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Catenae: serosal cancer stem cells

A cancer stem cell, serosa technique, applied in tumor/cancer cells, animal cells, nervous system cells, etc., can solve the problem of cloned pure cells that have not been reported

Inactive Publication Date: 2012-11-07
SLOAN KETTERING INST FOR CANCER RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Although some recent reports have reported subpopulations of cells isolated from ovarian cancer, which appeared to be enriched for tumor-inducing cells when transplanted into immunodeficient mice [Szotek, 2006; Zhang, 2008; Bapat, 2005], But there are no reports of clonal pure cells that can maintain their stem cell state in tissue culture systems

Method used

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  • Catenae: serosal cancer stem cells
  • Catenae: serosal cancer stem cells
  • Catenae: serosal cancer stem cells

Examples

Experimental program
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Effect test

preparation example Construction

[0087] 5. Preparation of Free-Floating Chains and Spheroids

[0088] The present invention relates to methods for preparing free floating chains and spheroids. This article describes two main methods. In one approach, serosal epithelial or mesenchymal cancer cells are injected intraperitoneally (ip) into an animal tumor model (preferably a mouse), preferably under conditions in which an inflammatory stimulus is added. After sufficient time for the development of ascites and / or solid tumors, ascites fluid is harvested from ip tumor bearing animals and separated into two or more size fractions, preferably two fractions. Fractions of smaller size contain free floating chains and single cells, usually leukocytes. Leukocytes can be easily removed and the remaining cells serially passaged in suspension culture to obtain a self-renewing population of free-floating strands of clonal plasma membranes. Larger fractions include spheroids retained on the filter. These spheroids were c...

Embodiment 1

[0172] Example 1: Development of an in vivo orthotopic ovarian cancer model

[0173] The Ovcar3 cell line (obtained from NCI, NCI-60 group) was originally derived from the ascites fluid of a patient with advanced ovarian adenocarcinoma with peritoneal metastases [Hamilton, 1983]. Cell lines were maintained in M5-FCS medium.

[0174] Ovcar3 expressing luciferase and green fluorescent protein was obtained by transduction with a retroviral vector expressing an eGFP-HSV-TK-luciferase (GTL) fusion gene [Ponomarev, 2004]. Transduction efficiency was ~10%. Transduced Ovcar3 cells were sorted for highest GFP expression by FACS at the Flow Cytometry Core Facility (MSKCC). GFP-sorted Ovcar3 cells were called Ovcar3-GTL. Ovcar3-GTL cells were maintained in M5-FCS medium. On tissue culture treated plates Forms an epithelial monolayer.

[0175] Bioluminescent imaging was performed by anesthetizing mice with isoflurane (Baxter Healthcare) and administering D-luciferin (Xenogen) in PB...

Embodiment 2

[0181] Example 2: Inflammatory responses stimulate tumor growth

[0182] When NSG mice were i.p. implanted with Ovcar3-GTL cells and i.p. injected with PBS (every 3 days for 13 weeks), intraperitoneal tumor growth reached "equilibrium", as figure 1 shown. In NSG mice, tumor size was maintained at the same level for several months after equilibration. However, in the ovarian NSG model, peritoneal tumor growth was always more rapid and extensive, and compared with the PBS-injected group, in the injection with lipidated N3'→P5' phosphoramidate oligonucleotide ("oligonucleotide ”) in the group with larger volume of ascites ( figure 1 ). The oligonucleotide is a 13mer with the following structure and sequence: 5'-palmitoyl-TAGGTGTAAGCAA-3'.

[0183] BLAST searches of the oligonucleotide sequences found matches to several murine and human genes. Thus, the tumor-promoting effect of oligonucleotides in vivo may be due to some changes in gene expression in tumor cells or in cells ...

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Abstract

The present invention relates to a clonally pure population of serosal cancer stem cells (CSCs) as well as methods of producing and culturing the CSCs and uses thereof. The CSCs form catenae (free floating chains of cells) which have a glycocalyx coat of hyaluronan and proteoglycans. This discovery has lead to the development of methods of treating serosal and ovarian cancers by targeting removal or inhibition of glycocalyx formation, including combination therapies using chemotherapeutics in conjunction with glycocalyx inhibitors. The invention also provides drug screening assays for identifying compounds effective against these CSCs as well as other serosal cancer cells. Methods to use catena gene signatures, protein and surface antigens are provided for monitoring patient samples for the presence of serosal cancer stem cells.

Description

[0001] This application claims priority to US Provisional Application Serial No. 61 / 258,570, filed November 5, 2009, and US Serial No. 61 / 293,113, filed January 7, 2010, the entire contents of each of which are incorporated herein by reference. technical field [0002] The present invention relates to clonal pure populations of serosa carcinoma stem cells (CSCs), methods of producing and culturing CSCs, and their use. CSCs form free-floating strands (free-floating strands of cells) with a glycocalyx coating of hyaluronic acid and proteoglycans. This discovery has led to the development of methods for the treatment of serosal and ovarian cancers by targeting the removal or inhibition of glycocalyx formation, including the use of chemotherapy in combination with glycocalyx inhibitors. The present invention also provides drug screening assays for identifying compounds effective against these CSCs as well as other serosal cancer cells. Provided are methods of applying free floati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0797G01N33/15C12N15/12C12Q1/68
CPCA01K2267/0331C12N5/0695A01K2207/12C12N9/1051A61P35/00
Inventor 马尔科姆·A·S·穆尔塞尔韦尔·A·埃特姆
Owner SLOAN KETTERING INST FOR CANCER RES
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