Isolation and Proteomic Analysis of Microvesicles and Exosomes from HT22 Cells and Primary Neurons

Witas, Richard; Chaput, Dale; Khan, Hirah; Stevens, Stanley M.; Kang, David E.

doi:10.1007/978-1-4939-6952-4_12

Cited by 10 publications

(14 citation statements)

References 11 publications

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“…Our proteomic analysis further revealed that exosomes-derived from GIST cells carry several common exosomal proteins, tumor specific markers, and previously unreported proteins indicating that GDE display shared proteomic features with exosomes-derived from other cells types, while conserving tumor identity. Many proteomic studies, aiming to identify tumor specific markers, took advantage of the homogeneous distribution profiles of exosomes produced by various cultured cell types (102)(103)(104)(105)(106)(107). Yet, translation and validation of these findings using in vivoderived exosomes are still very challenging.…”

Section: Discussionmentioning

confidence: 99%

Insights into the Proteome of Gastrointestinal Stromal Tumors-Derived Exosomes Reveals New Potential Diagnostic Biomarkers

Atay

Wilkey

Milhem³

et al. 2018

Molecular & Cellular Proteomics

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Developing tumors continuously release nano-sized vesicles that represent circulating "fingerprints" of the tumor's identity. In gastrointestinal stromal tumor (GIST), we have previously reported that these tumors release "oncosomes" carrying the constitutively activated tyrosine kinase (TK) receptor KIT. Despite the clinical utility of TK inhibitors, such as imatinib mesylate (IM), recurrence and metastasis are clinical problems that urge the need to identify new tumor-derived molecules. To this aim, we performed the first high quality proteomic study of GISTderived exosomes (GDEs) and identified 1,060 proteins composing the core GDE proteome (cGDEp). The cGDEp was enriched in diagnostic markers (e.g. KIT, CD34, ANO1, PROM1, PRKCQ, and ENG), as well as proteins encoded by genes previously reported expressed in GIST (e.g. DPP4, FHL1, CDH11, and KCTD12). Many of these proteins were validated using cell lines, patient-derived KIT ؉ exosomes, and GIST tissues. We further show that in vitro and in vivo-derived GDE, carry proteins associated with IM response, such as Sprouty homolog 4 (SPRY4), surfeit 4 (SURF4), ALIX, and the cGMP-dependent 3,5cyclic phosphodiesterase 2A (PDE2A). Additionally, we report that the total exosome levels and exosome-associated KIT and SPRY4 protein levels have therapeutic values. In fact, molecular characterization of in vivo-derived KIT ؉ exosomes indicate significant sorting of p-KIT Tyr719 , total KIT, and SPRY4 after IM-treatment of metastatic patients as compared with the pre-IM levels. Our data suggest that analysis of circulating exosomes levels

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Section: Discussionmentioning

confidence: 99%

Insights into the Proteome of Gastrointestinal Stromal Tumors-Derived Exosomes Reveals New Potential Diagnostic Biomarkers

Atay

Wilkey

Milhem³

et al. 2018

Molecular & Cellular Proteomics

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“…The cargoes contained within EVs reflect both the intracellular origin of the cargo as well as the cell type from which the vesicle was derived. We refer readers to the many profiling studies which have been published on this subject for further information . Prototypical exosome markers include tetraspannins such as CD9, CD63 and CD81, and ESCRT proteins Alix and TSG101 .…”

Section: Functional Cargoes In Microvesicles Derived From Tumor Cellsmentioning

confidence: 99%

“…We refer readers to the many profiling studies which have been published on this subject for further information. 36,66,[94][95][96][97][98][99][100] Prototypical exosome markers include tetraspannins such as CD9, CD63 and CD81, and ESCRT proteins Alix and TSG101. 37,101 Unique and universal MV markers are somewhat less well-defined.…”

Section: Functional Cargoes In Microvesicles Derived From Tumor Cellsmentioning

confidence: 99%

The biology of extracellular microvesicles

Sedgwick

D’Souza‐Schorey

2018

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The study of extracellular vesicles (EVs) is a rapidly evolving field, owing in large part to recent advances in the realization of their significant contributions to normal physiology and disease. Once discredited as cell debris, these membrane vesicles have now emerged as mediators of intercellular communication by interaction with target cells, drug and gene delivery, and as potentially versatile platforms of clinical biomarkers as a result of their distinctive protein, nucleic acid and lipid cargoes. While there are multiple classes of EVs released from almost all cell types, here we focus primarily on the biogenesis, fate and functional cargoes of microvesicles (MVs). MVs regulate many important cellular processes including facilitating cell invasion, cell growth, evasion of immune response, stimulating angiogenesis, drug resistance and many others.

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“…In these cells, hTERT mRNA was translated into an active form that transforms them into telomerase-positive cells [122]. Of interest, hTERT expression in hTERT negative cells affected their biological properties towards a tumor like-phenotype, characterized by extended or unlimited life span, prevention and reversal of senescence, resistance to DNA damage, and prevention of apoptosis [19,123]. …”

Section: Role Of Evs In the Malignancy-microenvironment Cross-talkmentioning

confidence: 99%

“…In some cases, EV cytoplasmic and surface content suggest a degree of specific cellular packaging into EVs [6,19]. …”

Section: Introductionmentioning

confidence: 99%

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

Caivano

Rocca

Laurenzana

et al. 2017

IJMS

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Extracellular vesicles (EVs) are a heterogeneous group of particles, between 15 nanometers and 10 microns in diameter, released by almost all cell types in physiological and pathological conditions, including tumors. EVs have recently emerged as particularly interesting informative vehicles, so that they could be considered a true “cell biopsy”. Indeed, EV cargo, including proteins, lipids, and nucleic acids, generally reflects the nature and status of the origin cells. In some cases, EVs are enriched of peculiar molecular cargo, thus suggesting at least a degree of specific cellular packaging. EVs are identified as important and critical players in intercellular communications in short and long distance interplays. Here, we examine the physiological role of EVs and their activity in cross-talk between bone marrow microenvironment and neoplastic cells in hematological malignancies (HMs). In these diseases, HM EVs can modify tumor and bone marrow microenvironment, making the latter “stronger” in supporting malignancy, inducing drug resistance, and suppressing the immune system. Moreover, EVs are abundant in biologic fluids and protect their molecular cargo against degradation. For these and other “natural” characteristics, EVs could be potential biomarkers in a context of HM liquid biopsy and therapeutic tools. These aspects will be also analyzed in this review.

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Isolation and Proteomic Analysis of Microvesicles and Exosomes from HT22 Cells and Primary Neurons

Cited by 10 publications

References 11 publications

Insights into the Proteome of Gastrointestinal Stromal Tumors-Derived Exosomes Reveals New Potential Diagnostic Biomarkers

Insights into the Proteome of Gastrointestinal Stromal Tumors-Derived Exosomes Reveals New Potential Diagnostic Biomarkers

The biology of extracellular microvesicles

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

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