CHOLINE TRANSPORTERS SPECIFICALLY TARGET NON-NEURONAL ACETYLCHOLINE SYNTHESIS

胆碱转运蛋白专门针对非神经元乙酰胆碱合成

• 批准号: 8173243
• 负责人: Pill-Soon Song
• 金额: $ 4.76万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173243
• 关键词: Acetylcholine; Asthma; Cell Proliferation; Cells; Choline; Computer Retrieval of Information on Scientific Projects Database; Coupled; Data; Epithelial Cells; Funding; Grant; Institution; Link; Lung; Lung diseases; Malignant Epithelial Cell; Malignant neoplasm of lung; Mediating; Modeling; Neuroendocrine Cell; Neuroglia; Neurons; Play; Proteins; Research; Research Personnel; Resources; Role; Signal Transduction; Sodium; Source; Squamous Cell Lung Carcinoma; Testing; United States National Institutes of Health; choline transporter; cholinergic; knock-down; lung Carcinoma; uptake

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Non-neuronal acetylcholine (Ach) synthesis and secretion may play a role in lung diseases such as lung cancer and asthma. In order to characterize and target non-neuronal signaling in the lung it is necessary to identify a step that differs between neuronal and non-neuronal cholinergic signaling. Choline uptake is a rate-limiting step for ACh synthesis and may provide this target. In neurons, the sodium-dependent, high-affinity choline transporter (CHT1) is absolutely required to transport choline into neurons for ACh synthesis. Our preliminary data shows that CHT1 is not required by some lung cells for ACh synthesis and instead a newly described class of choline transporters, the choline transporter-like proteins (CTL1 - 5), can mediate ACh synthesis. Therefore non-neuronal ACh synthesis in the lung may potentially be targeted without effecting neuronal ACh synthesis by targeting the correct CTL. To test this we propose these specific aims: (1) Identification of which choline transporter-like proteins are involved in choline-uptake in lung non-neuronal cells. (2) Identification of which CTL is coupled to non-neuronal ACh synthesis in the lung. (3) To determine if knockdown of the CTL linked to ACh synthesis can block lung cell proliferation. These aims will be implemented using specific siRNAs to knock down CTLs in H82 small cell lung carcinoma cells as a model for pulmonary neuroendocrine cells and H520 squamous cell lung carcinoma cells as a model for airway epithelial cells. These studies will allow the targeting of non-neuronal ACh synthesis as a first step in developing new therapies for lung diseases.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 非神经性乙酰胆碱（Ach）的合成和分泌可能在肺癌和哮喘等肺部疾病中发挥作用。为了表征和靶向肺中的非神经元信号传导，有必要确定神经元和非神经元胆碱能信号传导之间的不同步骤。胆碱摄取是乙酰胆碱合成的限速步骤，并可能提供这一目标。在神经元中，钠依赖性高亲和力胆碱转运蛋白（CHT 1）是将胆碱转运到神经元中进行ACh合成所必需的。我们的初步数据表明，CHT 1是不需要的一些肺细胞合成乙酰胆碱，而是一类新描述的胆碱转运蛋白，胆碱转运蛋白样蛋白（CTL 1 - 5），可以介导乙酰胆碱合成。因此，通过靶向正确的CTL，肺中的非神经元ACh合成可能潜在地被靶向而不影响神经元ACh合成。为了验证这一点，我们提出了这些具体的目标：（1）确定哪些胆碱转运蛋白样蛋白参与肺非神经元细胞的胆碱摄取。(2)鉴定哪种CTL与肺中的非神经元ACh合成偶联。(3)确定是否敲低与乙酰胆碱合成相关的CTL可以阻断肺细胞增殖。这些目标将使用特异性siRNA来实现，以敲低作为肺神经内分泌细胞模型的H82小细胞肺癌细胞和作为气道上皮细胞模型的H520鳞状细胞肺癌细胞中的CTL。这些研究将允许靶向非神经元乙酰胆碱合成作为开发肺部疾病新疗法的第一步。