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 医疗疾病。尽管进行了多次治疗，三分之一或更多的患者可能仍无法实现症状缓解 药物治疗；许多其他人根本无法或不愿意开始处方 药物或社会心理治疗。补充和替代药物（CAM）代表了 对于此类患者来说是重要的选择。除了了解哪些 CAM 抗抑郁策略之外 对抑郁症和压力有效，这是项目 1 的目标，了解这一点非常有价值 某些 CAM 治疗发挥治疗作用的机制。这种理解可以 提高当前治疗的可接受性，使患者能够更好地匹配有效的治疗，以及 促进新型 CAM 治疗方法的研究和开发。对于标准抗抑郁治疗， 已经提出了关于作用机制的多种假设。其中包括 (i) 促进 神经可塑性，（ii）炎症调节，以及（iii）促进神经发生。迄今为止，调查 后面这些假设因缺乏人类神经生物学的直接模型而受到阻碍——并且 特别是神经病理学 - 适合快速筛查和定量功能评估。也就是说，它 尚无法检验这些假设是否在患有以下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。干细胞技术与发育的进展 神经生物学提出了一种新颖的策略，该策略构成了项目 3 的焦点。来自 180 名患者的真皮成纤维细胞 项目一随机试验的参与者将被重新编程（转分化）以诱导 神经元（iN）。这些 iN 的转录谱将在暴露于 n-3 脂肪酸、S- 腺苷 L-甲硫氨酸 (SAMe)，或测试两种 CAM 是否调节相关基因的工具 神经可塑性（目标 1a），以及神经可塑性的调节程度是否与治疗相关 功效。与此同时，患者来源的成纤维细胞的子集（每个治疗组 10 个）将被重新编程为 诱导多能干细胞。然后这些细胞系将分化为神经元前体细胞 最终形成成熟的神经元。该小组和其他人的工作表明有可能产生这样的细胞 并将它们纳入高通量、定量功能测定中，以表征与相关的表型 抗抑郁机制。具体来说，n-3 脂肪酸和 SAMe 调节炎症的假设 神经谱系细胞上的标记（目标 2a）和促进神经发生（目标 2b）将使用经过验证的方法进行测试 化验。这些机制与治疗效果相关的假设也将得到检验。在 除了检验这些主要假设之外，该项目还将为未来建立关键资源 对 CAM 化合物的研究，这是一个包含 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，全部来自参与项目 1 安慰剂对照研究的 MDD 患者。 叙事 重度抑郁症是全球发病率的一个主要原因，现有的治疗方法无法产生效果 约 1/3 的患者症状缓解。而补充和替代医学 (CAM) 化合物 越来越多地用于治疗抑郁症，并被患者广泛接受，但其作用机制尚未明确 得到了充分的表征。拟议的调查将测试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.