title: Sensitivity and engineered resistance of myeloid leukemia cells to BRD9 inhibition.
abstract: Here we show that acute myeloid leukemia (AML) cells require the BRD9 subunit of the SWI-SNF chromatin-remodeling complex to sustain MYC transcription, rapid cell proliferation and a block in differentiation. Based on these observations, we derived small-molecule inhibitors of the BRD9 bromodomain that selectively suppress the proliferation of mouse and human AML cell lines. To establish these effects as on-target, we engineered a bromodomain-swap allele of BRD9 that retains functionality despite a radically altered bromodomain pocket. Expression of this allele in AML cells confers resistance to the antiproliferative effects of our compound series, thus establishing BRD9 as the relevant cellular target. Furthermore, we used an analogous domain-swap strategy to generate an inhibitor-resistant allele of EZH2. To our knowledge, our study provides the first evidence for a role of BRD9 in cancer and reveals a simple genetic strategy for constructing resistance alleles to demonstrate on-target activity of chemical probes in cells.
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.|Watson School of Biological Sciences, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Anja F
authors.initials: AF
authors.lastname: Hohmann
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Laetitia J
authors.initials: LJ
authors.lastname: Martin
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Minder
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Jae-Seok
authors.initials: JS
authors.lastname: Roe
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.|Molecular and Cellular Biology Program, Stony Brook University, Stony Brook, New York, USA.
authors.forename: Junwei
authors.initials: J
authors.lastname: Shi
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Steffen
authors.initials: S
authors.lastname: Steurer
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Gerd
authors.initials: G
authors.lastname: Bader
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Darryl
authors.initials: D
authors.lastname: McConnell
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Mark
authors.initials: M
authors.lastname: Pearson
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gerstberger
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Teresa
authors.initials: T
authors.lastname: Gottschamel
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Diane
authors.initials: D
authors.lastname: Thompson
authors.affiliation: Department of Medical Genome Sciences, University of Tokyo, Kashiwa, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: Boehringer Ingelheim Regional Center Vienna GmbH and Company KG, Vienna, Austria.
authors.forename: Manfred
authors.initials: M
authors.lastname: Koegl
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.|Watson School of Biological Sciences, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Vakoc
mesh_terms: D000483:Alleles; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D060846:Cell Engineering; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000092468:Bromodomain Containing Proteins
chemical_list: D000970:Antineoplastic Agents; D014157:Transcription Factors; C587454:BRD9 protein, human; D000092468:Bromodomain Containing Proteins