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Principal Investigator: Lori L O'Brien
Organization: UNIV OF NORTH CAROLINA CHAPEL HILL
Fiscal Year: 2024
Award: $421,067
Funding agency: Eunice Kennedy Shriver National Institute of Child Health and Human Development
Abstract
In utero exposures to insults such as pregestational diabetes can have significant repercussions for
proper growth and development. Exposure to the diabetic milieu can lead to diabetic embryopathy, a disruption
in normal cellular programs during critical stages of development which leads to congenital anomalies affecting
multiple organs. Kidney anomalies resulting from genetic and in utero perturbations are the leading cause of
chronic kidney disease in children, markedly affecting quality of life. However, there is a significant gap in our
understanding of how such insults affect these early developmental programs, precluding the generation of
interventional strategies. The limitations of mouse models and differences between human and mouse kidney
programs necessitate the development of alternate systems in which to study in utero exposures. Kidney
organoids represent an attractive model for their recapitulation of developmental programs in an easily
manipulatable environment. Yet, studies with kidney organoids thus far have almost exclusively focused on
genetic diseases and injury affecting cells of the mature nephron. To this end, we will exploit iPSC-derived
kidney organoids for their ability to recapitulate human developmental processes and test how in utero
exposures to the pregestational diabetic milieu affect kidney development programs. We will focus on
programs of nephron progenitors, the precursor of all the cells in the nephron, and podocytes, a nephron
progenitor descendant critical to establishing and maintaining proper blood filtration. We will mimic the
pregestational diabetic environment in culture and interrogate the effects on nephron progenitor programs by
single cell (sc)RNA-seq and the assay for transposase-accessible chromatin with sequencing (ATAC-seq) as
the organoids progress through this stage of differentiation (Aim 1). We will also assess the effects on maturing
podocytes utilizing similar methodology (Aim 2). We expect to identify alterations to the normal differentiation
programs of these cells and uncover critical pathways affected by the diabetic milieu. These findings will serve
as the foundations for future studies and aid the development of novel interventional strategies then help
mitigate the effects of in utero insults. Our studies will enable new directions for kidney organoid research,
laying the groundwork for future studies into in utero insults and congenital anomalies in this ex vivo,
manipulatable system.
Terms: <0-11 years old><ATAC sequencing><ATAC-seq><ATACseq><Affect><Amalgam><Assay for Transposase-Accessible Chromatin using sequencing><Birth Defects><Body System><Body Tissues><Cell Body><Cells><Child><Child Youth><Children (0-21)><Chromatin><Chronic Kidney Failure><Chronic Renal Disease><Chronic Renal Failure><Congenital Abnormality><Congenital Anatomical Abnormality><Congenital Cardiac Defects><Congenital Defects><Congenital Deformity><Congenital Heart Defects><Congenital Malformation><Critical Paths><Critical Pathways><D-Glucose><DNA Alteration><DNA Sequence Alteration><DNA mutation><Derivation><Derivation procedure><Development><Developmental Process><Dextrose><Diabetes Mellitus><Differentiation in cell culture><Endowment><Environment><Exposure to><Expression Signature><Fetal Kidney><Filtration><Filtration Fractionation><Foundations><Future><Gene Expression Profile><Generations><Genetic><Genetic Diseases><Genetic mutation><Glucose><Goals><Growth and Development><Growth and Development function><Health><Heart><Human><Human Biology><Hyperglycemia><In Vitro><In vitro cell differentiation><Injury><Intervention><Intervention Strategies><Investigation><Kidney><Kidney Diseases><Kidney Urinary System><Methodology><Mice><Mice Mammals><Modeling><Modern Man><Murine><Mus><Nephrons><Nephropathy><Organ><Organ System><Organoids><Physiologic><Physiological><Play><Population><Predisposition><Program Development><QOL><Quality of life><Renal Disease><Research><Risk><Role><Sequence Alteration><Structural defect><Structural malformation><Susceptibility><System><Techniques><Testing><Therapeutic><Therapeutic Intervention><Tissues><Uriniferous Tube><Visceral Epithelial Cell><assay for transposase accessible chromatin followed by sequencing><assay for transposase accessible chromatin seq><assay for transposase accessible chromatin sequencing><assay for transposase-accessible chromatin with sequencing><blood filtration><cell type><chronic kidney disease><congenital anomalies of the kidney><congenital anomaly><congenital kidney abnormalities><congenital renal abnormalities><congenital renal anomalies><develop therapy><developmental><diabetes><diabetic><diabetic embryopathy><differentiation in culture><differentiation in vitro><exposed in utero><fetal><fetal exposure><gene expression pattern><gene expression signature><genetic condition><genetic disorder><genomic alteration><glomerular visceral epithelial cell><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><hyperglycemic><iPS><iPSC><iPSCs><in utero><in utero exposure><in vitro cellular differentiation><in vivo><induced human pluripotent stem cells><induced pluripotent cell><induced pluripotent stem cell><inducible pluripotent stem cell><injuries><insight><intervention development><intervention therapy><interventional strategy><intra-uterine environmental exposure><intrauterine environmental exposure><kidney development><kidney disorder><kids><mortality><mouse model><murine model><nephrogenesis><nephron progenitor><novel><podocyte><prenatal exposure><prenatally exposed><prevent><preventing><progenitor><programs><renal><renal disorder><response><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><structural abnormalities><structural anomalies><therapy development><transcriptional profile><transcriptional signature><treatment development><treatment strategy><youngster>