Human enteric glia: A novel target for motility and postoperative ileus

肠神经胶质细胞(EGCs)调节肠道内环境平衡，可能参与嘌呤能信号转导和 运动性，但它们的确切作用尚不清楚，特别是在人类身上。在发炎的状态下，它们会转化为 “更多的反应表型”，释放炎症介质，这可能有助于病理生理。 问题：人类EGCs(HEGCs)在胃肠道的生理学和病理生理学中的作用不是 已知，解决知识中的这一关键差距对于确定hGCs如何调制GI非常重要 在健康和疾病方面的功能。我们的初步数据支持hEGCs中的嘌呤能信号的概念 调节细胞通讯、突触生理、运动行为和炎症信号。意义 这些翻译研究的突出表现是在神经胶质反应、机制上的显著物种差异 在人类胚胎中，神经胶质细胞与神经元的比率(与老鼠相比)高出7倍。开创性的-机械式的， 翻译研究是关于‘机械敏感性’、‘运动调节’和术后肠梗阻的。它 有必要开发和应用尖端的新技术来研究，即机械刺激 (压力、切应力、径向拉伸)hEGCs、电流、Vm、Ca~(2+)波，使用 ‘嗅觉细胞’，运动性和基因表达的时空成像(例如，RiboTag小鼠&107-定制 “反应性胶质细胞”的基因读数)。手术创伤/肠道操作、IL-1β或条件培养液的研究 为研究人类肠道中的“反应性神经胶质”提供了精确的方法。总体假设：“人类” EGCs是机械敏感性、细胞间通讯、突触生理学、 人类肠道中的神经肌肉传递和运动以及炎症破坏了它们的功能 正常活动。嘌呤能信号在“功能和功能障碍”中起着关键作用。EGC发挥作用 神经网络活性的抑制性调制。EGC可能在以下方面发挥不同的(相反)效果 两个肌肉层，通过作用于神经内窥镜下调节上行和下行时相 蠕动反射，协调或同步运动。神经胶质细胞的激活扰乱了脑内的运动 术后肠梗阻。研究得到了一个有合作记录的梦幻调查团队的支持， 一流的环境，强大的初步数据(图1-16)，《炎症性肠病》的2篇新出版物 以及关于“基本hEGC模式”的“GLIA”的一份修订出版物。我们的总体假设在4个月内得到了验证 最初的目标。目的1：探讨人脐静脉内皮细胞释放三磷酸腺苷的机械转导途径。目标2：实现 大鼠完整神经丛神经元力学感觉转导和神经胶质细胞通讯的研究 人的肠子。目的3：确定hEGCs在运动调节中的作用。目标4：确定 术后肠梗阻炎症对反应性胶质细胞和运动性的影响及对人体组织的影响 是有可能的。影响：翻译研究将填补关于小脑中神经胶质细胞知识的一个关键空白 人类肠道在健康和疾病中的作用，以及对动力障碍、肠道炎症和治疗的影响。

Enteric glial cells (EGCs) regulate intestinal homeostasis and may be involved in purinergic signaling and motility, but their precise role is not understood, especially in humans. In the inflamed state, they convert to a `more reactive phenotype', releasing inflammatory mediators that could contribute to pathophysiology. PROBLEM: The role of human EGCs (hEGCs) in the physiology and pathophysiology of the GI tract is not known, and addressing this critical gap in knowledge is important in determining how hEGCs modulate GI functions in health and disease. Our preliminary data support the concept that purinergic signals in hEGCs regulate cellular communication, synaptic physiology, motor behavior, and inflammatory signals. Significance of these translational studies is underscored by significant species differences in glial responses, mechanisms and a 7 fold higher glial to neuron ratio in human ENS (versus mouse). Groundbreaking – mechanistic, translational studies are proposed on `mechanosensitivity', `modulation of motility' and postoperative ileus. It became necessary to develop and apply sophisticated new technologies to study i.e. mechanical stimulation (pressure, shear stress, radial stretch) of hEGCs, currents, Vm, Ca2+waves, monitor ATP release using a `sniffer cell', spaciotemporal imaging of motility, and gene expression (i.e. RiboTag mouse & a 107-custom gene readout of `reactive glia'). Studies on surgical trauma/intestinal manipulation, IL1β or conditioned media from hEGCs provide precise ways to study `reactive glia' in human gut. OVERALL HYPOTHESIS: “Human EGCs are critical regulators of mechanosensitivity, cell-to-cell communication, synaptic physiology, neuromuscular transmission and motility in the human intestinal tract and inflammation disrupts their normal activity. Purinergic signaling plays a pivotal role in `function and dysfunction'. EGCs exert inhibitory modulation of neural network activity. EGCs may exert differential (opposing) effects in the two muscle layers by acting in the ENS to modulate ascending and descending phases of the peristaltic reflex and coordinate or synchronize motility. Glial activation disrupts motility in postoperative ileus. Studies are supported by a dream team of investigators with a record of collaboration, a superb environment, strong preliminary data (Figs 1-16), 2 new publications in `Inflammatory Bowel Diseases' in 2016 and one revised publication to `GLIA' on the `basic hEGC model'. Our overall hypothesis is tested in 4 original Aims. AIM 1: To investigate mechanotransduction pathways of ATP release in hEGCs. AIM 2: To evaluate mechanosensory transduction and glial communication in the ENS of intact neural plexus networks in human intestine. AIM 3: To determine the role of hEGCs in the modulation of motility. AIM 4: To determine the impact of inflammation in postoperative ileus on reactive glia and motility and use human tissue to the extent it is possible. IMPACT: Translational studies will fill a critical gap in knowledge of glia in the `little brain' of the human gut in health and disease, with implications for motility disorders, gut inflammation and therapeutics.