Mechanism of calcium phosphate stone formation in engineered 3D tubule

工程 3D 肾小管中磷酸钙结石形成机制

• 批准号: 9851212
• 负责人: Bidhan Chandra Bandyopadhyay
• 金额: $ 5.09万
• 依托单位: INSTITUTE FOR CLINICAL RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9851212
• 关键词: Mechanism; calcium; phosphate; stone; formation

Renal Calcium phosphate (CaP) and CaP plus Calcium Oxalate (CaOx) mixed crystal biomineralization results in diseases such as nephrolithiasis, which affect over 10% of adults in the United States, accounting for over $5 billion in economic costs in this country each year. The recurrence rate of nephrolithiasis is often high, reaching almost 50% over a 5-year period. Despite extensive research being conducted over the past century, the mechanism by which crystals nucleate, grow, and aggregate into stones is still poorly understood. This program seeks to improve our understanding of the origin and mechanism of such biologically controlled mineralization by developing a novel microfluidic-based workbench that can simulate the dynamical biological conditions of an in vivo renal tubular system. We hypothesize that mimicking the process of in vivo CaP deposition within this ex vivo tubular model created in microfluidics will enable us to systematically evaluate the multifactorial mechanism of CaP crystal formation by analyzing the contribution of each dynamic microenvironmental cue (e.g., cellular regulations, fluidic hydrodynamics and physiochemical interactions). Further, we believe that the combination of in situ characterization techniques and in vivo-like 3D tubular microenvironment generated in microfluidics will enable us to achieve a better understanding of the process of CaP stone formation at molecular and cellular level. Our program is novel in its approach to examine the mechanism of CaP stone formation in in vivo-like tubules with continuous renal fluidic flow. Unlike previous attempts, we propose to in situ map the compositions of renal fluids and solid mineral depositions in renal tubular structures with continuous flow of fluids, which will enable us to capture the most relevant molecular, cellular and hydrodynamic information that are not attainable by conventional approaches. Further, our specialized ex vivo tools can dissect the details of the complex in vivo event, revolutionizing our understanding of stone formation at the systems level. This research program involves two objectives: i) To understand the cellular interaction with CaP crystals under the influence of hydrodynamic renal fluid flow within 3D ex vivo renal tubular structure engineered in microfluidics, and ii) To understand the role of physiologically and clinically relevant molecules in the retention, dissolution, growth of CaP and/or CaOx crystals within the MF based ex vivo renal tubular structure. The proposed study will lead us to i) find strategies to prevent and treat calcium stone disease in kidney; ii) gain new insights on the abnormal CaP deposition in soft tissues (namely, extra-skeletal calcification); and iii) open up exciting research fronts in understanding the molecular and pharmacological basis of CaP stone formation in vascular and other similar cellular microenvironments. Because the approach is so new, we are requesting an exploratory grant to develop the enabling techniques required for the elucidation of the general mechanism of stone formation. Our objective is to demonstrate the feasibility of our approach from a fundamental science, engineering and biological perspective.

肾磷酸钙 (CaP) 和 CaP 加草酸钙 (CaOx) 混合晶体生物矿化结果 肾结石等疾病影响了美国 10% 以上的成年人，占 这个国家每年的经济损失达 50 亿美元。肾结石的复发率往往很高， 5年内达到近50%。尽管在过去的一个世纪里进行了广泛的研究， 晶体成核、生长和聚集成石头的机制仍然知之甚少。这 计划旨在提高我们对此类生物控制的起源和机制的理解 通过开发一种新型的基于微流体的工作台来模拟动态生物矿化 体内肾小管系统的条件。我们假设模仿体内 CaP 的过程 在微流体中创建的离体管状模型中的沉积将使我们能够系统地评估 通过分析各个动态的贡献，研究 CaP 晶体形成的多因素机制 微环境线索（例如细胞调节、流体流体动力学和理化相互作用）。 此外，我们相信原位表征技术和体内类 3D 管状结构的结合 微流控中产生的微环境将使我们能够更好地理解微流体的过程 分子和细胞水平上的 CaP 结石形成。我们的程序在检查方法上是新颖的 具有连续肾液流的体内样小管中 CaP 结石形成的机制。与之前不同 尝试，我们建议原位绘制肾液的成分和肾内固体矿物质沉积物 具有连续流体流动的管状结构，这将使我们能够捕获最相关的分子， 通过传统方法无法获得的细胞和流体动力学信息。此外，我们的 专门的离体工具可以剖析复杂的体内事件的细节，彻底改变我们的理解 系统层面的结石形成。该研究计划涉及两个目标： i) 了解 3D 离体肾液流体动力学影响下细胞与 CaP 晶体的相互作用 微流体工程设计的肾小管结构，以及 ii) 了解生理和功能的作用 MF 内 CaP 和/或 CaOx 晶体保留、溶解、生长的临床相关分子 基于离体肾小管结构。拟议的研究将引导我们 i) 找到预防和治疗的策略 肾脏钙结石病； ii) 获得关于软组织中异常 CaP 沉积的新见解（即， 骨外钙化）； iii) 开辟令人兴奋的研究前沿，以了解分子和 血管和其他类似细胞微环境中 CaP 结石形成的药理学基础。 由于该方法非常新，我们正在请求探索性拨款来开发支持技术 阐明结石形成的一般机制所需。我们的目标是展示 从基础科学、工程和生物学的角度来看我们的方法的可行性。