Hypertension affects 60 million adults in the United States, and is a major independent risk factor for stroke, myocardial infarction, and congestive heart failure, and a major cause of end-stage renal disease. More than half of hypertensive patients have a salt-sensitive component, and in approximately 30%, hypertension is predominantly due to abnormalities in sodium handling by the kidney. The kinases SPAK and OSR1 play important roles in the regulation of renal sodium transport. In vitro, both kinases activate the sodium transporters NKCC2 and NCC, but little is known about the pathways via which SPAK and OSR1 activate them in whole animals. Studies in mice indicate that OSR1 is more important for activation of NKCC2, while SPAK is the main activator of NCC. Recent evidence shows that multiple forms of SPAK and OSR1 exist in the kidney, some of which inhibit sodium transport, contrary to the prevailing view that SPAK and OSR1 only activate it. Stimuli that lower blood pressure (e.g. dietary salt restriction), or administration of hormones that increase blood pressure (e.g. aldosterone, vasopressin and angiotensin II), reduce the levels of inhibitory SPAK and OSR1, but increase levels of activating forms. Regulation of renal sodium transport and blood pressure by SPAK and OSR1 is thus more complex than previously believed. The objective of this proposal is to determine the mechanisms by which SPAK and OSR1 regulate renal sodium transport. The R01 Grant will provide the necessary resources for the principal investigator to test the hypothesis that SPAK and OSR1 isoforms differentially regulate renal sodium transport, and differentially mediate responses to physiological stimuli that alter blood pressure. To test this hypothesis, three specific aims are proposed.
Aim 1 is to examine the mechanisms by which SPAK and OSR1 isoforms differentially regulate renal sodium transport. The mechanisms by which SPAK and OSR1 isoforms inhibit activity of NKCC2 will be determined using Xenopus oocytes and mammalian cells.
Aim 2 is to identify the physiological factors that modulate isoform expression. Using wild-type, SPAK knockout and renal OSR1 knockout mice, the effects of sodium restriction, aldosterone infusion and induced-hypertension on the levels and renal localization of SPAK and OSR1 isoforms, as well as the mechanism and timing of these effects, will be determined.
Aim 3 is to examine how SPAK and OSR1 regulate renal sodium transport and blood pressure in whole animals. The effects of acute and chronic aldosterone, vasopressin or angiotensin II administration on NCC and NKCC2 phosphorylation and activity will be determined in wild-type, SPAK knockout and renal OSR1 knockout mice. Mice lacking both SPAK and OSR1 in the kidney will also be characterized. These studies will enable us to assign specific physiological functions to either SPAK or OSR1 isoforms, a significant advance towards understanding how two closely related kinases that activate the same targets in vitro have very different roles in vivo. In addition, we will gain insight into the pathways that activate cation cotransporters independently of SPAK/OSR1.

Public Health Relevance

Hypertension affects 60 million adults in the United States and is a major independent risk factor for stroke, myocardial infarction, and congestive heart failure, and a major cause of end-stage renal disease. The degree of sodium reabsorption by the kidney is the key determinant of blood pressure, but the mechanisms by which sodium reabsorption is regulated are incompletely understood. By studying the mechanisms by which the kidney reabsorbs sodium, we can learn more about the causes of hypertension, and design new therapies to treat the large number of American adults (about 6 million) whose hypertension is currently uncontrollable.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK098141-01A1
Application #
8629140
Study Section
Hypertension and Microcirculation Study Section (HM)
Program Officer
Ketchum, Christian J
Project Start
2014-04-01
Project End
2019-02-28
Budget Start
2014-04-01
Budget End
2015-02-28
Support Year
1
Fiscal Year
2014
Total Cost
$326,963
Indirect Cost
$109,463
Name
Oregon Health and Science University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
McCormick, James A; Yang, Chao-Ling; Zhang, Chong et al. (2014) Hyperkalemic hypertension-associated cullin 3 promotes WNK signaling by degrading KLHL3. J Clin Invest 124:4723-36
Terker, Andrew S; Yang, Chao-Ling; McCormick, James A et al. (2014) Sympathetic stimulation of thiazide-sensitive sodium chloride cotransport in the generation of salt-sensitive hypertension. Hypertension 64:178-84