Optogenetic and chemogenetic regulation of uterine vascular function

子宫血管功能的光遗传学和化学遗传学调控

基本信息

批准号：
10785667
负责人：
Ramon Lorca
金额：
$ 42.9万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-19 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10785667
关键词：
Acceleration Acute Address Adverse effects Affect Agonist Altitude Animal Genetics Animal Model Animals Area Arteries Blood Vessels Blood flow Bluetooth Cardiovascular Diseases Cardiovascular system Cell membrane Cervical Child Chronic Disease Contralateral Development Drug Design Elderly Endothelial Cells Endothelium Exposure to Fetal Growth Fetal Growth Retardation Fetal Reduction Fetus Fiber Foundations G alpha q Protein G-Protein-Coupled Receptors Genetic Genetic Models Genotype Goals Growth Halorhodopsins Human Hypertension Hypoxia Impairment Implant Invaded Knowledge Life Light Longevity Measures Microfluidics Molecular Morphology Mothers Mus Muscarinic Acetylcholine Receptor Muscarinic M3 Receptor Neonatal Mortality Nitric Oxide Opsin Optics Pericytes Physiological Placenta Population Positioning Attribute Predisposition Pregnancy Pregnancy Complications Pregnancy Outcome Pregnant Uterus Proliferating Pump Regulation Relaxation Reproducibility Reproductive Health Risk Signaling Protein Smooth Muscle Smooth Muscle Myocytes Stress Techniques Testing Therapeutic Time Uterus Vagus nerve structure Vasodilation Wild Type Mouse cardiovascular disorder risk design experimental study fetal fetal blood flexibility genetic technology genetically modified cells healthy pregnancy implantation improved in vivo infant morbidity/mortality innovation internal control microbial mouse model neonatal death novel novel therapeutics optogenetics perinatal period pharmacologic pregnant prevent receptor skull implant stillbirth subcutaneous therapeutically effective therapy development tool trophoblast vascular bed

项目摘要

PROJECT SUMMARY Fetal growth restriction (FGR) increases the risk of stillbirth and neonatal death. The adverse effects of being born FGR extend well beyond the perinatal period, increasing the risk of cardiovascular disease, among others, in later life. Impaired vascular adaptation to pregnancy is a predominant contributor to FGR. Hypoxic conditions also alter uteroplacental vascular function, reducing fetal growth, similarly to FGR. To understand the mechanisms by which an increase in uterine artery (UtA) blood flow can prevent hypoxia-dependent impaired uterine vasculature and reduced fetal growth, we propose using optogenetics and chemogenetics technology for manipulating uterine blood flow in live animals. Optogenetics is an innovative technique in which genetically modified cells express light-activated microbial opsins (e.g., halorhodopsin [NpHR]), which can then be selectively stimulated by light in vivo. Chemogenetics utilizes modified receptors such as the muscarinic M3 receptor (hM3Dq), which can be selectively activated by specific agonists (e.g., deschloroclozapine [DCZ]). Our goal is to develop novel and reliable murine models to prevent FGR via endothelium or smooth muscle-specific mechanisms in the UtA. To address this goal, we propose to conduct two scientific aims. In Aim 1, in mice expressing NpHR in the smooth muscle, we will address the capacity for light stimulation to vasodilate the UtA in vivo, increasing UtA blood flow and preventing FGR. Aim 2 will determine the effect of endothelium-dependent UtA vasodilation via expression of hM3Dq and its selective activation by DCZ locally applied through a microfluidic channel. Current murine models for increasing UtA blood flow in vivo require pharmacological activations that are non-tissue specific, and the timing of the activation cannot be controlled. Our optogenetic and chemogenetic models will provide better control of the degree of blood flow manipulation, enhanced reproducibility among experiments, improved selectivity of the stimulation, and the opportunity to test the timing of stimulation. This project will be the first to apply optogenetics and chemogenetics to the vasodilation of UtA in vivo. Importantly, our proposal can provide the foundation for applying optogenetics and chemogenetics to the study of other vascular beds in vivo.

项目概要胎儿生长受限（FGR）会增加死产和新生儿死亡的风险。存在的不利影响出生的 FGR 远远超出围产期，增加了心血管疾病等疾病的风险，在以后的生活中。血管对妊娠的适应受损是 FGR 的主要原因。缺氧条件与 FGR 类似，也会改变子宫胎盘血管功能，减少胎儿生长。要了解增加子宫动脉（UtA）血流量可预防缺氧依赖性受损的机制子宫脉管系统和胎儿生长减少，我们建议使用光遗传学和化学遗传学技术操纵活体动物的子宫血流。光遗传学是一种创新技术，其中基因修饰的细胞表达光激活的微生物视蛋白（例如盐视紫红质 [NpHR]），然后可以将其在体内受光选择性刺激。化学遗传学利用修饰受体，例如毒蕈碱 M3 受体 (hM3Dq)，可以被特定激动剂（例如去氯氯氮平 [DCZ]）选择性激活。我们的目标是开发新颖且可靠的小鼠模型，通过内皮或平滑肌特异性来预防 FGR UtA 中的机制。为了实现这一目标，我们建议实现两个科学目标。在目标 1 中，在小鼠中在平滑肌中表达 NpHR，我们将解决光刺激使 UtA 血管舒张的能力在体内，增加 UtA 血流量并预防 FGR。目标 2 将确定内皮依赖性的效果 UtA 血管舒张通过 hM3Dq 的表达及其通过 DCZ 局部应用的选择性激活来实现。微流体通道。目前增加体内 UtA 血流量的小鼠模型需要药理学非组织特异性的激活，并且激活的时间无法控制。我们的光遗传学化学遗传学模型将更好地控制血流操纵程度，增强实验之间的可重复性、改进的刺激选择性以及测试时机的机会的刺激。该项目将是第一个将光遗传学和化学遗传学应用于 UtA 血管舒张的项目体内。重要的是，我们的建议可以为应用光遗传学和化学遗传学提供基础体内其他血管床的研究。