title: Target Deconvolution Efforts on Wnt Pathway Screen Reveal Dual Modulation of Oxidative Phosphorylation and SERCA2.
abstract: Wnt signaling is critical for development, cell proliferation and differentiation, and mutations in this pathway resulting in constitutive signaling have been implicated in various cancers. A pathway screen using a Wnt-dependent reporter identified a chemical series based on a 1,2,3-thiadiazole-5-carboxamide (TDZ) core with sub-micromolar potency. Herein we report a comprehensive mechanism-of-action deconvolution study toward identifying the efficacy target(s) and biological implication of this chemical series involving bottom-up quantitative chemoproteomics, cell biology, and biochemical methods. Through observing the effects of our probes on metabolism and performing confirmatory cellular and biochemical assays, we found that this chemical series inhibits ATP synthesis by uncoupling the mitochondrial potential. Affinity chemoproteomics experiments identified sarco(endo)plasmic reticulum Ca2+-dependent ATPase (SERCA2) as a binding partner of the TDZ series, and subsequent validation studies suggest that the TDZ series can act as ionophores through SERCA2 toward Wnt pathway inhibition.
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Drug Discovery Program, Ontario Institute for Cancer Research, 661 University Avenue, Suite 510, Toronto, ON, M5G 0A3, Canada.
authors.forename: Matias
authors.identifier: 0000-0001-8514-0632
authors.initials: M
authors.lastname: Casás-Selves
authors.affiliation: Discovery Sciences-Chemical Biology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: Andrew X
authors.identifier: 0000-0003-0406-2404
authors.initials: AX
authors.lastname: Zhang
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: James E
authors.identifier: 0000-0002-3984-4169
authors.initials: JE
authors.lastname: Dowling
authors.affiliation: Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Discovery Unit, AstraZeneca, Pepparedsleden 1, Mölndal, 431 83, Sweden.
authors.forename: Stefan
authors.initials: S
authors.lastname: Hallén
authors.affiliation: Discovery Sciences-Chemical Biology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: Aarti
authors.initials: A
authors.lastname: Kawatkar
authors.affiliation: Department of Chemistry, Boston College, Chestnut Hill, MA, 02467, USA.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Pace
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Denz
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Pontz
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Synageva BioPharma Corp., 33 Hayden Avenue, Lexington, MA, 02421, USA.
authors.forename: Farzin
authors.initials: F
authors.lastname: Garahdaghi
authors.affiliation: Discovery Sciences-Computational Chemistry, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Ra Pharmaceuticals, Inc., 87 Cambridge Park Drive, Cambridge, MA, 02140, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Cao
authors.affiliation: Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Discovery Unit, AstraZeneca, Pepparedsleden 1, Mölndal, 431 83, Sweden.
authors.forename: Alan
authors.initials: A
authors.lastname: Sabirsh
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: Kumar
authors.initials: K
authors.lastname: Thakur
authors.affiliation: Discovery Sciences-Structure and Biophysics, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Nurix, Inc., 1700 Owens Street, Suite 290, San Francisco, CA, 94158, USA.
authors.forename: Nichole
authors.initials: N
authors.lastname: O'Connell
authors.affiliation: Discovery Sciences-Structure and Biophysics, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Shire, 300 Shire Way, Lexington, MA, 02421, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Hu
authors.affiliation: Discovery Sciences-Chemical Biology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Discovery Chemistry, Merck Research Laboratories, 33 Avenue Louis Pasteur, Boston, MA, 02115, USA.
authors.forename: Iván
authors.initials: I
authors.lastname: Cornella-Taracido
authors.affiliation: Department of Chemistry, Boston College, Chestnut Hill, MA, 02467, USA.
authors.forename: Eranthie
authors.initials: E
authors.lastname: Weerapana
authors.affiliation: Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Zinda
authors.affiliation: Discovery Sciences-Chemical Biology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.|Present address: Pharmaron, 303 Wyman Street, Room 322, Waltham, MA, 02451, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Goodnow
authors.affiliation: Discovery Sciences-Chemical Biology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
authors.forename: M Paola
authors.initials: MP
authors.lastname: Castaldi
mesh_terms: D004305:Dose-Response Relationship, Drug; D006801:Humans; D015394:Molecular Structure; D010085:Oxidative Phosphorylation / Q000187:drug effects*; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013830:Thiadiazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D060449:Wnt Signaling Pathway / Q000187:drug effects*
chemical_list: D013830:Thiadiazoles; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; C506946:ATP2A2 protein, human
keywords: Wnt signaling; chemical biology; chemoproteomics; target identification