title: Activation of the erythroid K-Cl cotransporter Kcc1 enhances sickle cell disease pathology in a humanized mouse model.
abstract: We used an N-ethyl-N-nitrosurea-based forward genetic screen in mice to identify new genes and alleles that regulate erythropoiesis. Here, we describe a mouse line expressing an activated form of the K-Cl cotransporter Slc12a4 (Kcc1), which results in a semi-dominant microcytosis of red cells. A missense mutation from methionine to lysine in the cytoplasmic tail of Kcc1 impairs phosphorylation of adjacent threonines required for inhibiting cotransporter activity. We bred Kcc1(M935K) mutant mice with a humanized mouse model of sickle cell disease to directly explore the relevance of the reported increase in KCC activity in disease pathogenesis. We show that a single mutant allele of Kcc1 induces widespread sickling and tissue damage, leading to premature death. This mouse model reveals important new insights into the regulation of K-Cl cotransporters and provides in vivo evidence that increased KCC activity worsened end-organ damage and diminished survival in sickle cell disease.
authors.affiliation: Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia;
authors.forename: Fiona C
authors.initials: FC
authors.lastname: Brown
authors.affiliation: Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia;
authors.forename: Ashlee J
authors.initials: AJ
authors.lastname: Conway
authors.affiliation: Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia;
authors.forename: Loretta
authors.initials: L
authors.lastname: Cerruti
authors.affiliation: Cancer and Haematology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia;
authors.forename: Janelle E
authors.initials: JE
authors.lastname: Collinge
authors.affiliation: The Alfred Hospital, Melbourne, Australia;
authors.forename: Catriona
authors.initials: C
authors.lastname: McLean
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, Australia; and.
authors.forename: James S
authors.initials: JS
authors.lastname: Wiley
authors.affiliation: Cancer and Haematology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia;
authors.forename: Ben T
authors.initials: BT
authors.lastname: Kile
authors.affiliation: Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia; The Alfred Hospital, Melbourne, Australia; Department of Medicine, Central Clinical School, Monash University, Melbourne, Australia.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Jane
authors.affiliation: Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia; The Alfred Hospital, Melbourne, Australia;
authors.forename: David J
authors.identifier: http://orcid.org/0000-0001-9497-0996
authors.initials: DJ
authors.lastname: Curtis
mesh_terms: D000755:Anemia, Sickle Cell / Q000235:genetics* / Q000473:pathology*; D000818:Animals; D004195:Disease Models, Animal; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D020125:Mutation, Missense; D020133:Reverse Transcriptase Polymerase Chain Reaction; D027981:Symporters / Q000235:genetics*; D000096922:K Cl- Cotransporters
chemical_list: D027981:Symporters; D000096922:K Cl- Cotransporters