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

 描述(申请人提供)：血清中的纳米颗粒在生物过程中发挥关键作用，包括细胞间信号、免疫反应和癌症等重要疾病的病理生理。一种这样的疾病是慢性肾脏疾病-矿物质和骨骼疾病(CKD-MBD)-这是一种临床实体，在大约2000万患有CKD的美国人中观察到了大多数。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的心血管发病率/死亡率。在CKD-MBD中，血清中形成的可溶纳米级钙蛋白颗粒(CPP)通过隔离钙磷晶体并阻止它们在血管中沉淀来减轻高磷血症的毒性。最近令人兴奋的发现表明，CPP浓度随着CKD的进展而增加，在肾功能正常的患者中并不常见。因此，CPPS可能是一种新的生物标志物，用于在血管系统开始不可逆矿化之前识别进行性血管钙化的高风险患者。然而，到目前为止，由于无法在临床样本中有效地测量这些纳米颗粒，CPP的表征一直受到阻碍。Spectradyne开发了一种功能强大的平台仪器nCS1，它可以精确测量血清等复杂基质中多分散纳米颗粒的大小和浓度。这项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，建立该仪器的一个版本，用于定量测定血清中的CPP。联合小组将共同研究利用血清样本中颗粒的大小分布来 预测小鼠和人类血管钙化的风险，长期目标是改进对这种情况的诊断和预防。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针对测量血清中的纳米颗粒进行优化。对于这项研究，弹药筒的产量也将增加。目的2将评估重新配置的仪器测量血管钙化高危人群和小鼠血清中CPP的能力。为了实现这一目标，我们将：A)在两个小鼠模型中量化血清CPP浓度和相关的血管钙化；以及B)比较CKD患者和健康对照组的血清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种用户友好的台式仪器，能够快速、高分辨率地分析血清样本中的颗粒。此外，还将评估血清CPPs定量仪器的实用性，并确定这一新的生物标志物在血管钙化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.