A Novel Tool for Detecting Amyloid Deposits in Systemic Amyloidosis In Vitro and In Vivo

Abstract

SUMMARY:We synthesized (trans,trans)-1-bromo-2,5-bis-(3-hydroxycarbonyl-4-hydroxy)styrylbenzene (BSB) and used this compound to detect amyloid fibrils in autopsy and biopsy samples from patients with localized amyloidosis, such as familial prion disease, and systemic amyloidosis, such as familial amyloidotic polyneuropathy, amyloid A (AA) amyloidosis, light chain (AL) amyloidosis, and dialysis-related amyloidosis. BSB showed reactions in all Congo red-positive and immunoreactive regions of the samples examined in the study, and some amyloid fibrils in the tissues could be detected more precisely with BSB than with the other methods. In the mouse model of AA amyloidosis, injected BSB reacted with amyloid in all regions in the serial sections in which Congo red staining was positive. A highly sensitive 27-MHz quartz crystal microbalance analysis revealed that BSB showed a significant affinity for amyloid fibrils purified from familial amyloidotic polyneuropathy and dialysis-related amyloidosis samples and suppressed formation of transthyretin amyloid in vitro. These results suggest that BSB may become a valuable tool for detection of amyloid deposits in amyloidosis and of the mechanism of amyloid formation. (Lab Invest 2003, 83:1751-1759. P rogress in molecular genetics and biochemical methodologies has led to the identification of various types of amyloidosis and their amyloidogenic precursor proteins (Benson, 1995;Ikeda et al, 2002;Saraiva, 2001;Tan and Pepys, 1994;Tan et al, 1995). Thus far, 24 different amyloidogenic proteins have been identified; however, the mechanism of amyloid formation has been elucidated in only several types of amyloidosis Benson et al, 1993;Benson and Uemichi, 1996;Pepys et al, 1993;Soutar et al, 1992;Westermark et al, 2002). One of the most important steps in the diagnosis of the specific type of amyloidosis is detection of amyloid deposits in tissues. It is not easy to predict the presence of these amyloid deposits on the basis of patients' clinical manifestations, that is, without histopathologic materials. Thus, amyloidosis is sometimes diagnosed after the death of the patients (Ishihara et al, 1989).In systemic amyloidosis, such as familial amyloidotic polyneuropathy (FAP), AA amyloidosis, AL amyloidosis, and dialysis-related amyloidosis (DRA), biopsy samples are often obtained from the gastrointestinal tract and abdominal fat to make the diagnosis (Guy and Jones, 2001;Kaplan et al, 1999;Masouye, 1997) because amyloid deposits are usually found in these tissues at the early stage of the disease. However, in certain cases it is sometimes difficult to make the diagnosis using only the biopsy samples, because the pattern of amyloid deposition in the body varies in each individual (Ishihara et al, 1989). In addition, biopsy for diagnostic purposes cannot usually be performed in patients with localized amyloidosis, such as in Alzheimer's disease and endocrine amyloidosis (Westermark and Westermark, 2000). Diagnosis at the early stage of amyloidosis might be possible if a tool were avai...

A novel age‐related venous amyloidosis derived from EGF‐containing fibulin‐like extracellular matrix protein 1

“…Although amyloid deposits are usually identified by means of CR staining, our histopathological studies showed that venous EFEMP1 amyloid deposits had faint CR staining. In contrast, these venous EFEMP1 amyloid deposits were confirmed by electron microscopy and were clearly detected by FSB fluorescent staining, which is reportedly a more sensitive method than CR staining to detect various kinds of amyloid deposits . We should note, therefore, that CR staining may be not sufficient to detect venous EFEMP1 amyloid deposits and may sometimes lead to misdiagnosis of EFEMP1 amyloidosis, so more sensitive methods, such as FSB staining and immunohistochemical staining, are preferred for a more accurate diagnosis of venous EFEMP1 amyloidosis.…”

Novel Cell- and Tissue-Based Assays for Detecting Misfolded and Aggregated Protein Accumulation Within Aggresomes and Inclusion Bodies

“…A fluorescent derivative of Congo Red, (trans, trans)-1-bromo-2,5-bis-(3-hydroxycarbonyl-4-hydroxy)styrylbenzene (BSB), has also been employed for the detection of amyloid deposits in tissue sections [19]. With an emission maximum of 520 nm, this probe offers improved performance with respect to the avoidance of background associated with intrinsic and fixative-induced autofluorescence in cells and tissues.…”

Diagnosis and Therapeutic Approaches to Transthyretin Amyloidosis

“…In the patients with small amyloid deposits, repeated biopsies should be required to confirm the diagnosis. In addition to the commonly used Congo red staining, (E,E)-1-bromo-2,5-bis-(3-hydroxycarbonyl-4-hydroxy) styrylbenzene (BSB) and (E,E)-1-fluoro-2,5-bis-(3-hydroxycarbonyl-4-hydroxy) styrylbenzene (FSB) can be employed to detect a small amount of amyloid deposits in biopsy specimens, because these two chemical agents bind to amyloid fibrils more strongly than Congo red [49,50].…”