Lead compound discovery from proprietary mycobacterial strains for treatment of PTSD

项目摘要 免疫调节，即，调节性T细胞和效应性T细胞的平衡表达，被认为是在 现代高收入环境，部分原因是减少了与商业和环境衍生的接触， 细菌，也被称为“老朋友”。失败的免疫调节被认为是导致最近 与压力有关的慢性炎症性疾病以及与压力有关的精神疾病的增加， 炎症是一个危险因素，如抑郁症，焦虑症和创伤后应激障碍（PTSD）， 高收入国家。这些“老朋友”之一是母牛分枝杆菌NCTC 11659，一种非致病性， 具有抗炎和免疫调节特性的环境无害物质。免疫接种的形式 一种M.母牛已经被证明可以增加小鼠的压力恢复能力， 预防压力引起的焦虑增加，预防压力引起的自发夸大 在炎性结肠炎模型中预防化学诱导的结肠炎的应激诱导的加重 肠道疾病。用M.接种疫苗，无论是在恐惧条件反射之前还是之后进行， 在大鼠恐惧强化惊吓模型中，在其他研究中， 与M.研究表明，牛痘可以防止压力引起的焦虑样防御行为的夸大。 大鼠习得性无助模型的反应与应激诱导的小胶质细胞抑制有关 引发和神经炎症。然而，在多大程度上，这些压力恢复力和压力保护 对其它分枝杆菌菌株的影响尚不清楚。在本STTR建议中，我们建议 使用小鼠骨髓来源的树突状细胞（BMDC）、人单核细胞来源的巨噬细胞 （THP-1细胞）和小胶质细胞系（BV 2细胞），20种环境分枝杆菌的专利菌株 用于抗炎和免疫调节特性。为了进行筛选，我们将针对符合 以下标准：它们可以容易地单独培养;它们与已知的分枝杆菌不密切相关 它们代表了跨越已知分枝杆菌系统发育宽度的不同谱系， 多样性（其中有>150种描述的分枝杆菌菌株）。我们将筛选出两名主要候选人， 使用雄性和雌性大鼠，以确定新的分枝杆菌菌株与压力保护作用， 通过防止不可避免的压力（IS）引起的焦虑，条件性恐惧和逃避的增加来衡量 缺陷，以及预防应激诱导的海马小胶质细胞引发和神经炎症。我们将 还在雄性和雌性大鼠的恐惧增强惊吓和线索恐惧模型中评估这些品系。在 完成这些研究后，我们期望我们将确定一个单一的铅菌株， 分枝杆菌，具有最好的性能，为进一步的临床前测试和临床开发。这些结果 有望对创伤和应激源相关疾病的治疗产生重要的积极影响 因为目前的药物治疗具有显著的局限性。

Project Summary Immunoregulation, i.e., balanced expression of regulatory and effector T cells, is thought to be compromised in modern high-income settings due in part to reduced contact with commensal and environmentally derived bacteria, also known as “old friends”. Failed immunoregulation is thought to be one factor contributing to recent increases in stress-related and chronic inflammatory disorders as well stress-related psychiatric disorders in which inflammation is a risk factor, such as depression, anxiety, and posttraumatic stress disorder (PTSD), in high-income countries. One of these “old friends” is Mycobacterium vaccae NCTC 11659, a nonpathogenic, environmental saprophyte with anti-inflammatory and immunoregulatory properties. Immunization in the form of a heat-killed preparation of M. vaccae has been shown to increase stress resilience in mice, as measured by prevention of stress-induced increases in anxiety, prevention of stress-induced exaggeration of spontaneous colitis, and prevention of stress-induced exaggeration of chemically induced colitis in a model of inflammatory bowel disease. Immunization with M. vaccae, when conducted either before or after fear conditioning, has also been shown to enhance fear extinction in a rat model of fear-potentiated startle. In other studies, immunization with M. vaccae has been shown to prevent stress-induced exaggeration of anxiety-like defensive behavioral responses in a rat model of learned helplessness in association with suppression of stress-induced microglial priming and neuroinflammation. However, the extent to which these stress resilience and stress-protective effects generalize to other strains of mycobacteria are not known. In this STTR proposal, we propose to screen, using murine bone marrow-derived dendritic cells (BMDCs), human monocyte-derived macrophages (THP-1 cells), and a microglial cell line (BV2 cells), twenty proprietary strains of environmental mycobacteria for anti-inflammatory and immunoregulatory properties. For screening, we will target strains that meet the following criteria: they can readily be cultivated in isolation; they are not closely related to known mycobacterial pathogens; and they represent distinct lineages spanning the phylogenetic breadth of known mycobacterial diversity (of which there are >150 described mycobacterial strains). We will then screen two lead candidates, using male and female rats, to identify novel strains of mycobacteria with stress-protective effects, as measured by prevention of inescapable stress (IS)-induced increases in anxiety, conditioned fear, and escape deficits, and prevention of stress-induced priming of hippocampal microglia and neuroinflammation. We will also evaluate these strains in models of fear-potentiated startle and cued fear in male and female rats. At the completion of these studies, it is our expectation that we will have identified a single lead strain of mycobacteria that has the best properties for further preclinical testing and clinical development. These results are expected to have an important positive impact for treatment of trauma- and stressor-related disorders because current pharmacological treatments have significant limitations.