Proteome profiling of exosomes derived from human primary and metastatic colorectal cancer cells reveal differential expression of key metastatic factors and signal transduction components

Ji, Hong; Greening, David W.; Barnes, Thomas W.; Lim, Jeremy; Tauro, Bow J.; Rai, Alin; Xu, Rong; Adda, Christopher G.; Mathivanan, Suresh; Zhao, Wei; Xue, Yanhong; Xu, Tao; Zhu, Hong; Simpson, Richard J.

doi:10.1002/pmic.201200562

Cited by 308 publications

(299 citation statements)

References 103 publications

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“…cholesterol and ceramide), membrane fusion-related proteins (Rab GTPases, flotillins and connexins), proteins involved in vesicle formation (Alix, Tsg 101), integral membrane proteins such as tetraspanins (CD9, CD63, CD 81) and major histocompatibility complex (MHC) class I and II as well as proteins related to the cytoskeleton and the cell metabolism have been identified (16). Also proteins involved in the pathogenesis of cancer such as oncoproteins MET and mutant KRAS have been found in exosomes (17,18). As nucleic acidrelated cargo, mRNA, miRNA, long non-coding RNAs as well as DNA have been detected (19).…”

Section: General Features Of Exosomesmentioning

confidence: 99%

The Multiple Roles of Exosomes in Metastasis

Weidle

Birzele

Kollmorgen

et al. 2017

CGP

168

133

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Abstract. Exosomes are important contributors to cellMetastases are responsible for the death of a large majority of cancer patients despite considerable progress in surgical techniques, radiotherapy, chemotherapy and targeted therapies including immuno-therapy (1). A dramatic reduction of metastatic burden has been observed, however, tumor elimination is almost always incomplete. This phenomenon is based on drug resistance, which is due to adaptation of intracellular pathways or on activation of survival-supporting autocrine and paracrine pathways and several secreted factors expressed by drug-sensitive tumor cells after therapy (2). Metastasis can occur through release of cancer cells from the primary tumor into body cavities that holds true for ovarian and CNS tumors, or via hematogeneous and lymphatic vessels of the circulatory system (3). Metastases of some tumors are directed besides to lymph nodes to mainly one type of organ only, such as prostate cancer to the bones, pancreatic cancer and uveal melanoma to the liver, whereas tumors such as melanoma, breast-and lung cancer can colonize several types of organs (3). Tumor cells have been found in the blood of patients with early-stage cancer and in some cases even before the primary tumor has been diagnosed (4, 5). Recently, making use of single-cell expression profiling, it has been shown that early metastatic cells possess a stem-like gene signature and give rise to heterogeneous metastases (6). The metastatic process is characterized by a defined sequence of events (7,8). Initial steps are detachment from the extracellular matrix (ECM), invasion into the surrounding tissue and proteolysis of the basement membrane. After intravasation, survival of circulating tumor cells (CTCs) is achieved by forming clusters, binding to platelets and immune evasion. Subsequently, they arrest in distal microvascular beds and extravasation can be achieved either by migration through intercellular junctions of endothelial cells (EC) or penetration of a single EC (9). CTCs can also colonize their tumors of origin, a process referred to as "tumor self-seeding", selecting for cancer cell populations more aggressive than those present in the primary tumor (10). After extravasation, colonization and outgrowth in the parenchyma of distant organs are the following steps. A common theme of the metastatic process is settlement of disseminated tumor cells (DTCs) into latency (dormancy), which can last from several months to decades (11). It has been observed that DTCs are recruited into pre-metastatic niches that support their survival by interactions with endothelial, myeloid cells, fibroblasts and other types of cells. After adaptation to the host microenvironment and suppression of an anti-tumoral immune response, DTCs are activated by not yet completely resolved mechanisms. Finally their outgrowth is based on an angiogenic switch mediated by pro-angiogenic factors and establishment of a vascular network to support the metabolic demands of colonizing tumor cells (12). In the follo...

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Section: General Features Of Exosomesmentioning

confidence: 99%

The Multiple Roles of Exosomes in Metastasis

Weidle

Birzele

Kollmorgen

et al. 2017

CGP

168

133

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“…For testing exosomes at the in situ protein level, we selected the ALG 2-interacting protein X (ALIX; also known as programmed cell death 6-interacting protein (PDCD6IP)), a multifunctional protein, which is involved in biogenesis of MVBs (as ESCRT I-III binding protein), lysosomal degradation and accumulates in exosomes. 3,[25][26][27] ALIX has been tested widely in exosome studies of colorectal carcinoma cell cultures 9,28 and has shown diagnostic and prognostic significance in different tumors. [29][30][31] Besides testing the transcript levels, here we determined ALIX protein expression in situ in the epithelial and stromal compartment including α-smooth muscle actin (SMA)-positive cells in low-and high-grade dysplastic adenomas as well as non-metastatic and metastatic (both lymph node and distant) colorectal carcinomas with particular focus on individual Musashi1-positive cancer stem cells at the tumor front.…”

mentioning

confidence: 99%

Exosomes in colorectal carcinoma formation: ALIX under the magnifying glass

et al. 2016

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Exosomes are small membrane vesicles that have important roles in transporting a great variety of bioactive molecules between epithelial compartment and their microenvironment during tumor formation including colorectal adenoma-carcinoma sequence. We tested the mRNA expression of the top 25 exosome-related markers based on ExoCharta database in healthy (n = 49), adenoma (n = 49) and colorectal carcinoma (n = 49) patients using Affymetrix HGU133 Plus2.0 microarrays. Most related genes showed significantly elevated expression including PGK1, PKM, ANXA5, ENO1, HSP90AB1 and MSN during adenoma-carcinoma sequence. Surprisingly, the expression of ALIX (ALG 2-interacting protein X), involved in multivesicular body (MVB) and exosome formation, was significantly reduced in normal vs adenoma (P = 5.02 × 10 − 13 ) and in normal vs colorectal carcinoma comparisons (P = 1.51 × 10 − 10 ). ALIX also showed significant reduction (Po 0.05) at the in situ protein level in the epithelial compartment of adenoma (n = 35) and colorectal carcinoma (n = 37) patients compared with 27 healthy individuals. Furthermore, significantly reduced ALIX protein levels were accompanied by their gradual transition from diffuse cytoplasmic expression to granular signals, which fell into the 0.6-2 μm diameter size range of MVBs. These ALIX-positive particles were seen in the tumor nests, including tumorstroma border, which suggest their exosome function. MVB-like structures were also detected in tumor microenvironment including α-smooth muscle actin-positive stromal cells, budding off cancer cells in the tumor front as well as in cancer cells entrapped within lymphoid vessels. In conclusion, we determined the top aberrantly expressed exosome-associated markers and revealed the transition of diffuse ALIX protein signals into a MVB-like pattern during adenoma-carcinoma sequence. These tumor-associated particles seen both in the carcinoma and the surrounding microenvironment can potentially mediate epithelial-stromal interactions involved in the regulation of tumor growth, metastatic invasion and therapy response.

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“…Cell-free nucleic acid has been identified in stable protein complexes, as well as encapsulated in microvesicles and exosomes (23)(24)(25). Exosomes are small (50-100 nm) vesicles that carry a unique composition of proteins (26), lipids (27), mRNA (28), and miRNA (29). Exosomes form in the endosomal multivesicular bodies of the cytoplasm of various cell types and are secreted into body fluids, including blood plasma (30).…”

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confidence: 99%

Telomeric repeat-containing RNA (TERRA) constitutes a nucleoprotein component of extracellular inflammatory exosomes

Wang

Deng

Dahmane

et al. 2015

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

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Telomeric repeat-containing RNA (TERRA) has been identified as a telomere-associated regulator of chromosome end protection. Here, we report that TERRA can also be found in extracellular fractions that stimulate innate immune signaling. We identified extracellular forms of TERRA in mouse tumor and embryonic brain tissue, as well as in human tissue culture cell lines using RNA in situ hybridization. RNA-seq analyses revealed TERRA to be among the most highly represented transcripts in extracellular fractions derived from both normal and cancer patient blood plasma. Cell-free TERRA (cfTERRA) could be isolated from the exosome fractions derived from human lymphoblastoid cell line (LCL) culture media. cfTERRA is a shorter form (∼200 nt) of cellular TERRA and copurifies with CD63-and CD83-positive exosome vesicles that could be visualized by cyro-electron microscopy. These fractions were also enriched for histone proteins that physically associate with TERRA in extracellular ChIP assays. Incubation of cfTERRAcontaining exosomes with peripheral blood mononuclear cells stimulated transcription of several inflammatory cytokine genes, including TNFα, IL6, and C-X-C chemokine 10 (CXCL10) Exosomes engineered with elevated TERRA or liposomes with synthetic TERRA further stimulated inflammatory cytokines, suggesting that exosome-associated TERRA augments innate immune signaling. These findings imply a previously unidentified extrinsic function for TERRA and a mechanism of communication between telomeres and innate immune signals in tissue and tumor microenvironments.TERRA | telomere | exosome | innate immunity | cytokine

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Proteome profiling of exosomes derived from human primary and metastatic colorectal cancer cells reveal differential expression of key metastatic factors and signal transduction components

Cited by 308 publications

References 103 publications

The Multiple Roles of Exosomes in Metastasis

The Multiple Roles of Exosomes in Metastasis

Exosomes in colorectal carcinoma formation: ALIX under the magnifying glass

Telomeric repeat-containing RNA (TERRA) constitutes a nucleoprotein component of extracellular inflammatory exosomes

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