Activation of the erythroid K-Cl cotransporter Kcc1 enhances sickle cell disease pathology in a humanized mouse model

Fiona C Brown, Ashlee J Conway, Loretta Cerruti, Janelle E Collinge, Catriona McLean, James S Wiley, Ben T Kile, Stephen M Jane, David John Curtis

Research output: Contribution to journalArticleResearchpeer-review

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 semidominant 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 Kcc1M935K 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.
Original languageEnglish
Pages (from-to)2863-2870
Number of pages8
JournalBlood
Volume126
Issue number26
DOIs
Publication statusPublished - 2015

Cite this

@article{272b0a3ff8b94ec38947ac040234a18a,
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 semidominant 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 Kcc1M935K 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.",
author = "Brown, {Fiona C} and Conway, {Ashlee J} and Loretta Cerruti and Collinge, {Janelle E} and Catriona McLean and Wiley, {James S} and Kile, {Ben T} and Jane, {Stephen M} and Curtis, {David John}",
year = "2015",
doi = "10.1182/blood-2014-10-609362",
language = "English",
volume = "126",
pages = "2863--2870",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "26",

}

Activation of the erythroid K-Cl cotransporter Kcc1 enhances sickle cell disease pathology in a humanized mouse model. / Brown, Fiona C; Conway, Ashlee J; Cerruti, Loretta; Collinge, Janelle E; McLean, Catriona; Wiley, James S; Kile, Ben T; Jane, Stephen M; Curtis, David John.

In: Blood, Vol. 126, No. 26, 2015, p. 2863-2870.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Activation of the erythroid K-Cl cotransporter Kcc1 enhances sickle cell disease pathology in a humanized mouse model

AU - Brown, Fiona C

AU - Conway, Ashlee J

AU - Cerruti, Loretta

AU - Collinge, Janelle E

AU - McLean, Catriona

AU - Wiley, James S

AU - Kile, Ben T

AU - Jane, Stephen M

AU - Curtis, David John

PY - 2015

Y1 - 2015

N2 - 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 semidominant 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 Kcc1M935K 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.

AB - 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 semidominant 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 Kcc1M935K 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.

UR - http://www.bloodjournal.org/content/126/26/2863?sso-checked=true

U2 - 10.1182/blood-2014-10-609362

DO - 10.1182/blood-2014-10-609362

M3 - Article

VL - 126

SP - 2863

EP - 2870

JO - Blood

JF - Blood

SN - 0006-4971

IS - 26

ER -