Project 1 Cowley, Jr., Allen Wilson INDIVIDUAL PROJECTS: PROJECT 1 MOLECULAR &FUNCTIONAL REGULATORY NETWORK IN HYPERTENSION Project Leader: Allen W. Cowley, Jr., Ph.D., Professor of Physiology Co-lnvestigators: Mingyu Liang, M.B., Ph.D., Associate Professor of Physiology Howard Jacob, Ph.D., Professor of Physiology Andrew Greene, Ph.D., Professor of Physiology ABSTRACT Essential hypertension affects more than 50 million Americans and increased blood pressure salt-sensitivity is a prominent feature in certain populations of hypertensive patients, especially African Americans. Although it is evident that the common forms of hypertension are multiifactorial (polygenic and environmental), the genetic basis of the frequent forms of this disease remain elusive. The Dahl salt-sensitive (SS) rat is a widely used animal model that recapitulates many aspects of human salt-sensitive hypertension and associated renal injury. The goal of Proiect 1 is to study the interplay of gene and protein expression using integrative systems approaches to determine the functionality of a single cell type of the kidney (renal medullary thick ascending limb of Henle = mTAL). The mTAL is known to play an important role in renal medullary function, sodium excretion, and hypertension in the SS rat and in human hypertension. We hypothesize that gene(s) sequence variants within a congenic region of chromosome 13 (SS.IS^*^^(R);BN alleles substituted for SS) alter molecular regulatory networks affecting function of the mTAL thereby contributing to salt-sensitive hypertension and renal injury.
In Aim 1, the congenic region (SS.IS^'^^(R);13.2 Mb) will be sequenced to obtain a finished high-quality assembly of this region and annotated to search for gene sequence variants. Subcongenic strains will be developed to identify a narrow region that attenuates salt-sensitivity to ~ 5 Mb and a sequence capture chip will then be utilized for follow-up sequencing of this region in 14 additional strains that have a different evolutionary history from the SS and BN. The sequence differences (SNPs and other genomic variants) will be filtered using the criteria that a casual variant in the SS should only be in common with other salt-sensitive strains.
Aim 2 will construct a molecular and physiological regulatory network (BayeN) of the mTAL epithelial cell and use that model to identify pathways and genes that may contribute to salt-sensitive hypertension and renal injury in SS rats.
Aim 3 will select a candidate gene based on results from Aims 1 and 2 and either knock the gene out using zinc finger nucleases (ZFN) and/or over-express the gene using transgenic approaches to study the impact of this gene upon the transcriptome/proteome/metabolome and associated functional network. Several technological leaps and conceptual approaches are unique to this Project. These include 1) Next generation sequencing to provide very high resolution sequencing of a 13.2 Mb region of SS and congenic SS.13^'^" strains coupled with a NimbleGen sequence capture chip for sequencing of additional rat strains;2) Transcriptome analysis (Affymetrix) and a mass spectrometry sub-proteome and metabolome analysis using isolated and purified mTAL cells comparing the SS and salt-resistant congenic rats;3) The use of a large-scale Bayesian analysis which is "knowledge-constrained" to seek yet unknown pathways hidden using the transcriptome and functional data;4) A novel technology (zinc finger nucleases) to efficiently knock out a gene found to contribute importantly to the mTAL regulatory network, providing a definitive way to validate and characterize functional relevance. Project 1 addresses a critical challenge facing the field of hypertension: to identify the complex components (genes, proteins and pathways) that are responsible for alterations in kidney function leading to salt-sensitive hypertension. It is highly synergistic with Project 2 which tests the innovative concept that non-protein-coding genes (microRNA) may play an important role in hypertension and renal injury and Project 3 which aims to Identify the mechanisms by which mutations distant from the renin gene on chr 13 regulate the activity of renin and angiogenesis in the SS rat. All projects benefit greatly from genomic, genetic, proteomic and computational technological advances provided by Cores 8 and C. 75 Program Director/Principal Investigator (Last, First, Middle);Cowley, Jr., Allen WllSOn Project 1 DETAILED BUDGET FOR INITIAL BUDGET PERIOD FROM THROUGH DIRECT COSTS ONLY 07/01/11 06/30/12 List

Agency
National Institute of Health (NIH)
Type
Research Program Projects (P01)
Project #
5P01HL082798-09
Application #
8691975
Study Section
Heart, Lung, and Blood Program Project Review Committee (HLBP)
Project Start
Project End
Budget Start
Budget End
Support Year
9
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Medical College of Wisconsin
Department
Type
DUNS #
City
Milwaukee
State
WI
Country
United States
Zip Code
Evans, Louise C; Ryan, Robert P; Broadway, Elizabeth et al. (2015) Null mutation of the nicotinamide adenine dinucleotide phosphate-oxidase subunit p67phox protects the Dahl-S rat from salt-induced reductions in medullary blood flow and glomerular filtration rate. Hypertension 65:561-8
Geurts, Aron M; Mattson, David L; Liu, Pengyuan et al. (2015) Maternal diet during gestation and lactation modifies the severity of salt-induced hypertension and renal injury in Dahl salt-sensitive rats. Hypertension 65:447-55
He, Xiaofeng; Liu, Yong; Usa, Kristie et al. (2014) Ultrastructure of mitochondria and the endoplasmic reticulum in renal tubules of Dahl salt-sensitive rats. Am J Physiol Renal Physiol 306:F1190-7
Cowley Jr, Allen W; Moreno, Carol; Jacob, Howard J et al. (2014) Characterization of biological pathways associated with a 1.37 Mbp genomic region protective of hypertension in Dahl S rats. Physiol Genomics 46:398-410
Prisco, Anthony R; Bukowy, John D; Hoffmann, Brian R et al. (2014) Automated quantification reveals hyperglycemia inhibits endothelial angiogenic function. PLoS One 9:e94599
Karcher, Jamie R; Greene, Andrew S (2014) Bone marrow mononuclear cell angiogenic competency is suppressed by a high-salt diet. Am J Physiol Cell Physiol 306:C123-31
Liu, Yong; Liu, Pengyuan; Yang, Chun et al. (2014) Base-resolution maps of 5-methylcytosine and 5-hydroxymethylcytosine in Dahl S rats: effect of salt and genomic sequence. Hypertension 63:827-38
Xu, Xialian; Kriegel, Alison J; Jiao, Xiaoyan et al. (2014) miR-21 in ischemia/reperfusion injury: a double-edged sword? Physiol Genomics 46:789-97
Yang, Chun; Stingo, Francesco C; Ahn, Kwang Woo et al. (2013) Increased proliferative cells in the medullary thick ascending limb of the loop of Henle in the Dahl salt-sensitive rat. Hypertension 61:208-15
Jia, Ping; Teng, Jie; Zou, Jianzhou et al. (2013) miR-21 contributes to xenon-conferred amelioration of renal ischemia-reperfusion injury in mice. Anesthesiology 119:621-30

Showing the most recent 10 out of 54 publications