Patient-Derived Cellular Models of Putative Antidepressants

项目3摘要：重度抑郁症（MDD）仍然是最普遍和最昂贵的疾病之一 医学疾病三分之一或更多的患者可能无法达到症状缓解，尽管有多种 药物治疗;许多其他人只是不能或不愿意开始处方 药物或心理治疗。补充和替代药物（CAM）是一种 这类患者的重要选择。除了了解哪些CAM抗抑郁药策略是 有效的抑郁症以及压力，项目1的目的，这将是非常有价值的了解 某些CAM治疗发挥其治疗效果的机制。这种理解可以 提高目前治疗的可接受性，使患者与有效治疗更好地匹配， 促进新型CAM治疗的研究和开发。对于标准的抗抑郁治疗， 关于作用机制的多种假说已经被提出。这些措施包括：（一）促进 神经可塑性，（ii）炎症的调节，和（iii）促进神经发生。迄今为止， 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， 特别是神经病理学-适合于快速筛查和定量功能评估。即它 目前还不可能检查这些假设是否在患有神经系统疾病的患者的神经组织中得到支持。 这些干预措施所针对的特定疾病。干细胞技术与发育 神经生物学允许一种新的策略，形成项目3的重点。180例患者的真皮成纤维细胞 参与项目一随机试验的受试者将被重新编程（转分化），以诱导 神经元（iNs）。将在暴露于n-3脂肪酸、S-脂肪酸和N-脂肪酸后比较这些iN的转录谱。 腺苷L-甲硫氨酸（SAMe）或载体，以测试两种CAM是否调节与以下相关的基因： 神经可塑性（目的1a），以及神经可塑性的调节程度是否与治疗相关 功效同时，将对患者来源的成纤维细胞亚组（每个治疗组10个）进行重编程， 诱导多能干细胞然后这些细胞系将分化成神经元前体， 最终发育成成熟的神经元。这个小组和其他人的工作表明， 并将其纳入高通量定量功能测定中，以表征与以下相关的表型： 抗抑郁机制具体地说，假设n-3脂肪酸和SAMe调节炎症反应， 神经谱系细胞上的标志物（Aim 2a）和促进神经发生（Aim 2b），将使用经过验证的 测定。还将检验这些机制与治疗疗效相关的假设。在 除了检查这些主要假设外，该项目还将为未来的研究建立一个关键资源。 CAM化合物的研究，180个成纤维细胞和30个多能干细胞和神经元的生物库 前体细胞，均来自参与项目1安慰剂对照研究的MDD患者。 叙事 重度抑郁症是世界范围内发病率的主要原因，现有的治疗方法无法产生 约1/3的患者症状缓解。补充和替代医学（CAM） 越来越多地被用于治疗抑郁症，并被患者所接受，但其作用机制还没有 被充分定性。拟议的调查将测试具体的假设， 使用独特的患者源性干细胞和诱导神经元，同时建立 生物标志物、生物特征和生物库，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.