Studies of Materials with Physiological Properties

这项研究的目标是最大限度地提高研究者的研究奖（MIRA）是发展变革性的 发现用于探索生物学和推进医学的新型小分子的能力。项目 旨在建立在NIGMS 038627 32年资助和HHMI 23年支持的研究基础上 （2018年11月结束）。尽管这项研究为生物医学的几项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存在重大差距。 克服这些局限性是拟议研究的主要目标。 在过去五年的GM 038627资助的研究中，我们使用了以下战略规划原则： 多样性导向合成（DOS）来合成一系列新颖的、排列的小分子。超过100个细胞- 进行了基于表型的筛选，产生了许多新的小分子探针和候选物， 新疗法。我们现在建议将现代不对称合成的应用扩展到发现 结合靶蛋白的化合物。发现这种“粘合剂”仍然是具有挑战性的第一步， 开发在结合后赋予靶特异性活性的化合物-这是 以人类遗传学和功能基因组学为指导的精准医学。为了推进小行星的发现- 分子粘合剂，我们建议开创DNA寡核苷酸编码的多样性导向合成 （DOSEDO）通过合成由DOS途径产生的DNA条形码化合物。最终的DNA- 编码的文库（DEL）将与标记的靶蛋白一起孵育以亲和纯化结合物，其可以是 使用DNA测序进行“解码”。该项目旨在将DOS的进步与DEL的进步融合在一起， 同时受益于当代合成化学的巨大进步。 第二个关键限制是发现新合成的小分子的活动需要很长时间。 分子。在过去的五年里，我们引入了小分子实时注释的概念， 使用细胞绘画，在几天内对新合成化合物进行超过1,000次细胞测量， 合成.我们提出的研究旨在开发一种同样廉价和互补的方法，用于真实的- 时间注释-测量化合物诱导的数千种细胞mRNA相对水平的变化 在单个RNA-Seq实验之前，通过条形码编码和汇集转录物（“HiTSeq”）来检测转录物。 最后，我们将测试DOSEDO，HiTSeq和其他在MIRA过程中开发的方法， 关注我们最近发现的癌细胞对化疗产生抵抗力的细胞状态， py，靶向治疗和免疫治疗。这种脂肪吞噬诱导的成肌纤维细胞状态及其 在我们过去五年的GM 038627资助的研究中发现了一个可药物化的漏洞。我们将 利用这种基于脂质的途径作为新方法的试验场，以发现小分子探针， 新的生物学见解，并发现可以用小分子靶向的新的依赖性。

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.