Abstract

More than 30 proteins and peptides have been found to form amyloid fibrils in human diseases. Fibrils formed by transthyretin (TTR) are associated with ATTR amyloidosis, affecting many vital organs, including the heart and peripheral nervous system. Congo red staining is the gold standard method for detection of amyloid deposits in tissue. However, Congo red staining and amyloid typing methods such as immunofluorescence labelling are limited to relatively large deposits. Detection of small ATTR deposits present at an early stage of the disease could enable timely treatment and prevent severe tissue damage. In this study, we developed an enhanced ATTR amyloid detection method that uses functionalised protein nanofibrils. Using this method, we achieved sensitive detection of monomeric TTR in a microplate immunoassay and immunofluorescence labelling of ex vivo tissue from two patients containing ATTR aggregates. The system's utility was confirmed on sections from a patient with AA amyloidosis and liver sections from inflamed mouse. These results suggest that the detection system constitutes important new technology for highly sensitive detection of microscopic amounts of ATTR amyloid deposited in tissue.

摘要

已发现 30 多种蛋白质和肽在人类疾病中形成淀粉样蛋白原纤维。转甲状腺素蛋白 (TTR) 形成的原纤维与 ATTR 淀粉样变性有关，影响许多重要器官，包括心脏和周围神经系统。刚果红染色是检测组织中淀粉样蛋白沉积的金标准方法。然而，刚果红染色和免疫荧光标记等淀粉样蛋白分型方法仅限于相对较大的沉积物。检测在疾病早期存在的小 ATTR 沉积物可以及时治疗并防止严重的组织损伤。在这项研究中，我们开发了一种增强的 ATTR 淀粉样蛋白检测方法，该方法使用功能化的蛋白质纳米纤维。使用这种方法，来自两名含有 ATTR 聚集体的患者的离体组织。该系统的实用性在来自 AA 淀粉样变性患者的切片和来自发炎小鼠的肝脏切片上得到证实。这些结果表明，该检测系统构成了用于高灵敏度检测组织中沉积的微量 ATTR 淀粉样蛋白的重要新技术。