title: Metabolic gatekeeper function of B-lymphoid transcription factors.
abstract: B-lymphoid transcription factors, such as PAX5 and IKZF1, are critical for early B-cell development, yet lesions of the genes encoding these transcription factors occur in over 80% of cases of pre-B-cell acute lymphoblastic leukaemia (ALL). The importance of these lesions in ALL has, until now, remained unclear. Here, by combining studies using chromatin immunoprecipitation with sequencing and RNA sequencing, we identify a novel B-lymphoid program for transcriptional repression of glucose and energy supply. Our metabolic analyses revealed that PAX5 and IKZF1 enforce a state of chronic energy deprivation, resulting in constitutive activation of the energy-stress sensor AMPK. Dominant-negative mutants of PAX5 and IKZF1, however, relieved this glucose and energy restriction. In a transgenic pre-B ALL mouse model, the heterozygous deletion of Pax5 increased glucose uptake and ATP levels by more than 25-fold. Reconstitution of PAX5 and IKZF1 in samples from patients with pre-B ALL restored a non-permissive state and induced energy crisis and cell death. A CRISPR/Cas9-based screen of PAX5 and IKZF1 transcriptional targets identified the products of NR3C1 (encoding the glucocorticoid receptor), TXNIP (encoding a glucose-feedback sensor) and CNR2 (encoding a cannabinoid receptor) as central effectors of B-lymphoid restriction of glucose and energy supply. Notably, transport-independent lipophilic methyl-conjugates of pyruvate and tricarboxylic acid cycle metabolites bypassed the gatekeeper function of PAX5 and IKZF1 and readily enabled leukaemic transformation. Conversely, pharmacological TXNIP and CNR2 agonists and a small-molecule AMPK inhibitor strongly synergized with glucocorticoids, identifying TXNIP, CNR2 and AMPK as potential therapeutic targets. Furthermore, our results provide a mechanistic explanation for the empirical finding that glucocorticoids are effective in the treatment of B-lymphoid but not myeloid malignancies. Thus, B-lymphoid transcription factors function as metabolic gatekeepers by limiting the amount of cellular ATP to levels that are insufficient for malignant transformation.
authors.affiliation: Department of Systems Biology, Beckman Research Institute and City of Hope National Medical Center, Pasadena, California 91016, USA.|City of Hope Comprehensive Cancer Center, Duarte, California 91010, USA.
authors.forename: Lai N
authors.initials: LN
authors.lastname: Chan
authors.affiliation: Department of Systems Biology, Beckman Research Institute and City of Hope National Medical Center, Pasadena, California 91016, USA.|City of Hope Comprehensive Cancer Center, Duarte, California 91010, USA.
authors.forename: Zhengshan
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Molecular and Medical Pharmacology, UCLA Metabolomics Center and Crump Institute for Molecular Imaging, University of California Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Braas
authors.affiliation: Department of Systems Biology, Beckman Research Institute and City of Hope National Medical Center, Pasadena, California 91016, USA.|City of Hope Comprehensive Cancer Center, Duarte, California 91010, USA.
authors.forename: Jae-Woong
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Systems Biology, Beckman Research Institute and City of Hope National Medical Center, Pasadena, California 91016, USA.|City of Hope Comprehensive Cancer Center, Duarte, California 91010, USA.
authors.forename: Gang
authors.initials: G
authors.lastname: Xiao
authors.affiliation: Departments of Laboratory Medicine and Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Huimin
authors.initials: H
authors.lastname: Geng
authors.affiliation: Department of Systems Biology, Beckman Research Institute and City of Hope National Medical Center, Pasadena, California 91016, USA.|City of Hope Comprehensive Cancer Center, Duarte, California 91010, USA.
authors.forename: Kadriye Nehir
authors.initials: KN
authors.lastname: Cosgun
authors.affiliation: Departments of Laboratory Medicine and Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Hurtz
authors.affiliation: Departments of Laboratory Medicine and Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Seyedmehdi
authors.initials: S
authors.lastname: Shojaee
authors.affiliation: Departments of Laboratory Medicine and Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Valeria
authors.initials: V
authors.lastname: Cazzaniga
authors.affiliation: Departments of Laboratory Medicine and Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Hilde
authors.initials: H
authors.lastname: Schjerven
authors.affiliation: Hämatologie/Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, 07743 Jena, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Ernst
authors.affiliation: Hämatologie/Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, 07743 Jena, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hochhaus
authors.affiliation: Department of Leukemia, The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, USA.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Kornblau
authors.affiliation: Department of Leukemia, The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Konopleva
authors.affiliation: Department of Biochemistry, University of Iowa, Iowa City, Iowa 52242, USA.
authors.forename: Miles A
authors.initials: MA
authors.lastname: Pufall
authors.affiliation: Centro Ricerca Tettamanti, Clinica Pediatrica, Università di Milano-Bicocca, Ospedale S. Gerardo, 20052 Monza MB, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Cazzaniga
authors.affiliation: Australian Centre for Blood Diseases, Monash University, Melbourne, Victoria 3004 Australia.
authors.forename: Grace J
authors.initials: GJ
authors.lastname: Liu
authors.affiliation: MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Milne
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Singapore 117599.|Division of Hematology/Oncology, Cedars-Sinai Medical Center, University of California School of Medicine, Los Angeles, California 90095, USA.
authors.forename: H Phillip
authors.initials: HP
authors.lastname: Koeffler
authors.affiliation: Department of Internal Medicine and Cancer Genetics, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
authors.forename: Theodora S
authors.initials: TS
authors.lastname: Ross
authors.affiliation: Experimental Therapeutics and Translational Oncology Program, Instituto de Biología Molecular y Celular del Cáncer, CSIC/Universidad de Salamanca and Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain.
authors.forename: Isidro
authors.initials: I
authors.lastname: Sánchez-García
authors.affiliation: Pediatric Oncology, Hematology and Clinical Immunology, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany.
authors.forename: Arndt
authors.initials: A
authors.lastname: Borkhardt
authors.affiliation: Departments of Laboratory Medicine and Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California 94143, USA.
authors.forename: Keith R
authors.initials: KR
authors.lastname: Yamamoto
authors.affiliation: Australian Centre for Blood Diseases, Monash University, Melbourne, Victoria 3004 Australia.
authors.forename: Ross A
authors.initials: RA
authors.lastname: Dickins
authors.affiliation: Department of Molecular and Medical Pharmacology, UCLA Metabolomics Center and Crump Institute for Molecular Imaging, University of California Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Graeber
authors.affiliation: Department of Systems Biology, Beckman Research Institute and City of Hope National Medical Center, Pasadena, California 91016, USA.|City of Hope Comprehensive Cancer Center, Duarte, California 91010, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Müschen
mesh_terms: D000091162:AMP-Activated Protein Kinase Kinases; D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D063646:Carcinogenesis / Q000235:genetics; D002352:Carrier Proteins / Q000819:agonists / Q000378:metabolism; D016923:Cell Death; D047369:Chromatin Immunoprecipitation; D002952:Citric Acid Cycle; D004195:Disease Models, Animal; D004734:Energy Metabolism / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D005938:Glucocorticoids / Q000494:pharmacology / Q000627:therapeutic use; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D051740:Ikaros Transcription Factor / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D051757:PAX5 Transcription Factor / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D019289:Pyruvic Acid / Q000378:metabolism; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D017423:Sequence Analysis, RNA; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: C588309:CNR2 protein, human; D002352:Carrier Proteins; D005938:Glucocorticoids; C497239:IKZF1 protein, human; C497713:NR3C1 protein, human; D051757:PAX5 Transcription Factor; C497281:PAX5 protein, human; C497282:Pax5 protein, mouse; D043885:Receptor, Cannabinoid, CB2; D011965:Receptors, Glucocorticoid; C486715:TXNIP protein, human; D014157:Transcription Factors; D051740:Ikaros Transcription Factor; D019289:Pyruvic Acid; D000255:Adenosine Triphosphate; D017346:Protein Serine-Threonine Kinases; C109790:STK11 protein, human; D000091162:AMP-Activated Protein Kinase Kinases; D055372:AMP-Activated Protein Kinases; D005947:Glucose