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The tetraspanins CD151 and Tspan8 are essential exosome components for the crosstalk between cancer initiating cells and their surrounding.

Yue S, Mu W, Erb U, Zöller M - Oncotarget (2015)

Bottom Line: Approaching to elaborate the underlying mechanism, we compared ASMLwt, -CD151kd and/or Tspan8kd clones.These effects are not seen or are weakened using ASML-CD151kd or -Tspan8kd exosomes, which is at least partly due to reduced binding/uptake of CD151- and/or Tspan8-deficient exosomes.Thus, CD151- and Tspan8-competent tumor exosomes support matrix degradation, reprogram stroma and hematopoietic cells and drive non-metastatic ASML-CD151/Tspan8kd cells towards a motile phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Tumor Cell Biology, University Hospital of Surgery, Heidelberg, Germany.

ABSTRACT
Tspan8 and CD151 are metastasis-promoting tetraspanins and a knockdown (kd) of Tspan8 or CD151 and most pronounced of both tetraspanins affects the metastatic potential of the rat pancreatic adenocarcinoma line ASML. Approaching to elaborate the underlying mechanism, we compared ASMLwt, -CD151kd and/or Tspan8kd clones. We focused on tumor exosomes, as exosomes play a major role in tumor progression and tetraspanins are suggested to be engaged in exosome targeting. ASML-CD151/Tspan8kd cells poorly metastasize, but regain metastatic capacity, when rats are pretreated with ASMLwt, but not ASML-CD151kd and/or -Tspan8kd exosomes. Both exosomal CD151 and Tspan8 contribute to host matrix remodelling due to exosomal tetraspanin-integrin and tetraspanin-protease associations. ASMLwt exosomes also support stroma cell activation with upregulation of cytokines, cytokine receptors and proteases and promote inflammatory cytokine expression in hematopoietic cells. Finally, CD151-/Tspan8-competent exosomes support EMT gene expression in poorly-metastatic ASML-CD151/Tspan8kd cells. These effects are not seen or are weakened using ASML-CD151kd or -Tspan8kd exosomes, which is at least partly due to reduced binding/uptake of CD151- and/or Tspan8-deficient exosomes. Thus, CD151- and Tspan8-competent tumor exosomes support matrix degradation, reprogram stroma and hematopoietic cells and drive non-metastatic ASML-CD151/Tspan8kd cells towards a motile phenotype.

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The impact of exosomal CD151 and Tspan8 on host cell adhesion molecule and protease expression(A,B) Flow cytometry analysis of protease expression in LnStr and LuFb after coculture with ASMLwt, -CD151kd and/or -Tspan8kd exosomes; mean percent±SD (3 assays) of stained cells; significant differences to untreated cells: *; (C,D) immunohistology of protease and adhesion molecule expression in draining LN after repeated ifp application of ASMLwt or -CD151kd and/or -Tspan8kd exosomes (scale bar: 150μm). Short term in vitro coculture of stroma cells with ASML exosomes hardly affected protease and adhesion molecule (data not shown) expression. Instead, after repeated exosome application in vivo, ASMLwt exosomes particularly promoted MMP2, MMP9 and TACE as well as CD49c, CD49d and CD104 expression. CD104 and TACE upregulation were weakened in ASML-CD151/Tspan8kd exosome treated rats and CD49c, CD49d, MMP2 and MMP9 expression were not supported.
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Figure 5: The impact of exosomal CD151 and Tspan8 on host cell adhesion molecule and protease expression(A,B) Flow cytometry analysis of protease expression in LnStr and LuFb after coculture with ASMLwt, -CD151kd and/or -Tspan8kd exosomes; mean percent±SD (3 assays) of stained cells; significant differences to untreated cells: *; (C,D) immunohistology of protease and adhesion molecule expression in draining LN after repeated ifp application of ASMLwt or -CD151kd and/or -Tspan8kd exosomes (scale bar: 150μm). Short term in vitro coculture of stroma cells with ASML exosomes hardly affected protease and adhesion molecule (data not shown) expression. Instead, after repeated exosome application in vivo, ASMLwt exosomes particularly promoted MMP2, MMP9 and TACE as well as CD49c, CD49d and CD104 expression. CD104 and TACE upregulation were weakened in ASML-CD151/Tspan8kd exosome treated rats and CD49c, CD49d, MMP2 and MMP9 expression were not supported.

Mentions: We first controlled the impact of exosomal CD151 and Tspan8 on protease expression. Only ASMLwt exosome uptake by LuFb and more pronounced by LnStr promoted TACE, MMP14, TIMP1 and TIMP2 expression. ADAMTS1, ADAMTS5 and uPA upregulation appeared largely CD151-dependent. Expression of MMP2 and MMP9 was not affected by short term in vitro coculture (Fig.5A,5B). However, distinct to the short term in vitro coculture, repeated ASML exosome application promoted TACE, but also MMP2 and MMP9 upregulation that was strongest in ASMLwt exosome-treated rats. ASML-CD151/Tspan8kd exosomes did not induce MMP9 and TACE upregulation; MMP2 expression was hardly increased in ASML-CD151kd exosome- and MMP9 and TACE expression in -Tspan8kd exosome-treated rats (Fig.5C). These findings fitted to the appearance of LN metastases with poor recovery of MMP2 in ASML-CD151kd tumors, poor recovery of MMP9 in both ASML-CD151kd and ASML-Tspan8kd tumors and very few ADAM17+ cells in ASML-Tspan8kd tumors. The reduced protease recovery was accompanied by a dense coll IV and LN332 matrix in ASML-CD151/Tspan8kd tumors (Suppl.Fig.4). Immunohistology of LN sections from ASMLwt exosome-treated rats additionally showed upregulated expression of CD49c and CD49d that was not seen in ASML-CD151/Tspan8kd exosome-treated rats. Upregulation of α6β4 was only seen in ASMLwt and ASML-CD151kd exosome-treated rats (Fig.5D).


The tetraspanins CD151 and Tspan8 are essential exosome components for the crosstalk between cancer initiating cells and their surrounding.

Yue S, Mu W, Erb U, Zöller M - Oncotarget (2015)

The impact of exosomal CD151 and Tspan8 on host cell adhesion molecule and protease expression(A,B) Flow cytometry analysis of protease expression in LnStr and LuFb after coculture with ASMLwt, -CD151kd and/or -Tspan8kd exosomes; mean percent±SD (3 assays) of stained cells; significant differences to untreated cells: *; (C,D) immunohistology of protease and adhesion molecule expression in draining LN after repeated ifp application of ASMLwt or -CD151kd and/or -Tspan8kd exosomes (scale bar: 150μm). Short term in vitro coculture of stroma cells with ASML exosomes hardly affected protease and adhesion molecule (data not shown) expression. Instead, after repeated exosome application in vivo, ASMLwt exosomes particularly promoted MMP2, MMP9 and TACE as well as CD49c, CD49d and CD104 expression. CD104 and TACE upregulation were weakened in ASML-CD151/Tspan8kd exosome treated rats and CD49c, CD49d, MMP2 and MMP9 expression were not supported.
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Figure 5: The impact of exosomal CD151 and Tspan8 on host cell adhesion molecule and protease expression(A,B) Flow cytometry analysis of protease expression in LnStr and LuFb after coculture with ASMLwt, -CD151kd and/or -Tspan8kd exosomes; mean percent±SD (3 assays) of stained cells; significant differences to untreated cells: *; (C,D) immunohistology of protease and adhesion molecule expression in draining LN after repeated ifp application of ASMLwt or -CD151kd and/or -Tspan8kd exosomes (scale bar: 150μm). Short term in vitro coculture of stroma cells with ASML exosomes hardly affected protease and adhesion molecule (data not shown) expression. Instead, after repeated exosome application in vivo, ASMLwt exosomes particularly promoted MMP2, MMP9 and TACE as well as CD49c, CD49d and CD104 expression. CD104 and TACE upregulation were weakened in ASML-CD151/Tspan8kd exosome treated rats and CD49c, CD49d, MMP2 and MMP9 expression were not supported.
Mentions: We first controlled the impact of exosomal CD151 and Tspan8 on protease expression. Only ASMLwt exosome uptake by LuFb and more pronounced by LnStr promoted TACE, MMP14, TIMP1 and TIMP2 expression. ADAMTS1, ADAMTS5 and uPA upregulation appeared largely CD151-dependent. Expression of MMP2 and MMP9 was not affected by short term in vitro coculture (Fig.5A,5B). However, distinct to the short term in vitro coculture, repeated ASML exosome application promoted TACE, but also MMP2 and MMP9 upregulation that was strongest in ASMLwt exosome-treated rats. ASML-CD151/Tspan8kd exosomes did not induce MMP9 and TACE upregulation; MMP2 expression was hardly increased in ASML-CD151kd exosome- and MMP9 and TACE expression in -Tspan8kd exosome-treated rats (Fig.5C). These findings fitted to the appearance of LN metastases with poor recovery of MMP2 in ASML-CD151kd tumors, poor recovery of MMP9 in both ASML-CD151kd and ASML-Tspan8kd tumors and very few ADAM17+ cells in ASML-Tspan8kd tumors. The reduced protease recovery was accompanied by a dense coll IV and LN332 matrix in ASML-CD151/Tspan8kd tumors (Suppl.Fig.4). Immunohistology of LN sections from ASMLwt exosome-treated rats additionally showed upregulated expression of CD49c and CD49d that was not seen in ASML-CD151/Tspan8kd exosome-treated rats. Upregulation of α6β4 was only seen in ASMLwt and ASML-CD151kd exosome-treated rats (Fig.5D).

Bottom Line: Approaching to elaborate the underlying mechanism, we compared ASMLwt, -CD151kd and/or Tspan8kd clones.These effects are not seen or are weakened using ASML-CD151kd or -Tspan8kd exosomes, which is at least partly due to reduced binding/uptake of CD151- and/or Tspan8-deficient exosomes.Thus, CD151- and Tspan8-competent tumor exosomes support matrix degradation, reprogram stroma and hematopoietic cells and drive non-metastatic ASML-CD151/Tspan8kd cells towards a motile phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Tumor Cell Biology, University Hospital of Surgery, Heidelberg, Germany.

ABSTRACT
Tspan8 and CD151 are metastasis-promoting tetraspanins and a knockdown (kd) of Tspan8 or CD151 and most pronounced of both tetraspanins affects the metastatic potential of the rat pancreatic adenocarcinoma line ASML. Approaching to elaborate the underlying mechanism, we compared ASMLwt, -CD151kd and/or Tspan8kd clones. We focused on tumor exosomes, as exosomes play a major role in tumor progression and tetraspanins are suggested to be engaged in exosome targeting. ASML-CD151/Tspan8kd cells poorly metastasize, but regain metastatic capacity, when rats are pretreated with ASMLwt, but not ASML-CD151kd and/or -Tspan8kd exosomes. Both exosomal CD151 and Tspan8 contribute to host matrix remodelling due to exosomal tetraspanin-integrin and tetraspanin-protease associations. ASMLwt exosomes also support stroma cell activation with upregulation of cytokines, cytokine receptors and proteases and promote inflammatory cytokine expression in hematopoietic cells. Finally, CD151-/Tspan8-competent exosomes support EMT gene expression in poorly-metastatic ASML-CD151/Tspan8kd cells. These effects are not seen or are weakened using ASML-CD151kd or -Tspan8kd exosomes, which is at least partly due to reduced binding/uptake of CD151- and/or Tspan8-deficient exosomes. Thus, CD151- and Tspan8-competent tumor exosomes support matrix degradation, reprogram stroma and hematopoietic cells and drive non-metastatic ASML-CD151/Tspan8kd cells towards a motile phenotype.

Show MeSH
Related in: MedlinePlus