Deciphering Mechanisms of Nitrogen-Containing Bisphosphonates

含氮双膦酸盐的破译机制

• 批准号: 10531297
• 负责人: Lauren Elizabeth Surface
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10531297
• 关键词: Advisory Committees; Affect; Age; American; Award; Binding; Biological Assay; Biology; Bone Density; Bone Diseases; Bone necrosis; CRISPR interference; Cells; Cellular Assay; Charge; Cytosol; Drug Prescriptions; Endocrine; Endocrinology; Epigenetic Process; Faculty; Femoral Fractures; Fright; Future; Genes; Genetic Screening; Genetic Transcription; Genetic study; Goals; Grant; Hospitals; Immersion; Impairment; In Vitro; Investigation; Jaw; Kidney; Knock-out; Knockout Mice; Leadership; Manuscripts; Mediating; Mentors; Mentorship; Modeling; Molecular; Molecular Target; Mus; NFIC gene; Nitrogen; Osteoclasts; Osteopenia; Osteoporosis; Ovariectomy; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Phase; Physiologic calcification; Physiological; Play; Population; Protein Biochemistry; Proteins; Publishing; Quality of life; Research; Research Institute; Research Personnel; Resistance; Risk; Role; Skeletal system; Technical Expertise; Time; Toxic effect; Training; Training Support; Treatment Efficacy; Treatment Side Effects; Wild Type Mouse; Work; Writing; base; bisphosphonate; bone; bone cell; bone loss; bone mass; career; dosage; drug mechanism; genome wide association study; genome wide screen; interest; member; mouse model; patient population; patient response; prevent; programs; public health relevance; response; side effect; skills; standard care; tenure track; therapy development; trafficking; transcription factor; treatment strategy

Project Summary/Abstract Osteoporosis and low bone mass (osteopenia) are estimated to affect 55 percent of the American population over the age of 50; over 50 million people in total, with major consequences for the patients' quality of life. The current standard treatment for osteoporosis is administration of nitrogen-containing bisphosphonates (NBPs). However, the mechanism by which these highly-charged drugs enter, traffic through, and reach their molecular targets and effect target cells is poorly understood. The long-term goal of this proposal is to deconstruct the molecular pathways essential for NBP response. To do this, I will build upon preliminary genetic studies by using cell assays and mouse models, as well as in vitro binding and functional assays to explore the interactions between NBPs and my identified targets. Our previous work utilized two distinct high-throughput genome-wide screens to identify over 200 genes required for the action of NBPs. In two recent manuscripts, I have initially focused on the role of two genes, ATRAID and SLC37A3, that strongly affect the response to NBPs, and found them likely to be required for the endocytic trafficking of these drugs. This proposal builds upon this preliminary work to i) characterize the physiological role of ATRAID and SLC37A3 in the organismal response to NBPs, ii) further examine their basal molecular function and how they facilitate NBP trafficking, and iii) investigate the role of two transcription factors, associated by GWAS with changes in BMD, that when depleted may sensitize cells to the effects of NBPs. Together, these studies generate a broader picture of the molecular pathways that NBP uses to affect cells by investigating other genes identified in our initial screens. While this proposal by necessity focuses on a subset of identified genes, I envision it will set the stage for my future work determining how genes identified in our screens may predict patient response to NBPs, including efficacy of treatment, dosage of NBPs needed, and adverse side effects. Moreover, this focus on understanding the mechanisms of an inexpensive, commonly prescribed drug will bring new perspectives and hypotheses to the development of treatment strategies for osteoporosis. During the early stage of this award, I will gain valuable technical skills, including in analysis of mouse models of osteoporosis, culture of primary bone cells, and biochemistry of protein interactions, as well as a deeper training and immersion in bone and endocrine biology, that will altogether enable me to develop a unique research program, which I intend to establish at a hospital-based research institute. Under the mentoring of my formal advisory committee, I will develop important soft skills, such as presentation skills, lab leadership, and grant writing. This combination of training, support and career mentoring will be instrumental in my transition to independence as a tenure-track faculty member.

项目概要/摘要 据估计，骨质疏松症和低骨量（骨质减少）影响着 55% 的美国人口 50岁以上；总共超过 5000 万人，对患者的生活质量产生了重大影响。这 目前骨质疏松症的标准治疗是给予含氮双膦酸盐（NBP）。 然而，这些高电荷药物进入、运输和到达其分子的机制 对靶标和作用靶细胞知之甚少。该提案的长期目标是解构 NBP 反应所必需的分子途径。为此，我将在初步遗传学研究的基础上 使用细胞测定和小鼠模型以及体外结合和功能测定来探索 NBP 和我确定的目标之间的相互作用。我们之前的工作利用了两种不同的高通量 全基因组筛选，以确定 NBP 发挥作用所需的 200 多个基因。在最近的两篇手稿中，我 最初关注的是两个基因 ATRAID 和 SLC37A3 的作用，它们强烈影响对 NBP，并发现它们可能是这些药物的内吞运输所必需的。该提案构建 根据这项初步工作，i) 描述 ATRAID 和 SLC37A3 在生物体中的生理作用 对 NBP 的反应，ii) 进一步检查它们的基础分子功能以及它们如何促进 NBP 贩运， iii) 研究两个转录因子的作用，通过 GWAS 与 BMD 变化相关，当 耗尽可能会使细胞对 NBP 的作用敏感。这些研究共同产生了更广泛的图景 NBP 通过研究我们初始筛选中发现的其他基因来影响细胞的分子途径。 虽然这个提议必然关注已识别基因的一个子集，但我想它将为我的研究奠定基础。 未来的工作将确定我们筛选中识别的基因如何预测患者对 NBP 的反应，包括 治疗效果、所需 NBP 剂量以及不良副作用。此外，本次重点关注 了解廉价、常用处方药物的机制将带来新的视角和 制定骨质疏松症治疗策略的假设。 在获得该奖项的早期阶段，我将获得宝贵的技术技能，包括小鼠模型的分析 骨质疏松症、原代骨细胞培养、蛋白质相互作用的生物化学以及更深入的研究 骨骼和内分泌生物学的培训和沉浸，这将使我能够开发出独特的 我打算在一家医院研究所建立一个研究计划。在我的指导下 在正式的咨询委员会中，我将培养重要的软技能，例如演讲技巧、实验室领导能力和 授予写作。这种培训、支持和职业指导的结合将有助于我过渡到 作为终身教授的独立性。