Viral vector targeting cancer stem cells

a cancer stem cell and vector technology, applied in the field of recombinant adenoviral vectors, can solve the problems of unknown activity of cd133 promoters in cancer stem cells, little progress in the study of biological analyses and cancer treatments (in particular, cancer stem cell-specific treatments) that target cancer stem cells, and achieve high infiltration/metastatic ability, efficient development, and the effect of determining the prognosis of a cancer patien

Inactive Publication Date: 2014-01-23
KAGOSHIMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent describes a special viral vector that targets cancer stem cells - the root causes of many types of cancer. By carrying genes or other tools designed to fight these cells, this vector could potentially help develop better ways to detect and treat cancer. It may even allow us to see if we have successfully treated all the cancer stem cells in a person's body, reducing the risk of relapse. Overall, this technology offers a promising tool for identifying and eliminating cancer stem cells, leading to improved outcomes for cancer patients.

Problems solved by technology

This patent discusses the problem of cancer stem cells causing disease and resisting traditional cancer treatments like chemotherapy and radiation. These cells often lead to new tumors and may contribute to the spread of cancer through their abilities to divide and repair damage caused by conventional therapies. To address this issue, researchers have developed ways to identify and target cancer stem cells using virus-based tools called conditionally replicating viruses (CRA). These viruses have shown promise in reducing cancer size and increasing patient response rates when combined with standard treatments. However, current techniques still struggle to improve the effectiveness of these approaches. Additionally, the literature suggests that certain genes found in cancer stem cells could potentially serve as targets for new therapies.

Method used

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  • Viral vector targeting cancer stem cells
  • Viral vector targeting cancer stem cells
  • Viral vector targeting cancer stem cells

Examples

Experimental program
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Effect test

example 1

Confirmation of the Cancer Stem Cell Properties of Human Glioblastoma Stem Cells and the Expression of a Cancer Stem Cell Surface Marker CD133 by Performing Immunostaining on the Cells

(1) Cells

[0117]Human glioblastoma stem cells described in Soeda A, et al., J Biol Chem 2008; 283; 10958-66 were used herein. X01 GBS cells are a fraction of cancer stem cells in X01 GB cells, and are a group of cells prepared by “concentration” of a cancer stem cell fraction using a medium under undifferentiation conditions according to a Sphere culture method. Thus, unless otherwise specified, hereinafter, the term “human glioblastoma stem cells (X01 GBS)” means human glioblastoma stem cells (or X01 GB-CSC) described in Soeda A, et al., J Biol Chem 2008; 283; 10958-66. As culture conditions, glioblastoma stem cells (X01 GBS) were cultured at 5% CO2, 37° C. in a Dulbecco's modified Eagle's medium / F-12 (D6421, Sigma) medium containing B-27 (Invitrogen), 10% FBS, to which recombinant human FGF-2 (20 ng / m...

example 2

Confirmation by Western Blotting of Expression of Cancer Stem Cell Surface Marker CD133 on Human Glioblastoma Stem Cells (X01 GBS)

(1) Cells

[0130]Human glioblastoma stem cells (X01 GBS), and as positive controls, colon cancer cells (Caco-2) and human iPS cells (201B7, purchased from RIKEN), were used.

(2) Antibodies

[0131]An anti-CD133 rabbit polyclonal antibody (Ab19898, Abcam, UK) was used as a primary antibody (dilution rate 1:1000), and a goat anti-rabbit polyclonal antibody IgG / HRP (Dako, Cytomation) (dilution rate 1:2000) was used as a secondary antibody.

(3) Western Blotting

[0132]A culture solution in a 10-cm dish containing the cultured cells was discarded, and the dish was then washed with PBS. Thereafter, 1 mL of a cell lysis RIPA buffer (0.5% NP40, 0.1% SDS, 0.5% sodium deoxycholate, 150 mM NaCl, and 50 mM Tris (pH7.5)), which contained a 0.5 mM protein protective agent PMSF and Protease inhibitor cocktail (which was added immediately before the reaction), was added, so as to...

example 3

Confirmation by Flow Cytometry (FCM) of the Number of CD133(+) Cells

(1) FCM (1st Time)

[0134]FCM analysis was carried out in accordance with protocols associated with the antibody used in the analysis, produced by the manufacturer Miltenyi Biotec. When human glioblastoma stem cells (X01 GBS) were used, the content of CD133(+) cells was determined.

[0135]In the figure, cells contained in the range enclosed with an ellipse were defined as total cells, and were subjected to the analysis.

(2) FCM (2nd Time)

[0136]FACS was carried out in the same manner as in the 1st FCM. A mouse anti-human CD133 / 2 (293C3)-PE and mouse IgG-PE (both of which were manufactured by Miltenyi Biotec) were used as antibodies. Detection was carried out using BD FACSAria™ II Flow Cytometer.

[0137]From among a plurality of experiments performed, representative results are shown in FIG. 3. In the figure, the horizontal axis (PE Log) indicates CD133(+) cells as logarithmic values, and the longitudinal axis (FITC Log) has...

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Abstract

[Problem] To provide a novel and effective method for treating cancer, method for preventing cancer, and method for suppressing metastasis by targeting cancer stem cells, the methods being capable of labeling and damaging cancer stem cells; and to provide a method for damaging and a method for identifying cancer stem cells. [Solution] The present invention provides a viral vector having a promoter that is specifically expressed in cancer stem cells and that can be used for treatment and diagnosis. Furthermore, the present invention provides a method for treating cancer, a method for preventing cancer, and a method for suppressing metastasis using this viral vector, and further provides a method for damaging and a method for identifying cancer stem cells. Furthermore, the present invention provides a labeling agent and a toxic agent for cancer stem cells containing the vector as the active ingredient, and further provides a diagnostic drug, a therapeutic drug, and a metastasis suppressant for cancer.

Description

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Claims

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Application Information

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Owner KAGOSHIMA UNIV
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