title: Functional RNAi screen targeting cytokine and growth factor receptors reveals oncorequisite role for interleukin-2 gamma receptor in JAK3-mutation-positive leukemia.
abstract: To understand the role of cytokine and growth factor receptor-mediated signaling in leukemia pathogenesis, we designed a functional RNA interference (RNAi) screen targeting 188 cytokine and growth factor receptors that we found highly expressed in primary leukemia specimens. Using this screen, we identified interleukin-2 gamma receptor (IL2Rγ) as a critical growth determinant for a JAK3(A572V) mutation-positive acute myeloid leukemia cell line. We observed that knockdown of IL2Rγ abrogates phosphorylation of JAK3 and downstream signaling molecules, JAK1, STAT5, MAPK and pS6 ribosomal protein. Overexpression of IL2Rγ in murine cells increased the transforming potential of activating JAK3 mutations, whereas absence of IL2Rγ completely abrogated the clonogenic potential of JAK3(A572V), as well as the transforming potential of additional JAK3-activating mutations such as JAK3(M511I). In addition, mutation at the IL2Rγ interaction site in the FERM domain of JAK3 (Y100C) completely abrogated JAK3-mediated leukemic transformation. Mechanistically, we found IL2Rγ contributes to constitutive JAK3 mutant signaling by increasing JAK3 expression and phosphorylation. Conversely, we found that mutant, but not wild-type JAK3, increased the expression of IL2Rγ, indicating IL2Rγ and JAK3 contribute to constitutive JAK/STAT signaling through their reciprocal regulation. Overall, we demonstrate a novel role for IL2Rγ in potentiating oncogenesis in the setting of JAK3-mutation-positive leukemia. In addition, our study highlights an RNAi-based functional assay that can be used to facilitate the identification of non-kinase cytokine and growth factor receptor targets for inhibiting leukemic cell growth.
authors.affiliation: 1] Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, OR, USA [2] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA.
authors.forename: A
authors.initials: A
authors.lastname: Agarwal
authors.affiliation: 1] Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, OR, USA [2] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA.
authors.forename: R J
authors.initials: RJ
authors.lastname: MacKenzie
authors.affiliation: 1] Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, OR, USA [2] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA [3] Howard Hughes Medical Institute, Portland, OR, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Eide
authors.affiliation: Department of Pediatrics, Oregon Health & Science University, Portland, OR, USA.
authors.forename: M A
authors.initials: MA
authors.lastname: Davare
authors.affiliation: 1] Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, OR, USA [2] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA.
authors.forename: K
authors.initials: K
authors.lastname: Watanabe-Smith
authors.affiliation: 1] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA [2] Howard Hughes Medical Institute, Portland, OR, USA.
authors.forename: C E
authors.initials: CE
authors.lastname: Tognon
authors.affiliation: 1] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA [2] Biostatistics Shared Resource, Oregon Health & Science University, Portland, OR, USA.
authors.forename: S
authors.initials: S
authors.lastname: Mongoue-Tchokote
authors.affiliation: 1] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA [2] Biostatistics Shared Resource, Oregon Health & Science University, Portland, OR, USA.
authors.forename: B
authors.initials: B
authors.lastname: Park
authors.affiliation: Department of Pathology, Oregon Health & Science University, Portland, OR, USA.
authors.forename: R M
authors.initials: RM
authors.lastname: Braziel
authors.affiliation: 1] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA [2] Department of Cell & Developmental Biology, Oregon Health & Science University, Portland, OR, USA.
authors.forename: J W
authors.initials: JW
authors.lastname: Tyner
authors.affiliation: 1] Division of Hematology and Medical Oncology, Oregon Health & Science University, Portland, OR, USA [2] Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA [3] Howard Hughes Medical Institute, Portland, OR, USA.
authors.forename: B J
authors.initials: BJ
authors.lastname: Druker
mesh_terms: D000818:Animals; D001665:Binding Sites; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D006801:Humans; D053631:Interleukin Receptor Common gamma Subunit / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D053616:Janus Kinase 3 / Q000235:genetics* / Q000378:metabolism; D007938:Leukemia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000494:pharmacology*; D015398:Signal Transduction
chemical_list: C507941:IL2RG protein, human; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; D034741:RNA, Small Interfering; C507933:JAK3 protein, human; D053616:Janus Kinase 3
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