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Principal Investigator: Lauren Elizabeth Parker
Organization: DUKE UNIVERSITY
Fiscal Year: 2024
Award: $53,974
Funding agency: National Heart Lung and Blood Institute
Abstract
Heart failure is a leading cause of death worldwide and there is a critical, unmet need for non-surgical
therapies. A key barrier to developing curative therapy is that the human heart, unlike that of zebrafish,
axolotls, and some neonatal mammals, lacks the ability to recover after acute or chronic injury. Our lab has
discovered that in neonatal mice, which can regenerate the heart after injury, cardiomyocytes and endothelial
cells proliferate in a spatiotemporally coordinated manner. When key cell-cell signaling factors are lost, such as
VEGFA or its receptor VEGFR2, we observe significantly decreased rates of cell cycling in both
cardiomyocytes and endothelial cells. This suggests that local signals between these cells are responsible for
inducing and maintaining a proliferative state in the neonatal period. For this information to be therapeutically
translational, however, we must understand whether these mechanisms are conserved in human cells. We
have devised a cardiac organoid (CO) platform enriched for endothelial cells that can be used to model human
myovascular interactions. With this model, we will use cardiomyocyte and endothelial cell reporters and live
imaging to investigate how modifying endothelial cell content within a human CO impacts organoid function
and regeneration after ischemia-reperfusion injury. In addition, we will use a CRISPRa lentiviral construct to
overexpress key factors secreted by cycling endothelial cells, with cognate receptors on cycling
cardiomyocytes, for effects on cardiomyocyte proliferation. This proposal details the use of a high-throughput
CO platform to disentangle the complexities of myovascular interactions in human cells, enabling discovery
and validation of novel therapeutic targets.
Terms: <21+ years old><7B4><Acceleration><Acute><Adult><Adult Human><Amblystoma><Amblystomas><Ambystoma><Ambystomas><Assay><Axolotl><Bioassay><Biologic Models><Biological Assay><Biological Models><Blood Vessels><Body Tissues><Bone Marrow><Bone Marrow Reticuloendothelial System><Brachydanio rerio><CDH5><CDH5 gene><CRISPR activation><CRISPR activator><CRISPR based activation><CRISPR gene activation><CRISPR transcription activation><CRISPR transcriptional activation><CRISPR-Cas-9-mediated gene activation><CRISPR-based gene activation><CRISPR-dCAS9 Activator><CRISPR-mediated transcriptional activation><CRISPR/CAS9 activation><CRISPR/CAS9 gene activation><CRISPR/dCas9 activation><CRISPR/dCas9-based transcriptional activation><CRISPRa><Cardiac><Cardiac Muscle Cells><Cardiac Myocytes><Cardiac development><Cardiac infarction><Cardiocyte><Cause of Death><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Cycle><Cell Division Cycle><Cell Growth in Number><Cell Interaction><Cell Multiplication><Cell Proliferation><Cell Signaling><Cell to Cell Communication and Signaling><Cell-Cell Signaling><Cell-to-Cell Interaction><Cells><Cellular Proliferation><Chronic><Complex><Coupling><Custom><Danio rerio><Data Set><Development><Disease><Disorder><Endothelial Cells><Endothelium><FLK1><Generalized Growth><Genetic><Genetic Models><Goals><Growth><Growth Agents><Growth Factor><Growth Substances><Guide RNA><Heart><Heart Injuries><Heart Muscle Cells><Heart failure><Heart myocyte><Human><Human Engineering><IGF2><IGF2 gene><Image><Injury><Intestinal><Intestines><Intracellular Communication and Signaling><Ischemia><Ischemia-Reperfusion Injury><KDR gene><LV Mass><Left Ventricular Mass><Length><Liver><Mammalia><Mammals><Mediating><Mendelian randomization><Mice><Mice Mammals><Model System><Modeling><Modern Man><Murine><Mus><Myocardial><Myocardial Infarct><Myocardial Infarction><Natural regeneration><Neonatal><Organism><Organoids><Physiologic><Physiological><Proliferating><Proteins Growth Factors><Protocol><Protocols documentation><Receptor Protein><Recovery><Regeneration><Reperfusion Damage><Reperfusion Injury><Reperfusion Therapy><Reporter><Reproducibility><Role><Signal Transduction><Signal Transduction Systems><Signaling><Testing><Tissue Growth><Tissues><United States><VEGF><VEGF Receptors><VEGFA><VEGFA gene><VEGFR><VEGFR-2><VEGFR2><VEGFs><VPF Receptor><Validation><Vascular Endothelial Cell Growth Factor Receptor><Vascular Endothelial Growth Factor A><Vascular Endothelial Growth Factor Receptor 2><Vascular Endothelial Growth Factors><Vascular Permeability Factor Receptor><Vasculotropin><Work><Zebra Danio><Zebra Fish><Zebrafish><activating CRISPR technology><adulthood><biological signal transduction><bone><bowel><cardiac failure><cardiac infarct><cardiac injury><cardiac regeneration><cardiogenesis><cardiomyocyte><combinatorial><coronary attack><coronary infarct><coronary infarction><crypt cell><curative intervention><curative therapeutic><curative therapy><curative treatments><customs><developmental><experiment><experimental research><experimental study><experiments><gRNA><heart attack><heart development><heart formation><heart infarct><heart infarction><heart regeneration><hepatic body system><hepatic organ system><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><human model><human subject><human tissue><imaging><induced human pluripotent stem cells><injuries><injury recovery><injury response><intercellular communication><intestinal crypt><live cell image><live cell imaging><live cellular image><live cellular imaging><living system><model of human><neonatal mice><neonatal period><new drug target><new drug treatments><new druggable target><new drugs><new pharmacological therapeutic><new pharmacotherapy target><new therapeutic target><new therapeutics><new therapy><new therapy target><next generation therapeutics><novel drug target><novel drug treatments><novel druggable target><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel pharmacotherapy target><novel therapeutic target><novel therapeutics><novel therapy><novel therapy target><ontogeny><overexpress><overexpression><progenitor cell niche><progenitor niche><receptor><recovery after injury><recovery following injury><recovery post injury><regenerate><regenerate new tissue><regenerate tissue><regenerating damaged tissue><regenerating tissue><regenerative><reperfusion><response to injury><scRNA-seq><self assembly><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><spatiotemporal><stem and progenitor cell niche><stem cell niche><tissue regeneration><tissue regrowth><tissue renewal><tissue specific regeneration><translational therapeutics><translational therapy><validations><vascular>