Developing Choroid Plexus-Based Tools and Drug Screens

虽然脉络丛以分泌脑脊液而闻名，但它吸引的血液相对较少。 基础和临床科学家的关注。每天，人脉络丛上皮细胞(CPECs)分泌 大约两杯富含蛋白质的脑脊液，它可以清除废物，并将有益的分子输送到 大脑和脊髓中的每一个细胞。中巴经济走廊的缺陷牵涉到大量主要的CNS 精神障碍，包括脑积水、精神疾病和神经退行性疾病。然而，几乎 由于缺乏针对中巴经济走廊的工具，所有与中巴经济走廊生物学和疾病有关的文献都是描述性的。 重要的是，如果存在这样的工具，将化合物输送到CPECs-无论是在实验动物中还是在患者中 -很简单，因为CPECs与外周循环自由交换。这极大地简化了药物 并增强了CPECs作为中枢神经系统研究和疾病治疗的新目标的吸引力。尽管 这一呼吁，中巴经济走廊目标药物筛查由于宣传和派生困难而无法实现 他们在文化中。然而，我们最近开发了一种从以下位置生成派生CPEC(DCPEC)的方法 小鼠和人类胚胎干细胞。这种方法第一次提供了一种可伸缩的方法来生产 高通量筛选(HTS)所需的大量dCPEC。 在这份R21提案中，我们将dCPEC技术发明者与化验开发专家结合起来 和药物筛选，开发高通量的DCPEC筛选平台。在初步研究中，我们 描述dCPEC的产生、要筛选的HTS兼容性候选细胞系以及灵敏的96井 分泌型人TTR的ELISA检测是检测CPEC分泌的理想指标，也是ITS的重要分子靶点 这是你的权利。TTRELISA将转换为与HTS兼容的AlphaLISA，将用于飞行员 筛查以确定化验准备情况。然后将筛选来自Pilot屏幕的点击和模拟，并 通过包括新的和已经建立的中巴经济走廊功能、机制、 和专一性。这一经过验证的高吞吐量dCPEC平台将为全面筛选奠定基础 提议为全新的基于中巴经济走廊的生物医学产生一流的工具和治疗线索。

Although famous for secreting the cerebrospinal fluid (CSF), the choroid plexus has drawn relatively little attention from basic and clinical scientists. Every day, human choroid plexus epithelial cells (CPECs) secrete about two cups of protein-rich CSF, which clears out waste products from and delivers beneficial molecules to every cell in the brain and spinal cord. CPEC defects have been implicated in a large number of major CNS disorders, including hydrocephalus, psychiatric conditions, and neurodegenerative diseases. However, almost all of the literature related to CPEC biology and disease is descriptive due to a lack of CPEC-targeted tools. Importantly, if such tools existed, delivery of compounds to CPECs - in either experimental animals or patients - is straightforward, because CPECs exchange freely with the peripheral circulation. This greatly simplifies drug delivery and strengthens the appeal of CPECs as a new target for CNS studies and disease therapies. Despite this appeal, CPEC-targeted drug screens have not been possible due to difficulties in propagating and deriving them in culture. However, we recently developed a method for generating derived CPECs (dCPECs) from mouse and human embryonic stem cells. This method provides, for the first time, a scalable means to produce dCPECs in the large numbers required for high-throughput screening (HTS). In this R21 proposal, we combine the dCPEC technology inventor with experts in assay development and drug screening to develop a high-throughput dCPEC screening platform. In preliminary studies, we describe dCPEC generation, candidate cell lines to be screened for HTS compatibility, and a sensitive 96-well ELISA for secreted human TTR, an ideal proxy for CPEC secretion and an important molecular target in its own right. The TTR ELISA will be converted into an HTS-compatible AlphaLISA, which will be used for a pilot screen to determine assay readiness. Hits and analogs from the pilot screen will then be screened and validated via "testing funnels" that include new and already-established assays of CPEC function, mechanism, and specificity. This validated high-throughput dCPEC platform will set the stage for full-scale screening proposals to generate first-in-class tool and therapy leads for a brand-new CPEC-based biomedicine.