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受给定疾病影响的患者组中的疗效和毒性，并在此过程中提供药物在人群中的效果估计。然而，它们通常无法让临床医生预测哪些特定患者可能受益或经历严重的毒性作用 被测试的药物。诱导多能干细胞（iPSC）的发现现在使得使用从患者中产生的iPSC的分化后代在体外模拟遗传疾病成为可能。例如，PiZZ iPS衍生的肝细胞已被证明重现了α-1抗胰蛋白酶缺乏症（AATD）相关肝病的关键特征，证明了疾病特异性iPS衍生细胞模拟这种和其他遗传疾病的潜力。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改善AATD相关肝病表型的有效性。将在iPS-衍生肝细胞中检测卡马西平，该肝细胞由入组正在进行的临床试验NCT 01379469（卡马西平治疗α-1抗胰蛋白酶缺乏所致重度肝病）的受试者产生，以进行“培养皿中的临床试验”。研究者将对体内临床试验的结果设盲，以创建“三盲”研究，其中iPS研究结果将用于预测体内研究的结果。然后在完成两项研究后评估这些预测的准确性，以确定这种iPS建模方法在多大程度上重现了实际患者对治疗的反应。该研究的总体目标是确定患者特异性iPS衍生的细胞类型是否可以作为患者体内类似细胞类型的忠实替代物。该研究的第二个目标是确定卡马西平是否能改善AATD相关肝病的影响。为实现这些目标，提出了三个具体目标。首先，将从入组临床试验NCT 01379469的受试者中创建iPSC文库。其次，这些iPSC的一个子集将经历基因组编辑，以纠正导致PiZZ AATD表型的单碱基对突变。然后，亲代患病（PiZZ）iPSC和基因校正的子代（PiMM）iPSC将与iPS肝细胞平行分化，以产生疾病的特征 这是由异常的AAT蛋白产生引起的。最后，在目标1中产生的iPSC将分化为iPS-肝细胞，并用卡马西平处理以概括临床 体外试验NCT 01379469。这两项研究之间的结果将相互关联，以确定是否iPS为基础的体外研究忠实地概括了体内临床试验结果，从而可以作为个性化医疗方法的基础，以及提供一个替代的未来方法，临床试验设计。

 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.