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Potassium channels, membrane potential, and CHD

钾通道、膜电位和 CHD

基本信息

批准号：
10439505
负责人：
Mustafa K Khokha
金额：
$ 55.37万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10439505
关键词：
Affect Biochemical Calcium Calcium Channel Calcium Signaling Candidate Disease Gene Cardiac Cardiac development Cell membrane Cell model Cells Chemicals Child Child Health Closure by clamp Collaborations Congenital Abnormality Data Defect Dependence Development Disease Ectoderm Ectoderm Cell Electrophysiology (science)Embryo Europe Exhibits Family member Fetus Fluorescence Gap Junctions Gastrula Genes Genetic Genetic Transcription Genomic approach Genomics Germ Cells Germ Layers Heart Image Individual Infant Mortality Ion Channel Kinetics Lead Left Ligands Measurement Measures Membrane Membrane Potentials Mesoderm Mesoderm Cell Methods Modeling Molecular Monitor Morbidity - disease rate Neurons Optics Paraxial Mesoderm Pathogenesis Pathway interactions Patients Pattern Phenotype Play Potassium Potassium Channel Property Reading Regulation Role Series Signal Pathway Signal Transduction Situs Inversus Structure Testing Transducers Voltage-Gated Potassium Channel Work Xenopus base blastocyst blastomere structure cardiogenesis congenital heart disorder electrical property embryo cell exome sequencing experimental study gastrulation genetic analysis genetic manipulation heart function infant death inhibitor intercellular communication mortality nodal myocyte pluripotency receptor structural heart disease transcription factor transcriptome sequencing voltage voltage clamp

项目摘要

Project Summary Congenital heart disease (CHD) leads to severe morbidity and mortality to children in the US and worldwide. Despite this impact on child health, we simply do not understand the genetic causes of CHD. Recently, trio based whole exome sequencing has identified a class of voltage-gated potassium channels (multiple KCNH family members) as candidates for CHD and, specifically heterotaxy, a disorder of left-right (LR) patterning that has a severe effect on cardiac function. However, a molecular role connecting potassium channels to structural heart disease and heterotaxy is unprecedented. We propose, and our preliminary data support, that KCNH6 defines a new paradigm for cell signaling in early embryonic cells. Our data support an electrophysiological model where specific germ layers fates (paraxial mesoderm and ectoderm) are dependent on an ion channel network. Our overarching hypothesis is that K+ channels define electrical membrane potential and regulate voltage gated Ca2+ channels that establish an exit from pluripotency towards specific cell fates, gastrulation, and LR patterning providing a plausible mechanism for our patients with Htx and CHD. Our electrophysiological pathway then integrates with biochemical signaling pathways that define specific cell fates in the embryo. In this proposal revision, we will focus on KCNH6 to see if gene depletion leads to LR patterning defects in Xenopus. In addition, we will test where in the LR patterning cascade, KCNH6 plays a role. Then, using a series of judiciously chosen chemical and ionic perturbations, we will test if membrane potential is indeed essential for pluripotency, cell fate, and calcium regulation. Due to the novelty of this project, we will also perform unbiased genomics (RNAseq) for discovery of transcriptional targets of  Vm. Finally, we will measure electrical properties electrophysiologically using both whole-cell voltage clamp and intracellular recordings and determine the various currents that define membrane potential in early germ cells. A major strength of our proposal is our expertise; we have forged a collaboration between Xenopus developmental biologists and electrophysiologists that will allow us to rigorously investigate membrane potential as an embryonic patterning mechanism.

项目总结