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The urgent need for translational research in each of these three focus areas will be addressed by the three Primary Research Projects as well as by the three Research Cores, all proposed by outstanding funded investigators with a track record of successful multidisciplinary collaborative efforts in the focus areas.
The major goal is to use point of care assessment of plasma neutrophil gelatinase associated lipocalin to direct randomization of children to fenoldopam vs. placebo after cardiopulmonary bypass to prevent cardiac surgery associated acute kidney injury. This will be a first in kind study to use a well validated AKI biomarker to enrich the population for study to those who have the highest and earliest signs of risk.
Aim 1. We propose a RNA-Seq/microarray dissection of pathogenic pathways in each major cell type of the glomerulus in Actn4 mutant mice. We propose to use transgenic-GFP mice that allow the FACS purification of podocytes, endothelial cells and mesangial cells from glomeruli of Actn4 mutant mice. In addition we will use laser capture microdissection to isolate their proximal tubules. We will then perform global gene expression profiling of each individual cellular compartment as a function of developmental progression of the disease using a combination of microarrays and RNA-Seq.
Aim 2. We propose a similar molecular dissection of the altered gene expression programs of each glomerular cell type in a bigenic mouse model of FSGS, namely the Cd2ap/Fyn mouse. We propose to purify each cellular component of the glomeruli of these bigenic mutants, as well as proximal tubules, and to globally define their perturbed gene expression patterns.
Aim 3. We propose a proteomics analysis of the glomeruli of the FSGS model Actn4 and Cd2ap/Fyn mutant mice. We will use sieve purified glomeruli, coupled with 2-D gels, MALDI-TOF/TOF analysis, as well as isotope tagging based quantitative mass spectrometry.
Aim 1. To prospectively validate the lupus nephritis (LN) Biomarker Panel as non-invasive markers of histologic activity in children with biopsy-proven LN.
Aim 2. To characterize the distribution of these biomarkers in kidney biopsy tissues, and assess the histologic association between biomarker expression and kidney architecture.
Aim 3. To prospectively test the LN Biomarker Panel as non-invasive markers of response to therapy in children with LN.
Aim 4. To use advanced proteomic techniques for discovering additional LN biomarkers that are synergistic with the LN Panel in measuring LN activity and LN damage.
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