One-compound, one-islet: A high-throughput platform for small-molecule discovery

一种化合物，一种胰岛：用于小分子发现的高通量平台

基本信息

批准号：
10450745
负责人：
ROHIT N. KULKARNI
金额：
$ 74.66万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-20 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10450745
关键词：
Apoptosis Autoimmune Diabetes Beta Cell Biological Biological Assay CASP3 gene Cell Count Cell Culture Techniques Cell Line Cell Proliferation Cell Survival Cell physiology Cells Cellular biology Chemicals Collection Communities Cytokine Suppression Deoxyuridine Ensure Fluorescence Glucose Goals Harmine Human Image Inbred NOD Mice Individual Inflammatory Institutes Insulin-Dependent Diabetes Mellitus Islet Cell Label Measurement Measures Methods Miniaturization Mission Monitor Mus Natural regeneration Nonesterified Fatty Acids Outcome Performance Permeability Phenotype Physiological Prodrugs Proteins Research Personnel Resort Resources Rodent S-nitro-N-acetylpenicillamine Sampling Ships Signal Transduction Specificity System Testing Thymidine Transplantation VAMP-2 Vesicle Zinc analog autoimmune pathogenesis base cell regeneration cell type cost cytokine endoplasmic reticulum stress high throughput screening in vivo insulin secretion islet novel response screening small molecule tool

项目摘要

PROJECT SUMMARY Methods to increase or protect beta-cell mass in vivo would have a substantial impact on T1D, where beta cells are selectively targeted for autoimmune destruction. The discovery of novel small molecules represents an attractive opportunity to protect and increase beta-cell mass in vivo, as chemical compounds provide temporal control, tunability, cell permeability, and reversibility. However, current methods to identify new compounds are relatively ad hoc and small in scale. These limitations have led to compromises in the number of compounds that can be screened. Here, we propose to leverage our expertise in chemical and cell biology to develop a high- throughput platform (“one-compound, one-islet”) for small-molecule discovery that enables the use of only a single islet to test each compound. Such an advance will allow us to screen many more compounds with each islet shipment, reducing the number of donors required, and increasing scientific rigor substantially by supporting multiple donors to be tested per compound as well. We will adapt three existing readouts for this purpose. First, we have previously developed a zinc-catalyzed prodrug system for beta cell-selective compound delivery. To measure proliferation, we will extend this synthetic system to the thymidine analog 5-ethynyl-2’-deoxyuridine (EdU). A beta cell-selective EdU will reduce the background from other proliferative cells and enable us to image intact islets after small-molecule treatment. For beta-cell survival, we will measure caspase-3 activation, which is specific to beta cells after cytokine treatment. Finally, to measure insulin secretion, we have developed a novel fluorescence-based assay that will allow us to monitor insulin secretion by imaging islets. The successful outcome of this proposal is a platform of small-molecule assays and chemical tools for understanding and promoting beta-cell regeneration and survival. A key advantage of this proposal is that the phenotypic readouts have already been fully developed, and will benefit tremendously from miniaturization to single-islet format. We are committed to making this platform available to the HIRN community, enabling investigators a) to probe their own compounds in this single-islet format, or b) to use a new readout to evaluate compounds screened in this project.

项目摘要在体内增加或保护β细胞质量的方法将对beta细胞的T1D产生重大影响选择性地针对自身免疫性破坏。新颖的小分子的发现代表在体内保护和增加β细胞质量的有吸引力的机会，因为化合物提供了临时性控制，可密钉性，渗透率和可逆性。但是，识别新化合物的当前方法是相关的临时规模和规模较小。这些限制导致化合物数量的妥协可以筛选。在这里，我们建议利用我们在化学和细胞生物学方面的专业知识来发展高用于小分子发现的吞吐量平台（“单一补充，一islet”），仅能使用单个胰岛测试每种化合物。这样的进步将使我们能够筛选更多的化合物胰岛发货，减少所需的捐助者数量，并通过支持大大增加科学严谨性每个化合物也要测试的多个供体。为此，我们将调整三个现有读数。第一的，我们以前已经开发了用于β细胞选择性化合物递送的锌催化前药系统。到测量增殖，我们将把这个合成系统扩展到胸苷类似物5-乙基-2'-脱氧尿苷（edu）。 Beta细胞选择性EDU将减少其他增殖物细胞的背景，并使我们能够形象小分子处理后完整的胰岛。对于β细胞存活，我们将测量caspase-3激活，这在细胞因子治疗后特定于β细胞。最后，为了衡量胰岛素分泌，我们开发了一种小说基于荧光的测定法可以使我们能够通过成像胰岛监测胰岛素分泌。成功该提案的结果是小型分子测定和化学工具的平台，用于理解和促进β细胞再生和生存。该建议的关键优势是表型读数已经完全开发了，并且将从小型化到单志式格式中受益。我们致力于使hirn社区提供此平台，使调查人员a）调查他们以这种单志式格式或b）使用新的读数来评估筛选的化合物项目。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Phenotypic Screening for Small Molecules that Protect β-Cells from Glucolipotoxicity.

DOI：
10.1021/acschembio.2c00052
发表时间：
2022-05-20
期刊：
ACS CHEMICAL BIOLOGY
影响因子：
4
作者：
Small, Jonnell C.;Joblin-Mills, Aidan;Carbone, Kaycee;Kost-Alimova, Maria;Ayukawa, Kumiko;Khodier, Carol;Dancik, Vlado;Clemons, Paul A.;Munkacsi, Andrew B.;Wagner, Bridget K.
通讯作者：
Wagner, Bridget K.