Design, development, and validation of a high-throughput drug-screening assay for targeting of human leukemia.
BACKGROUND
The authors developed an ex vivo methodology to perform drug library screening against human leukemia.


METHODS
The strategy for this screening relied on human blood or bone marrow cultures under hypoxia; under these conditions, leukemia cells deplete oxygen faster than normal cells, causing a hemoglobin oxygenation shift. Several advantages were observed: 1) partial recapitulation of the leukemia microenvironment, 2) use of native hemoglobin oxygenation as a real-time sensor/reporter, 3) cost-effectiveness, 4) species specificity, and 5) a format that enables high-throughput screening.


RESULTS
For a proof of concept, a chemical library (size, approximately 20,000 compounds) was screened against human leukemia cells. Seventy compounds were identified ("hit" rate, 0.35%; Z-factor = 0.71) that had activity, and 20 compounds were examined to identify 18 true-positive compounds (90%). Finally, the results demonstrated that carbonohydraxonic diamide group-containing compounds are potent antileukemia agents that induce cell death in leukemia cells and in patient-derived samples.


CONCLUSIONS
The current results indicated that this unique functional assay can identify novel drug candidates and can help with the development of future applications in personalized drug selection for patients with leukemia.
David H. Koch Center, The University of Texas MD Anderson Cancer Center, Houston, Texas; Department of Biological and Environmental Sciences, Division of Biochemistry, The University of Helsinki, Helsinki, Finland.
Katja
K
Karjalainen
Renata
R
Pasqualini
Jorge E
JE
Cortes
Steven M
SM
Kornblau
Benjamin
B
Lichtiger
Susan
S
O'Brien
Hagop M
HM
Kantarjian
Richard L
RL
Sidman
Wadih
W
Arap
Erkki
E
Koivunen
D004354:Drug Screening Assays, Antitumor; D057166:High-Throughput Screening Assays; D006801:Humans; D007938:Leukemia / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010100:Oxygen / Q000378:metabolism*; D054852:Small Molecule Libraries
D054852:Small Molecule Libraries; D010100:Oxygen
blood; bone marrow; chemical library; drug screening; hypoxia; leukemia targeting; tumor microenvironment