Discovery of molecular mechanisms of neuroprotection using cell-based bioassays and oligonucleotide arrays.
Oxidative injury and the resulting death of neurons is a major pathological factor involved in numerous neurodegenerative diseases. However, the development of drugs that target this mechanism remains limited. The goal of this study was to test a compound library of approved Food and Drug Administration drugs against a hydrogen peroxide-induced oxidant injury model in neuroblastoma cells. We identified 26 neuroprotective compounds, of which megestrol, meclizine, verapamil, methazolamide, sulindac, and retinol were examined in greater detail. Using large-scale oligonucleotide microarray analysis, we identified genes modulated by these drugs that might underlie the cytoprotection. Five key genes were either uniformly upregulated or downregulated by all six drug treatments, namely, tissue inhibitor of matrix metalloproteinase (TIMP1), ret-proto-oncogene, clusterin, galanin, and growth associated protein (GAP43). Exogenous addition of the neuropeptide galanin alone conferred survival to oxidant-stressed cells, comparable to that seen with the drugs. Our approach, which we term "interventional profiling," represents a general and powerful strategy for identifying new bioactive agents for any biological process, as well as identifying key downstream genes and pathways that are involved.
Sarang
Satinder S
SS
Biotechnology Center, Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts 02139, USA.
Yoshida
Takumi
T
Cadet
Rodolphe
R
Valeras
Andrew S
AS
Jensen
Roderick V
RV
Gullans
Steven R
SR
D001681:Biological Assay / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000378:metabolism; D008468:Meclizine / Q000494:pharmacology; D008535:Megestrol / Q000494:pharmacology; D008704:Methazolamide / Q000494:pharmacology; D009447:Neuroblastoma / Q000473:pathology; D019636:Neurodegenerative Diseases / Q000139:chemically induced / Q000517:prevention & control*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016877:Oxidants / Q000009:adverse effects / Q000037:antagonists & inhibitors; D000090063:Proto-Oncogene Mas; D013467:Sulindac / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014700:Verapamil / Q000494:pharmacology; D014801:Vitamin A / Q000494:pharmacology
C000715470:MAS1 protein, human; D018696:Neuroprotective Agents; D016877:Oxidants; D000090063:Proto-Oncogene Mas; D014801:Vitamin A; D013467:Sulindac; D008468:Meclizine; D006861:Hydrogen Peroxide; D014700:Verapamil; D008535:Megestrol; D008704:Methazolamide