Loss of exosomes in progranulin-associated frontotemporal dementia

Benussi, Luisa; Ciani, Maurizio; Tonoli, Elisa; Morbin, Michela; Palamara, Luisa; Albani, Diego; Fusco, Federica; Forloni, Gianluigi; Glionna, Michela; Baco, Monika; Paterlini, Anna; Fostinelli, Silvia; Santini, Benedetta; Galbiati, Elisabetta; Gagni, Paola; Cretich, Marina; Binetti, Giuliano; Tagliavini, Fabrizio; Prosperi, Davide; Chiari, Marcella; Ghidoni, Roberta

doi:10.1016/j.neurobiolaging.2016.01.001

Cited by 49 publications

(53 citation statements)

References 62 publications

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“…isolated from the supernatant of cells according to standard protocols 24,25 ( Figure 6A). After removal of cells and cellular debris, ectosomes were recovered by differential centrifugation in the P3 fraction (2.97310 7 particles per 1 ml medium) and smaller exosomes were recovered in the P4 fraction (5.89310 7 particles per 1 ml medium) as revealed by tunable resistive pulse sensing analysis with a qNano instrument ( Figure 6B).…”

Section: Tg2 Secretion Via Exosomes and Ectosomes In Established Epitmentioning

confidence: 99%

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Proteomic Profiling Reveals the Transglutaminase-2 Externalization Pathway in Kidneys after Unilateral Ureteric Obstruction

Furini

Schroëder

Huang

et al. 2018

JASN

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Increased export of transglutaminase-2 (TG2) by tubular epithelial cells (TECs) into the surrounding interstitium modifies the extracellular homeostatic balance, leading to fibrotic membrane expansion. Although silencing of extracellular TG2 ameliorates progressive kidney scarring in animal models of CKD, the pathway through which TG2 is secreted from TECs and contributes to disease progression has not been elucidated. In this study, we developed a global proteomic approach to identify binding partners of TG2 responsible for TG2 externalization in kidneys subjected to unilateral ureteric obstruction (UUO) using TG2 knockout kidneys as negative controls. We report a robust and unbiased analysis of the membrane interactome of TG2 in fibrotic kidneys relative to the entire proteome after UUO, detected by SWATH mass spectrometry. The data have been deposited to the ProteomeXchange with identifier PXD008173. Clusters of exosomal proteins in the TG2 interactome supported the hypothesis that TG2 is secreted by extracellular membrane vesicles during fibrosis progression. In established TEC lines, we found TG2 in vesicles of both endosomal (exosomes) and plasma membrane origin (microvesicles/ectosomes), and TGF-b1 stimulated TG2 secretion. Knockout of syndecan-4 (SDC4) greatly impaired TG2 exosomal secretion. TG2 coprecipitated with SDC4 from exosome lysate but not ectosome lysate. Ex vivo, EGFP-tagged TG2 accumulated in globular elements (blebs) protruding/retracting from the plasma membrane of primary cortical TECs, and SDC4 knockout impaired bleb formation, affecting TG2 release. Through this combined in vivo and in vitro approach, we have dissected the pathway through which TG2 is secreted from TECs in CKD. Significance StatementSecretion of the matrix crosslinking enzyme transglutaminase-2 (TG2) from tubular epithelial cells has been shown to contribute to fibrotic remodeling, a primary pathologic process in CKD. To discover the pathway for secretion of TG2, a comparative proteomic strategy was developed. Proteins that interact with TG2 were identified by TG2 immunoprecipitation from wild-type and TG2 knockout fibrotic kidney membranes followed by SWATH mass spectroscopy. The TG2 interactome was enriched in extracellular vesicle proteins, suggesting that TG2 is secreted in exosomes. Studies in cultured tubular epithelial cells support this conclusion and suggest that TG2 is a binding cargo of syndecan-4. The finding of TG2 in the urine of patients with CKD raises the possibility that block of vesicular TG2 could reduce TG2-driven matrix accumulation and diminish fibrosis.

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Section: Tg2 Secretion Via Exosomes and Ectosomes In Established Epitmentioning

confidence: 99%

“…24,25 Nanoparticle tracking analysis by tunable resistive pulse sensing was performed using qNano (Izon) to examine microparticle size and distribution. In some cases, cells were stimulated by recombinant TGF-b1 (10 ng/ml) to simulate conditions of fibrogenesis in vitro.…”

Section: Ev Isolation and Characterizationmentioning

confidence: 99%

Proteomic Profiling Reveals the Transglutaminase-2 Externalization Pathway in Kidneys after Unilateral Ureteric Obstruction

Furini

Schroëder

Huang

et al. 2018

JASN

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“…In particular, EVs are considered valuable for liquid biopsies in cancer diagnosis since they carry molecular and proteomic cargo from their tumour cell of origin10. In human CSF, EVs are rich reservoirs of biomarkers for neurological disorders and there is increasing evidence that deregulation of EVs secretion play a pathological role in neurodegenerative diseases such as Alzheimer’s disease (AD) and Frontotemporal dementia (FTD)1112131415.…”

mentioning

confidence: 99%

Digital Detection of Exosomes by Interferometric Imaging

Daaboul¹,

Gagni

Benussi

et al. 2016

Sci Rep

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236

214

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Exosomes, which are membranous nanovesicles, are actively released by cells and have been attributed to roles in cell-cell communication, cancer metastasis, and early disease diagnostics. The small size (30–100 nm) along with low refractive index contrast of exosomes makes direct characterization and phenotypical classification very difficult. In this work we present a method based on Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) that allows multiplexed phenotyping and digital counting of various populations of individual exosomes (>50 nm) captured on a microarray-based solid phase chip. We demonstrate these characterization concepts using purified exosomes from a HEK 293 cell culture. As a demonstration of clinical utility, we characterize exosomes directly from human cerebrospinal fluid (hCSF). Our interferometric imaging method could capture, from a very small hCSF volume (20 uL), nanoparticles that have a size compatible with exosomes, using antibodies directed against tetraspanins. With this unprecedented capability, we foresee revolutionary implications in the clinical field with improvements in diagnosis and stratification of patients affected by different disorders.

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“…GRN KO mice have a higher number of lysosomes due to decreased degradation (Tanaka et al, 2014). Fibroblasts from patients with progranuline haploinsufficiency have lysosomal dysfunction (Ward et al, 2017) Null mutation in GRN gene decreases the release of exosomes (Benussi et al, 2016) C9orf72…”

Section: Effect Of the Coded Protein On The Biogenesis/secretion/compmentioning

confidence: 99%

“…Rab7 regulates secretion of syntenin and syndecan-containing exosomes (Baietti et al, 2012) Neuronal ceroid lipofuscinoses (NCL) CLN-1&2 CLN1 and 2 encode lysosomal hydrolases (Hofmann et al, 2002) Undetermined GRN GRN encodes progranulin. Heterozygous mutations in progranulin cause NCL (Smith, Damiano, et al, 2012) Null mutation in GRN gene decreases the release of exosomes (Benussi et al, 2016) Gaucher disease GBA GBA encodes beta-glucosidase, a lysosomal enzyme. GBA mutations also increase the risk to PD (Gegg & Schapira, 2018) Undetermined TA B L E 1 (Continued) of CAG nucleotide repeats in the gene huntingtin.…”

Section: )mentioning

confidence: 99%

The interplay between aging‐associated loss of protein homeostasis and extracellular vesicles in neurodegeneration

Guix

2019

J of Neuroscience Research

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The finding of an effective cure or treatment for neurodegenerative diseases is one of the biggest challenges for this century. Although these diseases show different clinical manifestations, the presence of toxic protein aggregates in the brain of patients is a common feature to all of them, suggesting a loss of protein homeostasis.Aging, the primary risk factor for the majority of neurodegenerative disorders, is linked to the impairment of degradative compartments such as lysosomes and autophagosomes. Besides, many genetic factors for Alzheimer's disease, Parkinson's disease, or frontotemporal dementia, as examples of frequent neurodegenerative diseases, are causative of endo-lysosomal and autophagosomal dysfunctions. There is scientific evidence suggesting that neurons can counteract the accumulation of undegraded cellular material by the secretion of extracellular vesicles (EVs), which are vesicles with a size ranging from 50 to 100 nm generated in a type of endosomal compartment named multivesicular body. EVs play a crucial role in removing cellular waste, promoting protein aggregation, and spreading toxic protein aggregates in the brain of patients. In this review, the interplay between the impairment of degradative compartments, the secretion of EVs, and their pathological/beneficial role in neurodegeneration is described.

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Loss of exosomes in progranulin-associated frontotemporal dementia

Cited by 49 publications

References 62 publications

Proteomic Profiling Reveals the Transglutaminase-2 Externalization Pathway in Kidneys after Unilateral Ureteric Obstruction

Proteomic Profiling Reveals the Transglutaminase-2 Externalization Pathway in Kidneys after Unilateral Ureteric Obstruction

Digital Detection of Exosomes by Interferometric Imaging

The interplay between aging‐associated loss of protein homeostasis and extracellular vesicles in neurodegeneration

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