About This Project
Triple negative breast cancer (TNBC) lacks expression of estrogen receptors, progesterone receptors and the growth factor receptor HER-2. This category of breast cancer can be especially aggressive and difficult to treat. Recent studies have suggested that the chemokine CCL5 plays a role in TNBC. The activities of CCL5 are mediated primarily through the G protein coupled receptors CCR1, CCR3, and CCR5. A peptide antagonist for CCR1 and CCR5 known as Met-CCL5 has been shown to reduce tumor size in the 410.4 murine model of breast cancer. However, it is not known whether CCR1, CCR5, or both receptors were responsible for the effects observed with Met-CCL5. We propose examining several established aCCR1 selective antagonists using a TNBC cell line to establish if it plays a critical role in cell proliferation and/or cell invasion.
Ask the Scientists
Join The DiscussionWhat is the context of this research?
Using the TNBC cell line MDA-MB-231 we will evaluate the effects of established CCR1 selective antagonists (e.g. BX471, AZD-4818, CP-481715, MLN-3897, CCX354) on 1) cell proliferation and 2) cell invasion. In addition, we will test a collection of novel pyrrolidines based on a previously disclosed series of CCR1 antagonists.
What is the significance of this project?
TNBC accounts for up to 25 % of all breast cancers and is associated with aggressive growth, advanced stage at diagnosis, high tumor grade, high levels of Ki-67 (a nuclear protein associated with cellular proliferation), and metastasis. TNBC tends to affect younger patients, is more prevalent in African Americans, and occurs in a large percentage of women with germline BRCA1 mutations.
Unfortunately, patients with TNBC are unlikely to respond to hormone therapies or HER-2–targeted therapies and as a result, the therapy choices are limited. Thus, there is an unmet need for a specific targeted therapy for TNBC.
What are the goals of the project?
Purchase of chemokines, media for cell culture, MultiScreen®-MIC plates (Millipore) that allow us to measure cell invasion, the fluorescent dye Calcein AM, a cell permeable dye used to track cells, the dye AlamarBlue that allows us to determine cellular metabolic activity, and antibodies that determine how much CCR1 is on the surface of the cells.
Budget
Cell culture supplies: In addition to the media, fetal bovine serum, freezing media, and antibiotics for cell culture, it includes the AlamarBlue used to determine cellular metabolism.
Recombinant CCL5: This amount covers recombinant human CCL5 (R&D System) to be used in all the assays.
Disposables: This amount includes general glassware and disposable plasticware (pipettes, tips, microcentrifuge tubes, gloves, tubes, filtration units, etc) as well as chemicals and buffers (HEPES, Tris, EDTA) needed to carry out the proposed experiment. It also covers the purchase the MultiScreen®-MIC plates from Millipore and 96 well half volume black/clear plates for the metabolic activity assay from BioExpress.
Antibodies: Money is needed to purchase PE-conjugated anti-CCR1 and FITC conjugated anti-CCR5 from Biolegend for determining surface expression of the receptors using flow cytometry.
The only money being requested is for laboratory supplies.
Meet the Team
Affiliates
PhD in Immunology, University of Connecticut
Team Bio
G protein-coupled receptors (GPCRs) constitute the largest receptor family in the human genome and over the years they have proven themselves to be druggable targets. Binding of a ligand leads to subsequent activation of the GPCR with conformational changes that allow G protein coupling and ultimately cellular response driven through a variety of second messenger systems. Most GPCRs can couple to more than one G protein, and different agonists can produce selective G protein activation. Which G protein(s) gets activated by the GPCR ultimately determines the downstream cellular responses that occur (such as proliferation). y research focuses on functionally selective modulators of GPCRs in the context of cancer. My primary receptors of interest are the chemokine receptors CCR1 and CCR5.
Annette Gilchrist
G protein-coupled receptors (GPCRs) constitute the largest receptor family in the human genome and over the years they have proven themselves to be druggable targets. Binding of a ligand leads to subsequent activation of the GPCR with conformational changes that allow G protein coupling and ultimately cellular response driven through a variety of second messenger systems. Most GPCRs can couple to more than one G protein, and different agonists can produce selective G protein activation. Which G protein(s) gets activated by the GPCR ultimately determines the downstream cellular responses that occur (such as proliferation). y research focuses on functionally selective modulators of GPCRs in the context of cancer. My primary receptors of interest are the chemokine receptors CCR1 and CCR5.
Project Backers
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