemical function of a protein, such as biological activity.

[0042] As used herein, "gene" refers to a factor that determines a genetic trait. It is usually arranged in a specific order on a chromosome. A gene that determines the primary structure of a protein is called a structural gene, and a gene that controls its expression is called a regulatory gene. As used herein, "gene" can refer to "polynucleotide," "oligonucleotide," and "nucleic acid" (including DNA, RNA, etc.). "Gene product" refers to a substance produced based on a gene, such as a protein or mRNA. Therefore, mRNA can be included in the concept of a gene and can also be considered a gene product. Furthermore, "gene expression" often refers to the transcription level of mRNA, etc.

[0043] As used herein, the terms "protein," "polypeptide," "oligopeptide," and "peptide" are used interchangeably to refer to a polymer of amino acids of any length. The polymer may be linear, branched, or cyclic. The amino acids may be natural, non-natural, or modified. The term may also encompass multiple polypeptide chains assembled into complexes. The term also encompasses naturally occurring or artificially modified amino acid polymers. Such modifications include, for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification (e.g., conjugation with a labeling component). The definition also encompasses, for example, polypeptides containing one or more analogs of an amino acid (e.g., including non-natural amino acids), peptide-like compounds (e.g., peptoids), and other modifications known in the art.

[0044] As used herein, the terms "polynucleotide," "oligonucleotide," and "nucleic acid" are used interchangeably to refer to a polymer of nucleotides of any length. This term also includes "oligonucleotide derivatives" or "polynucleotide derivatives." "Oligonucleotide derivatives" or "polynucleotide derivatives" refer to oligonucleotides or polynucleotides that contain derivatives of nucleotides or have unusual internucleotide bonds, and are used interchangeably. Specific examples of such oligonucleotides include 2'-O-methyl-ribonucleotides, oligonucleotide derivatives in which the phosphodiester bond in the oligonucleotide has been converted to a phosphorothioate bond, oligonucleotide derivatives in which the phosphodiester bond in the oligonucleotide has been converted to an N3'-P5' phosphoramidate bond, oligonucleotide derivatives in which the ribose and phosphodiester bond in the oligonucleotide have been converted to a peptide nucleic acid bond, oligonucleotide derivatives in which the uracil in the oligonucleotide has been substituted with C-5 propynyl uracil, oligonucleotide derivatives in which the uracil in the oligonucleotide has been substituted with C-5 thiazole uracil, oligonucleotide derivatives in which the cytosine in the oligonucleotide has been substituted with C-5 propynyl cytosine, oligonucleotide derivatives in which the cytosine in the oligonucleotide has been substituted with phenoxazine-modified cytosine, oligonucleotide derivatives in which the ribose in the DNA has been substituted with 2'-O-propyl ribose, and oligonucleotide derivatives in which the ribose in the oligonucleotide has been substituted with 2'-methoxyethoxy ribose. Unless otherwise indicated, a particular nucleic acid sequence is also intended to encompass conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated.Specifically, degenerate codon substitutions can be achieved by creating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)). As used herein, "nucleic acid" is also used interchangeably with gene, cDNA, mRNA, oligonucleotide, and polynucleotide. As used herein, "nucleotide" may be natural or non-natural.

[0045] As used herein, "homology" of genes refers to the degree of identity between two or more gene sequences. Generally, "homology" refers to a high degree of identity or similarity. Thus, the higher the homology between two genes, the higher the identity or similarity between their sequences. Whether two genes are homologous can be determined by direct sequence comparison or, in the case of nucleic acids, by hybridization under stringent conditions. When two gene sequences are directly compared, the genes are homologous if their DNA sequences are typically at least 50% identical, preferably at least 70% identical, and more preferably at least 80%, 90%, 95%, 96%, 97%, 98%, or 99% identical. Therefore, as used herein, "homolog" or "homologous gene product" refers to a protein in another species, preferably a mammal, that performs the same biological function as a protein component of a complex described further herein. Such a homolog may also be referred to as an "orthologous gene product."

[0046] Amino acids may be referred to herein by either their commonly known three-letter symbols or the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides may also be referred to by their commonly accepted one-letter codes. Herein, comparisons of amino acid and nucleotide sequence similarity, identity, and homology are calculated using the sequence analysis tool BLAST with default parameters. Identity searches can be performed, for example, using NCBI's BLAST 2.2.9 (published May 12, 2004). The identity values ​​used herein generally refer to the values ​​obtained when aligned using the above-mentioned BLAST under default conditions. However, if a higher value is obtained by changing the parameters, the highest value is used as the identity value. When identity is evaluated in multiple regions, the highest value among them is used as the identity value. Similarity is a numerical value that takes into account not only identity but also similar amino acids.

[0047] As used herein, the term "polynucleotide that hybridizes under stringent conditions" refers to conditions commonly used in the art. Such polynucleotides can be obtained by colony hybridization, plaque hybridization, Southern blot hybridization, or other methods using a polynucleotide selected from the polynucleotides disclosed herein as a probe. Specifically, the term refers to a polynucleotide that can be identified by hybridizing a filter onto which colony- or plaque-derived DNA has been immobilized in the presence of 0.7 to 1.0 M NaCl at 65°C, followed by washing the filter at 65°C using 0.1 to 2x SSC (saline-sodium citrate) solution (1x SSC solution is 150 mM sodium chloride and 15 mM sodium citrate). Hybridization can be carried out in accordance with the methods described in laboratory manuals such as Molecular Cloning 2nd ed., Current Protocols in Molecular Biology, Supplements 1-38, and DNA Cloning 1: Core Techniques, A Practical Approach, Second Edition, Oxford University Press (1995). Low stringency conditions include hybridization for 18-20 hours at 40°C in a buffer containing 35% formamide, 5xSSC, 50mM Tris-HCl (pH 7.5), 5mM EDTA, 0.02% polyvinylpyrrolidone (PVP), 0.02% BSA, 100µg / ml denatured salmon sperm DNA, and 10% (weight / volume) dextran sulfate, followed by washing for 1-5 hours at 55°C in a buffer consisting of 2xSSC, 25mM Tris-HCl (pH 7.4), 5mM EDTA, and 0.1% SDS, and then washing for 1.5 hours at 60°C in a buffer consisting of 2xSSC, 25mM Tris-HCl (pH 7.4), 5mM EDTA, and 0.1% SDS.

[0048] As used herein, a "purified" substance or biological factor (e.g., a nucleic acid or protein) refers to a biological factor from which at least a portion of the factors naturally associated with the biological factor have been removed. Thus, the purity of the biological factor in a purified biological factor is typically higher (i.e., more concentrated) than in the state in which the biological factor normally exists. As used herein, the term "purified" means that preferably at least 75% by weight, more preferably at least 85% by weight, even more preferably at least 95% by weight, and most preferably at least 98% by weight of the same type of biological factor is present. The substance used in the present disclosure is preferably a "purified" substance.

[0049] As used herein, a "corresponding" amino acid or nucleic acid refers to an amino acid or nucleotide in a polypeptide or polynucleotide molecule that has or is predicted to have the same function as a given amino acid or nucleotide in a reference polypeptide or polynucleotide. In particular, in the case of an enzyme molecule, this refers to an amino acid that is located at a similar position in the active site and contributes similarly to catalytic activity. For example, in the case of an antisense molecule, this may be a similar portion in an orthologue corresponding to a specific portion of the antisense molecule. The corresponding amino acid may be, for example, a specific amino acid that is cysteinylated, glutathionylated, forms an S-type disulfide bond, oxidized (e.g., oxidation of the methionine side chain), formylated, acetylated, phosphorylated, glycosylated, myristylated, or the like. Alternatively, the corresponding amino acid may be an amino acid responsible for dimerization. Such a "corresponding" amino acid or nucleic acid may be a region or domain spanning a certain range. Therefore, in such cases, it is referred to herein as a "corresponding" region or domain.

[0050] As used herein, a "corresponding" gene (e.g., a polynucleotide sequence or molecule) refers to a gene (e.g., a polynucleotide sequence or molecule) that has or is predicted to have the same function in a given species as a given gene in a species used as a reference for comparison. When multiple genes with such function exist, the term refers to genes that have the same evolutionary origin. Thus, a gene corresponding to a given gene may be an ortholog of that gene. Such corresponding genes can be identified using techniques well known in the art. Thus, for example, a corresponding gene in a given animal (e.g., a mouse or rat) can be found by searching a sequence database for that animal using the sequence of a reference gene for the corresponding gene (e.g., a human gene) as a query sequence.

[0051] As used herein, the term "fragment" refers to a polypeptide or polynucleotide having a sequence length of 1 to n-1 relative to the full-length polypeptide or polynucleotide (length n). The length of the fragment can be varied appropriately depending on the purpose. For example, the lower limit of the length for a polypeptide can be 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, or more amino acids, and lengths represented by integers not specifically recited herein (e.g., 11) may also be suitable as the lower limit. Furthermore, for a polynucleotide, the lower limit can be 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 75, 100, or more nucleotides, and lengths represented by integers not specifically recited herein (e.g., 11) may also be suitable as the lower limit. As used herein, such fragments are understood to fall within the scope of the present disclosure, for example, if the full-length fragment functions as a marker, as long as the fragment itself also functions as a marker. In the present disclosure, a fragment of a molecule is a substance (typically a polypeptide) that includes any region of the molecule, and does not necessarily have the biological function of the native molecule, as long as it can be used for the purposes of the present disclosure (e.g., treatment, detection, diagnosis, etc.).

[0052] As used herein, the term "expression" of a gene, polynucleotide, polypeptide, etc. refers to the transformation of the gene, etc., into a different form as a result of a certain action in vivo. Preferably, this refers to the transcription and translation of a gene, polynucleotide, etc. into a polypeptide, although transcription to produce mRNA can also be a form of expression. More preferably, such a polypeptide form may be one that has undergone post-translational processing (a derivative as referred to herein). For example, the expression level of a molecule can be determined by any method. Specifically, the expression level can be determined by assessing the amount of mRNA, the amount of protein, or the biological activity of the protein of this molecule.

[0053] As used herein, "expression level" refers to the amount of a polypeptide or mRNA, etc., expressed in a cell, tissue, etc. of interest. Examples of such expression levels include the expression level of a polypeptide of the present disclosure at the protein level, assessed by any suitable method using an antibody of the present disclosure, including immunological assays such as ELISA, RIA, fluorescent antibody analysis, Western blotting, and immunohistochemistry, or the expression level of a polypeptide used in the present disclosure at the mRNA level, assessed by any suitable method, including molecular biological assays such as Northern blotting, dot blotting, and PCR. "Changes in expression level" refers to an increase or decrease in the expression level of a polypeptide used in the present disclosure at the protein or mRNA level, assessed by any suitable method, including the immunological or molecular biological assays described above. By measuring the expression level of a certain marker, various marker-based detections or diagnoses can be performed.

[0054] As used herein, the term "marker (substance, protein, or gene (nucleic acid))" refers to a substance that serves as an indicator for tracking whether a certain state (e.g., cell type, normal cell state, disease state, impaired state, or level of proliferation ability, differentiation state, etc.) is in existence or is at risk of being in existence. Examples of such markers include genes (nucleic acid = DNA level), gene products (mRNA, protein, etc.), metabolic substances, enzymes, etc. In the present disclosure, detection, diagnosis, preliminary detection, prediction, or pre-diagnosis of a certain state (e.g., diabetes) can be achieved using a drug, agent, factor, or means specific to a marker associated with that state, or a composition, kit, system, etc. containing them.

[0055] As used herein, the term "antibody" broadly encompasses polyclonal antibodies, monoclonal antibodies, multispecific antibodies, chimeric antibodies, and anti-idiotypic antibodies, as well as fragments thereof, such as Fv fragments, Fab' fragments, F(ab')2 and Fab fragments, and other recombinantly produced conjugates or functional equivalents (e.g., chimeric antibodies, humanized antibodies, multifunctional antibodies, bispecific or oligospecific antibodies, single-chain antibodies, scFV, diabodies, sc(Fv)2 (single chain (Fv)2), scFv-Fc). Furthermore, such antibodies may be covalently bound or recombinantly fused to enzymes, such as alkaline phosphatase, horseradish peroxidase, α-galactosidase, etc. Antibodies used in the present disclosure are not limited in their origin, type, or form. Specifically, known antibodies, such as non-human animal antibodies (e.g., mouse antibodies, rat antibodies, and camel antibodies), human antibodies, chimeric antibodies, and humanized antibodies, can be used. In the present disclosure, either monoclonal or polyclonal antibodies can be used, but monoclonal antibodies are preferred. Specific binding of the antibody to the target protein is preferred.

[0056] As used herein, the term "means" refers to any tool that can achieve a certain purpose (e.g., detection, diagnosis, treatment, prevention, migration), and in particular, as used herein, "means for selectively recognizing (detecting)" refers to a means that can recognize (detect) a certain object differently from others.

[0057] As used herein, "detection" or "quantification" of polynucleotide or polypeptide expression can be achieved using any suitable method, including, for example, measurement of mRNA and immunoassays involving binding or interaction with a marker detection agent. Examples of molecular biological assays include Northern blotting, dot blotting, and PCR. Examples of immunoassays include ELISA, RIA, fluorescent antibody testing, luminescence immunoassay (LIA), immunoprecipitation (IP), immunodiffusion (SRID), immunoturbidimetry (TIA), Western blotting, and immunohistochemistry using microtiter plates. Examples of quantification methods include ELISA and RIA. Genetic analysis can also be performed using arrays (e.g., DNA arrays and protein arrays). DNA arrays are extensively reviewed in "DNA Microarrays and the Latest PCR Methods," a special edition of Cell Engineering, edited by Shujunsha. Protein arrays are described in detail in Nat. Genet. 2002 Dec;32 Suppl:526-32. In addition to the above, gene expression analysis methods include, but are not limited to, RT-PCR, RACE, SSCP, immunoprecipitation, two-hybrid systems, in vitro translation, etc. Such additional analysis methods are described, for example, in Genome Analysis Experimental Methods, Nakamura Yusuke Lab Manual, edited by Nakamura Yusuke, Yodosha (2002), and the descriptions therein are all incorporated by reference herein.

[0058] The detection agent or detection means of the present disclosure may be a complex or complex molecule in which another substance (e.g., a label) is bound to a detectable moiety (e.g., an antibody, etc.). As used herein, a "complex" or "complex molecule" refers to any construct comprising two or more moieties. For example, if one moiety is a polypeptide, the other moiety may be either a polypeptide or another substance (e.g., a substrate, a sugar, a lipid, a nucleic acid, another carbohydrate, etc.). As used herein, the two or more moieties constituting a complex may be linked by a covalent bond or by other bonds (e.g., hydrogen bonds, ionic bonds, hydrophobic interactions, van der Waals forces, etc.). When two or more moieties are polypeptides, the complex may also be referred to as a chimeric polypeptide. Thus, as used herein, a "complex" includes molecules formed by linking multiple types of molecules, such as polypeptides, polynucleotides, lipids, sugars, and small molecules.

[0059] The detection agents or other pharmaceuticals of the present disclosure can take the form of probes and primers. The probes and primers of the present disclosure can specifically hybridize with target nucleic acid molecules. The probes and primers of the present disclosure can detect the expression of target nucleic acid molecules and can be polymers composed of multiple bases or base pairs, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Double-stranded cDNA is also known to be usable in tissue in situ hybridization, and the probes and primers of the present disclosure also include such double-stranded cDNA. RNA probes (riboprobes) are particularly preferred probes and primers for detecting RNA in tissues.

[0060] The primers and primer sets according to the present disclosure can be used as primers and primer sets according to standard methods in known methods for detecting target genes using nucleic acid amplification methods such as PCR, RT-PCR, real-time PCR, in situ PCR, and LAMP.

[0061] As used herein, the term "probe" refers to a substance used as a search tool in biological experiments such as in vitro and / or in vivo screening, and examples include, but are not limited to, nucleic acid molecules containing a specific base sequence, peptides containing a specific amino acid sequence, specific antibodies or fragments thereof, etc. As used herein, a probe is used as a means for detecting a marker.

[0062] Nucleic acid molecules typically used as probes include those having a nucleic acid sequence at least 8 contiguous nucleotides long that is homologous or complementary to the nucleic acid sequence of a gene of interest. Such nucleic acid sequences are preferably at least 9 contiguous nucleotides long, more preferably at least 10 contiguous nucleotides long, even more preferably at least 11 contiguous nucleotides long, at least 12 contiguous nucleotides long, at least 13 contiguous nucleotides long, at least 14 contiguous nucleotides long, at least 15 contiguous nucleotides long, at least 20 contiguous nucleotides long, at least 25 contiguous nucleotides long, at least 30 contiguous nucleotides long, at least 40 contiguous nucleotides long, or at least 50 contiguous nucleotides long. Nucleic acid sequences used as probes include nucleic acid sequences that are at least about 70% homologous, more preferably at least about 80% homologous, even more preferably at least about 90% homologous, or at least about 95% homologous to the above sequences.

[0063] In one embodiment, the detection agent of the present disclosure may be labeled, or may have a tag attached thereto.

[0064] As used herein, the term "label" refers to an entity (e.g., substance, energy, electromagnetic waves, etc.) that distinguishes a target molecule or substance from others. Examples of such labeling methods include the RI (radioisotope) method, the fluorescence method, the biotin method, and the chemiluminescence method. When labeling multiple markers of the present disclosure or factors or means for capturing them using the fluorescence method, labeling is carried out with fluorescent substances that have mutually different maximum fluorescence emission wavelengths. The difference in maximum fluorescence emission wavelength is preferably 10 nm or more. When labeling a ligand, any substance that does not affect its function can be used, but examples of fluorescent substances include Alexa TM Fluor is preferred. Alexa TM Fluor is a water-soluble fluorescent dye obtained by modifying coumarin, rhodamine, fluorescein, cyanine, etc., and is a series that corresponds to a wide range of fluorescent wavelengths. Compared to other fluorescent dyes of the corresponding wavelength, it is very stable, bright, and has low pH sensitivity. Fluorescent dye combinations with a maximum fluorescence wavelength of 10 nm or more include Alexa TM 555 and Alexa TM 633 combinations, Alexa TM 488 and Alexa TM When labeling nucleic acids, any substance can be used as long as it can bind to the base moiety. TM It is preferable to use the Cy5 series (Cy3, Cy5, etc.), rhodamine 6G reagent, N-acetoxy-N2-acetylaminofluorene (AAF), AAIF (an iodine derivative of AAF), etc. Fluorescent substances with a difference in maximum fluorescence emission wavelength of 10 nm or more include, for example, a combination of Cy5 and rhodamine 6G reagent, a combination of Cy3 and fluorescein, and a combination of rhodamine 6G reagent and fluorescein. In the present disclosure, such labels can be used to modify a target object so that it can be detected by the detection means used. Such modifications are known in the art, and those skilled in the art can carry out such methods as appropriate depending on the label and the target object.

[0065] As used herein, the term "tag" refers to a substance for selecting a molecule through a specific recognition mechanism such as receptor-ligand, more specifically, a substance that acts as a binding partner for binding a specific substance (e.g., having a relationship such as biotin-avidin or biotin-streptavidin), and can be included in the category of "label." Thus, for example, a specific substance bound to a tag can be selected by contacting it with a substrate to which a binding partner of the tag sequence is bound. Such tags or labels are well known in the art. Representative tag sequences include, but are not limited to, myc tags, His tags, HA, Avi tags, etc. Such tags may be bound to the markers or marker detection agents of the present disclosure.

[0066] In this specification, the terms "drug," "agent," or "factor" (all of which are equivalent to "agent" in English) are used. The terms "substance" and "substances") are used interchangeably in a broad sense and may refer to any substance or other element (e.g., energy such as light, radioactivity, heat, or electricity) as long as the intended purpose can be achieved. Examples of such substances include, but are not limited to, cells (e.g., T cells), proteins, polypeptides, oligopeptides, peptides, polynucleotides, oligonucleotides, nucleotides, nucleic acids (e.g., DNA such as cDNA and genomic DNA, and RNA such as mRNA), polysaccharides, oligosaccharides, lipids, small organic molecules (e.g., hormones, ligands, signaling substances, small organic molecules, molecules synthesized by combinatorial chemistry, small molecules that can be used as pharmaceuticals (e.g., small molecule ligands), etc.), and composite molecules thereof.

[0067] As used herein, the term "kit" refers to a unit in which the components to be provided (e.g., test agents, diagnostic agents, therapeutic agents, antibodies, labels, instructions, etc.) are provided, usually separated into two or more compartments. This kit form is preferred when the purpose is to provide a composition that, for reasons of stability, should not be provided in a mixed state, but is preferably mixed immediately before use. Such a kit advantageously includes instructions or instructions describing how to use the components to be provided (e.g., test agents, diagnostic agents, therapeutic agents) or how to handle the reagents. When the kit is used herein as a reagent kit, the kit usually includes instructions describing how to use the test agents, diagnostic agents, therapeutic agents, antibodies, etc.

[0068] As used herein, the term "instructions" refers to written instructions for a physician or other user on how to use the present disclosure. The instructions include instructions for the detection method, use of a diagnostic agent, or administration of a medicine or the like according to the present disclosure. The instructions may also include instructions for oral or esophageal administration (e.g., by injection) as the administration site. The instructions are prepared in accordance with a format specified by the regulatory agency of the country in which the present disclosure is implemented (e.g., the Ministry of Health, Labor and Welfare in Japan, the Food and Drug Administration (FDA) in the United States, etc.), and clearly state that they have been approved by the regulatory agency. The instructions are so-called package inserts, and are typically provided in paper form, but are not limited thereto and may also be provided in the form of electronic media (e.g., a website provided on the Internet, email, etc.).

[0069] As used herein, the "amount" of an analyte or the like in a sample generally refers to an absolute value that reflects the mass of the analyte that can be detected in a volume of the sample. However, the amount also contemplates a relative amount compared to the amount of another analyte. For example, the amount of an analyte in a sample may be an amount that is greater than a control value or normal value of the analyte that is normally present in the sample.

[0070] As used herein, the "level" of an analyte or the like in a sample generally refers to an absolute value that reflects the value of the analyte's activity, etc., when the analyte is the target of a function such as an enzyme. However, the level also contemplates a relative level compared to the level of another analyte. For example, the level of an analyte in a sample may be a level that is greater than a control value or normal value for the analyte that is normally present in the sample.

[0071] The term "about," as used herein, refers to the indicated value plus or minus 10%. Note that even when "about" is not explicitly stated, it can be interpreted as having the same meaning as "about."

[0072] (Summary of the Disclosure) The present inventors have discovered a new aspect in which the efficacy of diabetes treatment targeting abnormal stem cells characterized by the abnormal expression of CD106 and the like can be improved by combining it with stem cell migration. Based on this new diabetes treatment strategy, the present disclosure provides a new treatment strategy and diagnosis for diabetes and / or its related diseases.

[0073] (Preferred embodiment) Preferred embodiments of the present disclosure will be described below. The embodiments provided below are provided for a better understanding of the present disclosure, and it is understood that the scope of the present disclosure should not be limited to the following description. Therefore, it is clear that those skilled in the art can make appropriate modifications within the scope of the present disclosure in light of the description herein. It is also understood that the following embodiments of the present disclosure can be used alone or in combination.

[0074] (Characteristics of abnormal stem cells in diabetes and / or related diseases) In one embodiment, the abnormal stem cells of the present disclosure may be hematopoietic stem cells. In one embodiment, the abnormal stem cells of the present disclosure are characterized by not expressing and / or not functioning at normal levels at least one of CD106, CD34, TNF-α, proinsulin, and histone deacetylases (HDACs), and, if desired, may be further characterized by having other characteristics of abnormal stem cells described herein (e.g., c-Kit positivity, Sca-1 positivity, hematopoietic stem cell lineage marker negativity, the above-mentioned characteristics of short-term hematopoietic stem cells (e.g., CD38 negativity)). In one embodiment, the abnormal stem cells of the present disclosure may be characterized by not expressing CD106 at normal levels. In one embodiment, the abnormal stem cells of the present disclosure are characterized by (a) not expressing CD106 at normal levels, and (b) not expressing at normal levels at least one of CD34, TNF-α, proinsulin, and histone deacetylases (HDACs), and may optionally be further characterized by having other characteristics of abnormal stem cells described herein (e.g., c-Kit positivity, Sca-1 positivity, hematopoietic stem cell lineage marker negativity, short-term hematopoietic stem cell characteristics described above (e.g., CD38 negativity)).

[0075] In one representative embodiment, the abnormal stem cells (e.g., hematopoietic stem cells) of the present disclosure can be characterized by abnormal expression of CD106. In one embodiment, the abnormal stem cells of the present disclosure can be characterized by CD106 expression levels higher than the CD106 expression levels in total bone marrow cells or hematopoietic stem cells (e.g., CD34-positive Thy-1-positive cells) of a population of non-diabetic subjects ... 2 That's it, 2 x 10 2 That's it, 5 x 10 2 That's it, 1 x 10 3 That's it, 2 x 10 3 That's it, 5 x 10 3 That's it, 1 x 10 4 That's it, 2 x 10 4 That's it, 5 x 10 4 That's it, 1 x 105 That's it, 2 x 10 5 That's it, 5 x 10 5 That's it, 1 x 10 6 That's it, 2 x 10 6 That's it, 5 x 10 6 or more, or 1×10 7 In a specific embodiment, the abnormal stem cells of the present disclosure may be characterized by expressing at least about 1 x 10 CD106 molecules on the cell surface. 4 In one embodiment, when whole bone marrow cells or hematopoietic stem cells (e.g., CD34-positive Thy-1-positive cells) from a normal subject (e.g., human) are labeled with a fluorescent dye-conjugated antibody against CD106 and analyzed by FACS, the abnormal stem cells of the present disclosure can be indicated by an expression level that exhibits a fluorescent intensity that is 1-fold or more, 2-fold or more, 5-fold or more, 10-fold or more, 20-fold or more, 50-fold or more, 100-fold or more, 200-fold or more, 500-fold or more, or 1000-fold or more of the median fluorescent intensity from the fluorescent dye observed for whole bone marrow cells or hematopoietic stem cells. In one embodiment, the abnormal stem cells of the present disclosure may be characterized by CD106 mRNA expression levels that are 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 150% or more, or 200% or more higher than hematopoietic stem cells (e.g., CD34-positive cells) from a population of non-diabetic subjects.

[0076] In one embodiment, the abnormal stem cells of the present disclosure can be characterized by the expression of TNF-α. In one embodiment, the abnormal stem cells of the present disclosure can be characterized by being positive for the expression of TNF-α. In one embodiment, positive for the expression of TNF-α can be indicated by an expression level that exhibits a fluorescence intensity that is 1-fold or more, 2-fold or more, 5-fold or more, 10-fold or more, 20-fold or more, 50-fold or more, 100-fold or more, 200-fold or more, 500-fold or more, or 1000-fold or more than the median fluorescence intensity from the fluorescent dye observed for the whole bone marrow cells or hematopoietic stem cells when whole bone marrow cells or hematopoietic stem cells (e.g., CD34-positive Thy-1-positive cells) of a normal subject (e.g., human) are labeled with a fluorescent dye-conjugated antibody against TNF-α and analyzed by FACS. In one embodiment, the abnormal stem cells of the present disclosure may be characterized by TNF-α mRNA expression levels that are 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 150% or more, or 200% or more higher than hematopoietic stem cells (e.g., CD34-positive cells) from a population of non-diabetic subjects.

[0077] In one embodiment, the abnormal stem cells of the present disclosure can be characterized by abnormal expression of proinsulin. In one embodiment, the abnormal stem cells of the present disclosure can be characterized by being positive for proinsulin expression. In one embodiment, positive for proinsulin expression can be indicated by an expression level that exhibits a fluorescence intensity that is 1-fold or more, 2-fold or more, 5-fold or more, 10-fold or more, 20-fold or more, 50-fold or more, 100-fold or more, 200-fold or more, 500-fold or more, or 1000-fold or more than the median fluorescence intensity from the fluorescent dye observed for the whole bone marrow cells or hematopoietic stem cells when whole bone marrow cells or hematopoietic stem cells (e.g., CD34-positive Thy-1-positive cells) of a non-diabetic subject (e.g., human) are labeled with a fluorescent dye-conjugated antibody against proinsulin and analyzed by FACS. In one embodiment, the abnormal stem cells of the present disclosure may be characterized by insulin (or proinsulin) mRNA expression levels that are 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 150% or more, 200% or more, 300% or more, 400% or more, 500% or more, 700% or more, or 1000% or more higher than hematopoietic stem cells (e.g., CD34-positive cells) from a population of non-diabetic subjects.

[0078] In one embodiment, the abnormal stem cells of the present disclosure can be characterized by the expression of c-Kit. In one embodiment, the abnormal stem cells of the present disclosure can be characterized by being positive for the expression of c-Kit. In one embodiment, positive c-Kit expression can be indicated by an expression level that, when whole bone marrow cells or hematopoietic stem cells (e.g., CD34+Thy-1+ cells) from a non-diabetic subject (e.g., a human) are labeled with a fluorochrome-conjugated antibody against c-Kit and analyzed by FACS, exhibits a fluorescence intensity that is 0.1-fold or more, 0.2-fold or more, 0.5-fold or more, 1-fold or more, 2-fold or more, 5-fold or more, 10-fold or more, 20-fold or more, 50-fold or more, 100-fold or more, 200-fold or more, 500-fold or more, or 1000-fold or more of the median fluorescence intensity from the fluorochrome observed for whole bone marrow cells or hematopoietic stem cells, and / or exhibits a fluorescence intensity that is within the top 70%, 60%, 50%, 40%, 30%, 20%, or 10% of the cells when sorted in order of greatest fluorescence intensity from the fluorochrome.

[0079] In one embodiment, the abnormal stem cells covered by the present disclosure can be characterized by the expression of Sca-1. In one embodiment, the abnormal stem cells of the present disclosure can be characterized by being positive for the expression of Sca-1. In one embodiment, positive expression of Sca-1 can be indicated by an expression level that, when whole bone marrow cells from a non-diabetic subject (e.g., a human) are labeled with a fluorescent dye-conjugated antibody against Sca-1 and analyzed by FACS, exhibits a fluorescence intensity that is at least 0.1 times, at least 0.2 times, at least 0.5 times, at least 1 times, at least 2 times, at least 5 times, at least 10 times, at least 20 times, at least 50 times, at least 100 times, at least 200 times, at least 500 times, or at least 1000 times the median fluorescence intensity from the fluorescent dye observed for whole bone marrow cells, and / or exhibits a fluorescence intensity that is within the top 70%, 60%, 50%, 40%, 30%, 20%, or 10% of the whole bone marrow cells sorted in order of greatest fluorescence intensity from the fluorescent dye.

[0080] In one embodiment, the abnormal stem cells of the present disclosure can be characterized by the expression of lineage markers of hematopoietic stem cells, such as CD3 (T cells), CD19 (B cells), NK1.1 (NK cells), CD11c (dendritic cells), CD11b (monocytes), FcεRI (mast cells), and Gr-1 (granulocytes). In one embodiment, the abnormal stem cells of the present disclosure can be characterized by being negative for the expression of one or more (e.g., all) of CD3, CD19, NK1.1, CD11c, CD11b, FcεRI, and Gr-1. In one embodiment, negativity for each of CD3, CD19, NK1.1, CD11c, CD11b, FcεRI, and Gr-1 expression can be indicated by an expression level that, when whole bone marrow cells from a non-diabetic subject (e.g., a human) are labeled with a fluorochrome-conjugated antibody against CD3, CD19, NK1.1, CD11c, CD11b, FcεRI, or Gr-1 and analyzed by FACS, exhibits a fluorescence intensity that is 1000% or less, 500% or less, 200% or less, 100% or less, 50% or less, 20% or less, 10% or less, 5%, 2%, or 1% or less of the median fluorescence intensity from the fluorochrome observed for whole bone marrow cells, and / or that is within the bottom 50%, 20%, 10%, 5%, 2%, or 1% when whole bone marrow cells are sorted in descending order of fluorescence intensity from the fluorochrome. In one embodiment, the abnormal stem cells of the present disclosure may be characterized by CD34 mRNA expression levels that are 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 150% or more, or 200% or more higher than hematopoietic stem cells from a population of non-diabetic subjects.

[0081] In one embodiment, the abnormal stem cells covered by the present disclosure can be characterized as hematopoietic stem cells. In one embodiment, human hematopoietic stem cells can also be characterized by being CD34-positive, Thy-1-positive, or Lineage-negative, CD34-positive, CD38-negative, CD90-positive, CD45RA-negative. In one embodiment, mouse hematopoietic stem cells can be characterized by being c-kit-positive, Sca-1-positive, and lineage marker-negative (KSL). In one embodiment, hematopoietic stem cells can also be characterized by bone marrow cells stained with Hoechest 33342 dye being negative when excited with ultraviolet light (350 nm) and developed using two optical filters, Hoechst blue and Hoechst red.

[0082] In one embodiment, the abnormal stem cells covered by the present disclosure can be characterized by the cell stage of hematopoietic stem cells. In one embodiment, the abnormal stem cells of the present disclosure can be short-term hematopoietic stem cells.

[0083] In one embodiment, the abnormal stem cells of the present disclosure may be characterized by expression of one or more of histone deacetylase genes (HDACs) (e.g., Hdac3, Hdac4, Hdac8, Hdac9) that are 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 150% or more, 200% or more, 300% or more, 400% or more, or 500% or more higher than hematopoietic stem cells (e.g., c-Kit-positive, lineage marker-negative cells) from a population of non-diabetic subjects.

[0084] In one embodiment, proinsulin and / or TNF-α can be markers that indicate that the abnormal stem cells targeted by the present disclosure are localized in the bone marrow.

[0085] Treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders and / or symptoms by targeting abnormal stem cells in combination with stem cell migration In one aspect, the present disclosure provides treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders, and / or symptoms by inducing and inhibiting the migration of abnormal stem cells having the characteristics described herein. This treatment and / or prevention can be achieved by any means, such as by implementing a method, or by providing a composition, combination, kit, or system for this purpose. For example, when using an inhibitor such as an antibody, the inhibitory effect on cells present in the bone marrow may be limited (see, for example, the Examples). Therefore, as disclosed herein, allowing abnormal stem cells to migrate before inhibiting the abnormal stem cells may improve the inhibitory effect of the abnormal stem cells.

[0086] In one embodiment, the treatment and / or prevention disclosed herein can be carried out by inducing and inhibiting the migration of all cells that have at least one characteristic of the abnormal stem cells described herein. That is, in this embodiment, the cells that are migrated and inhibited are not limited to the abnormal stem cells described herein. It may be technically difficult to inducing and inhibiting only the abnormal stem cells described herein, and treatment and / or prevention methods may be selected taking into account the benefits of inducing and inhibiting the migration of abnormal stem cells and the effects of inducing and inhibiting the migration of other cells. On the other hand, because inducing and inhibiting the migration of only some of the abnormal stem cells described herein may be insufficient for the treatment and / or prevention of diabetes and / or diabetes-related diseases, disorders, and / or symptoms, it may be preferable to inducing and inhibiting the migration of all abnormal stem cells described herein. In the treatment and / or prevention disclosed herein, the characteristics of the abnormal stem cells used for migration and the characteristics of the abnormal stem cells used for inhibition may be the same or different. For example, in one embodiment, the treatment and / or prevention of the present disclosure may utilize the characteristic of abnormal stem cells being hematopoietic stem cells for migration (e.g., using a CXCR4 antagonist and / or a CXCR2 stimulator) and the characteristic of abnormal stem cells being abnormally expressing a specific cell surface protein (such as CD106) for inhibition (e.g., using an anti-CD106 antibody).

[0087] In one embodiment, the migration in the treatment and / or prevention of the present disclosure can be achieved by inducing the migration of abnormal stem cells having any of the characteristics described herein. In one embodiment, the migration of abnormal stem cells described herein can be achieved by a treatment that induces hematopoietic stem cell migration. In one embodiment, treatments that induce hematopoietic stem cell migration include, but are not limited to, CXCR4 inhibition (Future Oncol. 2007 Feb;3(1):19-27), epidermal growth factor receptor (EGFR) inhibition (Nature Medicine volume 16, pages 1141-1146 (2010)), granulocyte colony-stimulating factor (G-CSF) stimulation (Blood. 1995 Dec 15;86(12):4437-45), and CXCR2 stimulation (Cell. 2018 Jan 11;172(1-2):191-204.e10).

[0088] In one embodiment, the migration in the treatment and / or prevention of the present disclosure can be carried out using a chemotactic agent for abnormal stem cells having any of the characteristics described herein. Those skilled in the art can appropriately select a chemotactic agent that can be used in the migration in the treatment and / or prevention of the present disclosure. For example, a chemotactic agent that can be used in the treatment and / or prevention of the present disclosure can be selected by actually measuring the proportion of stem cells (e.g., abnormal stem cells having the characteristics described herein) in the bone marrow and / or blood before and after administration of the chemotactic agent and examining the migration effect. In one embodiment, the migration in the treatment and / or prevention of the present disclosure can be carried out using a chemotactic agent for hematopoietic stem cells. In one embodiment, the hematopoietic stem cell migration agent may have functions such as, but are not limited to, inhibition of CXCR4 (e.g., plerixafor (AMD3100)), inhibition of epidermal growth factor receptor (EGFR) (e.g., gefitinib, erlotinib, afatinib, osimertinib, etc.), induction of granulocyte colony-stimulating factor (G-CSF) stimulation (e.g., filgrastim, nartograstim, lenograstim, pegfilgrastim, etc.), and induction of CXCR2 stimulation (e.g., GROβ (MIP2)). In one embodiment, the hematopoietic stem cell migration agent may be used in any combination, for example, a combination of a CXCR4 inhibitor (e.g., plerixafor (AMD3100)) and a CXCR2 stimulation inducer (e.g., GROβ (MIP2)).

[0089] In one embodiment, the suppression of abnormal stem cells described herein can be achieved by targeting any of the characteristics of abnormal stem cells described herein, allowing the targeted cells to migrate, and then suppressing them. For example, when targeting molecules expressed on the cell surface by the abnormal stem cells described herein (e.g., CD106, CD34, TNF-α, proinsulin, c-Kit, Sca-1), molecules that specifically bind to the cell surface-expressed molecules (e.g., antibodies, T cell receptors, small molecules, etc.) can be used. Furthermore, when targeting cells that simultaneously express multiple types of molecules, molecules that bind to multiple types of molecules, such as multispecific antibodies, can be used to target the target cells.

[0090] In one embodiment, the suppression in the treatment and / or prevention of the present disclosure can be carried out by migrating and suppressing abnormal stem cells characterized by at least one of abnormal CD106 expression, abnormal CD34 expression, abnormal TNF-α expression, abnormal proinsulin expression, and abnormal histone deacetylase (HDAC) expression. In one embodiment, the suppression in the treatment and / or prevention of the present disclosure can be carried out by migrating and suppressing abnormal stem cells characterized by abnormal CD106 expression. In one embodiment, the suppression in the treatment and / or prevention of the present disclosure can be carried out by migrating and suppressing abnormal stem cells characterized by (a) abnormal CD106 expression and (b) at least one of abnormal CD34 expression, abnormal TNF-α expression, abnormal proinsulin expression, and abnormal histone deacetylase (HDAC) expression.

[0091] In one embodiment, the suppression in the treatment and / or prevention of the present disclosure may be carried out using one or more inhibitors that target at least one of CD106, CD34, TNF-α, proinsulin, and histone deacetylases (HDACs). In one embodiment, the suppression in the treatment and / or prevention of the present disclosure may be carried out using an inhibitor that targets CD106. In one embodiment, the suppression in the treatment and / or prevention of the present disclosure may be carried out using one or more inhibitors that target (a) CD106 and (b) at least one of CD34, TNF-α, proinsulin, and histone deacetylases (HDACs). The inhibitor can be any inhibitor described herein, for example, an antibody that binds to the above molecule. In one embodiment, the inhibitor targeting histone deacetylases (HDACs) may be an HDAC inhibitor, such as, but not limited to, TSA (trichostatin A), VPA (valproic acid), sodium butyrate (NaBu), SAHA (suberoylanilide hydroxamic acid or vorinostat), sodium phenylbutyrate, depsipeptide (FR901228, FK228), trapoxin (TPX), cyclic hydroxamic acid-containing peptide 1 (CHAP1), MS-275, LBH589, and PXD101.

[0092] In one embodiment, the treatment and / or prevention according to the present disclosure may be treatment and / or prevention of diabetes. In one embodiment, the treatment and / or prevention according to the present disclosure may be treatment and / or prevention of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorder, delayed fracture healing, eating disorder, or skin disorder.

[0093] The treatment and / or prevention of the present disclosure may be combined with any known therapeutic and / or preventive treatment or measure (e.g., therapeutic and / or preventive treatment or measure for diabetes and / or diabetes-related diseases, disorders and / or symptoms).

[0094] (Diagnosis of diabetes and / or diabetes-related diseases, disorders and / or symptoms, or risk thereof, based on abnormal stem cell migration and / or persistence) In one aspect, the present disclosure provides for the diagnosis of diabetes and / or diabetes-related diseases, disorders, and / or symptoms, or the risk thereof, based on the abnormal stem cell migration and / or persistence described herein, which diagnosis may be achieved by any means, including by practicing a method, providing a composition, combination, kit, or system for this purpose, or the like.

[0095] In one embodiment, the diagnosis of the present disclosure can be performed by detecting the presence and / or abundance of at least some cells having at least one characteristic of an abnormal stem cell described herein in a specific location. As used herein, unless otherwise specified, the "abundance" of a cell refers not only to the number of cells (or any indicator indicating the number of cells) but also to the proportion of a specific cell in a cell population (or any indicator indicating the proportion of a specific cell). In one embodiment, the diagnosis of the present disclosure can be performed by detecting the presence and / or abundance of an abnormal stem cell described herein in the bone marrow and / or bone marrow niche. In one embodiment, the diagnosis of the present disclosure can be performed by detecting the presence and / or abundance of an abnormal stem cell described herein in circulating blood. In one embodiment, the diagnosis of the present disclosure can be performed by detecting the presence and / or abundance of an abnormal stem cell in a specific location in a subject administered a migration agent described herein. For example, the diagnosis can be performed based on a change in the presence and / or abundance of an abnormal stem cell in a specific location (e.g., bone marrow) before and after administration of the migration agent. In this embodiment, for example, if the abundance of abnormal stem cells in the bone marrow of a subject is reduced after administration of the migration agent, the subject may be diagnosed as having, or at risk for, diabetes and / or diabetes-related diseases, disorders and / or symptoms, and in one embodiment, the subject may be subjected to the therapeutic and / or preventative treatment of the present disclosure using the migration agent.

[0096] In one embodiment, the diagnosis of the present disclosure can be performed based on a quantitative indicator of at least one characteristic of abnormal stem cells described herein (e.g., cell surface protein expression level, mRNA expression level). In one embodiment, when a sample (e.g., a blood sample, a bone marrow sample) from a subject or hematopoietic stem cells (e.g., CD34-positive cells) contained therein exhibits an mRNA expression level of CD106 that is 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 150% or more, or 200% or more higher than that of a corresponding sample or hematopoietic stem cells (e.g., CD34-positive cells) contained therein from a non-diabetic subject population, the subject can be determined to have, or be at risk for, diabetes and / or diabetes-related diseases, disorders, and / or symptoms, or to be suitable for the abnormal cell migration treatment of the present disclosure.

[0097] In one embodiment, the diagnosis of the present disclosure can be performed based on the percentage of cells that exhibit at least one characteristic (including a quantitative indicator) of abnormal stem cells described herein. In one embodiment, the diagnosis of the present disclosure can be performed based on the percentage of cells that exhibit abnormal levels of CD106 expression among the subject's hematopoietic stem cells, and can be performed by comparing the percentage of cells that exhibit abnormal levels of CD106 expression among the hematopoietic stem cells of a non-diabetic subject population or a diabetic subject population, as needed. In one embodiment, if the proportion of cells exhibiting abnormal levels of CD106 expression among the subject's hematopoietic stem cells at a particular site (e.g., bone marrow) is 1.1 times or more, 1.2 times or more, 1.3 times or more, 1.4 times or more, 1.5 times or more, 1.6 times or more, 1.7 times or more, 1.8 times or more, 1.9 times or more, 2 times or more, 2.1 times or more, 2.2 times or more, 2.3 times or more, 2.4 times or more, 2.5 times or more, 2.6 times or more, 2.7 times or more, 2.8 times or more, 2.9 times or more, 3 times or more, 3.5 times or more, 4 times or more, 4.5 times or more, or 5 times or more than the proportion of cells exhibiting abnormal levels of CD106 expression among the hematopoietic stem cells of a population of non-diabetic subjects, the subject can be determined to have, or be at risk of, diabetes and / or diabetes-related diseases, disorders and / or symptoms, or to be suitable for the abnormal cell migration treatment of the present disclosure.

[0098] In one embodiment, the diagnosis of the present disclosure can be performed by detecting any of the characteristics of abnormal stem cells described herein. In one embodiment, the diagnosis of the present disclosure can be performed by detecting cells having at least one of the characteristics of abnormal stem cells: abnormal CD106 expression, abnormal CD34 expression, abnormal TNF-α expression, abnormal proinsulin expression, and abnormal histone deacetylase (HDAC) expression, and, if necessary, other characteristics of abnormal stem cells described herein (e.g., c-Kit positivity, Sca-1 positivity, negative hematopoietic stem cell lineage markers, the above-mentioned characteristics of short-term hematopoietic stem cells (e.g., negative CD38)). In one embodiment, the diagnosis of the present disclosure can be performed by detecting cells characterized by abnormal CD106 expression. In one embodiment, the diagnosis of the present disclosure can be performed by detecting cells characterized by (a) abnormal expression of CD106 and (b) at least one of abnormal expression of CD34, abnormal expression of TNF-α, abnormal expression of proinsulin, and abnormal expression of histone deacetylases (HDACs), and optionally further having other characteristics of abnormal stem cells described herein (e.g., c-Kit positivity, Sca-1 positivity, negative lineage markers of hematopoietic stem cells, characteristics of the above-mentioned short-term hematopoietic stem cells (e.g., negative CD38)).

[0099] In one embodiment, the diagnosis of the present disclosure can be performed using one or more detection agents that target at least one of CD106, CD34, TNF-α, proinsulin, and histone deacetylases (HDACs). In one embodiment, the diagnosis of the present disclosure can be performed using a detection agent that targets CD106. In one embodiment, the diagnosis of the present disclosure can be performed using one or more detection agents that target (a) CD106 and (b) at least one of CD34, TNF-α, proinsulin, and histone deacetylases (HDACs). The detection agent can be any detection agent described herein, including, for example, an antibody that binds to the above molecules.

[0100] In one embodiment, the diagnosis of the present disclosure may be performed by administering a detection agent to a subject or by testing a sample from the subject. For example, the diagnosis of the present disclosure may be performed by administering a detection agent of the present disclosure to a subject to detect the presence and / or abundance of abnormal stem cells of the present disclosure in bone marrow (or a specific niche thereof). For example, the diagnosis of the present disclosure may be performed by obtaining a sample containing cells from the subject (e.g., a blood sample, a bone marrow sample) and determining whether cells having at least one characteristic of the abnormal stem cells of the present disclosure and / or a marker indicative of their presence in bone marrow (or a specific niche thereof) are present.

[0101] In one embodiment, the diagnosis of the present disclosure may be a diagnosis of diabetes, hi one embodiment, the diagnosis of the present disclosure may be a diagnosis of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorder, delayed fracture healing, eating disorder, or skin disorder, or the risk thereof.

[0102] The diagnosis of the present disclosure may be any known diagnosis (e.g., diabetes and / or diabetes-related The present invention may be combined with other methods, such as diagnosis of a disease, disorder and / or condition.

[0103] In one embodiment, the treatment and / or prevention of the present disclosure can be carried out based on the diagnosis of the present disclosure (e.g., for a subject in whom the diagnosis of the present disclosure predicts that abnormal stem cells are present in the bone marrow).

[0104] The therapeutic, prophylactic, and / or diagnostic methods of the present disclosure can be performed on any subject. In one embodiment, the subject is a mammal, such as a human, mouse, guinea pig, hamster, rat, mouse, rabbit, pig, sheep, goat, cow, horse, cat, dog, marmoset, monkey, or chimpanzee. In a specific embodiment, the subject is a human.

[0105] (drugs, dosage forms, etc.) The inhibitors, migration agents and detection agents for abnormal stem cells described herein can be provided as compositions or pharmaceuticals in various forms.

[0106] The administration route of the abnormal stem cell inhibitors, migration agents, and abnormal stem cell detectors described herein is preferably one that is effective in treating, preventing, or detecting diabetes and / or diabetes-related diseases, disorders, and / or symptoms, and may be, for example, intravenous, subcutaneous, intramuscular, intraperitoneal, or oral administration. The dosage form may be, for example, an injection, capsules, tablets, granules, or the like. Aqueous solutions for injection may be stored, for example, in vials or stainless steel containers. Furthermore, aqueous solutions for injection may contain, for example, physiological saline, sugars (e.g., trehalose), NaCl, or NaOH. Furthermore, therapeutic agents may contain, for example, buffers (e.g., phosphate buffer), stabilizers, and the like.

[0107] Generally, the compositions, medicaments, inhibitors, migration agents, detection agents, etc. of the present disclosure comprise a therapeutically effective amount of an active ingredient or a detectable amount of a detection means, and a pharmaceutically acceptable carrier or excipient. As used herein, "pharmaceutically acceptable" means approved by a government regulatory agency or listed in a Pharmacopoeia or other generally recognized pharmacopeia for use in animals, and more particularly, humans. As used herein, "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic or detection agent is administered. Such carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, including, but not limited to, peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a preferred carrier when medicaments are administered orally. Saline and aqueous dextrose are preferred carriers when pharmaceutical compositions are administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions are preferably used as liquid carriers for injectable solutions. Suitable excipients include light anhydrous silicic acid, crystalline cellulose, mannitol, starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, glycol, water, ethanol, carmellose calcium, carmellose sodium, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl acetal diethylaminoacetate, polyvinylpyrrolidone, gelatin, medium-chain triglycerides, polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethylcellulose, corn starch, inorganic salts, etc. The compositions can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired. These compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, etc. The compositions can also be formulated as suppositories, using traditional binders and carriers, such as triglycerides.Oral formulations can contain standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of suitable carriers are described in E.W. Martin, Remington's Pharmaceutical Sciences (Mark Publishing Company, Easton, USA). Such compositions contain a therapeutically effective amount of the therapeutic agent, preferably in purified form, together with an appropriate amount of carrier to provide a form that is properly administered to the patient. The formulation should be appropriate for the mode of administration. Other ingredients may include, for example, surfactants, excipients, colorants, flavoring agents, preservatives, stabilizers, buffers, suspending agents, isotonicity agents, binders, disintegrants, lubricants, flow enhancers, flavoring agents, etc.

[0108] In one embodiment of the present disclosure, the term "salt" refers to an anionic salt formed with any acidic (e.g., carboxyl) group or a cationic salt formed with any basic (e.g., amino) group. Salts include inorganic or organic salts, such as those described in Berge et al., J. Pharm. Sci., 1977, 66, 1-19. Examples of suitable salts include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, and salts with organic acids. In one embodiment of the present disclosure, a "solvate" refers to a compound formed by a solute and a solvent. For details of solvates, see, for example, J. Honig et al., The Van Nostrand Chemist's Dictionary, p. 650 (1953). When the solvent is water, the resulting solvate is a hydrate. Preferably, the solvent does not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water and various buffers.

[0109] In the present disclosure, various delivery systems can be used to administer the inhibitors and / or migration agents of the present disclosure to the appropriate site. Such systems include, for example, encapsulation in liposomes, microparticles, and microcapsules; use of receptor-mediated endocytosis; and construction of therapeutic nucleic acids as part of retroviral or other vectors. Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. Medicaments can be administered by any suitable route, such as by infusion, bolus injection, or absorption through epithelial or mucocutaneous linings (e.g., oral, rectal, and intestinal mucosa), or by inhaler or nebulizer using an aerosolizing agent, if necessary, and can be administered together with other biologically active agents. Administration can be systemic or local.

[0110] In a preferred embodiment, the composition can be formulated as a pharmaceutical composition adapted for administration to humans according to known methods. Such compositions can be administered by injection. Typically, compositions for injection administration are solutions in sterile isotonic aqueous buffer. If necessary, the composition can also include a solubilizing agent and a local anesthetic such as lidocaine to ease pain at the injection site. Generally, the ingredients are supplied separately or mixed together in unit dosage form, for example, as a lyophilized powder or water-free concentrate in a hermetically sealed container such as an ampoule or sachet indicating the quantity of active ingredient. If the composition is to be administered by injection, it can be dispensed using an infusion bottle containing sterile pharmaceutical-grade water or saline. If the composition is to be administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed prior to administration.

[0111] The compositions, pharmaceuticals, migration agents, and inhibitors of the present disclosure can be formulated in neutral or salt form or as other prodrugs (e.g., esters, etc.). Pharmaceutically acceptable salts include those formed with free carboxyl groups, such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, tartaric acid, etc.; those formed with free amine groups, such as those derived from isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc.; and those derived from sodium, potassium, ammonium, calcium, and ferric hydroxide, etc.

[0112] The amount of the disclosed inhibitor and / or migration agent may vary depending on the nature of the disorder or condition, but can be determined by one of skill in the art using standard clinical techniques based on the disclosure herein. In addition, in vitro assays may be used to help identify optimal dosage ranges. The exact dose to be used in the formulation may also vary depending on the route of administration and the severity of the disease or disorder, and should be determined according to the judgment of the attending physician and the circumstances of each patient. However, the dosage is not particularly limited, and may be, for example, 0.001, 1, 5, 10, 15, 100, or 1,000 mg / kg body weight per administration, or within any two of these ranges. The administration interval is not particularly limited, and may be, for example, once or twice every 1, 7, 14, 21, or 28 days, or once or twice within any two of these ranges. The dosage, administration interval, and administration method may be selected appropriately depending on the patient's age, weight, symptoms, target organ, etc. Therapeutic agents preferably contain a therapeutically effective amount, or an amount of an active ingredient effective to exert a desired effect. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0113] The pharmaceutical composition or therapeutic or prophylactic agent of the present disclosure can be provided as a kit.

[0114] In certain embodiments, the present disclosure provides pharmaceutical packs or kits comprising one or more containers filled with one or more ingredients of the compositions or medicaments of the present disclosure, optionally associated with such containers may be information indicating approval by a governmental agency to manufacture, use, or sell for human administration, in a manner prescribed by the governmental agency regulating the manufacture, use, or sale of pharmaceutical or biological products.

[0115] Procedures for formulating the therapeutic agents, prophylactic agents, etc. of the present disclosure as pharmaceuticals and the like are known in the art and are described, for example, in the Japanese Pharmacopoeia, the United States Pharmacopoeia, the Pharmacopoeias of other countries, etc. Therefore, those skilled in the art will be able to determine embodiments such as the amounts to be used without undue experimentation, given the description herein.

[0116] In this specification, "or" is used when "at least one or more" of the items listed in the sentence can be employed. The same applies to "alternative." In this specification, when "within a range" of "two values" is specified, the range includes the two values ​​themselves.

[0117] All references cited herein, including scientific literature, patents, patent applications, and the like, are incorporated by reference in their entirety to the same extent as if each were specifically set forth.

[0118] The present disclosure has been described above by showing preferred embodiments for ease of understanding. The present disclosure will be described below based on examples. However, the above description and the following examples are provided for illustrative purposes only and are not intended to limit the present disclosure. Therefore, the scope of the present disclosure is not limited to the embodiments or examples specifically described herein, but is limited only by the scope of the claims. [Example]

[0119] When necessary, animals used in the following examples were handled in accordance with the guidelines for animal experiments of Shiga University of Medical Science. The reagents used were specifically those listed in the examples, but equivalent products from other manufacturers (Sigma-Aldrich, Wako Pure Chemical Industries, Nakarai, R&D Systems, USCN Life Science Inc., etc.) can also be used.

[0120] In the following examples, the abbreviations have the following meanings. DM diabetes STZ-DM Streptozotocin-induced diabetes KSL cells c-Kit positive Sca-1 positive Lineage (lineage marker) negative cells SNCV sensory nerve conduction velocity

[0121] Example 1: Identification of abnormal stem cells involved in diabetes To identify abnormal cells involved in diabetes, we analyzed stem cells obtained from diabetic model mice.

[0122] method Obtaining mice and bone marrow cells C57BL / 6J wild-type mice (CLEA Japan, Osaka, Japan) were used for this study. Diabetes was induced by intravenous injection of streptozotocin (STZ) (150 mg / kg) (Nacalai Tesque, Kyoto, Japan) to create a type 2 diabetes model (STZ mice). Bone marrow cells were isolated from 8-week-old mice.

[0123] FACS Ficoll-Paque Plus (GE Healthcare Bio-Sciences AB, Uppsala Mononuclear cells were isolated from whole bone marrow using a centrifuge tube (Microfluidics Laboratory, University of Sweden).

[0124] To assess TNF-α and CD106 expression, mononuclear cells were immunoreacted with PE-Cy7-conjugated streptavidin antibody (BD Biosciences, San Jose, CA), biotin mouse lineage panel stain (BD Biosciences), APC-conjugated anti-c-kit antibody (BD Biosciences), APC-Cy7-conjugated anti-Ly6A / E (Sca-1) antibody (BD Biosciences), fluorescein isothiocyanate (FITC)-conjugated anti-CD106 antibody (BD Biosciences), and phycoerythrin (PE)-conjugated anti-TNF-α antibody (eBiosciences).

[0125] To assess proinsulin expression, mononuclear cells were incubated with PE-Cy7-conjugated streptavidin, APC-conjugated anti-c-kit, APC-Cy7-conjugated anti-Ly6A / E (Sca-1), and a panel of biotinylated mouse strains, followed by fixation with BD Cytofix / CytoPerm (BD Biosciences). Next, rabbit anti-insulin monoclonal antibody (Cell Signaling Technology) and PE-conjugated anti-rabbit IgG antibody (Cell Signaling Technology) were applied to the mononuclear cells. To deplete dead cells prior to staining, a LIVE / DEAD Fixable Dead Cell Blue Stain Kit (ThermoFisher Scientific Inc., Waltham, MA, USA) was used.

[0126] Four hours after staining, cells were analyzed using a FACS Canto II with FACS DIVA software (BD Biosciences).

[0127] result Proinsulin-positive cells were found in the KSL fraction of mononuclear cells from STZ-DM mice, but not in the KSL fraction from non-DM mice (Fig. 1). Similarly, TNF-α-positive cells were found in the KSL fraction of mononuclear cells from STZ-DM mice, but not in the KSL fraction from non-DM mice (Fig. 2). CD106-positive cells were found in the KSL fraction of mononuclear cells from both STZ-DM and non-DM mice, but were more abundant in STZ-DM mice (Fig. 3).

[0128] In diabetic mice, KSL cells contained TNF-α-positive cells and proinsulin-positive cells, and CD106-expressing cells were increased. It is thought that hematopoietic stem cells with these characteristics are responsible for the emergence of chronic diabetes mellitus characteristics.

[0129] Example 2: Identification of abnormal stem cells involved in type 1 diabetes As in Example 1, abnormal cells involved in diabetes were also characterized using stem cells obtained from type 1 diabetes model mice.

[0130] method Obtaining mice and bone marrow cells ICR mice (CLEA Japan, Osaka) and NOD mice (CLEA Japan, Osaka) were used in the test. Mononuclear cells were isolated in the same manner as in Example 1.

[0131] As in Example 1, mononuclear cells were stained for TNF-α, CD106, c-kit, Sca-1, and mouse lineage, and FACS analysis was performed.

[0132] result The proportion of KSL cells in NOD mice was reduced to approximately 25% compared with that in ICR mice (Fig. 4). The proportions of TNF-α-positive cells and CD106-expressing cells in the KSL cell population were significantly increased in NOD mice compared with that in ICR mice (Fig. 5).

[0133] The KSL cells of type 1 diabetic mice also showed an increase in TNF-α-positive cells and CD106-expressing cells, and it is thought that hematopoietic stem cells with these characteristics are responsible for the emergence of chronic diabetes characteristics.

[0134] Example 3: Cellular stages of abnormal stem cells involved in diabetes The stage of hematopoietic stem cells at which the abnormal stem cells were located was examined.

[0135] method The side population (SP) of KSL cells was analyzed by FACS according to the method reported by Goodell (see J Exp Med. 1996 Apr 1;183(4):1797-806; Nat Med. 1997 Dec;3(12):1337-45). Hoechest 33342 dye is cell-permeable and binds to DNA, but it can be efficiently exported from hematopoietic stem cells. Taking advantage of this property, it has been reported that when bone marrow cells stained with this dye were excited with ultraviolet light (350 nm) and developed using two optical filters, Hoechst blue and Hoechst red, the hematopoietic stem cell fraction was contained in the unstained cell population (SP cells). Whole bone marrow cells were stained with Hoechest 33342 dye (Sigma-Aldrich Japan KK, Tokyo, Japan) for exactly 90 minutes at 37°C. For gating, Hoechest 33342-positive cells were incubated with verapamil hydrochloride (Tocris Bioscience, Bristol, UK), and mononuclear cells were isolated using Ficoll-Paque Plus. After staining with a biotin mouse lineage panel, these cells were incubated with PE-Cy7-conjugated streptavidin, APC-conjugated anti-c-kit, APC-Cy7-conjugated anti-Ly6A / E (Sca-1), and PE-conjugated anti-TNF-α or FITC-conjugated anti-CD106. For dead cell depletion, propidium iodide (Sigma-Aldrich) was added to the samples to remove dead cells. Analysis was performed using a FACS Aria Fusion with FACS DIVA software (BD Biosciences). Cells stained with Hoechest 33342 were excited with 350 nm ultraviolet light, and two optical filters were used: 450BP20 (450 / 20 nm bandpass filter) (Hoechst blue) and 675EFLP (675 nm long-pass edge filter) (Hoechst red).

[0136] result The proportion of SP cells in KSL cells was increased and the proportion of non-SP cells was decreased in STZ-DM mice compared with non-DM mice (Fig. 6). Neither CD106-positive nor TNF-α-positive cells were detected in SP cells from either non-DM or STZ-DM mice (Fig. 7b, d). On the other hand, non-DM-derived TNF-α-positive cells were not detected in non-SP cells, but STZ-DM-derived TNF-α-positive cells were detected. More CD106-positive cells were observed in STZ-DM mice compared with non-DM mice (Fig. 7a, c). Therefore, CD106-positive and TNF-α-positive cells are present in the non-SP fraction of KSL cells in STZ-DM mice.

[0137] CD106- and TNF-α-positive cells, characteristic of hematopoietic stem cell abnormalities, were found in non-SP cells (short-term hematopoietic stem cells: ST-HSCs) among KSL cells, but not in SP cells (long-term hematopoietic stem cells: LT-HSCs) among KSL cells, which are at an earlier differentiation stage. This suggests that HSC abnormalities in diabetes are limited to the progenitor cell stage, and that abnormalities do not occur in the so-called "stem cells of stem cells." Eliminating progenitor cells may therefore be a potential treatment for diabetes and related diseases. While bone marrow transplantation may be necessary if "stem cells of stem cells" are targeted for treatment, elimination of progenitor cells may enable treatment, potentially enabling drug therapy. Thus, it became clear that the abnormal stem cells found in diabetes may be ST-HSCs.

[0138] Example 3: Further characterization of abnormal stem cells Further characterization of the abnormal stem cells involved in diabetes was carried out.

[0139] method RNA was extracted from KSL cells obtained by FACS sorting of non-DM and STZ-DM mice in the same manner as in Example 1, and gene expression of histone deacetylase genes (Hdacs) was analyzed using QT-PCR.

[0140] result The results are shown in Figure 8. Compared with non-DM mice, STZ-DM mice were found to have significantly increased mRNA expression of Hdac3, Hdac4, Hdac8, and Hdac9.

[0141] Diabetic KSL cells showed increased expression of epigenome-related genes (Hdacs), which are important for regulating gene expression through histone modifications, suggesting that hyperglycemia confers abnormalities on stem cells at the genetic level. It has been reported that cells exposed to transient hyperglycemia maintain abnormalities induced by hyperglycemia even after returning to a normal blood glucose environment (El-Osta et al. J Exp Med. 2008 Sep 29;205(10):2409-17). This suggests that the increased expression of histone deacetylase genes in KSL cells may be involved in the emergence of CD106-, TNF-α-, and proinsulin-positive cells. To clarify this, we will investigate whether Hdac inhibitors (e.g., trichostatin A) are therapeutic options.

[0142] Example 5: Diabetes caused by abnormal stem cells We examined whether the abnormal stem cells found above cause diabetes.

[0143] method Figure 9 shows an outline of the experiment in which non-DM or STZ-DM KSL cells were transplanted into normoglycemic mice. GFP-Tg mice (The Jackson Laboratory, Bar Harbor, ME) were treated with STZ (STZ-DM GFP) or a control treatment (intravenous citrate buffer injection) as described in Example 1. Three months later, KSL cells obtained from non-DM or STZ-DM mice were transplanted into normoglycemic wild-type C57BL / 6J mice (Claire, Osaka) that had been lethally irradiated with 9 Gy (non-DM-derived KSL-T or STZ-DM-derived KSL-T, respectively) (Figure 10, left). Blood glucose levels were monitored 3 months after transplantation (non-DM-derived KSL-T (n = 9), STZ-DM-derived KSL-T (n = 10)). Three months after transplantation, SNCV in the sciatic nerve was measured (non-DM-derived KSL-T (n=5), STZ-DM-derived KSL-T (n=6)) (Figure 10, right). Three months after transplantation, dorsal root ganglia (DRGs) were obtained from each mouse and immunofluorescently stained for nuclei, GFP, MAP2, proinsulin, and TNF-α (PE-conjugated) (Figures 11, 12, 13).

[0144] The SNCV measurements were performed as follows. Sensory nerve conduction tests were performed on mice under anesthesia (sodium pentobarbital, 5 mg / kg i.p.) at 30–32°C using a Medelec Sapphire EMG (Medelec, Woking, UK). Skin was removed from the left dorsal surface of the thigh to expose the sciatic nerve. Sensory nerve action potentials (SNAPs) were recorded, and sensory nerve conduction velocity (SNCV) was calculated by dividing the distance from the stimulation site to the recording site by the initial latency of the SNAP.

[0145] result The KSL cell fraction from STZ-DM-derived KSL-T mice showed abnormal stem cell characteristics, such as TNF-α and proinsulin, but the KSL cell fraction from non-DM-derived KSL-T mice did not show abnormal stem cell characteristics.

[0146] When abnormal KSL cells found in diabetic mice were transplanted into normal mice, they developed symptoms similar to diabetic neuropathy, even though their blood glucose levels were normal. This revealed that even when blood glucose levels in diabetic mice were normalized, the abnormal stem cells did not disappear. This indicates that the abnormal cells remained abnormal and established in the bone marrow, i.e., as pathological stem cells. Furthermore, it is understood that cells derived from these abnormal stem cells migrated to neural tissue and caused neuropathy. Therefore, it was revealed that as long as these stem cells remain, diabetes cannot be cured even by blood glucose control. Furthermore, when bone marrow containing abnormal stem cells from these diabetic mice was transplanted into normal mice, impaired glucose tolerance was observed, and the glucose metabolism abnormalities that are the core of diabetes were reproduced by bone marrow transplantation.

[0147] This is consistent with the clinical symptoms of diabetes and diabetic complications not being cured even when hyperglycemia is improved, suggesting that the ultimate therapeutic target for diabetes is abnormal hematopoietic stem cells. Thus, by transplanting diabetic hematopoietic stem cells into normal mice, we were able to reproduce abnormal glucose metabolism and diabetic neuropathy in normal mice.

[0148] Example 6: Characteristics of abnormal stem cells in human diabetic patients Patients admitted to the intensive care unit at Shiga University of Medical Science Hospital were surveyed. Samples were collected from three patients diagnosed with diabetes (three males / zero females, mean age 72.0±14.2 years) and four healthy subjects (three males / one female, mean age 76.5±8.5 years).

[0149] Blood samples were obtained from these subjects, and RNA was extracted from the blood samples. Specifically, 1.5 ml of blood was collected, DNA was digested with DNAse (Qiagen, Hilden, Germany), and RNA was extracted using a QIAamp RNA blood mini kit (Qiagen, Hilden, Germany). cDNA was prepared from the obtained RNA using the SuperScript® III First-Strand Synthesis System (Invitrogen / ThermoFisher Scientific, MA, USA). Quantitative PCR was performed using Power SYBR Green PCR Master Mix in a real-time PCR system (Applied Biosystems / ThermoFisher Scientific, MA, USA) to quantify expressed mRNA. Comparisons between the two groups were performed using t-tests. The results are shown in Figure 14.

[0150] Similar to the mouse results, we also confirmed that the expression of insulin, TNF-α, and CD106 tended to increase in diabetic patients. Furthermore, we also confirmed that the expression of CD34 tended to increase in diabetic patients. Because CD34 is a stem cell characteristic in humans, this suggests that abnormal stem cells exist in diabetic patients, similar to the mouse results.

[0151] Example 7: Treatment of Type 1 Diabetes by Targeting Abnormal Stem Cells In NOD mice with elevated blood glucose levels, insulin pellets will be implanted subcutaneously in the back to normalize blood glucose levels, and then anti-CD106 antibody will be administered via the tail vein at a dose of 250 μg / mouse once a week to determine whether normal blood glucose levels can be maintained independently of insulin pellets.

[0152] Diabetic mice with insulin pellet implants that normalize blood glucose levels were irradiated to kill bone marrow stem cells (both LT-HSCs and ST-HSCs). Instead, LT-HSCs from normal mice were transplanted into the bone marrow. This procedure resulted in the de novo generation of ST-HSCs from LT-HSCs in the bone marrow of the transplanted mice. The abnormal ST-HSCs originally present in the diabetic mice, as well as the T cell progenitors (abnormalities observed in STZ mice and diabetic NOD mice) and B cell progenitors (abnormalities observed in prediabetic NOD mice), were killed, preventing organ damage. Similar diabetic mice were also transplanted with LT-HSCs but maintained at elevated blood glucose levels without insulin pellet implantation. Because hyperglycemia persisted in these mice, abnormal ST-HSCs would develop from LT-HSCs, potentially damaging various organs and potentially preventing the cure of diabetes and its complications.

[0153] Treatment with insulin pellets plus LT-HSCs can eliminate abnormal ST-HSCs, normalize damaged pancreatic islets, and eliminate complications.

[0154] Example 8: Combination Treatment of Stem Cell Inhibitors and Migration Agents in Streptozotocin-Induced Diabetic Mice The therapeutic effect of combining an inhibitor of abnormal stem cells with a migration agent was tested in streptozotocin-induced diabetic mice.

[0155] method Streptozotocin-induced diabetic mice (3 months after the onset of diabetes) were prepared as described above and subcutaneously injected with AMD3100 (5 mg / kg, Abcam) and GROβ (0.1 mg / kg, Peprotech) together with saline. 15 minutes later, anti-CD106 antibody (250 μg / mouse) was injected via the tail vein (GA-CD106 group). This treatment was repeated weekly. Similarly, a group administered anti-CD106 antibody without AMD3100 or GROβ (CD106 group) and a group administered neither AMD3100 nor GROβ nor anti-CD106 antibody (untreated group) were also tested. Blood glucose levels and body weights were measured daily (Figure 15), and mice were perfused and fixed 20 days after the start of treatment.

[0156] After 24 hours of fixation, the fixed mice were transferred to 15% sucrose, 0.1M PB solution, and then the pancreas was harvested from the mice. After preparing a frozen block of the pancreas, 8 μm frozen sections were prepared using a cryostat.

[0157] The prepared frozen sections were then immunostained according to the following procedure (FIG. 16). Wash three times with PBS(-) for 10 minutes each. Inactivate endogenous peroxidase by immersing in 0.3% H2O2 in PBS(-) at room temperature for 30 minutes. Wash three times with PBS(-) for 5 minutes each. Incubate with blocking buffer (5% normal goat serum + 0.3% Triton-X100 in PBS) for 1 hour at room temperature. Add primary antibody (anti-insulin antibody: CST) and incubate overnight at 4°C. Wash three times with PBS(-) for 5 minutes each. Add ImmPRESS Reagent (anti-rabbit: VECTOR Laboratories) and incubate at room temperature for 30 minutes. Wash three times with PBS(-) for 5 minutes each. Add ImmPACT DAB substrate (VECTOR Laboratories) and incubate at room temperature for 30 seconds. Add dH20 to stop the DAB reaction. Counterstain with hematoxylin for 30 seconds. Wash with tap water. Dehydrate and infuse.

[0158] The prepared section samples were observed under a microscope, and approximately 8 to 10 pancreatic islets were randomly selected, and the proportion of insulin-positive islets relative to the size of the islets was calculated using Image J software (FIG. 17).

[0159] result In the group in which stem cells were allowed to migrate and treated with anti-CD106 antibody, a decrease in blood glucose levels was observed (Figure 15), and immunohistochemistry showed an increase in insulin-positive areas compared to the untreated group (Figure 17).

[0160] After stem cell migration, treatment to suppress abnormal stem cells restored the insulin-positive areas of pancreatic islets, which had remained at only about 10% after STZ administration, to about 30-40%. This suggests that abnormal stem cells released from the bone marrow niche to the periphery by administration of a stem cell migration agent are destroyed by anti-CD106 antibodies, while normally functioning stem cells may regenerate pancreatic islets.

[0161] Example 9: Combination Treatment of Stem Cell Inhibitors and Migration Agents in NOD Mice The therapeutic effect of combining an inhibitor of abnormal stem cells with a migration agent was examined in spontaneous type 1 diabetes model mice (NOD mice).

[0162] method NOD mice, a model of spontaneous type 1 diabetes, were purchased from CLEA Japan (Osaka Prefecture), and blood glucose levels and body weight were measured every two weeks. Animals showing elevated blood glucose levels were treated with a subcutaneous injection of AMD3100 (5 mg / kg: Abcam) and GROβ (0.1 mg / kg: Peprotech) in saline. 15 minutes later, anti-CD106 antibody (250 μg / mouse) was administered via the tail vein (administration interval: once a week). Blood glucose levels and body weight were measured daily after the start of treatment (Figure 18).

[0163] In addition, before starting treatment, to confirm the state of the pancreatic islets in ICR mice, NOD mice that had not developed diabetes, and NOD mice that had developed diabetes (all mice were the same age), perfusion fixation was performed and immunostaining was performed as described below (Figure 19). Wash three times with PBS(-) for 10 minutes each. Inactivate endogenous peroxidase by immersing in 0.3% H2O2 in PBS(-) at room temperature for 30 minutes. Wash three times with PBS(-) for 5 minutes each. Incubate with blocking buffer (5% normal goat serum + 0.3% Triton-X100 in PBS) for 1 hour at room temperature. Add primary antibody (anti-insulin antibody: CST) and incubate overnight at 4°C. Wash three times with PBS(-) for 5 minutes each. Add ImmPRESS Reagent (anti-rabbit: VECTOR Laboratories) and incubate at room temperature for 30 minutes. Wash three times with PBS(-) for 5 minutes each. Add ImmPACT DAB substrate (VECTOR Laboratories) and incubate at room temperature for 30 seconds. Add dH20 to stop the DAB reaction. Counterstain with hematoxylin for 30 seconds. Wash with tap water. Dehydrate and infuse. Images were acquired from each prepared slide.

[0164] result In NOD mice, the inflammatory response in the pancreatic islets was very strong even before the onset of diabetes, and insulin staining was very weak (Figure 19). Furthermore, the above treatment was confirmed to reduce blood glucose levels in diabetic mice (Figure 18). The combined therapy of the chemotactic agent and the antibody showed a significant hypoglycemic effect, which is thought to be due to the restoration of islet function by the antibody. It has been suggested that removal of abnormal hematopoietic stem cells may also be a useful treatment for type 1 diabetes.

[0165] Example 10: Abnormal cells in the bone marrow of human diabetic patients We also confirmed that abnormal cells exist in the bone marrow of human diabetic patients.

[0166] We examined the presence of proinsulin-positive cells in the bone marrow of patients with type 2 diabetes mellitus (DM) admitted to Shiga University of Medical Science and autopsied between January 1, 2000, and December 31, 2010. Tissues were embedded in paraffin at the Anatomy Center of Shiga University of Medical Science. Five-micrometer-thick sections of the paraffin-embedded specimens were processed for immunohistochemistry using the avidin-biotin peroxidase complex (ABC) method and the diaminobenzidine (DAB)-nickel reaction. After deparaffinization in xylene and alcohol, the sections were microwaved (10 mmol / L citrate buffer, pH 6.0, 0.5 kW, 10 min), then incubated overnight with a 1:1,000 dilution of a proinsulin antibody (mouse monoclonal, Abcam, UK) in 0.1% PBS containing 0.3% Triton X-100 (PBST) at 4°C and then processed for immunohistochemistry. After DAB-nickel reaction, sections were counterstained with nuclear fast red solution.

[0167] The results are shown in Figure 20. Proinsulin expression was not observed in bone marrow cells derived from patients without DM, but was observed in bone marrow cells derived from patients with DM. The findings of abnormal stem cells observed in mice are thought to be applicable to humans.

[0168] Example 11: Application to humans We will identify abnormal bone marrow-derived hematopoietic stem cells in human diabetic patients and target them to treat diabetes.

[0169] Research plan 1. Identification of abnormal hematopoietic stem cells derived from bone marrow in diabetic patients (subject) For the non-diabetic group, volunteers with no history of impaired glucose tolerance and no current diabetes treatment will be recruited from among staff at Shiga University of Medical Science and its affiliated hospital. Random blood glucose and HbA1c measurements will be conducted. Those with a random blood glucose level of <140 mg / dl and HbA1c <6.0% will be defined as non-diabetic, and 20 people will be enrolled as a control group. For the diabetic group, HbA1c levels will be 6.5% or higher or those currently undergoing diabetes treatment. A list of outpatients from the Department of Diabetes and Endocrinology at Shiga University of Medical Science Hospital, matched by sex and age to the non-diabetic group, will be created from electronic medical records, and 80 people (40 with type 1 diabetes and 40 with type 2 diabetes) will be randomly selected and enrolled.

[0170] (Research method) Subjects were interviewed, their height and weight were measured, and blood and urine tests were performed to determine the presence or absence of diabetes. Mononuclear cells were extracted from the collected blood, and CD34-labeled bone marrow progenitor cells were collected and fixed. Morphology and expressed proteins were identified by immunostaining. mRNA was extracted, cDNA was prepared, and mRNA expression levels were quantified by quantitative PCR. Specifically, TNF-α mRNA and insulin mRNA expression levels were measured in peripheral blood CD34-positive and CD106-positive (CD34 / CD106) bone marrow progenitor cells. Protein expression in these cells was also measured. Furthermore, the relationship between the presence or absence of diabetic complications and glycemic control in diabetic patients was analyzed. To identify the presence or absence of typical diabetic complications, such as diabetic neuropathy, retinopathy, nephropathy, fatty liver, and dyslipidemia, as well as macrovascular disease, nerve conduction velocity tests, electrocardiogram RR interval tests, fundus retinal examinations, urinary albumin excretion rates, abdominal CT scans to quantify intraperitoneal fat mass, blood lipid tests, electrocardiograms, carotid artery ultrasound examinations, and lower limb arterial ultrasound examinations were performed.

[0171] (Prediction of results) (1) In both non-diabetic and type 2 diabetic groups, expression of TNF-α mRNA and insulin mRNA was observed in CD34 / CD106 bone marrow progenitor cells in peripheral blood, but expression levels were increased in type 2 diabetic groups. On the other hand, in type 1 diabetic groups, expression of TNF-α mRNA was increased in CD34 / CD106 bone marrow progenitor cells compared to non-diabetic groups, but insulin mRNA was not expressed at all. (2) In non-diabetic patients, few CD34 / CD106 bone marrow progenitor cells in peripheral blood express TNF-α and proinsulin proteins. In contrast, in type 2 diabetes, the number of cells expressing both proteins increases. In type 1 diabetes, on the other hand, the number of cells expressing TNF-α protein increases, but no cells expressing proinsulin. (3) In type 1 diabetes patients, the onset of typical complications such as diabetic neuropathy, retinopathy, nephropathy, fatty liver, and dyslipidemia is associated with an increase in TNF-α protein-positive cells among CD34 / CD106 bone marrow progenitor cells in peripheral blood. On the other hand, in type 2 diabetes patients, the onset of diabetic neuropathy, retinopathy, nephropathy, fatty liver, and dyslipidemia is associated with an increase in TNF-α protein-positive cells and proinsulin-positive cells among CD3 / CD106 bone marrow progenitor cells in peripheral blood.

[0172] (Considerations and predictions of conclusions) 1) In non-diabetic individuals, there are small numbers of CD34 / CD106 bone marrow progenitor cells expressing insulin mRNA and TNF-α mRNA in the blood, which may function as endothelial cells presenting autoantigens when homing to pancreatic islets. 2) In type 2 diabetes, hyperglycemia causes these cells (CD34 / CD106 bone marrow progenitor cells that express insulin mRNA and TNF-α mRNA) to be present in the bone marrow and blood, and it is thought that they may already express proinsulin and TNF-α proteins, and differentiate into vascular endothelial cells with abnormal functions, or have abnormal cell fusion ability, which may cause insulin resistance and various complications.

[0173] Research plan 2. Treatment of diabetes by targeting bone marrow-derived abnormal hematopoietic stem cells (subject) According to Study Plan 1, 280 diabetic patients (140 with type 1 diabetes and 140 with type 2 diabetes) will be enrolled at Shiga University of Medical Science and collaborating research facilities. It is known that once diabetes develops, it cannot be cured, regardless of whether it is type 1 or type 2. However, in type 1 diabetes, a honeymoon period of temporary remission can occur with strict glycemic control using insulin. The duration of this period varies widely, but generally ranges from 1 month to 13 years (Wallensteen M, Dahlquist G, Persson B, Landin-Olsson M, Lernmark A, Sundkvist G, Thalme B (1988) Factors influencing the magnitude, duration, and rate off all of β-cell function in type 1 (insulin-dependent) diabetic children followed for two years from their clinical diagnosis. Diabetologia 31: 664-669). Therefore, it may be difficult to distinguish whether this treatment plan has initiated a honeymoon phase or cured the disease itself. Furthermore, it has been reported that strict control of insulin resistance in type 2 diabetes can result in milder insulin resistance (HE Lebovitz (2001) Insulin resistance: definition and consequences. Clin Endocrinol Diabetes 109 Suppl 2: S135-S148). This may result in improved insulin and oral medication dosages, as well as improved endogenous insulin secretion. Therefore, in both type 1 and type 2 diabetes, treatment studies should include a group treated with insulin alone and a group receiving a novel treatment, and then compare these groups.

[0174] (Research method) Subjects were interviewed, their height and weight were measured, and blood and urine tests were performed to determine the presence or absence of diabetes. Mononuclear cells were extracted from the collected blood, and CD34-labeled bone marrow progenitor cells were collected and fixed. Morphology and expressed proteins were identified by immunostaining. mRNA was extracted, cDNA was prepared, and mRNA expression levels were quantified by quantitative PCR. Specifically, TNF-α mRNA and insulin mRNA expression levels were measured in CD34-positive and CD106-positive bone marrow progenitor cells in peripheral blood. Protein expression in these cells was also measured. Furthermore, the relationship between the presence or absence of diabetic complications and glycemic control in diabetic patients was analyzed. To identify the presence or absence of typical diabetic complications, such as diabetic neuropathy, retinopathy, nephropathy, fatty liver, and dyslipidemia, as well as macrovascular disease, nerve conduction velocity tests, electrocardiogram RR interval tests, fundus retinal examinations, urinary albumin excretion rates, abdominal CT scans to quantify intraperitoneal fat mass, blood lipid tests, carotid artery ultrasound, and lower limb arterial ultrasound were performed.

[0175] Type 1 patients were randomly assigned to seven groups of 20 patients each, and type 2 patients were assigned to seven groups of 20 patients each. Twenty patients from each group were controlled with insulin treatment alone for three months (control group). Of the remaining 120 patients, 60 patients were initiated on insulin treatment, with 20 patients each starting one of the following three treatments (treatment group without chemoattractant): 1) anti-TNF-α antibody (adalimumab 40 mg or infliximab 3 mg / kg), 2) anti-CD106 antibody (0.8 mg / kg), or 3) trichostatin (0.5 mg / kg) administered intravenously once weekly for 12 weeks. The remaining 60 patients were initiated on insulin treatment, with 20 patients each starting one of the following three treatments (treatment group with chemoattractant). Along with the cell migration agent Groβ (100μg / kg) + plerixafor (0.24mg / kg), 1) anti-TNF-α antibody (adalimumab 40mg or infliximab 3mg / kg), 2) anti-CD106 antibody (0.8mg / kg), and 3) trichostatin (0.5mg / kg) will be administered intravenously once a week for 12 weeks.

[0176] (Method for assessing therapeutic effect) Patients will measure their own blood glucose six times a day. The goal of glycemic control is to achieve a pre-meal blood glucose level of 140 mg / dL or less and a 2-hour post-meal blood glucose level of 200 mg / dL or less through insulin treatment. The insulin dose will be adjusted to meet the control criteria, and the treatment study will end with the insulin required for glycemic control at week 12. The therapeutic effect will be assessed by following up the following evaluation items before treatment, at the end of treatment, and 3, 6, 9, and 12 months after the end of treatment.

[0177] (Judgment item) (A) Elimination of abnormal hematopoietic stem cells The expressed proteins (CD34, proinsulin, TNF-α, CD106, etc.) and expressed genes (CD34 mRNA, insulin mRNA, TNF-α mRNA, CD106 mRNA, etc.) in CD34 / CD10 bone marrow progenitor cells in peripheral blood are quantified. (B) Curing diabetes Blood glucose level, HbA1c, urinary CPR, lipids, insulin secretion function using glucagon tolerance test will be measured before and after treatment. Pancreatic islet-associated antibodies will be measured and whether they have disappeared will be clarified. (C) Treatment effects on complications The presence or absence of typical complications such as diabetic neuropathy, retinopathy, nephropathy, fatty liver, dyslipidemia, and macroangiopathy, as well as changes in these, will be compared before and after treatment.

[0178] (Prediction of results) 1) Treatment with insulin alone reveals the following: (A) In both type 1 and type 2 diabetes, abnormal hematopoietic stem cells do not disappear. (B) There is no evidence that it can cure diabetes. (C) There is some therapeutic benefit to treating complications, but there is no cure.

[0179] 2) Novel treatments with or without cell migration agents reveal the following: (A) In both type 1 and type 2 diabetes, abnormal hematopoietic stem cells disappear. (B) Both type 1 and type 2 diabetes are cured temporarily. (C) The progression of complications in both type 1 and type 2 diabetes is halted, and clear therapeutic effects are observed.

[0180] 3) The following will become clear through the new treatment that includes a cell migration agent: (A) Compared with treatment without a chemotactic agent, the new treatment with a chemotactic agent results in earlier disappearance of abnormal hematopoietic stem cells in both type 1 and type 2 diabetes. (B) Compared to treatment without a chemotactic agent, the new treatment with a chemotactic agent improves the cure rate for both type 1 and type 2 diabetes. (C) Compared to treatment without a chemotactic agent, the new treatment with a chemotactic agent improves the cure rate of complications in both type 1 and type 2 diabetes.

[0181] (Considerations and predictions of conclusions) 1) In type 1 diabetes, even with insulin treatment alone, a honeymoon period may occur due to the blood glucose control effect. However, in this case, remission will eventually end and insulin secretion will once again become insufficient. 2) In type 2 diabetes, insulin treatment alone may improve blood sugar control and make insulin treatment unnecessary, but since abnormal hematopoietic stem cells do not disappear, diabetes will not be cured. 3) Even if type 1 diabetes is cured by a new treatment that includes a cell migration agent, there is still a possibility that the autoimmune system that caused the production of autoantibodies may relapse. However, this can be prevented by administering the new treatment that includes a cell migration agent again. 4) Even if type 2 diabetes is completely cured by a new treatment that includes a cell migration agent, if hyperglycemia occurs again due to excessive energy intake or lack of exercise, abnormal hematopoietic stem cells may appear. In this case, treatment is possible by repeating the new treatment that includes a cell migration agent.

[0182] (Note) While the present disclosure has been illustrated using preferred embodiments thereof, it is understood that the scope of the present disclosure should be construed solely in terms of the claims that follow. It is understood that the patents, patent applications, and other documents cited herein are incorporated by reference into this specification in their entirety as if the contents themselves were specifically set forth herein.

[0183] This application claims the benefit of priority to Japanese Patent Application No. 2019-191369, filed with the Japan Patent Office on October 18, 2019, the entire contents of which are incorporated herein by reference. [Industrial Applicability]

[0184] The present disclosure provides improved diabetes treatments that target stem cells, and based on this finding, new approaches are provided for treating and preventing diabetes and / or diabetes-related diseases, disorders and / or conditions, as well as diagnosing diabetes and / or diabetes-related diseases, disorders and / or conditions, or the risk thereof.

Claims

1. A composition for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising an inhibitor of abnormal hematopoietic stem cells (HSCs), wherein the inhibitor is administered in combination with a stem cell migration agent; wherein the inhibitor comprises at least one selected from the group consisting of an anti-CD106 antibody or a functional variant thereof; The composition, wherein the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, pegfilgrastim, and GROβ.

2. The composition of claim 1, wherein the abnormal HSC is one in which a gene or protein selected from the group consisting of CD106 and its functional equivalents is not expressed and / or does not function at normal levels.

3. The composition of claim 2, wherein the expression is not at a normal level is overexpression.

4. The composition of claim 2 or 3, wherein the abnormal HSC further lacks normal levels of expression of a gene or protein selected from the group consisting of tumor necrosis factor alpha (TNF-α), histone deacetylase (HDAC), and proinsulin.

5. The composition of any one of claims 1 to 4, wherein the disease, disorder and / or condition comprises a diabetic complication.

6. The composition according to any one of claims 1 to 4, wherein the disease, disorder and / or symptom is selected from the group consisting of neuropathy, nephropathy, hepatopathy, retinopathy, fatty liver, gastrointestinal disorder, delayed fracture healing, eating disorder, and skin disorder.

7. The composition according to any one of claims 1 to 6, wherein the stem cell migration agent has the ability to migrate the abnormal HSC from a niche.

8. The composition of any one of claims 1 to 6, wherein the inhibitor comprises an anti-CD106 antibody and the stem cell migration agent comprises plerixafor and GROβ.

9. The composition according to any one of claims 1 to 6, wherein the inhibitor is an anti-CD106 antibody and the stem cell migration agent is plerixafor and GROβ.

10. A pharmaceutical for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising a combination of an inhibitor of abnormal hematopoietic stem cells (HSC) and a stem cell migration agent, wherein the inhibitor comprises at least one selected from the group consisting of anti-CD106 antibodies or functional variants thereof, and the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, pegfilgrastim, and GROβ.

11. A composition for treating and / or preventing diabetes and / or diabetes-related diseases, disorders and / or symptoms, comprising a stem cell migration agent, characterized in that the stem cell migration agent is administered in combination with an inhibitor of abnormal hematopoietic stem cells (HSCs), wherein the inhibitor comprises at least one selected from the group consisting of anti-CD106 antibodies or functional variants thereof, and the stem cell migration agent comprises at least one selected from the group consisting of plerixafor, gefitinib, erlotinib, afatinib, osimertinib, filgrastim, nartograstim, lenograstim, pegfilgrastim, and GROβ.

12. The composition according to any one of claims 1 to 9 for alleviating diabetes.

13. The composition according to any one of claims 1 to 9 for curing diabetes.

14. The composition according to any one of claims 1 to 9, 12 to 13 for alleviating type 2 diabetes.

15. The composition according to any one of claims 1 to 9 and 12 to 14, characterized in that it is administered to a subject in which abnormal HSCs have been detected.

16. 16. The composition of claim 15, wherein the subject has had the abnormal HSC detected by testing bone marrow cells obtained from the subject.

17. The composition of claim 15 or 16, wherein the subject has the abnormal HSCs detected by increased expression of CD106 compared to normal subjects.

18. The composition of any one of claims 15 to 17, wherein the subject has the abnormal HSC detected by elevated expression of histone deacetylase (HDAC) compared to a normal subject.

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