Direct detection of nanoparticles in blood: a novel diagnostic for Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD)

 描述（由申请人提供）：血清中的纳米级颗粒在生物过程中发挥关键作用，包括细胞间信号传导、免疫应答和重要疾病（如癌症）的病理生理学。一种这样的疾病是慢性肾脏病-矿物质和骨骼疾病（CKD-MBD）-在约2000万患有CKD的美国人中的大多数中观察到的临床实体。CKD-MBD促进血管钙化，导致进行性心血管功能障碍，并显著导致在该人群中观察到的极高心血管发病率/死亡率。在CKD-MBD中，血清中可溶性纳米级钙蛋白颗粒（CPP）的形成通过螯合磷酸钙晶体并防止其在血管系统中沉淀来减轻高磷酸盐血症的毒性。令人兴奋的最新发现表明，CPP浓度随着CKD进展而增加，并且在肾功能正常的患者中未常规观察到。因此，CPP可能代表一种新的生物标志物，用于在血管系统发生不可逆矿化之前识别具有进行性血管钙化高风险的患者。然而，迄今为止，CPP的表征一直受到无法有效测量临床样品中这些纳米颗粒的阻碍。Spectradyne开发了一种功能强大的平台仪器nCS 1，可精确测量血清等复杂基质中多分散纳米颗粒的尺寸和浓度。该技术非常适合将生物颗粒表征为健康和疾病的指标。Spectradyne建议与堪萨斯大学医学中心（KUMC）的研究人员合作，建立一个版本的这种仪器，用于定量血清中的CPP。联合团队将共同研究使用血清样本中颗粒的大小分布的潜力， 预测小鼠和人类血管钙化的风险，长期目标是改善这种疾病的诊断和预防。为了实现这些目标，将实现两个具体目标。在目标1中，仪器将被配置用于测量血清纳米颗粒：样品制备、仪器本身、分析盒和数据分析软件都将被优化用于测量血清中的纳米颗粒。本研究还将增加笔芯生产。目标2将评估重新配置的仪器测量血管钙化高风险的人和小鼠血清中CPP的能力。为此，我们将：A）在两种鼠模型中定量血清CPP浓度和相关的血管钙化;和B）比较CKD患者和健康对照中的血清CPP水平，并与传统的矿物质代谢标志物相关联。这项工作的完成将产生一个用户友好的台式仪器能够快速，高分辨率的血清样品中的粒子分析。此外，将评估用于定量血清CPP的仪器的实用性，并确定这种新的生物标志物在血管钙化的病理生理学中的作用。

 DESCRIPTION (provided by applicant): Nanoscale particles in serum play critical roles in biological processes, including intercellular signaling, immune response, and the pathophysiology of important diseases such as cancer. One such disease is Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD)-a clinical entity that is observed in a majority of the ~20 million Americans with CKD. CKD-MBD promotes vascular calcification that leads to progressive cardiovascular dysfunction, and significantly contributes to the exceedingly high rate of cardiovascular morbidity/mortality observed in this population. In CKD-MBD, the formation of soluble nanoscale calciprotein particles (CPPs) in serum mitigates the toxicity of hyperphosphatemia by sequestering calcium phosphate crystals and preventing their precipitation in the vasculature. Exciting recent discoveries have shown that CPP concentrations increase with CKD progression, and are not routinely observed in patients with normal kidney function. CPPs may thus represent a novel biomarker for identifying patients at high risk for progressive vascular calcification prior to the onset of irreversible mineralization f the vasculature. However, characterization of CPPs to date has been hindered by an inability to efficiently measure these nanoparticles in clinical samples. Spectradyne has developed a powerful platform instrument, the nCS1, which accurately measures the size and concentration of polydisperse nanoparticles in complex matrices such as serum. The technology is ideally suited to characterize biological particles as indicators of health and disease. Spectradyne proposes to collaborate with researchers at the University of Kansas Medical Center (KUMC) to build a version of this instrument for quantification of CPPs in serum. Together the combined team will investigate the potential of using the size distribution of particles in serum samples to predict the risk for vascular calcification in mice and humans, with the long-term goal of improving diagnosis and prevention of this condition. To accomplish these goals, two specific aims will be met. In Aim 1, the instrument will be configured for measurements of serum nanoparticles: Sample preparation, the instrument itself, the analysis cartridge, and the data analysis software will all be optimized for measuring nanoparticles in serum. Cartridge production will also be increased for the study. Aim 2 will evaluate the ability of the reconfigure instrument to measure CPPs in sera from humans and mice at high risk for vascular calcification. To achieve this we will: A) Quantify serum CPP concentrations and associated vascular calcification in two murine models; and B) Compare serum CPP levels in CKD patients and healthy controls, and associate with traditional mineral metabolism markers. Completion of this work will yield a user-friendly bench top instrument capable of rapid, high-resolution particl analysis in serum samples. Moreover, the utility of the instrument for the quantification of serum CPPs will be evaluated, and the role of this novel biomarker in the pathophysiology of vascular calcification determined.