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WiDr xenograft model
The WiDr cell line is a human colon cancer cell line that was originally derived from the colon adenocarcinoma of a patient in 1971. WiDr cells are commonly used in cancer research to study the biology of colon cancer and test new therapies. Xenotransplantation is a well-known research tool for assessing the effectiveness of drugs against colon cancer. The WiDr cell line was established from the HT-29 cell line, taken from a colon adenocarcinoma of a 44-year-old Caucasian female. WiDr is tumorigenic in nude mice and expresses carcinoembryonic antigen (CEA), colon-specific antigen (CSAp), transforming growth factor beta and epidermal growth factor (EGF). In addition to being negative for colon antigen 3 expression, this cell line is positive for keratin by immunoperoxidase staining and expresses p53 antigen. Much research has benefitted from the WiDr cell model including a 2008 by Shindoh et al. used the WiDr xenograft model to study the relationship between antitumor effects and pharmacokinetic and pharmacodynamics properties of a novel histone deacetylase (HDAC) inhibitor known as YM753, a bacterially derived cyclic-peptide based molecule. Results demonstrated a selective tumor-cell death, tumor growth inhibition and an accumulation of acetylated histones that occurred in tumor but not normal cells. Extracellular plasma did not retain YM753 however its reduced form was retained by tumor tissues, suggesting clinical potential for this antitumor agent. A 1997 study used the cell line and a WiDr derived murine xenograft model to examine differences in sensitivity between the semisynthetic Camptothecin derivative CPT-11 and its metabolite SN-38, a known antineoplastic drug. Results showed DNA topoisomerase-1, critical for replication, activity rather than carboxylesterase, which metabolizes CPT-11, activity was a predictor for antitumor activity; higher topoisomerase activity correlated to higher sensitivity to both drugs which can be used to predict efficacy in a clinical setting. CPT-11 is now a chemotherapy drug (Camptosar®, camptothecin-11) used to treat colorectal cancer. A 2005 Oncology Reports study by Yorozuya et al. used the WiDr xenograft model to test TSU-68 (SU-6668), an anti-receptor tyrosine kinase drug that inhibits angiogenesis and cell proliferation and affects VEGF, bFGF and PDGF signaling in vitro. In vivo results showed TSU-68 significantly inhibited metastasis and tumor growth. The WiDr xenograft model is known to contain high expression levels of EGFR, thus making it an ideal model to study EGFR-TKIs. The EGFR status of this model is exceptionally beneficial when studying combinatorial effects (e.g. gefitinib, irinotecan). Also, antitumor efficacy has been shown using HDAC inhibitors (e.g. YM753).
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Basic study design
- WiDr cells are collected with trypsin-EDTA. Viable cells are counted using trypan blue exclusion. The mice (athymic BALB/C or NOD/SCID, 11 to 13 weeks) are injected subcutaneously in the flank of a hind leg. One million cells are inoculated into the mice (vol = 140-180 microliters of matrigel plus WiDr cells suspension).
- The injection are continuously observed until tumors are established. Tumors are measured until they reach an average of 100-150 mm3. Animals are sorted into treatment cohorts and the in-life portion of the study begins.
- Injections of test material is performed following the dosing schedule. Tumors are continually measured and body weights recorded.
- At the end of the study necropsies and tissue collections are followed according to the study design. Tumors are excised and weighed. All tissues collected can be placed in RNA-Later, snap frozen or added to 10% NBF for histology.
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WiDr Xenograft Model
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). 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).
Following options are available for the WiDr xenograft model:
- WiDr Tumor Growth Delay (TGD; latency)
- WiDr 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)
- WiDr 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)