This project will examine how calcineurin inhibitors, such as tacrolimus and cyclosporine, cause hypertension with potassium retention. In preliminary published studies, we showed that tacrolimus activates the thiazide-sensitive Na- Cl cotransporter (NCC) in the kidney. This effect was found to be essential for the resulting hypertension and potassium retention. Here we will examine the mechanisms involved. Calcineurin is a protein phosphatase and we will test whether it removes phosphates from NCC directly, to inhibit its actions, or whether the pathway involves calcineurin acting on intermediar proteins. We will determine whether other protein phosphatases, including protein phosphatases 1A, 2A, and 4 remove phosphates from NCC. Protein phosphatase inhibitor 1 (PPP1r1a) is a canonical target of calcineurin and is highly expressed along the distal nephron. In preliminary experiments by our collaborator, genetic deletion of this inhibitor was found to reduce the abundance of phosphorylated NCC in vivo. Therefore, we will test whether calcineurin regulates NCC activity via PPP1r1a. PPP1r1a is known to inhibit the actions of protein phosphatase 1A, which can act on the kinase SPAK. SPAK is the canonical NCC activating protein. Thus, we will test the over arching hypothesis that calcineurin inhibitors activate PPP1r1a in the distal nephron, leading to inhibition of PP1A, SPAK activation, and enhanced NCC activity. This model will be tested both in vitro, using a novel cell system that we developed recently, and in vivo, by knocking out key components of the signaling pathway. The proposal has substantial clinical implications, both in terms of patient treatment today, and in terms of drug development for tomorrow.

Public Health Relevance

Calcineurin inhibitors are immunosuppressive that commonly cause hypertension and kidney failure. We recently showed that these drugs cause hypertension, in large part, by activating kidney salt transport. Here, we will determine the mechanism by which salt transport is activated to guide treatment and the future drug development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK095841-02
Application #
8549218
Study Section
Special Emphasis Panel (KMBD)
Program Officer
Ketchum, Christian J
Project Start
2012-09-21
Project End
2016-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
2
Fiscal Year
2013
Total Cost
$356,830
Indirect Cost
$115,580
Name
Oregon Health and Science University
Department
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Cornelius, Ryan J; Zhang, Chong; Erspamer, Kayla J et al. (2018) Dual gain and loss of cullin 3 function mediates familial hyperkalemic hypertension. Am J Physiol Renal Physiol 315:F1006-F1018
Blankenstein, K I; Borschewski, A; Labes, R et al. (2017) Calcineurin inhibitor cyclosporine A activates renal Na-K-Cl cotransporters via local and systemic mechanisms. Am J Physiol Renal Physiol 312:F489-F501
Lazelle, Rebecca A; McCully, Belinda H; Terker, Andrew S et al. (2016) Renal Deletion of 12 kDa FK506-Binding Protein Attenuates Tacrolimus-Induced Hypertension. J Am Soc Nephrol 27:1456-64
Borschewski, Aljona; Himmerkus, Nina; Boldt, Christin et al. (2016) Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na?-K?-2Cl? Cotransporter. J Am Soc Nephrol 27:107-19
McCormick, James A; Ellison, David H (2015) Distal convoluted tubule. Compr Physiol 5:45-98
Subramanya, Arohan R; Ellison, David H (2014) Distal convoluted tubule. Clin J Am Soc Nephrol 9:2147-63
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
Picard, Nicolas; Trompf, Katja; Yang, Chao-Ling et al. (2014) Protein phosphatase 1 inhibitor-1 deficiency reduces phosphorylation of renal NaCl cotransporter and causes arterial hypotension. J Am Soc Nephrol 25:511-22
Ellison, David H (2013) Ubiquitylation and the pathogenesis of hypertension. J Clin Invest 123:546-8
Hoorn, Ewout J; Walsh, Stephen B; McCormick, James A et al. (2012) Pathogenesis of calcineurin inhibitor-induced hypertension. J Nephrol 25:269-75

Showing the most recent 10 out of 11 publications