Deciphering Mechanisms of Nitrogen-Containing Bisphosphonates

含氮双膦酸盐的破译机制

• 批准号: 10553672
• 负责人: Lauren Elizabeth Surface
• 金额: $ 24.39万
• 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/10553672
• 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; Population; Protein Biochemistry; Proteins; Publishing; Quality of life; Research; Research Institute; Research Personnel; Resistance; Risk; Role; Skeletal system; Technical Expertise; Time; Toxic effect; Training; Treatment Efficacy; Treatment Side Effects; Wild Type Mouse; Work; Writing; 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; 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; Population; Protein Biochemistry; Proteins; Publishing; Quality of life; Research; Research Institute; Research Personnel; Resistance; Risk; Role; Skeletal system; Technical Expertise; Time; Toxic effect; Training; Treatment Efficacy; Treatment Side Effects; Wild Type Mouse; Work; Writing; 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.

项目摘要/摘要 骨质疏松症和低骨骼质量（骨质减少）估计会影响55％的美国人群 超过50岁；总共超过5000万人，对患者的生活质量产生了重大影响。这 当前用于骨质疏松症的标准治疗方法是给药含氮的双膦酸盐（NBPS）。 但是，这些高电荷药物进入，流量并到达分子的机制 靶标细胞尚不清楚靶细胞。该提议的长期目标是解构 NBP反应必不可少的分子途径。为此，我将以初步的遗传研究为基础 使用细胞测定和小鼠模型，以及体外结合和功能测定 NBP与我确定的目标之间的相互作用。我们以前的工作利用了两个不同的高通量 全基因组筛选以识别NBPS作用所需的200多个基因。在最近的两个手稿中，我 最初侧重于两个基因的作用，即Atraid和Slc37a3，这强烈影响对对的反应 NBP，发现这些药物的内吞贩运可能需要它们。该建议建立 在这项初步工作中 对NBP的反应，ii）进一步检查其基础分子功能以及它们如何促进NBP运输， iii）研究了两个转录因子的作用，该因子与BMD的变化相关联，即 耗尽的可能使细胞对NBP的影响敏感。这些研究在一起产生了更广泛的了解 NBP通过研究我们初始筛选中鉴定出的其他基因来影响细胞的分子途径。 虽然该提案必要的重点是确定基因的一部分，但我设想它将为我的舞台奠定 确定屏幕中鉴定基因的未来工作可能会预测患者对NBP的反应，包括 治疗的功效，需要的NBP剂量和不良副作用。此外，关注 了解廉价，常规药物的机制将带来新的观点，并且 假设骨质疏松症的治疗策略的发展。 在该奖项的早期阶段，我将获得宝贵的技术技能，包括分析鼠标模型 骨质疏松症，原代骨细胞的培养和蛋白质相互作用的生物化学以及更深层次的 训练和沉浸在骨骼和内分泌生物学中，这将使我能够发展独特 我打算在医院的研究所建立的研究计划。在我的指导下 正式的咨询委员会，我将发展重要的软技能，例如演示技巧，实验室领导和 授予写作。培训，支持和职业指导的这种结合将有助于我过渡到 独立作为终身教师。