TACL Leukemia Preclinical Testing Lab
The leukemia preclinical testing laboratory in USC-CHLA Institute for Pediatric Clinical Research (IPCR) Developmental Therapeutics Program provides preclinical testing for the Therapeutic Advances in Childhood Leukemia (TACL) consortium.
The Leukemia Preclinical Testing lab for TACL is under the direction of Min Kang , Pharm D, who works closely with C Patrick Reynolds, MD PhD and Barry J. Maurer, MD PhD also at the USC-CHLA IPCR. In addition, a collaboration with the ALL preclinical testing laboratory directed by Richard Lock PhD of the Childrens Cancer Institute of Australia in Sydney provides additional testing of new drugs and drug combinations in the nod/SCID mouse models of childhood acute lymphoblastic leukemia (ALL) developed by Dr. Lock.
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 leukemia cell lines (3). All in vitro testing is carried out in bone marrow-level oxygen tension (5 % O2) (4). 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.
Xenograft testing is carried out in nod/SCID mice using cell lines or directly established xenografts injected intravenously (5, 6). Mice are treated with drugs after engraftment of the leukemia cells. The mouse leukemia burden is monitored using flow cytometry to detect human CD-45-positive cells (5, 6).
Pre-B ALL Cell Lines
NALM-6
NALL
SMS-SB
COG-LL-319
RS;4-11
T Cell ALL Cell Lines
Molt-3
Molt-4
CEM
COG-LL-317
COG-LL-332
Acute myelogenous leukemia Cell Lines
HL-60
CHLA-221
Kasmi 1
Pre-B ALL Xenografts
COG-LL-319
T Cell ALL Xenografts
COG-LL-317
B Lymphoma
Ramos
Clinical Trials Enabled by this Laboratory
Leukemia and Lymphoma Xenograft Drug Testing
A high fraction of human acute lymphoblastic leukemia (ALL) cells can be engrafted in nod/SCID mice (1, 2). As demonstrated by Lock et al, ALL xenografts show responses to anti-leukemia drugs in mice that are consistent with the sensitivity of the leukemia cells to chemotherapy in the patients (1,2).
To increase the number of ALL preclinical models available for preclinical testing and biological studies, the USC-CHLA IPCR has established a lab devoted to initiating and characterizing new animal models of childhood leukemia. The lab focuses primarily on establishing and characterizing ALL xenografts in nod/SCID mice. ALL samples are obtained in collaboration with the Children's Oncology Group (COG) and the Therapeutic Advances in Childhood Leukemia (TACL) consortia.
The USC-CHLA IPCR Leukemia Animal Model Lab is also under the direction of Min Kang , Pharm D.
Xenografts Established
| Cell line name | Xenograft name | |
| none | COG-LL-329x | T Cell |
| none | COG-LL-335x | pending |
| none | COG-LL-330x | Pre-B |
| COG-LL-319 | COG-LL-319x | Pre-B |
| COG-LL-317 | COG-LL-317x | T Cell |
| none | COG-LL-336x | pending |
| none | COG-LL-337x | pending |
| none | COG-LL-338x | pending |
| none | COG-LL-339x | pending |
| COG-LL-332 | COG-LL-332x | T Cell |
Laboratory Funding
The USC-CHLA Institute for Pediatric Clinical Research
Laboratory Publications
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.
- O'Donnell PH, Guo WX, Reynolds CP, Maurer BJ: N-(4-hydroxyphenyl)retinamide increases ceramide and Is cytotoxic to acute lymphoblastic leukemia cell lines, but not to non-malignant lymphocytes. Leukemia 16:902-910, 2002.
- 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.
- Liem, NLM, Papa, RA, Milross, CG, Schmid, MA, Tajbakhsh, M, Choi, S, Ramirez, CD, Rice, AM, Haber, M, Norris, MD, MacKenzie, KL, and Lock, RB Characterization of childhood acute lymphoblastic leukemia xenograft models for the preclinical evaluation of new therapies. Blood, 103: 3905-3914, 2004.
- Look RB, Liem NL, Papa RA:Preclinical testing of antileukemic drugs using an in vivo model of systemic disease Methods Mol Med. ;111:323-34, 2005.