MV4-11 xenograft model
Acute myeloid leukemia (AML) is a rapidly progressing cancer characterized by uncontrolled proliferation of bone marrow cells as well as loss of normal differentiation. The Fms-like tyrosine kinase 3 (FLT3) receptor plays an essential role in proliferation and differentiation of stem cells. FLT3 genes are the most commonly mutated genes in AML that is often associated with poor overall survival. The MV4-11 cell line was derived by Rovera and associates from the blast cells of a 10-year-old male patient with biphenotypic B-myelomonocytic leukemia and is frequently used in preclinical oncology studies. A 2017 study published in Investigational New Drugs, evaluated the ability of a novel FLT3/AXL inhibitor, gilteritinib, to inhibit mutated FLT3 in both cellular and animal models of AML. According to the article, gilteritinib demonstrates inhibitory activity against the internal tandem duplication (FLT3-ITD) and FLT3-D835Y point mutations in vitro. Also, it shows tumor regression and improved survival in the MV4-11 xenograft model in vivo. The results indicate that gilteritinib could be a next-generation FLT3 inhibitor that is beneficial for the treatment of AML patients. The MV4-11 cell line (human leukemia) is used to create the CDX (Cell Line Derived Xenograft) MV4-11 xenograft mouse model. The MV4-11 xenograft model is an AML preclinical mouse model enables to study inhibitors of FLT3 (e.g. SU11248, CHIR-258, silvestrol).
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Basic study design
- MV4-11 cell viability is confirmed using trypan blue (>98% viability req’d). The final concentration of the cell suspension is adjusted to the appropriate starting density for injections.
- Inoculations consist of each mouse receiving a single, s.c. injection of 1 x 10^6 cells (100uL vol) in the flank of one hind leg. The injection is a mixture of matrigel plus MV4-11 cells.
- Animals are continually observed and injection sites are palpated until tumors are established. Tumors are calipered and the study begins when average tumor size is in the range of 50-150 mm3.
- Mice are randomized into study groups to generate consistent animal groupsing across cohorts. The administration of test materials is performed consitent with the treatment schedule.
- Tumors and mouse weights are continually measured throughout the in-life portion of the study.
- When the tumor size limit is reached, animals are euthanized humanely. Tumors are excised from the animals and weighed. optional digital imaging for tumors is avaiilable.
- Standard gross necropsies for the collection of tissues is performed and tissues/tumors can be snap frozen in Liq-N2, nucleic acids isolated or prepared for histology.
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 MV4-11 xenograft model:
- MV4-11 Tumor Growth Delay (TGD; latency)
- MV4-11 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)
- MV4-11 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