Immune profiling of plasma-derived extracellular vesicles identifies Parkinson disease

Vacchi, Elena; Burrello, Jacopo; Silvestre, Dario Di; Burrello, Alessio; Bolis, Sara; Mauri, Pierluigi; Vassalli, Giuseppe; Cereda, Carlo; Farina, Cinthia; Barile, Lucio; Kaelin‐Lang, Alain; Melli, Giorgia

doi:10.1212/nxi.0000000000000866

Cited by 50 publications

(47 citation statements)

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“…The exome marker curves for PD and MSA were rather congruent and different from those of corticobasal degeneration and progressive supranuclear palsy. In the panels shared by PD and MSA was a transcription factor playing, SP1, which is an important regulator of neuroinflammation in multiple sclerosis ( Vacchi et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Inflammatory Diseases Among Norwegian LRRK2 Mutation Carriers. A 15-Years Follow-Up of a Cohort

Aasly

2021

Front. Neurosci.

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The first families with LRRK2 related Parkinson’s disease (PD) were presented around 15 years ago and numerous papers have described the characteristics of the LRRK2 phenotype. The prevalence of autosomal dominant PD varies around the world mainly depending on local founder effects. The highest prevalence of LRRK2 G2019S PD in Norway is located to the central part of the country and most families could be traced back to common ancestors. The typical Norwegian LRRK2 phenotype is not different from classical PD and similar to that seen in most other LRRK2 families. The discovery of LRRK2 PD has allowed us to follow-up multi-incident families and to study their phenotype longitudinally. In the Norwegian LRRK2 families there has been a significantly higher incidence of inflammatory diseases like multiple sclerosis and rheumatoid arthritis that seen in other PD populations. Recent studies in LRRK2 mechanisms have indicated that this protein may be crucial in initiating disease processes. In this short survey of 100 Norwegian mutation carriers followed through more than 15 years are presented. The prevalence of inflammatory diseases among these cases is highlighted. The role of LRRK2 in the conversion process from carrier status to PD phenotype is still unknown and disease generating mechanisms important for initiating LRRK2 PD are still to be identified.

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Section: Discussionmentioning

confidence: 99%

Inflammatory Diseases Among Norwegian LRRK2 Mutation Carriers. A 15-Years Follow-Up of a Cohort

Aasly

2021

Front. Neurosci.

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“…MuSIC probes have a wide variety of applications for flow cytometry, one of which is immune profiling. Immune profiling is typically performed using flow cytometry [45][46][47] , which has previously limited multiplexing to roughly a dozen analytes (depending on the capabilities of the instrument) as a result of spectral overlap 48,49 . Mass cytometry has been transformative for immune profiling [50][51][52] , but is slower than flow cytometry and is destructive so prevents further use of the cells after analysis 48 .…”

Section: Discussionmentioning

confidence: 99%

Fluorescent Antibody Multiplexing with Oligo-Based Combinatorial Labeling

McCarthy

Anglin

Pear

et al. 2020

Preprint

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Fluorescent antibodies are a workhorse of biomedical science, but fluorescence multiplexing has been notoriously difficult due to spectral overlap between fluorophores. We recently established proof-of-principal for fluorescence Multiplexing using Spectral Imaging and Combinatorics (MuSIC), which uses combinations of existing fluorophores to create unique spectral signatures for increased multiplexing. However, a method for labeling antibodies with MuSIC probes has not yet been developed. Here, we present a method for labeling antibodies with MuSIC probes. We conjugate a DBCO-Peg5-NHS ester linker to antibodies, a single stranded DNA docking strand to the linker, and finally, hybridize two MuSIC-compatible, fluorescently-labeled oligos to the docking strand. We validate the labeling protocol with spin-column purification and absorbance measurements, which show a degree of labeling of ~9.66 linker molecules / antibody. We demonstrate the approach using (i) Cy3, (ii) Tex615, and (iii) a Cy3-Tex615 combination as three different MuSIC probes attached to three separate batches of antibodies. We incubated MuSIC probe-labeled antibodies with protein A beads to create single and double positive beads that are analogous to single cells. Spectral flow cytometry experiments demonstrate that each MuSIC probe can be uniquely distinguished, and the fraction of beads in a mixture with different staining patterns is accurately measured. The approach is general and might be more broadly applied to cell type profiling or tissue heterogeneity studies in clinical, biomedical, and drug discovery research.

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“…We characterized distinctive EV subpopulations in plasma by simultaneously immunophenotyping 37 different membrane proteins using a flow cytometry multiplex bead-based platform [12,13] and then a diagnostic model based on the distinctive EV surface proteins profile was generated via supervised machine learning algorithms. The resulting diagnostic model was able to correctly classify 88.9% of patients, with reliable diagnostic performance after internal and external validations [14]. Indeed, recent studies trying to dissect the cell/tissue origin of circulating EVs, showed that the majority of plasma-derived EVs come from hematopoietic cells, in particular platelets, B cells and T cells [15].…”

Section: Introductionmentioning

confidence: 93%

Profiling Inflammatory Extracellular Vesicles in Plasma and Cerebrospinal Fluid: An Optimized Diagnostic Model for Parkinson’s Disease

Vacchi

Burrello

et al. 2021

Biomedicines

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Extracellular vesicles (EVs) play a central role in intercellular communication, which is relevant for inflammatory and immune processes implicated in neurodegenerative disorders, such as Parkinson’s Disease (PD). We characterized and compared distinctive cerebrospinal fluid (CSF)-derived EVs in PD and atypical parkinsonisms (AP), aiming to integrate a diagnostic model based on immune profiling of plasma-derived EVs via artificial intelligence. Plasma- and CSF-derived EVs were isolated from patients with PD, multiple system atrophy (MSA), AP with tauopathies (AP-Tau), and healthy controls. Expression levels of 37 EV surface markers were measured by a flow cytometric bead-based platform and a diagnostic model based on expression of EV surface markers was built by supervised learning algorithms. The PD group showed higher amount of CSF-derived EVs than other groups. Among the 17 EV surface markers differentially expressed in plasma, eight were expressed also in CSF of a subgroup of PD, 10 in MSA, and 6 in AP-Tau. A two-level random forest model was built using EV markers co-expressed in plasma and CSF. The model discriminated PD from non-PD patients with high sensitivity (96.6%) and accuracy (92.6%). EV surface marker characterization bolsters the relevance of inflammation in PD and it underscores the role of EVs as pathways/biomarkers for protein aggregation-related neurodegenerative diseases.

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Immune profiling of plasma-derived extracellular vesicles identifies Parkinson disease

Cited by 50 publications

References 50 publications

Inflammatory Diseases Among Norwegian LRRK2 Mutation Carriers. A 15-Years Follow-Up of a Cohort

Inflammatory Diseases Among Norwegian LRRK2 Mutation Carriers. A 15-Years Follow-Up of a Cohort

Fluorescent Antibody Multiplexing with Oligo-Based Combinatorial Labeling

Profiling Inflammatory Extracellular Vesicles in Plasma and Cerebrospinal Fluid: An Optimized Diagnostic Model for Parkinson’s Disease

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