Sickle cell disease and thalassemia are characterized by pathological red cell dehydration. The mean corpuscular hemoglobin concentration of sickle erythrocytes critically determines the lag time preceding the rapid phase of deoxygenation-induced polymerization of hemoglobin S. Several erythroid ion transporters and channels are believed on the basis of pharmacological and physiological studies to regulate red cell hemoglobin concentration secondary to regulation of red cell volume. Among these activities already studied as therapeutic targets in sickle cell disease are the KCNN/IK1/SK4 Ca2+-activated K+ channel of intermediate conductance (Gardos channel), several types of KCC K-CI cotransporters, at least two types of erythroid CI- conductance, and at least one type of Ca2+permeable cation conductance. The K-CI cotransporters have also been tested as therapeutic targets for thalassemia. The genes encoding these ion-transporting polypeptides are strong candidate risk modifier genes for the hemoglobinopathies. This application proposes the general hypothesis that genetic modulation of these transporter and channel activities will modulate disease severity in mouse models of hemoglobinopathies. This general hypothesis will be tested by experiments designed to pursue the following Specific Aims:
Aim 1. We will test the hypothesis that genetic deficiency of the erythroid IK1/Gardos channel will decrease pathologic red cell dehydration and will ameliorate clinical severity in mouse models of sickle cell disease.
Aim 2. We will test the hypothesis that genetic deficiency of erythroid KCC K-CI cotransporters will decrease pathologic red cell dehydration and will ameliorate clinical severity in mouse models of sickle cell disease and of beta-thalassemia intermedia.
Aim 3. We will test the hypothesis that combined genetic deficiency of erythroid IK1/Gardos channel and of erythroid KCC K-CI cotransporters will further ameliorate clinical severity in mouse models of sickle cell disease. The proposed experiments will increase understanding of sickle cell disease and thalassemia by providing mouse models for genetic tests of new drug therapies under development for near-term clinical testing. ? ?

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL077765-02
Application #
7245127
Study Section
Erythrocyte and Leukocyte Biology Study Section (ELB)
Program Officer
Evans, Gregory
Project Start
2006-06-15
Project End
2010-05-31
Budget Start
2007-06-01
Budget End
2008-05-31
Support Year
2
Fiscal Year
2007
Total Cost
$390,245
Indirect Cost
Name
Beth Israel Deaconess Medical Center
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02215
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Shmukler, Boris E; Hsu, Ann; Alves, Jessica et al. (2013) N-ethylmaleimide activates a Cl(-)-independent component of K(+) flux in mouse erythrocytes. Blood Cells Mol Dis 51:9-16
Huang, Junwei; Shan, Jiajie; Kim, Dusik et al. (2012) Basolateral chloride loading by the anion exchanger type 2: role in fluid secretion by the human airway epithelial cell line Calu-3. J Physiol 590:5299-316
Shmukler, Boris E; Rivera, Alicia; Vandorpe, David H et al. (2012) Cation-leak stomatocytosis in standard schnauzers does not cosegregate with coding mutations in the RhAG, SLC4A1, or GLUT1 genes associated with human disease. Blood Cells Mol Dis 48:219-25
Stewart, Andrew K; Shmukler, Boris E; Vandorpe, David H et al. (2011) Loss-of-function and gain-of-function phenotypes of stomatocytosis mutant RhAG F65S. Am J Physiol Cell Physiol 301:C1325-43
Stewart, Andrew K; Kedar, Prabhakar S; Shmukler, Boris E et al. (2011) Functional characterization and modified rescue of novel AE1 mutation R730C associated with overhydrated cation leak stomatocytosis. Am J Physiol Cell Physiol 300:C1034-46
Reimold, Fabian R; Heneghan, John F; Stewart, Andrew K et al. (2011) Pendrin function and regulation in Xenopus oocytes. Cell Physiol Biochem 28:435-50
Stewart, Andrew K; Chebib, Fouad T; Akbar, Syed W et al. (2011) Interactions of mouse glycophorin A with the dRTA-related mutant G719D of the mouse Cl-/HCO3- exchanger Ae1. Biochem Cell Biol 89:224-35
Telio, David; Bailey, Denis; Chen, Christine et al. (2010) Two distinct syndromes of lymphoma-associated AL amyloidosis: a case series and review of the literature. Am J Hematol 85:805-8
Vandorpe, David H; Xu, Chang; Shmukler, Boris E et al. (2010) Hypoxia activates a Ca2+-permeable cation conductance sensitive to carbon monoxide and to GsMTx-4 in human and mouse sickle erythrocytes. PLoS One 5:e8732

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