Preclinical Diagnostic Imaging of Amyloid

淀粉样蛋白的临床前诊断成像

• 批准号: 7727182
• 负责人: JONATHAN S WALL
• 金额: $ 46.4万
• 依托单位: UNIVERSITY OF TENNESSEE KNOXVILLE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7727182
• 关键词: Acute Lymphocytic Leukemia; Affinity; Alzheimer' s Disease; Amyloid; Amyloidosis; Amyotrophic Lateral Sclerosis; Animals; Antibodies; Autoradiography; B lymphoid malignancy; Binding; Biodistribution; Biological Assay; Biological Markers; Blood Circulation; Cardiac; Cerebrum; Chemistry; Chronic; Chronic Myeloid Leukemia; Clinic; Clinical; Clinical Trials; Data; Deposition; Diagnosis; Diagnostic Imaging; Dialysis procedure; Disease; Disease Progression; Enzyme-Linked Immunosorbent Assay; Equilibrium; Evaluation; Excision; Exposure to; Familial Mediterranean Fever; Generic Drugs; Goals; Heparan Sulfate Proteoglycan; Heparin; Heparitin Sulfate; Hepatic; High Dose Chemotherapy; Human; I125 isotope; Image; In Vitro; Incidence; Inflammation; Inflammatory; Inherited; Inorganic Sulfates; Investigation; Kidney; Kinetics; Label; Lesion; Liver; Methods; Modeling; Modification; Monitor; Mus; Non-Insulin-Dependent Diabetes Mellitus; Normal tissue morphology; Organ; Pathology; Patients; Peptides; Peripheral; Phage Display; Physicians; Plasma Cell Neoplasm; Positron-Emission Tomography; Probability; Proteins; Proteoglycan; Radiolabeled; Reagent; Research; Rheumatoid Arthritis; Severities; Surface Plasmon Resonance; Techniques; Testing; Therapeutic; Time; Tissues; Tracer; Transgenic Organisms; Translating; Translations; Tuberculosis; Unspecified or Sulfate Ion Sulfates; Validation; Work; alternative treatment; amyloid formation; amyloid imaging; base; in vivo; man; mouse model; novel; oncology; outcome forecast; pre-clinical; programs; protein misfolding; public health relevance; radiotracer; research clinical testing; response; single photon emission computed tomography; whole body imaging

DESCRIPTION (provided by applicant): Amyloid is associated with a diverse group of often fatal hereditary and sporadic protein misfolding disorders, characterized by the deposition of fibrils and heparan sulfate proteoglycan in vital organs and tissues. Although the most common manifestation is in patients with Alzheimer's disease, the incidence of peripheral (non-cerebral) amyloid diseases in the USA, is estimated to be ~ 5300 per yr which exceeds that for acute lymphocytic leukemia (~ 4,000 per yr). Treatment options for peripheral amyloidosis are limited and focus on reducing synthesis of the amyloidogenic protein e.g., with high dose chemotherapy and there are currently 31 new trials for treating AL, AA, and ATTR amyloidosis underway (clinicaltrials.gov). Unfortunately, there are no methods available in the USA to visualize response to these anti-amyloid therapies directly, nor to determine the extent and severity of amyloid deposits or the target organ in patients. This kind of information can be obtained by imaging, using an amyloid-specific radiotracer. There exists therefore an urgent need to identify tracers that target amyloid for whole body imaging that can be used for monitoring disease progression and response to therapy both within the clinic and as part of clinical trials. The aim of this proposal is to target the amyloid biomarker heparan sulfate proteoglycan and test antibody-derived and peptide tracer molecules that bind specifically to this constituent of all known amyloid deposits. Our approach is based on the fact that all amyloid deposits contain heparan sulfate proteoglycan at levels as high as 250 5g per gram of diseased tissue. We have begun testing a panel of antibody-derived proteins (scFv) known to bind heparan sulfate, for their ability to bind heparan sulfate in amyloid deposits. We have shown that scFvs to hypersulfated heparin sulfate successfully imaged amyloid in vivo even though heparan sulfate was expressed in some normal tissues. Novel heparan sulfate-binding peptides will also be generated and tested. In vitro characterization of the amyloid-reactive scFv and peptides will be performed by surface plasmon resonance and ELISA to identify those scFv with equilibrium binding affinity of < 1 5M - suitable for imaging in patients. Small animal SPECT, PET, and CT imaging will be used to compare the co-localization of the available and novel heparan sulfate- binding tracers and identify those with acceptable (3:1) target to background ratios. We will use an established murine model of systemic peripheral (AA) amyloidosis that recapitulates many aspects of human amyloid disease including the deposition of proteoglycans which enhances the probability that we will identify tracers for imaging amyloidosis that translate favorably into the clinic. Identifying and validating molecules that specifically bind heparan sulfate proteoglycan found in tissue amyloid will provide new reagents for the clinical evaluation of patients with this devastating disease. PUBLIC HEALTH RELEVANCE: Peripheral amyloidosis is a protein-misfolding condition associated with type 2 diabetes, chronic inflammatory disorders such as tuberculosis and rheumatoid arthritis, as well as certain B-cell malignancies. Currently here are no methods available in the USA that can detect the extent of amyloid deposits in patients with peripheral amyloidosis, nor to monitor their progression, or document their removal in response to therapy. This kind of information can be obtained by imaging, using an amyloid-specific radiotracer. To this end, we aim to target the amyloid biomarker heparan sulfate proteoglycan and test antibody-derived and peptide tracer molecules that bind specifically to this constituent of all known amyloid deposits for the purpose of whole body imaging that can be used to monitor disease progression and response to therapy.

描述（由申请人提供）：淀粉样蛋白与一群经常致命的遗传性和零星蛋白质错误折叠式疾病有关，其特征是在重要的器官和组织中的原纤维和乙酰肝素蛋白聚糖的沉积。尽管最常见的表现是在阿尔茨海默氏病的患者中，但据估计，在美国，外周（非脑）淀粉样蛋白疾病的发生率估计约为每年约5300，超过了急性淋巴细胞性白血病（每年约4,000）。外周淀粉样变性的治疗方案受到限制，专注于减少淀粉样蛋白的合成，例如高剂量化疗，目前有31项用于治疗AL，AA和Attryloilisoiss正在进行的新试验（ClinicalTrialtrials.gov）。不幸的是，美国没有直接可视化对这些抗淀粉样蛋白疗法的反应的方法，也不能确定淀粉样蛋白沉积物的程度和严重程度或患者中的靶器官。可以使用淀粉样蛋白特异性放射性示踪剂来通过成像获得此类信息。因此，迫切需要鉴定瞄准淀粉样蛋白用于全身成像的示踪剂，这些示踪剂可用于监测诊所内的疾病进展和对治疗的反应，也是临床试验的一部分。该提案的目的是靶向淀粉样生物标志物硫酸乙酰肝素蛋白聚糖和测试抗体衍生和肽示踪剂分子，该分子专门与所有已知淀粉样蛋白沉积物的组成部分结合。我们的方法是基于这样一个事实，即所有淀粉样蛋白沉积物都含有硫酸乙酰肝素蛋白聚糖的水平高达每克组织250 5G。我们已经开始测试一组已知可以结合硫酸乙酰肝素的抗体衍生蛋白（SCFV），因为它们结合淀粉样蛋白沉积物中硫酸乙酰肝素的能力。我们已经证明，尽管在某些正常组织中表达了硫酸乙酰肝素，但硫酸肝素超硫酸盐的SCFV仍成功成像淀粉样蛋白。新型的硫酸肝素结合肽也将被生成和测试。淀粉样蛋白反应性SCFV和肽的体外表征将由表面等离子体共振和ELISA进行，以鉴定具有<1 5M的平衡结合亲和力的SCFV，适用于患者的成像。小型动物SPECT，PET和CT成像将用于比较可用硫酸乙酰肝素结合示踪剂的共定位，并鉴定具有可接受（3：1）靶标与背景比率的人。我们将使用已建立的全身外围（AA）淀粉样变性的鼠模型，该模型概括了人淀粉样蛋白疾病的许多方面，包括蛋白聚糖的沉积，这增强了我们将识别出对淀粉样变化的示踪剂的可能性，从而使淀粉样蛋白变成有利地转化为诊所。识别和验证分子在组织淀粉样蛋白中发现的特异性结合硫酸肝素蛋白聚糖的分子将为这种毁灭性疾病的患者提供新试剂。公共卫生相关性：周围淀粉样变性是一种与2型糖尿病有关的蛋白质不满意疾病，慢性炎症性疾病，例如结核病和类风湿关节炎以及某些B细胞恶性肿瘤。目前，在美国尚无可用的方法，可以检测外周淀粉样变性患者的淀粉样蛋白沉积程度，也可以监测其进展或记录其对治疗的响应。可以使用淀粉样蛋白特异性放射性示踪剂来通过成像获得此类信息。为此，我们旨在靶向淀粉样蛋白酶硫酸乙酰肝素蛋白聚糖和测试抗体衍生和肽示踪剂分子，这些分子特异性地与所有已知淀粉样蛋白沉积物的该组成部分结合，以用于全身成像，用于监测疾病进展和对治疗的反应。