Tetraspanin CD63 acts as a pro-metastatic factorviaβ-catenin stabilization

High-affinity IgE receptor (FcεRI) cross-linking on mast cells (MCs) induces secretion of preformed allergy mediators (degranulation) and synthesis of lipid mediators and cytokines. Degranulation produces many symptoms of immediate-type allergic reactions and is modulated by adhesion to surfaces coated with specific extracellular matrix (ECM) proteins. The signals involved in this modulation are mostly unknown and their contribution to allergic reactions in vivo is unclear. Here we report the generation of monoclonal antibodies that potently suppress FcεRI-induced degranulation, but not leukotriene synthesis. We identified the antibody target as the tetraspanin CD63. Tetraspanins are membrane molecules that form multimolecular complexes with a broad array of molecules including ECM protein-binding β integrins. We found that anti-CD63 inhibits MC adhesion to fibronectin and vitronectin. Furthermore, anti-CD63 inhibits FcεRI-mediated degranulation in cells adherent to those ECM proteins but not in nonadherent cells. Thus the inhibition of degranulation by anti-CD63 correlates with its effect on adhesion. In support of a mechanistic linkage between the two types of inhibition, anti-CD63 had no effect on FcεRI-induced global tyrosine phosphorylation and calcium mobilization but impaired the Gab2–PI3K pathway that is known to be essential for both degranulation and adhesion. Finally, we showed that these antibodies inhibited FcεRI-mediated allergic reactions in vivo. These properties raise the possibility that anti-CD63 could be used as therapeutic agents in MC-dependent diseases.

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DNA-Directed Patterning for Versatile Validation and Characterization of a Lipid-Based Nanoparticle Model of SARS-CoV-2

10.26434/chemrxiv.14208455 ◽

2021 ◽

Author(s):

Molly Kozminsky ◽

Thomas Carey ◽

Lydia L. Sohn

Keyword(s):

High Throughput ◽

Glass Substrate ◽

Rapid Development ◽

Neutralizing Antibody ◽

S Protein ◽

Biological Relevance ◽

The Face ◽

Great Control ◽

Tetraspanin Cd63

Lipid-based nanoparticles have risen to the forefront of the COVID-19 pandemic—from encapsulation of vaccine components to modeling the virus, itself. Their rapid development in the face of the volatile nature of the pandemic requires high-throughput, highly flexible methods for characterization. DNA-directed patterning is a versatile method to immobilize and segregate lipid-based nanoparticles for subsequent analysis. DNA-directed patterning selectively conjugates oligonucleotides onto a glass substrate and then hybridizes them to complementary oligonucleotides tagged to the liposomes, thereby patterning them with great control and precision. The power of this method is demonstrated by characterizing a novel recapitulative lipid-based nanoparticle model of SARS-CoV-2 —S-liposomes— which present the SARS-CoV-2 spike (S) protein on their surfaces. Patterning of a mixture of S-liposomes and liposomes that display the tetraspanin CD63 into discrete regions of a substrate is used to show that ACE2 specifically binds to S-liposomes. Importantly, DNA-directed patterning of S-liposomes is used to verify the performance of a commercially available neutralizing antibody against the S protein. Ultimately, the introduction of S-liposomes to ACE2-expressing cells demonstrates the biological relevance of DNA-directed patterning. Overall, DNA-directed patterning enables a wide variety of custom assays for the characterization of any lipid-based nanoparticle.

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Tetraspanin CD63 Promotes Vascular Endothelial Growth Factor Receptor 2-β1 Integrin Complex Formation, Thereby Regulating Activation and Downstream Signaling in Endothelial Cellsin Vitroandin Vivo

Journal of Biological Chemistry ◽

10.1074/jbc.m113.468199 ◽

2013 ◽

Vol 288 (26) ◽

pp. 19060-19071 ◽

Cited By ~ 30

Author(s):

Sònia Tugues ◽

Satoshi Honjo ◽

Christian König ◽

Narendra Padhan ◽

Jeffrey Kroon ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Complex Formation ◽

Growth Factor Receptor ◽

Β1 Integrin ◽

Vascular Endothelial ◽

Downstream Signaling ◽

Endothelial Growth Factor ◽

Tetraspanin Cd63

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Abstract 5444: Clinical significance of tetraspanin CD63 in human ESCC

10.1158/1538-7445.am2020-5444 ◽

2020 ◽

Author(s):

Yasunori Matsumoto ◽

Masayuki Kano ◽

Kentaro Murakami ◽

Satoshi Endo ◽

Takeshi Toyozumi ◽

...

Keyword(s):

Clinical Significance ◽

Tetraspanin Cd63

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RPN2-mediated glycosylation of tetraspanin CD63 regulates breast cancer cell malignancy

Molecular Cancer ◽

10.1186/1476-4598-13-134 ◽

2014 ◽

Vol 13 (1) ◽

pp. 134 ◽

Cited By ~ 36

Author(s):

Naoomi Tominaga ◽

Keitaro Hagiwara ◽

Nobuyoshi Kosaka ◽

Kimi Honma ◽

Hitoshi Nakagama ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Tetraspanin Cd63 ◽

Cell Malignancy

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The tetraspanin CD63 is involved in granule targeting of neutrophil elastase

Blood ◽

10.1182/blood-2007-10-116285 ◽

2008 ◽

Vol 112 (8) ◽

pp. 3444-3454 ◽

Cited By ~ 25

Author(s):

Linda Källquist ◽

Markus Hansson ◽

Ann-Maj Persson ◽

Hans Janssen ◽

Jero Calafat ◽

...

Keyword(s):

Neutrophil Elastase ◽

Adaptor Protein ◽

State Level ◽

Integral Membrane Protein ◽

Cos Cells ◽

Large Extracellular Loop ◽

Constitutive Secretion ◽

Primary Granules ◽

Secretory Lysosomes ◽

Tetraspanin Cd63

Abstract Targeting mechanisms of neutrophil elastase (NE) and other luminal proteins stored in myeloperoxidase (MPO)–positive secretory lysosomes/primary granules of neutrophils are unknown. These granules contain an integral membrane protein, CD63, with an adaptor protein-3–dependent granule delivery system. Therefore, we hypothesized that CD63 cooperates in granule delivery of the precursor of NE (proNE). Supporting this hypothesis, an association was demonstrated between CD63 and proNE upon coexpression in COS cells. This also involved augmented cellular retention of proNE requiring intact large extracellular loop of CD63. Furthermore, depletion of CD63 in promyelocytic HL-60 cells with RNA interference or a CD63 mutant caused reduction of cellular NE. However, the proNE steady-state level was similar to wild type in CD63-depleted clones, making it feasible to examine possible effects of CD63 on NE trafficking. Thus, depletion of CD63 led to reduced processing of proNE into mature NE and reduced constitutive secretion. Furthermore, CD63-depleted cells showed a lack of morphologically normal granules, but contained MPO-positive cytoplasmic vacuoles with a lack of proNE and NE. Collectively, our data suggest that granule proteins may cooperate in targeting; CD63 can be involved in ER or Golgi export, cellular retention, and granule targeting of proNE before storage as mature NE.

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Palmitoylation-dependent association with CD63 targets the Ca2+ sensor synaptotagmin VII to lysosomes

The Journal of Cell Biology ◽

10.1083/jcb.201003021 ◽

2010 ◽

Vol 191 (3) ◽

pp. 599-613 ◽

Cited By ~ 45

Author(s):

Andrew R. Flannery ◽

Cecilia Czibener ◽

Norma W. Andrews

Keyword(s):

Plasma Membrane ◽

Golgi Complex ◽

Membrane Repair ◽

Particle Uptake ◽

Dependent Process ◽

Synaptotagmin Vii ◽

Plasma Membrane Repair ◽

Lysosomal Targeting ◽

Tetraspanin Cd63

Syt VII is a Ca2+ sensor that regulates lysosome exocytosis and plasma membrane repair. Because it lacks motifs that mediate lysosomal targeting, it is unclear how Syt VII traffics to these organelles. In this paper, we show that mutations or inhibitors that abolish palmitoylation disrupt Syt VII targeting to lysosomes, causing its retention in the Golgi complex. In macrophages, Syt VII is translocated simultaneously with the lysosomal tetraspanin CD63 from tubular lysosomes to nascent phagosomes in a Ca2+-dependent process that facilitates particle uptake. Mutations in Syt VII palmitoylation sites block trafficking of Syt VII, but not CD63, to lysosomes and phagosomes, whereas tyrosine replacement in the lysosomal targeting motif of CD63 causes both proteins to accumulate on the plasma membrane. Complexes of CD63 and Syt VII are detected only when Syt VII palmitoylation sites are intact. These findings identify palmitoylation-dependent association with the tetraspanin CD63 as the mechanism by which Syt VII is targeted to lysosomes.

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The Tetraspanin CD63/lamp3 Cycles between Endocytic and Secretory Compartments in Human Endothelial Cells

Molecular Biology of the Cell ◽

10.1091/mbc.11.5.1829 ◽

2000 ◽

Vol 11 (5) ◽

pp. 1829-1843 ◽

Cited By ~ 179

Author(s):

Toshihide Kobayashi ◽

Ulrich M. Vischer ◽

Corinne Rosnoblet ◽

Cécile Lebrand ◽

Margaret Lindsay ◽

...

Keyword(s):

Endothelial Cells ◽

Plasma Membrane ◽

Late Endosome ◽

Human Endothelial Cells ◽

Late Endosomes ◽

Lysobisphosphatidic Acid ◽

Type C ◽

Niemann Pick ◽

Tetraspanin Cd63

In the present study, we show that in human endothelial cells the tetraspanin CD63/lamp3 distributes predominantly to the internal membranes of multivesicular–multilamellar late endosomes, which contain the unique lipid lysobisphosphatidic acid. Some CD63/lamp3 is also present in Weibel–Palade bodies, the characteristic secretory organelle of these cells. We find that CD63/lamp3 molecules can be transported from late endosomes to Weibel–Palade bodies and thus that CD63/lamp3 cycles between endocytic and biosynthetic compartments; however, movement of CD63/lamp3 is much slower than that of P-selectin, which is known to cycle between plasma membrane and Weibel–Palade bodies. When cells are treated with U18666A, a drug that mimics the Niemann-Pick type C syndrome, both proteins accumulate in late endosomes and fail to reach Weibel–Palade bodies efficiently, suggesting that P-selectin, like CD63/lamp3, cycles via late endosomes. Our data suggest that CD63/lamp3 partitions preferentially within late endosome internal membranes, thus causing its accumulation, and that this mechanism contributes to CD63/lamp3 retention in late endosomes; however, our data also indicate that the protein can eventually escape from these internal membranes and recycle toward Weibel–Palade bodies to be reused. Our observations thus uncover the existence of a selective trafficking route from late endosomes to Weibel–Palade bodies.

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