Regulation of Leukocytes by TspanC8 Tetraspanins and the “Molecular Scissor” ADAM10

Matthews, Alexandra L.; Koo, Chek Ziu; Szyroka, Justyna; Harrison, Neale; Kanhere, Aditi; Tomlinson, Michael G.

doi:10.3389/fimmu.2018.01451

Cited by 24 publications

(26 citation statements)

References 66 publications

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“…The TspanC8s comprise Tspan5, 10, 14, 15, 17 and 33, and are so-called because of eight cysteines within their large extracellular loop. Different cell types express distinct TspanC8 repertoires (9,10) and emerging evidence suggests that each TspanC8 may cause ADAM10 to cleave specific substrates (6,(11)(12)(13)(14)(15)(16). However, there is an urgent need to generate monoclonal antibodies (mAbs) to all TspanC8s, to define fully their expression profiles and functional mechanisms.…”

mentioning

confidence: 99%

The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Koo

Harrison

Noy

et al. 2020

Journal of Biological Chemistry

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A disintegrin and metalloprotease 10 (ADAM10) is a transmembrane protein essential for embryonic development, and its dysregulation underlies disorders such as cancer, Alzheimer’s disease and inflammation. ADAM10 is a “molecular scissor” that proteolytically cleaves the extracellular region from >100 substrates, including Notch, amyloid precursor protein, cadherins, growth factors, and chemokines. ADAM10 has been recently proposed to function as six distinct scissors with different substrates, depending on its association with one of six regulatory tetraspanins, termed TspanC8s. However, it remains unclear to what degree ADAM10 function critically depends on a TspanC8 partner, and a lack of monoclonal antibodies specific for most TspanC8s has hindered investigating this question. To address this knowledge gap, here we designed an immunogen to generate the first monoclonal antibodies targeting Tspan15, a model TspanC8. The immunogen was created in an ADAM10-knockout mouse cell line stably overexpressing human Tspan15, because we hypothesized that expression in this cell line would expose epitopes that are normally blocked by ADAM10. Following immunization of mice, this immunogen strategy generated four Tspan15 antibodies. Using these antibodies, we show that endogenous Tspan15 and ADAM10 co-localize on the cell surface, that ADAM10 is the principal Tspan15-interacting protein, that endogenous Tspan15 expression requires ADAM10 in cell lines and primary cells, and that a synthetic ADAM10/Tspan15 fusion protein is a functional scissor. Furthermore, two of the four antibodies impaired ADAM10/Tspan15 activity. These findings suggest that Tspan15 directly interacts with ADAM10 in a functional scissor complex.

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confidence: 99%

The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Koo

Harrison

Noy

et al. 2020

Journal of Biological Chemistry

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“…ADAM10 is a transmembrane protease that cleaves extracellular regions of several transmembrane target proteins, including Notch . Furthermore, other Tspans have been shown to regulate ADAM10 maturation and activity, and Tspan3 itself has been shown to regulate ADAM10 expression . Our studies show that the Cldn‐11 effect on Notch signaling may be mediated via Tspan3 regulation of ADAM10 function.…”

Section: Discussionmentioning

confidence: 66%

“…ADAM10 is a transmembrane protein that cleaves extracellular regions of several transmembrane target proteins, including Notch, and we previously showed that osteoblasts express ADAM10 . Further, it was recently found that mutation of Tspan14 reduced ADAM10 surface expression and activity in human umbilical vein endothelial cells, and a number of other Tspans, including Tspan5, Tspan10, Tspan15, Tspan17, and Tspan33, have been shown to regulate ADAM10 activity . Notably, Tspan3 has also been found to regulate ADAM10 expression .…”

Section: Resultsmentioning

confidence: 93%

Novel Role for Claudin-11 in the Regulation of Osteoblasts via Modulation of ADAM10-Mediated Notch Signaling

Lindsey

Xing

Pourteymoor

et al. 2019

Journal of Bone and Mineral Research

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The claudin (Cldn) family comprises 27 members of 20 to 34 kDa transmembrane tight junction proteins. In addition to Cldns' established canonical role as barriers controlling paracellular flow of molecules, a distinct noncanonical role for them as mediators of cell signaling is now emerging. In our studies evaluating Cldn family expression levels during osteoblast differentiation, Cldn-11 showed the largest increase (60-fold). Immunohistochemistry studies revealed high Cldn-11 expression in trabecular (Tb) bone lining cells. Micro-CT analysis of femurs and vertebrae of Cldn-11 knock-out (KO) mice at 12 weeks of age exhibited a 40% (p < 0.01) reduction in Tb bone volume adjusted for tissue volume compared with control mice, a change caused by significant reductions in Tb number and thickness and increase in Tb separation. Histomorphometry and serum biomarker studies revealed that reduced bone formation, not increased resorption, is the cause for reduced Tb bone volume in the Cldn-11 KO mice. Cldn-11 KO osteoblasts expressed reduced ALP and BSP, whereas Cldn-11 overexpression in MC3T3-E1 cells increased expression of ALP and BSP. Mechanistically, Cldn-11 interacted with tetraspanin (Tspan)3 in osteoblasts, and Tspan3 knockdown reduced osteoblast differentiation. Because members of the Tspan family regulate cell functions via Notch signaling, we evaluated whether Cldn-11/ Tspan3 regulates Notch signaling in osteoblasts. Accordingly, Notch targets Hey1 and Hey2 were significantly upregulated in Cldn-11 overexpressing cultures but downregulated in both Cldn-11 KO and Tspan3 knockdown osteoblasts. Because ADAM10 has been shown to interact with Tspan family members to regulate Notch signaling, we evaluated whether Cldn-11 regulates ADAM10 expression. Cldn-11 overexpressing cells express more mature ADAM10, and an ADAM10 inhibitor blocked the Cldn-11 effect on osteoblast differentiation. Based on these data, we propose Cldn-11 as a novel component of an osteoblast cell surface protein complex, comprising Tspan3 and ADAM10, which regulates Notch signaling and cell differentiation.

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“…Due to the ease and simplicity of sample handling, QPM techniques can aid predictive models of thrombus formation, contraction, and stability across a range of shear rates and are ideal modalities for development for point-of-care devices to assess platelet function and thrombosis and bleeding risk in at risk patients. These imaging approaches can address research questions targeting mechanisms involved in the regulation of thrombus size, for example the respective roles of metalloproteinases (34,198) or tetraspanins (199,200) in modulating thrombus size and stability. However, as QPM techniques rely on phase information in transmitted light, they can often be limited by strongly scattering media.…”

Section: Beyond Fluorescence Imaging: Quantitative Imaging Without Flmentioning

confidence: 99%

Imaging Platelet Processes and Function—Current and Emerging Approaches for Imaging in vitro and in vivo

et al. 2020

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Platelets are small anucleate cells that are essential for many biological processes including hemostasis, thrombosis, inflammation, innate immunity, tumor metastasis, and wound healing. Platelets circulate in the blood and in order to perform all of their biological roles, platelets must be able to arrest their movement at an appropriate site and time. Our knowledge of how platelets achieve this has expanded as our ability to visualize and quantify discreet platelet events has improved. Platelets are exquisitely sensitive to changes in blood flow parameters and so the visualization of rapid intricate platelet processes under conditions found in flowing blood provides a substantial challenge to the platelet imaging field. The platelet's size (∼2 µm), rapid activation (milliseconds), and unsuitability for genetic manipulation, means that appropriate imaging tools are limited. However, with the application of modern imaging systems to study platelet function, our understanding of molecular events mediating platelet adhesion from a single-cell perspective, to platelet recruitment and activation, leading to thrombus (clot) formation has expanded dramatically. This review will discuss current platelet imaging techniques in vitro and in vivo, describing how the advancements in imaging have helped answer/expand on platelet biology with a particular focus on hemostasis. We will focus on platelet aggregation and thrombus formation, and how platelet imaging has enhanced our understanding of key events, highlighting the knowledge gained through the application of imaging modalities to experimental models in vitro and in vivo. Furthermore, we will review the limitations of current imaging techniques, and questions in thrombosis research that remain to be addressed. Finally, we will speculate how the same imaging advancements might be applied to the imaging of other vascular cell biological functions and visualization of dynamic cell-cell interactions.

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Regulation of Leukocytes by TspanC8 Tetraspanins and the “Molecular Scissor” ADAM10

Cited by 24 publications

References 66 publications

The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Novel Role for Claudin-11 in the Regulation of Osteoblasts via Modulation of ADAM10-Mediated Notch Signaling

Imaging Platelet Processes and Function—Current and Emerging Approaches for Imaging in vitro and in vivo

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