Chemical tools for developmental biology

发育生物学化学工具

• 批准号: 10623101
• 负责人: JAMES K CHEN
• 金额: $ 62.5万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10623101
• 关键词: ALDH1A2 gene; Ablation; Adult; Biology; Bone Morphogenetic Proteins; Cells; Chemicals; Chemistry; Clinical; Colorectal Cancer; Cytoplasm; Development; Developmental Biology; Enzymes; Erinaceidae; Evolution; GLI gene; Genes; Genetic; Glioblastoma; Growth; High-Throughput Nucleotide Sequencing; Individual; Intestines; Knowledge; Life; Light; Male Contraceptive Agents; Malignant Neoplasms; Malignant neoplasm of pancreas; Mitochondria; Modeling; Molecular; Motor Neurons; Oncogenic; Oxygen; Pancreas; Pathway interactions; Pattern; Photoreceptors; Protein Engineering; Protein Isoforms; Reagent; Research; Role; Science; Signal Pathway; Signaling Protein; Specific qualifier value; Spermatogenesis; System; Technology; Tissues; Translating; Zebrafish; aldehyde dehydrogenase 1; aldehyde dehydrogenases; chemical synthesis; contraceptive target; empowerment; genetic technology; human disease; inhibitor; innovation; insight; malignant breast neoplasm; medulloblastoma; melanoma; motor control; multipotent cell; novel; optogenetics; programs; reverse genetics; small molecule; small molecule inhibitor; stem cells; stemness; tool; transcription factor; tumor initiation; tumor progression; tumorigenesis; voltage

Developmental biology is now both a molecular and systems-level science. Forward- and reverse-genetic technologies have identified individual genes that regulate tissue formation and contribute to their oncogenic transformation later in life. High-throughput sequencing has revealed the “omic” features that differentiate cellular states. Translating this knowledge into mechanistic understanding will require new scientific approaches, and chemistry can help bridge this gap. Chemical synthesis and protein engineering can empower us to interrogate tissue biology in new ways, and the resulting technologies and insights can lead to innovative treatments for human disease. With these goals in mind, our research group has explored the interface of chemistry and developmental biology. Over the past four years, we have developed an optogenetic system for targeted cell ablation, identified novel small-molecule inhibitors of Gli transcription factor function, and discovered the first specific inhibitors of aldehyde dehydrogenase 1B1 (ALDH1B1), a mitochondrial enzyme that is expressed in multipotent cells of the adult intestine and pancreas and promotes colorectal and pancreatic cancer. We have also used high-throughput and systems-level approaches to establish a regulatory model for ARHGAP36, a non- canonical Gli activator that controls motor neuron specification and can induce medulloblastoma. We now seek to build upon these accomplishments and explore new scientific directions. One focus of our research program will be the creation of new optogenetic tools that act through inducible allostery rather than proximity. To facilitate the discovery of such constructs, we have established a transposon-based platform that recapitulates the evolution of natural photoreceptors. Using this approach, we will develop optogenetic regulators of the Hedgehog pathway, focusing on light-oxygen-voltage (LOV) domain-functionalized forms of Smoothened and GLI1. We will elucidate the mechanistic basis of their light-dependent activities, optimize their functionality, and apply these reagents to study Hedgehog pathway-dependent patterning in zebrafish models. We will also extend this platform to other developmental pathways such as bone morphogenetic protein signaling. Our second research focus will be ALDH1 isoforms that are highly expressed in normal stem cells and integral to tumor initiation and progression. We will investigate the roles of ALDH1B1 in pancreatic cancer and develop specific inhibitors of ALDH1A3, a cytoplasmic enzyme that promotes breast cancer, melanoma, and glioblastoma, and other malignancies. In addition, we will pursue small molecules that target ALDH1A1 and/or ALDH1A2, motivated by the roles of these enzymes in spermatogenesis and their potential as non-hormonal male contraceptive targets. Collectively, our studies will open new windows into developmental biology and new doors to clinical therapies.

发育生物学现在既是分子级科学，又是一种分子级科学。正向和反向遗传 技术已经确定了调节组织形成并有助于其致癌的单个基因 后来的转变。高通量测序已揭示了区分细胞的“ OMIC”特征 国家。将这些知识转化为机械理解将需要新的科学方法，并且 化学可以帮助弥合这一差距。化学合成和蛋白质工程可以使我们能够审问 组织生物学以新的方式以及由此产生的技术和见解会导致创新的治疗方法 人类疾病。考虑到这些目标，我们的研究小组探索了化学的界面和 发展生物学。在过去的四年中，我们开发了一个用于靶向细胞的光遗传系统 消融，鉴定出新型的GLI转录因子功能的小分子抑制剂，并发现了第一个 醛脱氢酶1B1（ALDH1B1）的特异性抑制剂，这是一种线粒体酶 成人肠和胰腺的多态细胞，并促进结直肠癌和胰腺癌。我们有 还使用高通量和系统级别的方法来建立ARHGAP36的监管模型，一种非 - 控制运动神经元规范并诱导髓母细胞瘤的典型GLI激活剂。 我们现在寻求基于这些成就并探索新的科学方向。我们的一个重点 研究计划将创建新的光遗传学工具，这些工具通过可诱导的变构而不是 接近。为了促进这种结构的发现，我们建立了一个基于转座的平台 概括自然感受器的演变。使用这种方法，我们将开发光遗传调节器 刺猬途径，专注于光氧 - 电压（LOV）域功能化的平滑形式 和Gli1。我们将阐明其光依赖性活动的机械基础，优化其功能， 并应用这些试剂在斑马鱼模型中研究刺猬途径依赖性图案。我们也会 将该平台扩展到其他发育途径，例如骨形态发生蛋白信号传导。我们的 第二个研究重点将是在正常干细胞中高度表达的ALDH1同工型，并且不可或缺 肿瘤的启动和进展。我们将研究Aldh1b1在胰腺癌中的作用并发展 ALDH1A3的特异性抑制剂，一种促进乳腺癌，黑色素瘤和 胶质母细胞瘤和其他恶性肿瘤。此外，我们将追求针对aldh1a1和/或的小分子 Aldh1a2，这些酶在精子发生中的作用及其作为非激素的潜力而成熟 男性避孕目标。总的来说，我们的研究将为发展生物学和新的新窗口开放新窗口 临床疗法的门。