Patient-Derived Cellular Models of Putative Antidepressants

项目3摘要：重度抑郁障碍(MDD)仍然是最普遍和最昂贵的疾病之一 医学上的障碍。三分之一或更多的患者可能无法实现症状缓解，尽管 药物治疗；许多其他人根本不能或不愿意开始开处方 药物治疗或心理社会治疗。补充和替代药物(CAM)代表着一种 对于这类患者来说，这是一个重要的选择。除了了解哪些CAM抗抑郁药物策略 对抑郁症和压力有效，这是项目1的目标，了解这一点将非常有价值 某些CAM治疗发挥其治疗效果的机制。这种理解可能会 提高当前治疗的可接受性，使患者与有效治疗更好地匹配，以及 促进新型CAM治疗方法的研究和开发。对于标准的抗抑郁药物治疗， 关于作用机制的多种假说已经被提出。这些措施包括(I)推广 神经可塑性，(Ii)炎症的调节，和(Iii)促进神经发生。到目前为止，对 后一种假说由于缺乏人类神经生物学的直接模型而受到阻碍--以及 尤其是神经病理学--适用于快速筛查和定量功能评估。就是它 一直无法检验这些假说是否在患有脑血管疾病的患者的神经组织中得到支持 这些干预措施针对的特定疾病。干细胞技术与发展的进展 神经生物学允许一种新的策略形成项目3的焦点。180名患者的皮肤成纤维细胞 在项目一的随机试验中，参与者将被重新编程(转分化)为诱导 神经元(INS)。S说，在接触n-3脂肪酸后，这些蛋白的转录谱将被比较。 腺苷L-蛋氨酸(同)，或载体，以测试这两个CAM是否调节相关基因 神经可塑性(目标1a)，以及神经可塑性的改变程度是否与治疗有关 功效。同时，患者来源的成纤维细胞的一个子集(每个治疗组10个)将被重新编程以 诱导的多能干细胞。这些细胞系随后将被分化为神经元前体细胞 最终到达成熟的神经元。这个研究小组和其他人的研究表明，有可能产生这样的细胞 并将它们纳入高通量、定量的功能分析中，以表征与 抗抑郁机制。具体地说，n-3脂肪酸及其类似物调节炎症的假设 神经谱系细胞的标记物(目标2a)和促进神经发生(目标2b)将使用VALIDATED进行测试 化验。这些机制与治疗效果相关的假设也将得到检验。在……里面 除了检验这些基本假设外，这个项目还将为未来建立一个关键的资源 含180个成纤维细胞和30个多能干细胞和神经元的生物库--CAM化合物的研究 前体细胞，全部来自参与项目1‘S安慰剂对照研究的MDD患者。 叙事 严重的抑郁障碍是全球发病率的主要原因，现有的治疗方法无法奏效 症状缓解率为1/3。而补充和替代医学(CAM)化合物 越来越多地被用于治疗抑郁症，并被患者很好地接受，但它们的作用机制尚未 已经被完全定性了。拟议的调查将检验扩展初步数据的具体假设 使用独特的患者来源的干细胞和诱导的神经元，同时建立 生物标志物、生物签名和生物库，以促进未来的CAM研究。

PROJECT 3 ABSTRACT: Major depressive disorder (MDD) remains one of the most prevalent and costly medical disorders. A third or more of patients may not achieve symptomatic remission despite multiple medication treatments; many other individuals are simply unable or unwilling to initiate prescription pharmacologic or psychosocial treatment. Complementary and alternative medications (CAM) represent an important option for such patients. In addition to understanding which CAM antidepressant strategies are efficacious for depression as well as for stress, the aim of Project 1, it would be highly valuable to understand the mechanisms by which certain CAM treatments exert their therapeutic effect. This understanding could increase the acceptability of current treatments, allow better matching of patients with effective treatments, and facilitate the investigation and development of novel CAM treatments. For standard antidepressant treatments, multiple hypotheses regarding mechanisms of action have been developed. These include (i) promotion of neuroplasticity, (ii) modulation of inflammation, and (iii) promotion of neurogenesis. To date, investigation of these latter hypotheses has been hampered by a lack of direct models of human neurobiology – and particularly neuropathology - amenable to rapid screening and quantitative functional assessment. That is, it has not been possible to examine whether these hypotheses are supported in neural tissue from patients with the particular disease targeted by these interventions. Progress in stem cell technology and developmental neurobiology allows a novel strategy that forms the focus of Project 3. Dermal fibroblasts from 180 patient participants in the randomized trial of Project One will be reprogrammed (transdifferentiated) to induced neurons (iNs). Transcriptional profiles of these iN's will be compared after exposure to n-3 fatty acids, S- adenosyl L-methionine (SAMe), or vehicle to test whether the two CAMs regulate genes related to neuroplasticity (Aim 1a), and whether degree of modulation of neuroplasticity is associated with treatment efficacy. In parallel, a subset (10 per treatment arm) of patient-derived fibroblasts will be reprogrammed to induced pluripotent stem cells. These cell lines will then be differentiated into neuronal precursors and ultimately to mature neurons. Work by this group and others indicates that it is possible to generate such cells and incorporate them in high-throughput, quantitative functional assays to characterize phenotypes relevant to antidepressant mechanism. Specifically, the hypothesis that n-3 fatty acids and SAMe modulate inflammatory markers on neural-lineage cells (Aim 2a), and promote neurogenesis (Aim 2b), will be tested using validated assays. The hypothesis that these mechanisms are associated with treatment efficacy will also be tested. In addition to examining these primary hypotheses, this project will establish a critical resource for future investigation of CAM compounds, a biobank of 180 fibroblasts and 30 pluripotent stem cells and neuronal precursor cells, all derived from patients with MDD participating in Project 1's placebo-controlled investigations. NARRATIVE Major depressive disorder is a major contributor to morbidity worldwide, and existing treatments fail to yield symptomatic remission in ~1/3 of patients. While Complementary and Alternative Medicine (CAM) compounds are increasingly used to treat depression, and well-accepted by patients, their mechanisms of effect have not been fully characterized. The proposed investigation will test specific hypotheses extending preliminary data using unique patient-derived stem cells and induced neurons, while at the same timing establishing biomarkers, biosignatures, and a biobank to facilitate future CAM studies.