α-Synuclein molecular behavior and nigral proteomic profiling distinguish subtypes of Lewy body disorders

Martínez-Valbuena, Iván; Swinkin, Emily; Santamaría, Enrique; Fernández‐Irigoyen, Joaquín; Sackmann, Valerie; Kim, Ain; Li, Jun; Gonzàlez‐Latapi, Paulina; Kuhlman, G. M.; Bhowmick, Suvorit Subhas; Visanji, Naomi P.; Lang, Anthony E.; Kovács, Gábor G.

doi:10.1007/s00401-022-02453-0

Cited by 23 publications

(41 citation statements)

References 69 publications

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“…We did not observe seeding in the control subject. This is consistent with our previous findings in the post-mortem frozen human brain tissue [20,21].…”

Section: Replication Of Disease-specific Seeding Patterns In Ffpe Hum...supporting

confidence: 94%

“…To evaluate the differential seeding behavior between LBD and MSA subjects, two SAA protocols were performed [20,21]. In the first protocol, an LBD-favoring SAA, the plate was incubated at 42°C with cycles of 1-minute shaking and 1-minute rest.…”

Section: Protein Extraction and Quantificationmentioning

confidence: 99%

“…Different temperature, pH, salt and buffer composition can alter the in vitro fibril growth and sensitivity of the assay, further enabling a selective detection of pathogenic seeds [17][18][19]. Recent studies from our group have identified optimal assay conditions that favor either LBD or MSA seeding activity, and demonstrated that not only seeding differs between synucleinopathies but also between regions within subjects [20,21].…”

Section: Introductionmentioning

confidence: 99%

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Disease-Specific α-Synuclein Seeding and Structural Properties in Lewy Body Disease and Multiple System Atrophy are Preserved in Formaldehyde-Fixed Paraffin-Embedded Human Brain

Kim¹,

Martínez-Valbuena²,

Li³

et al. 2023

Preprint

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Recent studies have been able to detect α-synuclein (αSyn) seeding in formaldehyde-fixed paraffin-embedded (FFPE) tissues from patients with synucleinopathies using seed amplification assays (SAAs), but with relatively low sensitivity due to limited protein extraction efficiency. With the aim of introducing an alternative option to frozen tissues, we developed a streamlined protein extraction protocol for evaluating disease-specific seeding in FFPE human brain. We evaluated the protein extraction efficiency of different tissue preparations, deparaffinizations, and protein extraction buffers using formaldehyde-fixed and FFPE tissue of a single Lewy body disease (LBD) subject. Alternatively, we incorporated heat-induced antigen-retrieval and dissociation using a commercially available kit. Our novel protein extraction protocol has been optimized to work with 10 sections of 4.5-µm-thickness or 2-mm-diameter micro-punch of FFPE tissue that can be used to seed SAAs. We demonstrated that extracted proteins from FFPE still preserve seeding potential and further show disease-specific seeding in LBD and multiple system atrophy. To the best of our knowledge, our study is the first to recapitulate disease-specific αSyn seeding behavior in FFPE human brain, with structural validation. Our findings open new perspectives in re-evaluating archived human brain tissue, extending the disease-specific seeding assays to larger cohorts to facilitate molecular subtyping of synucleinopathies.

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“…We did not observe seeding in the control subject. This is consistent with our previous findings in the post-mortem frozen human brain tissue [20,21].…”

Section: Replication Of Disease-specific Seeding Patterns In Ffpe Hum...supporting

confidence: 94%

Section: Protein Extraction and Quantificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Disease-Specific α-Synuclein Seeding and Structural Properties in Lewy Body Disease and Multiple System Atrophy are Preserved in Formaldehyde-Fixed Paraffin-Embedded Human Brain

Kim¹,

Martínez-Valbuena²,

Li³

et al. 2023

Preprint

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“…3c). To evaluate the cytotoxic effect of primary motor cortex-derived homogenates, we used two different approaches 21 . First, at day in vitro (DIV) 8 and DIV 10 we measured the concentration of lactate dehydrogenase (LDH), as damage of the plasma membrane results in a release of this cytosolic enzyme into the surrounding cell culture medium.…”

Section: Resultsmentioning

confidence: 99%

“…These assays capitalize on the property of self-propagation to monitor a seed's propensity to induce aggregation by amplifying a signal that can sensitively detect minute amounts of these protein seeds using real-time detection of thioflavin T (ThT) fluorescence at multiple time points 20 . We have recently demonstrated the power of this approach to detect differences in the seeding capacity of soluble α-synuclein species isolated from distinct patients with Lewy body disorders (LBD) 21 . In that study, despite similar amounts of α-synuclein pathology in the substantia nigra (SN) of LBD patients with distinct dominant clinical symptoms (motor versus cognitive), the seeding capacity of α-synuclein was dissimilar between LBD subtypes.…”

Section: Introductionmentioning

confidence: 99%

4R-Tau seeding activity unravels molecular subtypes in patients with Progressive Supranuclear Palsy

Martinez-Valbuena,

Lee,

Santamaria

et al. 2023

Preprint

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Progressive Supranuclear palsy (PSP) is a 4-repeat (4-R) tauopathy. We hypothesized that the molecular diversity of tau could explain the heterogeneity seen in PSP disease progression. To test this hypothesis, we performed an extensive biochemical characterisation of the high molecular weight tau species (HMW-Tau) in 20 different brain regions of 25 PSP patients. We found a correlation between the HMW-Tau species and tau seeding capacity in the primary motor cortex, where we confirmed that an elevated 4R-Tau seeding activity correlates with a shorter disease duration. To identify factors that contribute to these differences, we performed proteomic and spatial transcriptomic analysis that revealed key mechanistic pathways, in particular those involving the immune system, that defined patients demonstrating high and low tau seeding capacity. These observations suggest that differences in the tau seeding activity may contribute to the considerable heterogeneity seen in disease progression of patients suffering from PSP.

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Misfolded α‐Synuclein Seeding Is Detected in Suspected LRRK2‐Parkinson's Disease without Immunohistochemically Detectable α‐Synuclein Pathology

Kim,

Martinez‐Valbuena,

Keith

et al. 2023

Movement Disorders

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α-Synuclein molecular behavior and nigral proteomic profiling distinguish subtypes of Lewy body disorders

Cited by 23 publications

References 69 publications

Disease-Specific α-Synuclein Seeding and Structural Properties in Lewy Body Disease and Multiple System Atrophy are Preserved in Formaldehyde-Fixed Paraffin-Embedded Human Brain

Disease-Specific α-Synuclein Seeding and Structural Properties in Lewy Body Disease and Multiple System Atrophy are Preserved in Formaldehyde-Fixed Paraffin-Embedded Human Brain

4R-Tau seeding activity unravels molecular subtypes in patients with Progressive Supranuclear Palsy

Misfolded α‐Synuclein Seeding Is Detected in Suspected LRRK2‐Parkinson's Disease without Immunohistochemically Detectable α‐Synuclein Pathology

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