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至30，000个基因被认为编码复杂的人类蛋白质组。然而，挑战仍然是了解来自这些看似少量的基因的大量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中明显更高的复杂性。从基因组到蛋白质组的这种复杂性飞跃，在人体中没有任何地方比大脑更明显。因此，在大脑中发现新的蛋白质和蛋白质功能存在巨大的机会，具有新型治疗和诊断产品的商业潜力。我们在这里提出，以确定新的神经内分泌脑蛋白的治疗意义。我们的方法将是挖掘下丘脑分泌组，蛋白质组的一部分，编码分泌和膜结合蛋白。这类蛋白质是药物靶标和基于蛋白质的治疗剂的最丰富的来源之一，包括肽激素、生长因子和细胞因子，它们通过细胞-细胞通讯在维持健康中起关键作用。我们将使用一种新的噬菌体颗粒展示策略，旨在从更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。在我们的第一阶段研究中，我们将对下丘脑进行概念验证实验，因为它是影响体内多个器官系统的调节激素的重要来源。我们的第一个目标将是确定分泌和膜结合的蛋白质从人类下丘脑cDNA文库使用的信号序列陷阱。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。一个片段化的分泌组cDNA文库将展示在噬菌体颗粒上，我们将使用靶细胞系上的功能筛选从噬菌体颗粒中选择活性配体。在第二阶段，我们将描述的表达模式，以阐明我们的铅配体的功能意义，我们将挖掘更多的配体从下丘脑。我们将从基于配体的研究试剂的商业化中获得近期收入，如生长因子，抗体和基于所发现配体的ELISA试剂盒。我们的长期目标是选择最有前途的先导配体用于治疗和诊断开发以及III期商业化。 公共卫生相关性：在后基因组时代，挑战是充分了解人类基因组的蛋白质产物以及这些产物在健康和疾病中的作用。一类在治疗上重要的蛋白质是所谓的“分泌组”，即从细胞分泌并对细胞间通讯重要的那些蛋白质。我们建议确定下丘脑中产生的新的和新颖的分泌蛋白，下丘脑是一个复杂的大脑区域，将身体对其环境的感知与其对身体功能的调节（如温度，血压，食欲，消化和睡眠）联系起来。它调节身体的主腺，脑垂体，并影响情绪和行为。我们将使用新的蛋白质展示和选择技术从下丘脑分泌蛋白组中鉴定新的蛋白质和活性蛋白片段。我们将开发和商业化的抗体和检测试剂盒的新蛋白质，并评估其诊断和治疗潜力。

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.