Development of Controlled Release Mitochondrial Protonophore (CRMP) as a Novel Treatment for Type-2 Diabetes and Non-Alcoholic Steatohepatitis in Dysmetabolic Non-Human Primates

据估计，非酒精性脂肪性肝病（NAFLD）发生在三分之一的普通人群中，是肝脏胰岛素抵抗和2型糖尿病（T2 D）发病机制的主要诱发因素。当肝脏的脂质供应超过脂质氧化和脂质输出速率时，发生NAFLD。已经采用了许多疗法来减少肝脏中的异位脂肪积累和肝脏胰岛素抵抗，然而这些方法在长期中的成功有限，需要新的疗法。为了满足这一巨大的未满足的需求，我们的实验室最近开发了一种控释线粒体质子载体（CRMP），其功能性肝靶向，并通过促进肝脏线粒体解偶联活性的微妙持续增加来增加线粒体脂肪氧化。重要的是，我们之前已经证明CRMP在NAFLD/NASH/肝纤维化和T2 D的啮齿动物模型中安全地逆转高脂血症、脂肪肝、脂肪性肝炎和肝纤维化，而不诱导高热、体重减轻或任何相关的肝/全身毒性。总之，这些啮齿动物研究为在高度相关的非人灵长类动物NAFLD和代谢异常综合征模型中进一步评估CRMP作为治疗NAFLD/NASH和T2 D的新型治疗策略提供了重要的概念证据。因此，在本提案中，我们的目的是确定慢性CRMP治疗在代谢异常的自发性肥胖恒河猴中逆转高脂血症、NAFLD以及肝脏和肌肉胰岛素抵抗的安全性和有效性。此外，我们将使用最先进的液相色谱-串联质谱法和核磁共振光谱法进行一组全面的肝代谢通量测量，以直接评估慢性CRMP治疗对该肥胖代谢异常NAFLD猴模型中肝线粒体脂肪酸氧化、肝异生、丙酮酸脱氢酶通量、丙酮酸激酶通量和肝生酮速率的影响。我们假设慢性CRMP治疗将逆转NAFLD，并通过促进肝线粒体脂肪酸氧化速率的增加而不依赖于食物摄入量/体重的变化，以安全有效的方式改善肥胖代谢异常恒河猴的全身胰岛素反应性。总的来说，这项研究的结果将是非常有影响力的，因为它们将在肥胖代谢异常的非人灵长类动物的高度相关临床模型中提供重要的概念验证和安全性数据，以支持开发用于治疗人类NAFLD/NASH和T2 D的新型肝脏靶向线粒体解偶联剂。

Non-alcoholic fatty liver disease (NAFLD) is estimated to occur in one third of the general population and is a major predisposing factor in the pathogenesis of hepatic insulin resistance and type 2 diabetes (T2D). NAFLD occurs when lipid supply to the liver exceeds rates of lipid oxidation and lipid export. A number of therapies have been employed to reduce ectopic-fat accumulation in liver and hepatic insulin resistance, however these approaches have been met with limited success in the long-term and new therapies are required. In order to meet this great unmet need, our lab has recently developed a Controlled Release Mitochondrial Protonophore (CRMP) that is functionally liver-targeted and causes increased mitochondrial fat oxidation by promoting a subtle sustained increase in hepatic mitochondrial uncoupling activity. Importantly, we have previously demonstrated that CRMP safely reverses hypertriglyceridemia, fatty liver, steatohepatitis and liver fibrosis in rodent models of NAFLD/NASH/liver fibrosis and T2D without inducing hyperthermia, weight loss or any associated hepatic/systemic toxicities. Taken together, these rodent studies provide important proof of concept for the further assessment of CRMP as a novel therapeutic strategy for the treatment of NAFLD/NASH and T2D in a highly relevant non-human primate model of NAFLD and the dysmetabolic syndrome. Therefore, in the present proposal, we aim to determine the safety and efficacy of chronic CRMP treatment on the reversal of hypertriglyceridemia, NAFLD, and liver and muscle insulin resistance in dysmetabolic, spontaneously obese Rhesus monkeys. In addition, we will perform a comprehensive set of hepatic metabolic flux measurements using state-of-the-art liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance spectroscopy methods to directly assess the impact of chronic CRMP treatment on rates of hepatic mitochondrial fatty acid oxidation, gluconeogenesis, pyruvate dehydrogenase flux, pyruvate kinase flux and hepatic ketogenesis in this obese dysmetabolic monkey model of NAFLD. We hypothesize that chronic CRMP treatment will reverse NAFLD and improve whole-body insulin responsiveness in obese dysmetabolic Rhesus monkeys in a safe and effective manner by promoting increased rates of hepatic mitochondrial fatty acid oxidation independent of changes in food intake/body weight. Collectively, the results of this research will be highly impactful in that they will provide important proof-of-concept and safety data in a highly relevant clinical model of obese dysmetabolic non-human primates to support the development of novel liver-targeted mitochondrial uncoupling agents for the treatment of NAFLD/NASH and T2D in humans.