A bioluminescent-based imaging probe for noninvasive longitudinal monitoring of CoQ10 uptake in vivo

项目摘要 辅酶Q10（CoQ 10）是一种有效的能量传递分子，也是氧化磷酸化的核心辅因子 存在于细胞膜和线粒体中。辅酶Q10也被称为一种强大的抗炎和 抗氧化剂，保护组织免受炎症激活引起的损伤。 信号通路和自由基。它在肝脏、心脏和肾脏中以高水平自然合成。 然而，身体合成自身辅酶Q10的能力随着年龄的增长而显著下降。除了有 在正常的衰老过程中，许多人的辅酶Q10水平显着下降， 病理学因此，辅酶Q10的营养补充提供了一个强大的工具，以维持其足够的 我们的目标是改善人体健康，降低患慢性病的风险。在这一背景下， 多项临床试验揭示了辅酶Q10补充剂在心血管疾病、神经元疾病、 和肌肉退行性疾病，他汀类药物引起的肌病，高血压，偏头痛，癌症，以及许多 他人然而，许多此类临床试验结果的可变成功被认为是由于 缺乏对决定细胞摄取、胃肠道吸收的生物学过程的了解， 辅酶Q10的后续生物利用度。这些未解决的问题大大减缓了临床翻译， 进一步扩大辅酶Q10的治疗应用。因此，了解 调节辅酶Q10摄取并优化其口服生物利用度的生物过程， 是辅酶Q10补充剂的主要挑战之一。然而，研究外源基因的进展 辅酶Q10通量由于缺乏工具而受到严重阻碍，并且目前没有成像技术用于测量辅酶Q10通量。 CoQ 10摄取的无创纵向监测。解决营养摄取成像的未满足需求 工具，我们目前正在开发一种新的分析平台的基础上结合多功能的“点击”化学 与非侵入性和超灵敏度生物发光成像的反应（功能纵向光学平台 体内代谢物摄取成像，1-R 01-EB 034607 - 01 A1）。该方法不依赖于放射性和/或 寿命短的同位素，成本较低，并允许纵向监测代谢物吸收。而第一 我们使用葡萄糖和烟酰胺核苷成功地验证了这种方法的应用， 以维生素B3为例（Nat Methods，2019和Biosens.生物电子。2023年），我们现在正在扩大这一点 技术来研究不同的氨基酸，脂肪酸和核苷的吸收，这些都发挥着核心作用， 许多人类病理学。由于探头的设计是基于非侵入性成像，因此可以获得更多的数据。 从单个动物收集，从而能够更好地再现。在这里，我们建议应用这种通用的方法， 开发一种新的成像工具，用于非侵入性纵向测量CoQ 10摄取， 体外和体内。这种新型探针将为了解辅酶Q10的摄取、组织 分布，并将成为优化其口服生物利用度的有价值的工具。

PROJECT SUMMARY Coenzyme Q10 (CoQ10) is a potent energy transfer molecule and a central cofactor in oxidative phosphorylation present in both cell membranes and mitochondria. CoQ10 is also known as a powerful anti-inflammatory and antioxidant agent that protects the tissues from the damage resulting from the activation of inflammatory signaling pathways and free radicals. It is naturally synthesized at high levels in the liver, heart, and kidneys. However, the ability of the body to synthesize its own CoQ10 significantly decreases with age. In addition to the regular aging process, significant decrease in CoQ10 levels have been identified in a number of human pathologies. Therefore, nutritional supplementation with CoQ10 provides a powerful tool to maintain its sufficient levels in the body with the goal to improve human health and reduce the risk of chronic diseases. In this context, multiple clinical trials revealed potential benefits of CoQ10 supplementation in cardiovascular diseases, neuronal and muscular degenerative diseases, statin-induced myopathy, high blood pressure, migraine, cancer, and many others. However, the variable success in the outcomes of many of such clinical trials is thought to be due to the lack of knowledge of the biological process determining cellular uptake, gastrointestinal absorption, and subsequent bioavailability of CoQ10. These unresolved issues significantly slow down clinical translation and further expansion of CoQ10 therapeutic applications. Thus, it is of critical importance to understand the biological processes that regulate CoQ10 uptake and to optimize its oral bioavailability, which remains to be one of the main challenges in CoQ10 supplementation. However, the progress of studying exogenous CoQ10 fluxes is significantly hampered by the lack of tools and no imaging techniques currently exist for noninvasive longitudinal monitoring of CoQ10 uptake. To address the unmet need for nutrient uptake imaging tools, we are currently developing a novel analytical platform based on a combination of versatile “click” chemistry reactions with noninvasive and ultrasensitive bioluminescent imaging (Optical platform for functional longitudinal imaging of metabolite uptake in vivo, 1-R01-EB034607-01A1). The method is independent of radioactive and/or short-lived isotopes, less costly, and allows longitudinal monitoring of metabolite absorption. While the first application of this approach was successfully validated by us using glucose and nicotinamide riboside, a form of vitamin B3, as an example (Nat Methods, 2019 and Biosens. Bioelectron. 2023), we are now expanding this technology to study uptake of different amino acids, fatty acids, and nucleosides, which all play a central role in many human pathologies. Since the design of the probes is based on noninvasive imaging, more data could be collected from a single animal enabling better reproducibility. Here, we propose to apply this versatile method to develop a novel imaging tool for noninvasive, longitudinal measurements of CoQ10 uptake both in vitro and in vivo. This novel probe will provide a unique opportunity for understanding CoQ10 uptake, tissue distribution, and will become a valuable tool for optimizing its oral bioavailability.