title: Englerin A Agonizes the TRPC4/C5 Cation Channels to Inhibit Tumor Cell Line Proliferation.
abstract: Englerin A is a structurally unique natural product reported to selectively inhibit growth of renal cell carcinoma cell lines. A large scale phenotypic cell profiling experiment (CLiP) of englerin A on ¬over 500 well characterized cancer cell lines showed that englerin A inhibits growth of a subset of tumor cell lines from many lineages, not just renal cell carcinomas. Expression of the TRPC4 cation channel was the cell line feature that best correlated with sensitivity to englerin A, suggesting the hypothesis that TRPC4 is the efficacy target for englerin A. Genetic experiments demonstrate that TRPC4 expression is both necessary and sufficient for englerin A induced growth inhibition. Englerin A induces calcium influx and membrane depolarization in cells expressing high levels of TRPC4 or its close ortholog TRPC5. Electrophysiology experiments confirmed that englerin A is a TRPC4 agonist. Both the englerin A induced current and the englerin A induced growth inhibition can be blocked by the TRPC4/C5 inhibitor ML204. These experiments confirm that activation of TRPC4/C5 channels inhibits tumor cell line proliferation and confirms the TRPC4 target hypothesis generated by the cell line profiling. In selectivity assays englerin A weakly inhibits TRPA1, TRPV3/V4, and TRPM8 which suggests that englerin A may bind a common feature of TRP ion channels. In vivo experiments show that englerin A is lethal in rodents near doses needed to activate the TRPC4 channel. This toxicity suggests that englerin A itself is probably unsuitable for further drug development. However, since englerin A can be synthesized in the laboratory, it may be a useful chemical starting point to identify novel modulators of other TRP family channels.
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Carson
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Pichai
authors.initials: P
authors.lastname: Raman
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Tullai
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Martin
authors.initials: M
authors.lastname: Henault
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Emily
authors.initials: E
authors.lastname: Thomas
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Sarita
authors.initials: S
authors.lastname: Yeola
authors.affiliation: Genomics Institute of the Novartis Research Foundation, San Diego, California, United States of America.
authors.forename: Jianmin
authors.initials: J
authors.lastname: Lao
authors.affiliation: Novartis Institutes for Biomedical Research, Horsham, United Kingdom.
authors.forename: Mark
authors.initials: M
authors.lastname: McPate
authors.affiliation: Novartis Institutes for Biomedical Research, Horsham, United Kingdom.
authors.forename: J Martin
authors.initials: JM
authors.lastname: Verkuyl
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: George
authors.initials: G
authors.lastname: Marsh
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Jason
authors.initials: J
authors.lastname: Sarber
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Adam
authors.initials: A
authors.lastname: Amaral
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Scott
authors.initials: S
authors.lastname: Bailey
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Danuta
authors.initials: D
authors.lastname: Lubicka
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Helen
authors.initials: H
authors.lastname: Pham
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Nicolette
authors.initials: N
authors.lastname: Miranda
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Jian
authors.initials: J
authors.lastname: Ding
authors.affiliation: Novartis Institutes for Biomedical Research, East Hanover, New Jersey, United States of America.
authors.forename: Hai-Ming
authors.initials: HM
authors.lastname: Tang
authors.affiliation: Novartis Institutes for Biomedical Research, East Hanover, New Jersey, United States of America.
authors.forename: Haisong
authors.initials: H
authors.lastname: Ju
authors.affiliation: Novartis Institutes for Biomedical Research, Horsham, United Kingdom.
authors.forename: Pamela
authors.initials: P
authors.lastname: Tranter
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Nan
authors.initials: N
authors.lastname: Ji
authors.affiliation: Novartis Institutes for Biomedical Research, Basel, Switzerland.
authors.forename: Philipp
authors.initials: P
authors.lastname: Krastel
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Rishi K
authors.initials: RK
authors.lastname: Jain
authors.affiliation: Genomics Institute of the Novartis Research Foundation, San Diego, California, United States of America.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Schumacher
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Loureiro
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: George
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Giuliano
authors.initials: G
authors.lastname: Berellini
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Nathan T
authors.initials: NT
authors.lastname: Ross
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Simon M
authors.initials: SM
authors.lastname: Bushell
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Gül
authors.initials: G
authors.lastname: Erdemli
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, United States of America.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Solomon
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002292:Carcinoma, Renal Cell / Q000188:drug therapy; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D057809:HEK293 Cells; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007680:Kidney Neoplasms / Q000188:drug therapy; D051379:Mice; D008819:Mice, Nude; D010880:Piperidines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering; D051381:Rats; D045789:Sesquiterpenes, Guaiane / Q000494:pharmacology*; D050052:TRPC Cation Channels / Q000819:agonists* / Q000037:antagonists & inhibitors / Q000235:genetics; D014162:Transfection
chemical_list: D000970:Antineoplastic Agents; D007211:Indoles; C567966:ML 204; D010880:Piperidines; D034741:RNA, Small Interfering; D045789:Sesquiterpenes, Guaiane; D050052:TRPC Cation Channels; C489571:TRPC4 ion channel; C491780:TRPC5 protein, human; C534971:englerin A; C551614:englerin B