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

项目总结 辅酶Q10(CoQ10)是一种强有力的能量转移分子，也是氧化磷酸化的中心辅因子 存在于细胞膜和线粒体中。辅酶Q10也被认为是一种强大的抗炎和 一种抗氧化剂，可保护组织免受因炎症激活而造成的损害 信号通路和自由基。它在肝脏、心脏和肾脏中自然合成水平很高。 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 在有规律的衰老过程中，已发现许多人的辅酶Q10水平显著下降 病理学。因此，辅酶Q10的营养补充为维持其充足的营养提供了有力的工具 其目的是改善人体健康，降低慢性病的风险。在这方面， 多项临床试验显示辅酶Q10补充剂在心血管疾病、神经元疾病中的潜在益处 以及肌肉退行性疾病、他汀类药物引起的肌病、高血压、偏头痛、癌症和许多 其他。然而，许多这样的临床试验结果的可变成功被认为是由于 对决定细胞摄取、胃肠道吸收的生物过程缺乏了解，以及 辅酶Q10的后续生物利用度。这些悬而未决的问题大大减慢了临床翻译和 辅酶Q10治疗应用的进一步扩大。因此，理解 调节辅酶Q10摄取并优化其口服生物利用度的生物学过程 成为辅酶Q10补充的主要挑战之一。然而，外源物质的研究进展 由于缺乏工具，CoQ10的通量严重受阻，而且目前还没有成像技术 辅酶Q10摄取的无创纵向监测。为了解决营养摄取成像未得到满足的需求 工具，我们目前正在开发一种新的分析平台，该平台基于多功能的“点击”化学 非侵入性和超灵敏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-R01-EB034607-01A1)。该方法不受放射性和/或 短寿命的同位素，成本较低，并允许对代谢物吸收进行纵向监测。而第一个 我们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方法，2019和Biosens。生物电子。2023)，我们现在正在扩大这一范围 研究不同氨基酸、脂肪酸和核苷摄取的技术，这些都在 很多人类的病态。由于探头的设计是基于非侵入性成像，因此可以获得更多数据 从单一动物采集，具有更好的重复性。在这里，我们建议应用这个通用的方法 开发一种新的成像工具，用于无创性、纵向测量辅酶Q10摄取 体外和体内。这一新的探针将为理解CoQ10摄取、组织 这将成为优化其口服生物利用度的宝贵工具。

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.