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

• 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
• 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

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.

项目摘要 胎儿生长受限（FGR）会增加死胎和新生儿死亡的风险。被认为是 出生的FGR远远超过围产期，增加了心血管疾病的风险，除其他外， 在以后的生活中。血管对妊娠的适应性受损是FGR的主要原因。缺氧条件 也改变子宫胎盘血管功能，减少胎儿生长，类似于FGR。了解 子宫动脉（UtA）血流增加可防止缺氧依赖性受损的机制 子宫血管和胎儿生长减少，我们建议使用光遗传学和化学遗传学技术， 在活体动物中控制子宫血流。光遗传学是一种创新技术， 修饰的细胞表达光激活的微生物视蛋白（例如，[001 pdf 1st-31 files]盐视紫红质（Halorhodopsin，NpHR） 选择性地在体内受到光的刺激。化学遗传学利用修饰的受体，如毒蕈碱M3 受体（hM 3 Dq），可以被特定激动剂（例如，去氯氯氮平[DCZ]）。我们 目的是开发新的和可靠的小鼠模型，以防止FGR通过内皮或平滑肌特异性 在UTA的机制。为了实现这一目标，我们提出了两个科学目标。在Aim 1中，小鼠中 在平滑肌中表达NpHR，我们将讨论光刺激血管舒张UtA的能力， 在体内，增加UtA血流量并防止FGR。目的2将确定内皮依赖性 UtA血管舒张通过hM 3Dq的表达及其通过DCZ局部应用的选择性激活， 微流体通道目前用于增加体内UtA血流的鼠模型需要药理学 非组织特异性的激活，并且激活的时间不能被控制。我们的光遗传学 和化学发生模型将提供对血流操纵程度的更好控制， 实验之间的可重复性，刺激的选择性提高，以及测试时间的机会 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， vivo.重要的是，我们的建议可以为将光遗传学和化学遗传学应用于肿瘤提供基础。 体内其他血管床的研究。