Studies of Materials with Physiological Properties

这个最大化调查人员研究奖(MIRA)的研究目标是开发变革性的 发现用于探索生物学和推进医学的新型小分子的能力。该项目 旨在建立在NIGMS 038627 32年的资助和HHMI 23年的支持下实现的研究 (截至2018年11月)。尽管这项研究对生物医学的几个重要进展做出了贡献，但它也 揭示了我们将人类生物学的见解转化为公共卫生新药的能力方面的重大差距。 克服这些限制是拟议研究的主要目标。 在过去五年由GM038627资助的研究中，我们使用了战略规划原则 以多样性为导向的合成(DOS)，以合成一系列新颖的排列的小分子。超过100个手机- 基于表型的筛选产生了许多新的小分子探针和候选的 新的治疗方法。我们现在建议将现代不对称合成的这种用途扩展到发现 结合目标蛋白的化合物。发现这样的“活页夹”仍然是迈向 开发在结合后赋予目标特定活性的化合物--这是 以人类遗传学和功能基因组学为指导的精准医学。为了推进小型-- 分子结合剂，我们建议开创DNA寡核苷酸编码的多样性合成的先河 (DOSEDO)通过合成由DOS途径产生的DNA条码化合物。由此产生的DNA- 编码库(Del)将与标记的目标蛋白孵育，以亲和纯化结合子，这可以 用DNA测序法“破译”。该项目旨在将DOS的进步与DELS的进步相融合，但 同时受益于当代合成化学令人印象深刻的进步。 第二个关键限制是需要很长时间才能发现新合成的Small的活性 分子。在过去的五年里，我们引入了小分子实时注释的概念， 使用细胞涂色在合成化合物的几天内进行&gt；1000个细胞测量 综合。我们建议的研究旨在开发一种同样廉价和互补的真实- 时间注释-测量化合物引起的数千个细胞mRNA相对水平的变化 在单一的RNA-Seq实验(`HiTSeq‘)之前，通过条形码和汇集转录本获得转录本。 最后，我们将通过以下方式测试DOSEDO、HiTSeq和其他在本次Mira过程中开发的方法 聚焦于我们最近展示的一种细胞状态，癌细胞采用这种状态来赋予对化疗药物的抵抗力- PY、靶向治疗和免疫治疗。这种脂噬诱导的肌成纤维细胞状态及其 在我们GM038627资助的研究的过去五年中发现了可药物漏洞。我们会 利用这种基于脂质的途径作为发现小分子探针的新方法的试验场，以获得 新的生物学洞察力，并发现可以用小分子靶向的新依赖关系。

The goal of the research in this Maximizing Investigators' Research Award (MIRA) is to develop transformative capabilities for discovering novel small molecules used to explore biology and advance medicine. The project aims to build on research enabled by 32 years of funding by NIGMS 038627 and 23 years of support by HHMI (ending Nov 2018). Although this research contributed to several important advances in biomedicine, it also revealed major gaps in our ability to translate insights from human biology into new medicines for public health. Overcoming these limitations is a primary aim of the proposed research. During the past five years of our GM038627-funded research, we used strategic planning principles of diversity-oriented synthesis (DOS) to synthesize a collection of novel, arrayed small molecules. Over 100 cell- based phenotypic screens were performed that yielded many novel small-molecule probes and candidates for novel therapeutics. We now propose to extend this use of modern, asymmetric synthesis to the discovery of compounds that bind target proteins. Discovering such `binders' remains a challenging first step towards developing compounds that confer specific activities on the target following binding – a key shortcoming in precision medicine guided by human genetics and functional genomics. To advance the discovery of small- molecule binders, we propose to pioneer diversity-oriented synthesis encoded by DNA oligonucleotides (DOSEDO) by synthesizing DNA bar-coded compounds resulting from DOS pathways. The resulting DNA- encoded libraries (DELs) will be incubated with tagged target proteins to affinity purify binders, which can be `decoded' using DNA sequencing. This project aims to meld advances in DOS with those of DELs, but benefitting simultaneously from the impressive advance of contemporary synthetic chemistry. A second key limitation has been the long time required to uncover the activities of newly synthesized small molecules. During the past five years, we introduced the concept of real-time annotation of small molecules, using cell painting to make > 1,000 cellular measurements of novel synthetic compounds within days of their synthesis. Our proposed research aims to develop an equally inexpensive and complementary method of real- time annotation – measuring compound-induced changes in the relative levels of thousands of cellular mRNA transcripts by bar-coding and pooling transcripts prior to a single RNA-Seq experiment (`HiTSeq'). Lastly, we will test the methods of DOSEDO, HiTSeq and others developed in the course of this MIRA by focusing on a cell state we recently showed to be adopted by cancer cells to confer resistance to chemothera- py, targeted therapeutics and immunotherapy. This lipophagy-induced, myofibroblastic cell state and its druggable vulnerability was discovered during the past five years of our GM038627-funded research. We will use this lipid-based pathway as a testing ground for new methods to discover small-molecule probes, to gain new biological insights, and to uncover novel dependencies that can be targeted with small molecules.