Rapid Diagnostics for Mucormycosis

毛霉菌病的快速诊断

• 批准号: 8832062
• 负责人: Marwan Nasralla
• 金额: $ 15.11万
• 依托单位: VITALEX BIOSCIENCES, LLC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-09 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8832062
• 关键词: Acidosis; Adrenal Cortex Hormones; Age; Amino Acids; Amphotericin B; Angioinvasion; Animal Model; Animals; Antibodies; Antifungal Agents; Antifungal Therapy; Antigens; Applications Grants; Aspergillus fumigatus; Binding; Biological; Biological Assay; Biological Markers; Blood Vessels; Blood specimen; Bronchoalveolar Lavage; Candida; Cell Line; Cell Surface Proteins; Clinical; Colony-forming units; Data; Detection; Diabetes Mellitus; Diabetic Ketoacidosis; Diagnosis; Diagnostic; Diagnostic tests; Disease; Disease Outcome; Disease Progression; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Feasibility Studies; Goals; Heat shock proteins; Hematogenous; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Human; Immune system; Immunocompromised Host; Incidence; Infection; Invaded; Iron; Life; Liposomes; Malignant Neoplasms; Mammalian Cell; Medical center; Methods; Modeling; Monoclonal Antibodies; Mucorales; Mucormycosis; Mus; Mycoses; Neutropenia; Operative Surgical Procedures; Organ Transplantation; Organism; Oryza; Outcome; Pathogenesis; Patients; Penetration; Pharmaceutical Preparations; Phase; Population; Predictive Value; Prevalence; Proteins; Reporting; Rhizopus; Risk; Risk Factors; Sampling; Self-Sustained Sequence Replication; Sensitivity and Specificity; Serum; Signal Transduction; Small Business Technology Transfer Research; Techniques; Testing; Thrombosis; Tissues; United States; Urine; Western Blotting; Zygomycosis; base; clinically relevant; fungus; glucose-regulated proteins; improved; mortality; mouse model; progression marker; public health relevance; rapid detection; rapid diagnosis; response; usability

 DESCRIPTION (provided by applicant): Mucormycosis, most commonly caused by Rhizopus oryzae, is a life-threatening infection that occurs in patients immunocompromised by diabetic ketoacidosis (DKA), neutropenia, corticosteroid use, and/or increased serum iron. Because of the rising prevalence of these risk factors, the incidence of mucormycosis has risen. Despite disfiguring surgery and aggressive antifungal therapy, the mortality of mucormycosis remains >40%, and approaches 100% in patients with disseminated disease and prolonged neutropenia. A key factor which contributes to these abysmal mortality rates of mucormycosis is the current lack of rapid diagnostic tests. This deficiency often results in delayed therapy. Thus, a rapid diagnostic test for mucormycosis is likely to improve the outcome of the disease. Clinical hallmarks of R. oryzae infection include its remarkable angiotropism. We recently made the important discovery that the fungal cell surface proteins encoded by CotH facilitate disease progression by allowing R. oryzae to invade mammalian cells via binding to Glucose Regulated Protein 78 (GRP78), a heat shock protein expressed on endothelial cells lining blood vessels during mucormycosis. Importantly, CotH proteins were found to be conserved among Mucorales (organisms that cause mucormycosis) with amino acid identity ranging from 55-98%. Equally important CotH proteins are unique to Mucorales since they are absent from any other known organisms. Finally, CotH proteins were found to be expressed in clinically relevant animal models of mucormycosis including the DKA mouse model. These features strongly indicate that CotH and their gene products can be utilized for rapid detection of mucormycosis. Indeed our preliminary data show PCR-related methods using specific primers to CotH can amplify signals in blood samples spiked with different Mucorales but not Aspergillus fumigatus or Candida. Moreover, anti-CotH antibodies can recognize a band similar to the predicted size of CotH proteins in serum samples collected from infected mice. We propose to build on these exciting data to further establish CotH and/or their gene products as biomarkers for diagnosis of mucormycosis, progression of the disease and response to therapy. Our goal in this Phase I feasibility study is to use our established and clinically relevant DKA and neutropenic mucormycosis mouse models to develop a PCR-based assay and/or an antigen detection test targeting CotH in biological samples collected from mice infected with Mucorales. Phase II of this STTR application will focus on establishing a PCR-based kit, sandwiched ELISA and/or dipstick assays for the rapid detection of CotH or circulating CotH antigens by testing using human clinical samples. Establishing a rapid detection test will improve mucormycosis outcome by early initiation of proper therapy and inform the response to antifungal therapy.

 描述（由适用提供）：最常见的是由根瘤菌引起的粘膜菌病，是一种威胁生命的感染，发生在糖尿病性酮症酸中毒（DKA），中性粒细胞减少，皮质类固醇和/或血清铁的患者免疫受损的患者中。由于这些危险因素的患病率上升，粘膜菌病的发生的发生率增加了。尽管手术和侵略性抗真菌疗法毁容，但粘膜菌病的死亡率仍> 40％，并且在散布疾病和延长中性减少症的患者中接近100％。导致粘膜细胞增多的这些糟糕死亡率的关键因素是目前缺乏快速诊断测试。这种缺乏通常会导致治疗延迟。这是对粘膜细胞增多的快速诊断测试可能会改善疾病的结果。 R. oryzae感染的临床标志包括其显着的血管形主义。我们最近提出了一个重要发现，即通过允许蛋黄酱r. oryzae通过与葡萄糖调节蛋白78（GRP78）结合来侵袭哺乳动物细胞的真菌细胞表面蛋白是通过在粘液酸中毒过程中在内皮血管内衬上血管内皮血管上表达的热休克蛋白来侵袭哺乳动物细胞的。重要的是，发现Coth蛋白在粘膜（引起粘膜菌病的生物）中配置，氨基酸同一性范围为55-98％。同样重要的Coth蛋白是粘膜独有的，因为它们不存在任何其他已知生物。最后，发现COTH蛋白在包括DKA小鼠模型在内的临床相关动物模型中表达。这些特征强烈表明COTH及其基因产物可用于快速检测粘膜细胞增多。实际上，我们的初步数据表明，使用特定引物的PCR相关方法可以放大带有不同粘膜的血液样本中的信号，但不能放大烟草烟草或曲霉或念珠菌的信号。此外，抗Coth抗体可以识别类似于从感染小鼠收集的血清样品中的Coth蛋白预测尺寸的条带。我们建议以这些令人兴奋的数据为基础，以进一步建立COTH和/或其基因产品作为诊断粘膜菌病，疾病进展和对治疗反应的生物标志物。我们在这一阶段的I可行性研究中，我们的目标是使用我们已建立的且与临床相关的DKA和中性肉类菌病小鼠模型来开发基于PCR的测定法和/或靶向COTH的抗原检测测试，该测试针对从感染粘膜的小鼠收集的生物样品中的COTH。该STTR应用的II阶段将着重于建立基于PCR的试剂盒，夹心ELISA和/或量强壮的测定法，以快速检测COTH或通过使用人类临床样品进行测试，以快速检测COTH或循环的Coth抗原。建立快速检测测试将通过早期开始适当的治疗来改善粘胶菌病结果，并告知对抗真菌疗法的反应。