Olfactory receptor-based sensors for biomedical applications

7.项目摘要/摘要 嗅觉受体是人类最大的化学细胞表面受体家族之一， 异位表达的嗅觉受体在16个不同的组织中被发现，包括肺， 肾脏和结肠。在非嗅觉组织中，Exor并不能检测到“气味”，而是驱动了许多生物 过程，包括趋化、发育和血管生成。理解外遇在人类中的作用 健康-包括哪些外泄与医学相关，以及哪些化合物激活体内的外泄 内源性组织--是一个具有挑战性的、缓慢和费力的过程。Exor的下游目标是 通常是未知的，读出exOR的激活通常是不可能的。最近，佩拉尔塔-叶海亚组织 在酵母中开发了基于OR的传感器，用于检测生物技术应用环境中的化学物质， 例如检测水介质中的化学污染物和生物燃料。通过在上面表达人类的OR 酵母细胞表面与连接或配体结合到荧光蛋白表达，这一传感器技术 为OR激活提供快速读数。在这个Mira提案中，我们阐述了基于OR的传感器如何 技术可以加快和简化外源性物质在人类健康中的作用的研究。我们将重点关注 基于OR的传感器技术在研究骨骼肌细胞中存在的Exors中的应用 冒号。在骨骼肌细胞中，我们将确定肌肉细胞中存在的激活内源性化学物质 Exor MOR23和导致肌肉修复时肌原纤维分支减少，导致更强的 修复了肌肉。一旦确定，这种化学物质就可以作为治疗药物合成的先导化合物。 可以促进肌肉愈合。此外，这样的治疗方法也可能在治疗 神经肌肉疾病，可见大量分枝的肌原纤维。在冒号中，我们假设 肠道微生物区系通过Exors与人类宿主沟通，而健康和患病的结肠 可能存在不同的OR激活指纹，可以作为生物标志物来诊断生物状况。 对激活的特定ORs的鉴定将提供分子靶标，以在未来阐明它们的 进一步了解ORs在肠道疾病中的作用。此外，已识别的OR可能 也可以作为治疗靶点；大约40%的药物靶点与G蛋白偶联受体结合 (GPCRs)，其中OR是其中的子集。更广泛地说，知道哪些肠道代谢物与人类相互作用 受体，如ORs，将填补理解微生物群-宿主相互作用的关键空白。作为该计划的一部分 Mira获奖后，我们还将把基于OR的传感器技术与微流体相结合，以使高 微生物产生的非比色/非荧光化学品的吞吐量筛选(&gt；107个样品/天)。 这一筛选能力将允许基因组和进化工程策略的应用，这 已被证明能显著增加比色化学物质的微生物产量。具体来说，我们 将应用该技术来增加微生物生产先进的医药中间体。

7. Project Summary/Abstract Olfactory receptors are one of the largest family of chemical cell-surface receptors in humans, and ectopically expressed olfactory receptors (exORs) have been found in 16 different tissues, including the lungs, kidneys and colon. In non-olfactory tissue, exORs do not detect “odors”, but drive a number of biological process, including chemotaxis, development, and angiogenesis. Understanding the role of exORs in human health—including which exORs are medically relevant, and what compounds activate exORs in the endogenous tissues—is a challenging, slow and laborious process. The downstream targets of exORs are often not known and reading out exORs activation is often not possible. Recently, the Peralta-Yahya group has developed OR-based sensors in yeast to detect chemicals in the environment for biotechnology applications, such as the detection of chemical pollutants and biofuels in aqueous medium. By expressing human ORs on the yeast cell surface and linking OR ligand binding to fluorescent protein expression, this sensor technology provides a rapid readout for OR activation. In this MIRA proposal, we set forth how the OR-based sensor technology can expedite and streamline the study of the role of exORs in human health. We will focus on applying the OR-based sensor technology to the study of exORs present in skeletal muscle cells and in the colon. In skeletal muscle cells, we will identify the endogenous chemical present in muscle cells that activates the exOR mOR23 and results in decreased myofibril branching upon muscle repair, resulting in a stronger repaired muscle. Once identified, the chemical can serve as a lead compound for the synthesis of therapeutics that improve muscle healing. Further, such a therapeutic may also have implication in the treatment of neuromuscular diseases where a large number of branched myofibrils are seen. In the colon, we hypothesize that the gut microbiota communicates with the human host via exORs and that healthy and diseased colons may different OR activation fingerprints that can be used as biomarkers to diagnose biological conditions. Identification of the specific ORs activated will provide molecular targets to, in the future, elucidate their downstream effectors to further understand the role of ORs in gut disease. Additionally, identified ORs could also function as therapeutic targets; roughly 40% of pharmaceutical targets bind G-protein coupled receptors (GPCRs), of which ORs are a subset. More generally, knowing which gut metabolites interact with human receptors, such as ORs, will fill a crucial gap in understanding the microbiome-host interactions. As part of the MIRA award, we will also interphase the OR-based sensor technology with microfluidics to enable the high- throughput screening (>107 samples/day) of non-colorimetric/non-fluorescent chemicals produced by microbes. This screening throughput will allow the application of genome and evolutionary engineering strategies, which have been shown to dramatically increase the microbial production of colorimetric chemicals. Specifically, we will apply the technology to increase the microbial production of advanced pharmaceutical intermediates.