Chemical tools for developmental biology

发育生物学化学工具

基本信息

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
项目类别：
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

项目摘要

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水平的科学，也是系统水平的科学。技术已经确定了调节组织形成并有助于其致癌的个体基因高通量测序揭示了区分细胞的“组学”特征。指出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探究这一问题。以新的方式进行组织生物学，由此产生的技术和见解可以带来创新的治疗方法考虑到这些目标，我们的研究小组探索了化学与人类疾病的界面。在过去的四年里，我们开发了一种针对靶细胞的光遗传学系统。消融，鉴定了 Gli 转录因子功能的新型小分子抑制剂，并发现了第一个乙醛脱氢酶 1B1 (ALDH1B1) 的特异性抑制剂，醛脱氢酶 1B1 是一种线粒体酶，在成人肠道和胰腺的多能细胞并促进结直肠癌和胰腺癌。还使用高通量和系统级方法建立了 ARHGAP36 的监管模型，ARHGAP36 是一种非经典的 Gli 激活剂，控制运动神经规范并可诱导髓母细胞瘤。我们现在寻求在这些成就的基础上探索新的科学方向。研究计划将是创建新的光遗传学工具，通过诱导变构而不是为了促进此类构建体的发现，我们建立了一个基于转座子的平台利用这种方法，我们将开发光遗传学调节剂。 Hedgehog 通路的研究，重点关注 Smoothened 的光氧电压 (LOV) 域功能化形式我们将阐明其光依赖性活动的机制基础，优化其功能，并应用这些试剂来研究斑马鱼模型中的 Hedgehog 通路依赖性模式。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。第二个研究重点是 ALDH1 亚型，它在正常干细胞中高度表达，并且是我们将研究 ALDH1B1 在胰腺癌中的作用并进行开发。 ALDH1A3 的特异性抑制剂，ALDH1A3 是一种细胞质酶，可促进乳腺癌、黑色素瘤和此外，我们将寻找针对 ALDH1A1 和/或的小分子。 ALDH1A2，受到这些酶在精子发生中的作用及其作为非激素的潜力的启发总的来说，我们的研究将为发育生物学和新的研究打开新的窗口。通往临床治疗的大门。