Role of Nanobacteria in Human Calcifying Disease

描述（由申请人提供）：炎症是人类钙化疾病的特征，包括肾结石和动脉粥样硬化。虽然有证据表明感染因子与动脉粥样硬化中存在的炎症有关，但导致钙化的因素仍有待确定。初步数据表明，纳米细菌，一种新的，缓慢生长的微生物，形成矿化外壳，可以从人类和实验动物的病变，钙化组织中分离出来。因此，本提案的中心假设是纳米细菌是这些人类疾病中先前意想不到的致病因素。纳米细菌在科学界仍然存在争议，因为遗传物质尚未被确定。因此，这项为期两年的R-21提案的主要目标是克隆和表征独特的纳米细菌DNA序列，以便明确地识别和分类这种有机体，并为评估人类和实验组织开发明确的基因测试。一个多学科团队已经成立，其长期目标是从人类和实验动物的组织中分离出纳米细菌，在培养皿中培养分离物，然后用这种有机体接种实验动物和细胞来重现钙反应，这是实现科赫假设所必需的。这项拨款申请的明显优势包括在提取、浓缩和纯化核酸和开发传染病病原体快速分子诊断测试的实验室医学方面的丰富经验，以及使用低温微机断层扫描（CT），该技术允许对单个样本进行3D成像、组织形态学/免疫学成像和遗传探测。这是一个高风险的建议，但如果纳米细菌在病变和钙化的肾脏和动脉粥样硬化组织中被常规识别，我们对这些常见人类疾病的理解将会发生革命性的变化，并可能产生高影响的后果，包括针对这些疾病和其他钙化疾病的新治疗方法。

DESCRIPTION (provided by applicant): Inflammation is characteristic of human calcifying diseases, including nephrolithiasis and atherosclerosis. While evidence has linked infectious agents to the inflammation present in atherosclerosis, factors that contribute to the calcification remain to be identified. Preliminary data are presented demonstrating that nanobacteria, a novel, slow-growing microorganism which forms a mineralized shell, can be isolated from diseased, calcified tissues of humans and experimental animals. Therefore, the central hypothesis of this proposal is that nanobacteria are a previously unexpected pathogenic factor in these human diseases. Nanobacteria remain controversial in the scientific community because genetic material has not yet been identified. Therefore, the major goal of this two-year R-21 proposal is to clone and characterize unique nanobacterial DNA sequences in order to unequivocally identify and classify this organism, and to develop definitive genetic tests for evaluating human and experimental tissues. A multidisciplinary team has been created with the long-term goal to isolate nanobacteria from tissue of humans and experimental animals, grow the isolates in culture, and then inoculate experimental animals and cells with the organism to reproduce a calcific response, as required to fulfill Koch's postulates. Distinct advantages of this grant application include the vast experience in Laboratory Medicine for extracting, concentrating, and purifying nucleic acid and developing rapid molecular diagnostic tests for infectious disease agents, as well as the use of cryogenic microcomputer tomography (CT) which allows 3D imaging, histomorphometric/immunological imaging, and genetic probing of a single sample. This is a HIGH-RISK proposal, but if nanobacteria are routinely identified in diseased and calcified renal and atherosclerotic tissue, our understanding of these common human diseases would be revolutionized and HIGH-IMPACT consequences would likely ensue, including new treatment approaches for these and possibly other calcifying diseases.