Cryo-EM structure of a transthyretin-derived amyloid fibril from a patient with hereditary ATTR amyloidosis

Schmidt, Matthias; Wiese, Sebastian; Adak, Volkan; Engler, Jonas; Agarwal, Shalabh; Fritz, Günter; Westermark, Per; Zacharias, Martin; Fändrich, Marcus

doi:10.1038/s41467-019-13038-z

Cited by 151 publications

(208 citation statements)

References 56 publications

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“…For example, α-synuclein aggregates isolated from different types of synucleinopathies (e.g., PD and multiple system atrophy) exhibited drastically different structure (52,53) and seeding capacity, and induced different patterns of pathology spreading in in vitro neuronal cell models and animal models of synucleinopathies (16). Similar findings have been made for Aβ aggregates from AD brains and other amyloidogenic proteins (20,(54)(55)(56). While these differences have been attributed to differences in the structural properties of the fibrils present in these samples, direct visualization and characterization of some of the tissue-derived aggregates has not been possible.…”

Section: Discussionsupporting

confidence: 62%

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Cendrowska

Silva

Ait-Bouziad

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Increasing evidence suggests that amyloid polymorphism gives rise to different strains of amyloids with distinct toxicities and pathology-spreading properties. Validating this hypothesis is challenging due to a lack of tools and methods that allow for the direct characterization of amyloid polymorphism in hydrated and complex biological samples. Here, we report on the development of 11-mercapto-1-undecanesulfonate-coated gold nanoparticles (NPs) that efficiently label the edges of synthetic, recombinant, and native amyloid fibrils derived from different amyloidogenic proteins. We demonstrate that these NPs represent powerful tools for assessing amyloid morphological polymorphism, using cryogenic transmission electron microscopy (cryo-EM). The NPs allowed for the visualization of morphological features that are not directly observed using standard imaging techniques, including transmission electron microscopy with use of the negative stain or cryo-EM imaging. The use of these NPs to label native paired helical filaments (PHFs) from the postmortem brain of a patient with Alzheimer’s disease, as well as amyloid fibrils extracted from the heart tissue of a patient suffering from systemic amyloid light-chain amyloidosis, revealed a high degree of homogeneity across the fibrils derived from human tissue in comparison with fibrils aggregated in vitro. These findings are consistent with, and strongly support, the emerging view that the physiologic milieu is a key determinant of amyloid fibril strains. Together, these advances should not only facilitate the profiling and characterization of amyloids for structural studies by cryo-EM, but also pave the way to elucidate the structural basis of amyloid strains and toxicity, and possibly the correlation between the pathological and clinical heterogeneity of amyloid diseases.

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Section: Discussionsupporting

confidence: 62%

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Cendrowska

Silva

Ait-Bouziad

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…As thermodynamic stability of fibrils is determined by intermolecular bonds, this information may be complementary to the structural observations on fibrils obtained by solid-state NMR (27) and cryo-EM, which have recently been applied to a number of natural pathologic filaments (28) and fibrils (14,15,29). Ultrastructural polymorphism of amyloid fibrils has been observed among fibrils derived from the different monoclonal immunoglobulin light chains and different mutations; different sources of natural fibrils may lead to significant structural differences.…”

Section: Mechano-enzymatic Fibrils Amentioning

confidence: 93%

“…The heterogeneity in structure may correlate with the thermodynamic stability of fibrils and subsequently with their kinetics of growth and resistance to solubilization. Recently, Fändrich's team has released the first cryo-EM structure of natural fibrils extracted from the heart of a patient with V30M variant TTR amyloidosis (29). In this case, apparently, the fibrils were composed of a mixture of full-length monomers and C-terminal truncated fragments plus the N-terminal cleaved polypeptide.…”

Section: Mechano-enzymatic Fibrils Amentioning

confidence: 99%

Comparative study of the stabilities of synthetic in vitro and natural ex vivo transthyretin amyloid fibrils

Raimondi

Mangione

Verona

et al. 2020

Journal of Biological Chemistry

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Systemic amyloidosis caused by extracellular deposition of insoluble fibrils derived from the pathological aggregation of circulating proteins, such as transthyretin, is a severe and usually fatal condition. Elucidation of the molecular pathogenic mechanism of the disease and discovery of effective therapies still represents a challenging medical issue. The in vitro preparation of amyloid fibrils that exhibit structural and biochemical properties closely similar to those of natural fibrils is central to improving our understanding of the biophysical basis of amyloid formation in vivo and may offer an important tool for drug discovery. Here, we compared the morphology and thermodynamic stability of natural transthyretin fibrils with those of fibrils generated in vitro using either the common acidification procedure or primed by limited selective cleavage by plasmin. The free energies for fibril formation were −12.36 kcal mol-1, −8.10 kcal mol-1 and −10.61 kcal mol-1, respectively. The fibrils generated via plasmin cleavage were more stable than those prepared at low pH and were thermodynamically and morphologically similar to natural fibrils extracted from human amyloidotic tissue. Determination of thermodynamic stability is an important tool that is complementary to other methods for structural comparison between ex vivo fibrils and fibrils generated in vitro. Our finding that fibrils created via an in vitro amyloidogenic pathway are structurally similar to ex vivo human amyloid fibrils does not necessarily establish that the fibrillogenic pathway is the same for both, but it narrows the current knowledge gap between in vitro models and in vivo pathophysiology.

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“…Currently, anti-inflammatory drugs or colchicine are typically used in the therapy of AA amyloidosis to reduce SAA levels in blood circulation [ 10 , 18 ]. Although the mechanism of AA amyloidosis is currently unclear, the structure and basic composition of SAA have gradually become clear, including the use of cryo-electron microscopy (cryo-EM), allowing a better understanding of the pathogenesis [ 24 , 25 ]. As cryo-EM technology improves, this will provide a better understanding of how polymorphism is related to the disease phenotype and how fibril structure is affected by the cellular environment.…”

Section: Introductionmentioning

confidence: 99%

Dietary Intake of Rosmarinic Acid Increases Serum Inhibitory Activity in Amyloid A Aggregation and Suppresses Deposition in the Organs of Mice

Lin

Watanabe

Kuragano

et al. 2020

IJMS

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Serum amyloid A (SAA) is one of the most important precursor amyloid proteins and plays a vital step in AA amyloidosis, although the underlying aggregation mechanism has not been elucidated. Since SAA aggregation is a key step in this pathogenesis, inhibitors are useful to prevent and treat AA amyloidosis, serving as tools to investigate the pathogenic mechanism. In this study, we showed that rosmarinic acid (RA), which is a well-known inhibitor of the aggregation of amyloid β (Aβ), displayed inhibitory activity against SAA aggregation in vitro using a microliter-scale high-throughput screening (MSHTS) system with quantum-dot nanoprobes. Therefore, we evaluated the amyloid aggregation inhibitory activity of blood and the deposition of SAA in organs by feeding mice with Melissa officinalis extract (ME) containing RA as an active substance. Interestingly, the inhibitory activity of ME-fed mice sera for SAA and Aβ aggregation, measured with the MSHTS system, was higher than that of the control group. The amount of amyloid deposition in the organs of ME-fed mice was lower than that in the control group, suggesting that the SAA aggregation inhibitory activity of serum is associated with SAA deposition. These results suggest that dietary intake of RA-containing ME enhanced amyloid aggregation inhibitory activity of blood and suppressed SAA deposition in organs. This study also demonstrated that the MSHTS system could be applied to in vitro screening and to monitor comprehensive activity of metabolized foods adsorbed by blood.

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Cryo-EM structure of a transthyretin-derived amyloid fibril from a patient with hereditary ATTR amyloidosis

Cited by 151 publications

References 56 publications

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Comparative study of the stabilities of synthetic in vitro and natural ex vivo transthyretin amyloid fibrils

Dietary Intake of Rosmarinic Acid Increases Serum Inhibitory Activity in Amyloid A Aggregation and Suppresses Deposition in the Organs of Mice

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