MDA-MB-453 xenograft model
Breast cancer contributes to over 458,000 deaths annually and 1.4 million newly diagnosed cases worldwide, as per the American Cancer Society. Xenograft murine models accurately mimic human tumor aiming to predict a patient’s response to chemotherapy. The MDA-MB-435 epithelial cell line was isolated in 1976 from a pericardial effusion of a 48-year old Caucasian female with metastatic breast carcinoma by Cailleau at M. D. Anderson Hospital and Tumor Institute, according to a 1980 study in Cancer Research. These cells are not tumorigenic and overexpress fibroblast growth factor (FGF). Also, MDA-MB-453 cells have an active glycerol 3-phosphate shuttle and express high amounts of functional androgen receptor (AR).They are not quite a triple negative breast cancer (TNBC) as sometimes reported in that they are estrogen receptor-alpha negative and progesterone receptor negative however they show Her-2/neu protein activity (Vranic et al., 2011). Studies that have used MDA-MB-453 cells include O-Brien et al. (2014) who published data supporting the use for combination therapy of PI3K/mTOR inhibition with tratsuzumab (a common chemotherapy drug for treating HER2+ cancers) for tratsuzumab-resistant breast cancer. Naderi et al. (2011) released a study demonstrating a synergistic effect between AR and extracellular signal-regulated kinase (ERK) inhibitors that also overcomes trastuzumab resistance that was verified in vitro and in vivo; this is particularly relevant for ER-/AR+ breast cancers. Finally, a 2011 Cancer Cell study (Ni et al.) studied endocrine therapies for breast cancer and used MDA-MB-453 for their ER-/AR+/HER2+ profile; their results deciphered the AR contribution to breast cancer as an activator of Wnt and HER pathways thereby stimulating tumor growth. The MDA-MB-453 cell line is used to create the CDX (Cell Line Derived Xenograft) MDA-MB-453 xenograft mouse model. The MDA-MB-453 xenograft model exhibits tumor growth inhibition from small molecules (e.g. enzalutamide, flutamide, CI-1040, DHT).
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Basic study design
- MDA-MB-453 cells are trypsinized and total cell viability is determined. The trypan blue assay results must minimally exhibit 98% cell viability.
- After adjusting the cell suspension concentration to be one million cells per 100 µL (Matrigel plus MDA-MB-453), 10 to 12 week old mice (athymic BALB/C or NOD/SCID) receive a subcutaneous injection. Injections are in the flank of the rear hind leg.
- All injection sites are observed and palpated until tumors are established. Tumors are continually measured with calipers until tumor averages are 50-150 mm3.
- Randomization into the client determined treatment groups and administration of client supplied test compound is performed. Daily tumor measurements and mouse weights (3 times weekly) are logged.
- At the end of the study, tissues are collected and can be snap frozen in liquid N2, prepared for histological analysis or nucleic acids isolated.
Xenograft animal models are used to assess the effectiveness of drugs against specific types of cancer. New medicines are tested on staged tumor growths that have been engrafted via subcutaneous or orthotopic inoculation in an immunocompromised mouse or rat model. All clinically approved anti-cancer agents have been evaluated with conventional preclinical in vivo models. Xenograft studies can be highly complex, starting with the selection of the appropriate animal model, choice of tumorigenic cell line, administration method, dosing, analysis of tumor growth rates and tumor analysis (histology, mRNA and protein expression levels).
Altogen Labs provides an array of laboratory services using over 30 standard Cell Line Derived Xenograft (CDX) models and over 20 PDX models. Researchers investigating the role of specific proteins or gene products in regulating tumor growth can benefit from development of protein overexpression (genetically engineered to ectopically express proteins, tumor suppressors, or oncogenes) and RNAi cell lines with long term gene silencing. Altogen Labs provides quantitative gene expression analysis of mRNA expression (RT-PCR) and protein expression analysis using the WES system (ProteinSimple).
The dosing of the experimental compound of interest is initiated, for a staged study, when the mean tumor size reaches a specified volume (typically 50-100 mm3). In an unstaged study, the dosing of the compound of interest is initiated immediately after xenografting. Mice are dosed once or twice a day for 28 days (or other desired study duration) via the chosen route of administration. Tumor volume (mm3) is calculated via the “(W x W x L) / 2” formula, where W is tumor width and L is tumor length.
Animal handling and maintenance at the Altogen Labs facility is IACUC-regulated and GLP-compliant. Following acclimation to the vivarium environment, mice are sorted according to body mass. The animals are examined daily for tumor appearance and clinical signs. We provide detailed experimental procedures, health reports and data (all-inclusive report is provided to the client that includes methods, results, discussion and raw data along with statistical analysis). Additional services available include collection of tissue, histology, isolation of total protein or RNA and analysis of gene expression. Our animal facilities have the flexibility to use specialized food or water systems for inducible gene expression systems.
Following options are available for the MDA-MB-453 xenograft model:
- MDA-MB-453 Tumor Growth Delay (TGD; latency)
- MDA-MB-453 Tumor Growth Inhibition (TGI)
- Dosing frequency and duration of dose administration
- Dosing route (intravenous, intratracheal, continuous infusion, intraperitoneal, intratumoral, oral gavage, topical, intramuscular, subcutaneous, intranasal, using cutting-edge micro-injection techniques and pump-controlled IV injection)
- MDA-MB-453 tumor immunohistochemistry
- Alternative cell engraftment sites (orthotopic transplantation, tail vein injection and left ventricular injection for metastasis studies, injection into the mammary fat pad, intraperitoneal injection)
- Blood chemistry analysis
- Toxicity and survival (optional: performing a broad health observation program)
- Gross necropsies and histopathology
- Positive control group employing cyclophosphamide, at a dosage of 50 mg/kg administered by intramuscular injection to the control group daily for the study duration
- Lipid distribution and metabolic assays
- Imaging studies: Fluorescence-based whole body imaging, MRI