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

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.

项目总结/摘要 据估计，骨质疏松症和低骨量（骨质减少）影响了55%的美国人口 年龄在50岁以上;总共超过5000万人，对患者的生活质量产生重大影响。的 目前骨质疏松症的标准治疗是施用含氮的二膦酸盐（NBP）。 然而，这些高电荷药物进入、穿过并到达其分子的机制是不确定的。 靶点和作用靶细胞的认识很少。这项提案的长期目标是解构 NBP反应所必需的分子途径。为了做到这一点，我将建立在初步的遗传研究基础上， 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。我们以前的工作利用了两个不同的高通量 全基因组筛选，以确定NBP作用所需的200多个基因。在最近的两份手稿中，我 他们最初关注的是两个基因的作用，ATRAID和SLC 37 A3，这两个基因强烈影响对 NBP，并发现他们可能需要这些药物的内吞贩运。这一建议建立 i）表征ATRAID和SLC 37 A3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本分子功能以及它们如何促进NBP的运输， 和iii）研究两种转录因子的作用，这两种转录因子与GWAS和BMD的变化相关， 耗尽的蛋白质可以使细胞对NBP的作用敏感。总之，这些研究产生了一个更广泛的图片， NBP通过研究我们最初筛选中鉴定的其他基因来影响细胞的分子途径。 虽然这个建议的必要性集中在一个子集的确定基因，我设想它将设置阶段，我的 未来的工作，确定如何在我们的屏幕上识别的基因可以预测患者对NBP的反应，包括 治疗效果、所需NBP剂量和不良副作用。此外，这种专注于 了解一种廉价的、常用的处方药的机制将带来新的视角， 骨质疏松症的治疗策略的发展假设。 在这个奖项的早期阶段，我将获得宝贵的技术技能，包括在小鼠模型的分析 骨质疏松症，原代骨细胞的培养，蛋白质相互作用的生物化学，以及更深层次的 培训和沉浸在骨骼和内分泌生物学，这将使我能够开发一个独特的 研究计划，我打算在医院的研究所建立。在我的指导下， 正式咨询委员会，我将发展重要的软技能，如演讲技巧，实验室领导， 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 作为终身教职员工的独立性。