Functional Selection of Novel Ligands from the Neuroendocrine Secretome

描述（由申请人提供）：在后基因组时代，估计有25,000到30,000个基因被认为编码复杂的人类蛋白质组。然而，挑战仍然是要了解从这些看似少数的基因中衍生的大量rna和蛋白质，并了解它们在健康和疾病中的相互作用和功能。RNA剪接和蛋白质加工的改变导致比人类转录组和蛋白质组明显更高的复杂性。在人体中，从基因组到蛋白质组的这种复杂性的飞跃在大脑中表现得最为明显。因此，有巨大的机会发现新的蛋白质和蛋白质在大脑中的功能，具有新的治疗和诊断产品的商业潜力。我们在此建议鉴定新的具有治疗意义的神经内分泌脑蛋白。我们的方法是挖掘下丘脑分泌组，这是蛋白质组中编码分泌蛋白和膜结合蛋白的部分。这类蛋白质是药物靶点和基于蛋白质的治疗方法最丰富的来源之一，包括肽激素、生长因子和细胞因子，它们通过细胞间通讯在维持健康方面发挥关键作用。我们将使用一种新的噬菌体颗粒展示策略，旨在从较大的蛋白质前体中识别活性配体，以识别新的配体及其靶细胞。在我们的一期研究中，我们将对下丘脑进行概念验证实验，因为它是影响人体多个器官系统的调节激素的重要来源。我们的第一个目标是使用信号序列陷阱从人类下丘脑cDNA文库中识别分泌和膜结合蛋白。在我们的第二个目标中，我们将使用噬菌体展示从分泌组中选择活性配体。将在噬菌体颗粒上显示一个片段化的分泌组cDNA文库，我们将在目标细胞系上使用功能筛选从中选择活性配体。在第二阶段，我们将描述表达模式以阐明我们的铅配体的功能意义，我们将从下丘脑中挖掘更多的配体。我们将从基于配体的研究试剂的商业化中获得近期收入，如基于发现的配体的生长因子、抗体和ELISA试剂盒。我们的长期目标是为治疗和诊断开发和III期商业化选择最有前途的铅配体。

DESCRIPTION (provided by applicant): In the post-genomic era, an estimated 25,000 to 30,000 genes are thought to encode the complex human proteome. However, the challenge remains to understand the multitude of RNAs and proteins derived from this deceptively small number of genes and to understand their interactions and function in health and in disease. Alterations in RNA splicing and protein processing result in the much higher complexity than is evident in the human transcriptome and in turn the proteome. Nowhere in the human body is this leap in complexity from genome to proteome more evident than in the brain. There is therefore tremendous opportunity for discovering new proteins and protein function in the brain with commercial potential for novel therapeutic and diagnostic products. We propose here to identify novel neuroendocrine brain proteins of therapeutic significance. Our approach will be to mine the hypothalamic secretome, the portion of the proteome that encodes secreted and membrane bound proteins. This class of proteins, which is one of the most abundant sources of drug targets and protein based therapeutics, includes peptide hormones, growth factors and cytokines which play a critical role in maintaining health through cell-cell communication. We will use a novel phage particle display strategy designed to identify active ligands from within larger protein precursors to identify novel ligands and their target cells. In our phase one study, we will perform proof of concept experiments on the hypothalamus as it is an important source of regulatory hormones that affect multiple organ systems in the body. Our first aim will be to identify secreted and membrane bound proteins from a human hypothalamic cDNA library using a signal sequence trap. In our second aim we will select active ligands from the secretome using phage display. A fragmented secretome cDNA library will be displayed on phage particles from which we will select the active ligands using functional screens on target cell lines. In phase II, we will characterize the expression pattern to elucidate the functional significance of our lead ligands and we will mine additional ligands from the hypothalamus. We will develop near term revenue from commercialization of ligand based research reagents, such as growth factors, antibodies, and ELISA kits based on the discovered ligands. Our long term goal will be to select the most promising lead ligands for therapeutic and diagnostic development and phase III commercialization. PUBLIC HEALTH RELEVANCE: In the post-genomic era the challenge is to fully understand the protein products of the human genome and how these function in health and disease. One therapeutically important class of proteins is the so called "secretome", those proteins that are secreted from cells and are important for cell to cell communication. We propose to identify new and novel secreted proteins made in the hypothalamus, a complex brain region that links the body's perception of its environment with its regulation of body functions such as temperature, blood pressure, appetite, digestion, and sleep. It regulates the master gland of the body, the pituitary, and effects emotions and behavior. We will identify new proteins and active protein fragments from the hypothalamic secretome using novel protein display and selection technology. We will develop and commercialize antibodies and detection kits for the novel proteins and assess their diagnostic and therapeutic potential.