title: Cell based screening of inhibitors of transthyretin aggregation.
abstract: The amyloidoses are the extracellular subset of a group of diseases in which in vivo protein misfolding leads to a pathologic gain of function, i.e., aggregation leading to protein deposition, with subsequent tissue damage. Wild-type and mutant transthyretins (TTR) are the etiologic agents in prototypic systemic amyloidoses. We describe a cell-based assay that measures the cytotoxicity of physiologic concentrations of the amyloidogenic Val30Met TTR variant (V30M TTR) using cells of the same lineage as the in vivo tissue target of amyloid deposition. We have utilized the assay to screen small molecules for their capacity to inhibit the TTR-induced cell damage. We compared the inhibitory activity of each compound with its ability to prevent TTR fibril formation in vitro. Our results emphasize the importance of screening compounds under physiologic conditions. Moreover, if a common conformational intermediate is responsible for cell death in all the amyloid diseases, the cell-based assay has the potential to aid in the discovery of compounds useful in the treatment of amyloidoses caused by other misfolded proteins as well as those caused by TTR.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Natàlia
authors.initials: N
authors.lastname: Reixach
authors.forename: Sara L
authors.initials: SL
authors.lastname: Adamski-Werner
authors.forename: Jeffery W
authors.initials: JW
authors.lastname: Kelly
authors.forename: James
authors.initials: J
authors.lastname: Koziol
authors.forename: Joel N
authors.initials: JN
authors.lastname: Buxbaum
mesh_terms: D000686:Amyloidosis / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D045744:Cell Line, Tumor; D019070:Cell Lineage / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004061:Diflunisal / Q000031:analogs & derivatives / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D006801:Humans; D008715:Methionine / Q000235:genetics; D020894:Microfibrils / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D011228:Prealbumin / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology; D014633:Valine / Q000235:genetics
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D011228:Prealbumin; D013267:Stilbenes; D004061:Diflunisal; D008715:Methionine; D014633:Valine; D000077185:Resveratrol