Role of Nanobacteria in Human Calcifying Disease

描述（由申请人提供）：炎症是人类钙化性疾病的特征，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，一种新型的，生长缓慢的微生物，形成一个矿化的外壳，可以从人类和实验动物的患病，钙化组织分离。因此，这一提议的中心假设是，纳米细菌是这些人类疾病中以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生物体，并开发用于评估人类和实验组织的确定性遗传测试。一个多学科的团队已经创建了长期目标，从人类和实验动物的组织中分离纳米细菌，在培养物中培养分离物，然后用生物体培养实验动物和细胞，以重现钙化反应，这是满足科赫假设所需的。这项拨款申请的独特优势包括在实验室医学方面的丰富经验，用于提取，浓缩和纯化核酸，开发传染病病原体的快速分子诊断测试，以及使用低温微计算机断层扫描（CT），允许3D成像，组织形态学/免疫学成像和单个样本的遗传探测。这是一个高风险的提议，但如果纳米细菌在患病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发生革命性变化，高影响的后果可能会随之而来，包括这些和其他可能的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.