Noninvasive urinary monitoring of NASH by activity-based nanosensors

项目概要/摘要 非酒精性脂肪性肝炎（NASH）被称为“隐形流行病”，仅在美国，20%的 据估计，1600万NASH患者将进展并发展为危及生命的疾病 包括纤维化、肝硬化、肝细胞癌和肝衰竭。到2020年，NASH预计将 成为肝移植的主要指征，超过丙型肝炎病毒（HCV）和酒精相关的肝移植。 到2025年，NASH药物市场总规模估计将达到150亿美元。目前， 肝活检是诊断和监测NASH的“金标准”，但它是侵入性的，昂贵的， （约3，000美元），并与显著的患者发病率相关。重要的是，单次活检仅提供 组织结构的瞬时“快照”，并且不能识别“快速”的NASH患者， 因此迫切需要治疗的患者，或者那些正在接受治疗的“应答者”。由于这些 尽管存在局限性，但肝活检仍然是NASH安全、准确诊断和监测的关键瓶颈。 该提案旨在开发一种非侵入性尿液诊断方法，即甘精酶肝脏检测（GLT）， 驱动NASH的肝蛋白酶的活性。GLT由可注射的纳米传感器组成， 蛋白酶与炎症和纤维化有关，并产生在尿液中检测到的信号。的 检测信号通过肝脏中的酶促作用放大，然后在患者的尿液中浓缩， 通过质谱法定量。因为酶的活性可以在分子水平上测量， 由于活检的微观变化是明显的，因此该测试是超灵敏和预测性的。我们将 证明了在惰性聚合物支架上配制GLT纳米传感器以感测基质的可行性 金属蛋白酶（MMPs）-参与NASH进展和消退的关键酶家族。 然后，我们将验证GLT在小鼠模型中预测疾病进展和消退的能力。 NASH，并进行临床前毒理学研究，为IND提交做准备。GLT将提供 临床医生快速确定患者肝脏的状态和轨迹，而没有活检的发病率， 将诊断成本从3,000美元降低到500美元，并将灵敏度提高40%。对医疗保健的益处 每名患者的系统估计可减少活检相关费用48，000美元和药物费用145，000美元 在患者的一生中，每年为20%的NASH患者节省超过120亿美元， 疾病独自采用GLT的影响将跨越多个选区-增加患者 护理和舒适，改善医生的决策，加速药物试验，降低 医疗费用。

Project Summary/Abstract Nonalcoholic steatohepatitis (NASH) is dubbed the “stealth epidemic” and in the U.S. alone, 20 percent of the 16 million people who are estimated to have NASH will progress and develop life-threatening conditions including fibrosis, cirrhosis, hepatocellular carcinoma, and liver failure. By the year 2020, NASH is projected to be the leading indication for liver transplantation surpassing hepatitis C virus (HCV) and alcoholic related cirrhosis, and by 2025, the total NASH pharmaceutical market size is estimated to reach $15 billion. Currently, the liver biopsy is the “gold standard” for diagnosing and monitoring NASH, yet it is invasive, expensive (~$3,000), and associated with significant patient morbidities. Importantly, a single biopsy provides only an instantaneous `snapshot' of tissue architecture and is unable to identify NASH patients who are `rapid progressors' and thus urgently require treatment, or those on therapy that are `responders'. As a result of these limitations, liver biopsy remains the key bottleneck in the safe, accurate diagnosis and monitoring of NASH. This proposal aims to develop the Glympse liver test (GLT), a noninvasive urine diagnostic that detects the activity of hepatic proteases that drive NASH. GLT consists of an injectable panel of nanosensors that detect proteases involved with inflammation and fibrosis, and produce signals that are detected in urine. The detection signals are amplified by enzymatic action in the liver and then concentrated in the patient's urine for quantification by mass spectrometry. Because enzyme activity is measurable at the molecular level before microscopic changes are evident by biopsy, the test is thus both ultrasensitive and predictive. We will demonstrate the feasibility of formulating GLT nanosensors on an inert polymer scaffold to sense matrix metalloproteinases (MMPs) – a key family of enzymes that are involved in NASH progression and regression. We will then validate the ability of GLT to predict disease progression and regression in a mouse model of NASH, and conduct preclinical toxicology studies in preparation for an IND submission. The GLT will offer clinicians a rapid determination of the state and trajectory of the patient's liver without the morbidity of biopsy, reduce cost of diagnosis from $3,000 to $500, and raise sensitivity by 40%. The benefit to the healthcare system per patient is estimated to be $48,000 in reduced biopsy-associated costs and $145,000 in drug costs over a patient's lifetime, saving more than $12 billion annually for the 20% of NASH patients with progressive disease alone. The impact of the adoption of GLT will reach across multiple constituencies – increasing patient care and comfort, improving physician decision making, accelerating pharmaceutical drug trials, and lowering healthcare costs.