Personalized therapy for AATD-associated liver disease via IPS modeling

 描述(由申请人提供)：人类临床药物试验测试药物在特定疾病患者群体中的疗效和毒性，并在这样做的过程中提供对药物在人群中的影响的估计。然而，它们往往不能让临床医生预测哪些特定的患者可能受益或经历严重的毒性反应。 从正在测试的药物中。诱导多能干细胞(IPSCs)的发现使利用患者产生的IPSCs的分化后代在体外建立遗传病模型成为可能。例如，PIZIPS来源的肝细胞已被证明概括了与α-1抗胰蛋白酶缺乏症(AATD)相关的肝病的关键特征，展示了疾病特异性iPS来源细胞模拟该疾病和其他遗传性疾病的潜力。本申请中概述的研究测试了一种新颖的、个性化的临床药物试验方法，使用基于iPS的模型系统来测试药物卡马西平在改善AATD相关肝病表型方面的有效性。卡马西平将在从正在进行的临床试验NCT01379469(卡马西平治疗由于α-1抗胰蛋白酶缺乏引起的严重肝病)的受试者中产生的iPS来源的肝细胞中进行测试，以进行一项“培养皿临床试验”。研究人员将对体内临床试验的结果视而不见，以创建一项“三盲”研究，其中iPS研究结果将用于预测体内研究的结果。这些预测的准确性将在两项研究完成后进行评估，以确定这种iPS建模方法对实际患者的治疗反应有多好。这项研究的总体目标是确定患者特有的iPS来源的细胞类型是否可以忠实地替代创造它们的患者体内类似的细胞类型。这项研究的第二个目标是确定卡马西平是否可以改善AATD相关性肝病的影响。为了实现这些目标，提出了三个具体目标。首先，将从参加临床试验NCT01379469的受试者中创建IPSCs库。其次，这些IPSCs的一个子集将接受基因组编辑，以纠正导致PIZAATD表型的单碱基突变。然后，亲代、患病(PIZZ)IPSCs和基因校正的子代(PIMM)IPSCs将与iPS-肝细胞平行分化，以产生疾病的标志 这特别是由于AAT蛋白产生异常所致。最后，在AIM 1中创造的IPSCs将被分化为iPS-肝细胞，并用卡马西平治疗总结临床 体外试验NCT01379469。这两项研究之间的结果将相互关联，以确定基于iPS的体外研究是否忠实地概括了体内临床试验结果，清华大学可以作为个性化药物方法的基础，并为未来的临床试验设计提供一种替代方法。

 DESCRIPTION (provided by applicant): Human clinical drug trials test the efficacy and toxicity of drugs in groups of patients affected with a given disease and in doing so provide an estimate of a drug's effects across a population. However, they often do not give clinicians the ability to predict which specific patients are likely either to benefit or to experience serious toxic effects from the drug being tested. The discovery of induced pluripotent stem cells (iPSCs) has now made it possible to model genetic diseases in vitro using the differentiated progeny of iPSCs created from patients. For example, PiZZ iPS-- derived hepatocytes have been shown to recapitulate key features of alpha-- 1 antitrypsin deficiency (AATD) associated liver disease, demonstrating the potential of disease-- specific iPS-- derived cells to model this and other genetic diseases. The studies outlined in this application test a novel, personalized approach to clinical drug trials, using an iPS-- based model system to test the effectiveness of the drug carbamazepine in ameliorating an AATD-- associated liver disease phenotype. Carbamazepine will be tested in iPS-- derived hepatocytes created from subjects enrolled in the ongoing clinical trial NCT01379469 (Carbamazepine in Severe Liver Disease Due to Alpha-- 1 Antitrypsin Deficiency) to perform a "clinical trial in a dish". The investigators will be blinded to the resuls of the in vivo clinical trial to create a "triple-- blinded" study in which the iPS study results willbe used to predict outcomes of the in vivo study. The accuracy of these predictions will then be assessed upon completion of the two studies to determine how well this iPS modeling approach recapitulates response to treatments in actual patients. The overall goal of the research is to determine whether patient-- specific, iPS-- derived cell types serve as faithful surrogates for analogous in vivo cell types in the patient from whom they were created. A second goal of the research is to determine whether carbamazepine ameliorates the affects of AATD-- associated liver disease. To achieve these goals three specific aims are proposed. First, a library of iPSCs will be created from subjects enrolled in clinical trial NCT01379469. Second, a subset of these iPSCs will undergo genome editing to correct the single base pair mutation responsible for the PiZZ AATD phenotype. Parental, diseased (PiZZ) iPSCs and gene-- corrected, daughter (PiMM) iPSCs will then be differentiated in parallel to iPS-- hepatocytes to create a signature of disease that results specifically from aberrant AAT protein production. Finally, iPSCs created in aim 1 will be differentiated to iPS-- hepatocytes and treated with carbamazepine to recapitulate clinical trial NCT01379469 in vitro. The results between the two studies will be correlated to determine whether iPS-- based in vitro studies faithfully recapitulate in vivo clinical trial results and thu could serve as the basis for personalized medicine approaches as well as provide an alternative future approach to clinical trial design.