The research objective of this application is to develop a detailed molecular and mechanistic understanding of the heterogeneity in the rheological and adherence properties of circulating sickle red cells and define the in vivo pathologic sequelae of these cellular changes. To achieve our objective we propose the following series of studies: 1) Determine the structural basis for the heterogeneous rheological properties of distinct subpopulations of circulating sickle red cells and study the in vivo flow behavior and survival of these subpopulations. 2) Determine the contributions of the K-CI cotransporter and the Gardos channel to the generation of theologically compromised dense sickle cells. 3) Evaluate the contributions of specific adhesive receptors, ligands, and plasma proteins in mediating adhesion of sickle red cells to endothelial cells in vitro. 4) Validate the contribution of various adhesive ligands and receptors to sickle cell adhesion by performing adhesion studies using cells from sickle mice in which genes encoding specific adhesive ligands or receptors are inactivated. For the proposed studies, we will use multiple techniques in biophysics, molecular biology, cell biology, mouse genetics and circulatory physiology. The sickle mice that we developed expressing exclusively human sickle hemoglobin and exhibiting several clinical features of human disease are a key component of the proposed experimental strategy. Our application of novel experimental strategies and exploration of innovative biological concepts offers promise for elucidating the causes of painful vaso-occlusion and chronic organ damage. We anticipate that our findings will lead to the identification of useful therapeutic strategies for the effective clinical management of this debilitating human disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL031579-30
Application #
7858332
Study Section
Erythrocyte and Leukocyte Biology Study Section (ELB)
Program Officer
Goldsmith, Jonathan C
Project Start
1989-10-01
Project End
2012-05-31
Budget Start
2010-06-01
Budget End
2012-05-31
Support Year
30
Fiscal Year
2010
Total Cost
$355,022
Indirect Cost
Name
New York Blood Center
Department
Type
DUNS #
073271827
City
New York
State
NY
Country
United States
Zip Code
10065
Koshino, Ichiro; Mohandas, Narla; Takakuwa, Yuichi (2012) Identification of a novel role for dematin in regulating red cell membrane function by modulating spectrin-actin interaction. J Biol Chem 287:35244-50
Safeukui, Innocent; Buffet, Pierre A; Deplaine, Guillaume et al. (2012) Quantitative assessment of sensing and sequestration of spherocytic erythrocytes by the human spleen. Blood 120:424-30
Mohandas, Narla; An, Xiuli (2012) Malaria and human red blood cells. Med Microbiol Immunol 201:593-8
Liu, Jing; Mohandas, Narla; An, Xiuli (2011) Membrane assembly during erythropoiesis. Curr Opin Hematol 18:133-8
Chen, Lixiang; Hughes, Richard A; Baines, Anthony J et al. (2011) Protein 4.1R regulates cell adhesion, spreading, migration and motility of mouse keratinocytes by modulating surface expression of beta1 integrin. J Cell Sci 124:2478-87
An, Xiuli; Guo, Xinhua; Yang, Yang et al. (2011) Inter-subunit interactions in erythroid and non-erythroid spectrins. Biochim Biophys Acta 1814:420-7
Yang, Shaomin; Weng, Haibo; Chen, Lixiang et al. (2011) Lack of protein 4.1G causes altered expression and localization of the cell adhesion molecule nectin-like 4 in testis and can cause male infertility. Mol Cell Biol 31:2276-86
Gauthier, Emilie; Guo, Xinhua; Mohandas, Narla et al. (2011) Phosphorylation-dependent perturbations of the 4.1R-associated multiprotein complex of the erythrocyte membrane. Biochemistry 50:4561-7
Blanc, Lionel; Salomao, Marcela; Guo, Xinhua et al. (2010) Control of erythrocyte membrane-skeletal cohesion by the spectrin-membrane linkage. Biochemistry 49:4516-23
Manno, Sumie; Mohandas, Narla; Takakuwa, Yuichi (2010) ATP-dependent mechanism protects spectrin against glycation in human erythrocytes. J Biol Chem 285:33923-9

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