Norris Cancer Center Preclinical Testing Laboratory
Tthe USC Norris Cancer Center Developmental Therapeutics Program has, via a grant from the Whitier Foundation, conduct testing of drugs in cell culture and xenograft models of adult cancers. This preclinical testing is carried out by USC-CHLA Institute for Pediatric Clinical Research (IPCR) Developmental Therapeutics Program investigators C Patrick Reynolds, MD PhD and Barry J. Maurer, MD PhD.
Trainee: Younglem Kim, PhD
Methods employed:
The primary cytotoxicity assay for the lab is the DIMSCAN 96 well assay. DIMSCAN employs fluorescein diacetate (FDA) to identify viable cells, with quenching of fluorescence by eosin Y, and viable cells quantified using a DIMSCAN digital image microscopy system. (1,2). The DIMSCAN assay has a 4 log dynamic range when testing solid tumor cell lines (3, 4). Testing is carried out under standard culture conditions (20% O2), but drugs shown to be active in initial testing are also assessed in bone marrow-level oxygen tension (5 % O2) and in 2% O2, which approximates the oxygen levels found in many tumors (5). Assessing drug interactions (additivity, synergy, antagonism) is done employing fixed-ratios of drug concentrations and the combination index approach (4,6) All cell lines used are human, have been tested and shown to be free of mycoplasma, and have been demonstrated to be unique using short tandem repeats (7).
Xenograft testing is carried out using cell lines established as xenograft tumors. Primary testing employs standard subcutaneous tumor models in nu/nu mice (8). Testing against disseminated disease models in SCID mice (created by tail vein injection) is also employed (8). Bone metastases in disseminated disease models are monitored using a Faxitron radiograph system (8).
Bibliography
- Proffitt RT, Tran JV, Reynolds CP: A fluorescence digital image microscopy system for quantitating relative cell numbers in tissue culture plates. Cytometry 24:204-213,1996.
- Keshelava N, Frgala T, .Krejsa J, Kalous O, Reynolds, CP: DIMSCAN: a microcomputer fluorescence-based cytotoxicity assay suitable for pre-clinical testing of combination chemotherapy. Methods in Molecular Medicine Chemosensitivity Vol 1 ed. Blumenthal RD, Totowa: Humana Press pp 139-154, 2005.
- Keshelava N, Seeger RC, Groshen S, Reynolds CP: Drug resistance patterns of human neuroblastoma cell lines derived from patients at different phases of therapy. Cancer Research 58:5396-5405, 1998.
- Maurer BJ, Cabot MC, Reynolds CP: Syngergism of N-(4-hydroxypheynl)retinamide cytotoxcity by modulators of ceramide metabolism in solid tumor cell lines. J Natl Cancer Inst 92:1897-1908, 2000.
- Grigoryan R, Keshelava N, Anderson C, Reynolds, CP: In vitro testing of chemosensitivity in physiological hypoxia. Methods in Molecular Medicine Chemosensitivity Vol 1 ed. Blumenthal RD, Totowa: Humana Press pp 87-100, 2005.
- Reynolds, CP, Maurer BJ: Assessing response to anti-neoplastic drug combinations in tissue culture models. Methods in Molecular Medicine Chemosensitivity Vol 1 ed. Blumenthal RD, Totowa: Humana Press pp 173-184, 2005.
- In preparation
- Reynolds, CP, Sun BC, DeClerck YA, Moats RA: Assessing growth and response to therapy in murine tumor models. Methods in Molecular Medicine Chemosensitivity Vol 2 ed. Blumenthal RD, Totowa: Humana Press pp 335-350, 2005.
Cell Culture Models
Prostate Cancer:
- LNCAP
- PC3
Ovarian Cancer
- Ovacar 3
Leukemia
- SMS-SB
- HL-60
Xenograft Models
Clinical Trials Enabled by this Laboratory
California Cancer Consortium Phase II study of oral capsular fenretnide in ovarian cancer (completed)
PhI-42 California Cancer Consortium Phase I trial of intravenous fenretinide in hematological malignancies
California Cancer Consortium Phase II trial of oral (capsule) fenretinide in men with prostate cancer and asymptomatic rising PSA
California Cancer Consortium Phase I trial of intravenous fenretinide in refractory solid tumors
Laboratory Publications
Laboratory Funding
The Whittier Foundation