Extracellular cadherin repeat domains EC1 and EC5 of T-cadherin are essential for its ability to stimulate angiogenic behavior of endothelial cells

Manjunath B. Joshi, Emmanouil Kyriakakis, Dennis Pfaff, Katharina Rupp, Maria Philippova, Paul Erne, Thérèse J. Resink

    Research output: Contribution to journalArticle

    7 Citations (Scopus)

    Abstract

    T-cadherin (T-cad) promotes survival, proliferation, and migration of endothelial cells and induces angiogenesis. We aimed to identify domains of T-cad functionally relevant to its effects on endothelial cell behavior. To specifically target the functional properties of the 5 cadherin repeat domains (EC1-EC5) of T-cad, endothelial cells were transduced with lentivectors containing specific T-cad-domain-deletion mutant constructs (ΔI, ΔII, ΔIII, ΔIV, ΔV). Empty (E) lentivector-transduced cells served as control. Similarly to overexpression of native T-cad, cells expressing ΔII, ΔIII, or ΔIV displayed elevated levels of p-Akt and p-GSK3β and increased proliferation rates (for ΔII, ΔIII) vs. E. ΔI- and ΔV-transduced cells exhibited reduced levels of p-Akt and p-GSK3β and retarded growth rates vs. E. Stimulatory effects of native T-cad overexpression on Akt and GSK3β phosphorylation were dose dependently inhibited by coexpression of ΔI or ΔV. Subsequent functional analyses compared only ΔI-, ΔII-, and ΔV-mutant constructs with E as a negative control. Unlike ΔII cells, ΔI and ΔV cells failed to exhibit homophilic ligation and deadhesion responses on a substratum of T-cad protein. In the wound assay, migration was increased for ΔII cells but impaired for ΔI and ΔV cells. In endothelial cell-spheroid assay, angiogenic sprouting was augmented for ΔII cells but inhibited for ΔI and ΔV cells. We conclude that EC1 and EC5 domains of T-cad are essential for its proangiogenic effects. ΔI and ΔV constructs may serve as dominant-negative mutants and as potential tools targeting excessive angiogenesis.

    Original languageEnglish
    Pages (from-to)4011-4021
    Number of pages11
    JournalFASEB Journal
    Volume23
    Issue number11
    DOIs
    Publication statusPublished - 2009

    Fingerprint

    Endothelial cells
    Cadherins
    Endothelial Cells
    Assays
    Phosphorylation
    Cells
    H-cadherin
    Ligation
    Wounds and Injuries
    Growth

    All Science Journal Classification (ASJC) codes

    • Biochemistry
    • Biotechnology
    • Genetics
    • Molecular Biology

    Cite this

    Joshi, Manjunath B. ; Kyriakakis, Emmanouil ; Pfaff, Dennis ; Rupp, Katharina ; Philippova, Maria ; Erne, Paul ; Resink, Thérèse J. / Extracellular cadherin repeat domains EC1 and EC5 of T-cadherin are essential for its ability to stimulate angiogenic behavior of endothelial cells. In: FASEB Journal. 2009 ; Vol. 23, No. 11. pp. 4011-4021.
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    abstract = "T-cadherin (T-cad) promotes survival, proliferation, and migration of endothelial cells and induces angiogenesis. We aimed to identify domains of T-cad functionally relevant to its effects on endothelial cell behavior. To specifically target the functional properties of the 5 cadherin repeat domains (EC1-EC5) of T-cad, endothelial cells were transduced with lentivectors containing specific T-cad-domain-deletion mutant constructs (ΔI, ΔII, ΔIII, ΔIV, ΔV). Empty (E) lentivector-transduced cells served as control. Similarly to overexpression of native T-cad, cells expressing ΔII, ΔIII, or ΔIV displayed elevated levels of p-Akt and p-GSK3β and increased proliferation rates (for ΔII, ΔIII) vs. E. ΔI- and ΔV-transduced cells exhibited reduced levels of p-Akt and p-GSK3β and retarded growth rates vs. E. Stimulatory effects of native T-cad overexpression on Akt and GSK3β phosphorylation were dose dependently inhibited by coexpression of ΔI or ΔV. Subsequent functional analyses compared only ΔI-, ΔII-, and ΔV-mutant constructs with E as a negative control. Unlike ΔII cells, ΔI and ΔV cells failed to exhibit homophilic ligation and deadhesion responses on a substratum of T-cad protein. In the wound assay, migration was increased for ΔII cells but impaired for ΔI and ΔV cells. In endothelial cell-spheroid assay, angiogenic sprouting was augmented for ΔII cells but inhibited for ΔI and ΔV cells. We conclude that EC1 and EC5 domains of T-cad are essential for its proangiogenic effects. ΔI and ΔV constructs may serve as dominant-negative mutants and as potential tools targeting excessive angiogenesis.",
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    Extracellular cadherin repeat domains EC1 and EC5 of T-cadherin are essential for its ability to stimulate angiogenic behavior of endothelial cells. / Joshi, Manjunath B.; Kyriakakis, Emmanouil; Pfaff, Dennis; Rupp, Katharina; Philippova, Maria; Erne, Paul; Resink, Thérèse J.

    In: FASEB Journal, Vol. 23, No. 11, 2009, p. 4011-4021.

    Research output: Contribution to journalArticle

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    AU - Joshi, Manjunath B.

    AU - Kyriakakis, Emmanouil

    AU - Pfaff, Dennis

    AU - Rupp, Katharina

    AU - Philippova, Maria

    AU - Erne, Paul

    AU - Resink, Thérèse J.

    PY - 2009

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    N2 - T-cadherin (T-cad) promotes survival, proliferation, and migration of endothelial cells and induces angiogenesis. We aimed to identify domains of T-cad functionally relevant to its effects on endothelial cell behavior. To specifically target the functional properties of the 5 cadherin repeat domains (EC1-EC5) of T-cad, endothelial cells were transduced with lentivectors containing specific T-cad-domain-deletion mutant constructs (ΔI, ΔII, ΔIII, ΔIV, ΔV). Empty (E) lentivector-transduced cells served as control. Similarly to overexpression of native T-cad, cells expressing ΔII, ΔIII, or ΔIV displayed elevated levels of p-Akt and p-GSK3β and increased proliferation rates (for ΔII, ΔIII) vs. E. ΔI- and ΔV-transduced cells exhibited reduced levels of p-Akt and p-GSK3β and retarded growth rates vs. E. Stimulatory effects of native T-cad overexpression on Akt and GSK3β phosphorylation were dose dependently inhibited by coexpression of ΔI or ΔV. Subsequent functional analyses compared only ΔI-, ΔII-, and ΔV-mutant constructs with E as a negative control. Unlike ΔII cells, ΔI and ΔV cells failed to exhibit homophilic ligation and deadhesion responses on a substratum of T-cad protein. In the wound assay, migration was increased for ΔII cells but impaired for ΔI and ΔV cells. In endothelial cell-spheroid assay, angiogenic sprouting was augmented for ΔII cells but inhibited for ΔI and ΔV cells. We conclude that EC1 and EC5 domains of T-cad are essential for its proangiogenic effects. ΔI and ΔV constructs may serve as dominant-negative mutants and as potential tools targeting excessive angiogenesis.

    AB - T-cadherin (T-cad) promotes survival, proliferation, and migration of endothelial cells and induces angiogenesis. We aimed to identify domains of T-cad functionally relevant to its effects on endothelial cell behavior. To specifically target the functional properties of the 5 cadherin repeat domains (EC1-EC5) of T-cad, endothelial cells were transduced with lentivectors containing specific T-cad-domain-deletion mutant constructs (ΔI, ΔII, ΔIII, ΔIV, ΔV). Empty (E) lentivector-transduced cells served as control. Similarly to overexpression of native T-cad, cells expressing ΔII, ΔIII, or ΔIV displayed elevated levels of p-Akt and p-GSK3β and increased proliferation rates (for ΔII, ΔIII) vs. E. ΔI- and ΔV-transduced cells exhibited reduced levels of p-Akt and p-GSK3β and retarded growth rates vs. E. Stimulatory effects of native T-cad overexpression on Akt and GSK3β phosphorylation were dose dependently inhibited by coexpression of ΔI or ΔV. Subsequent functional analyses compared only ΔI-, ΔII-, and ΔV-mutant constructs with E as a negative control. Unlike ΔII cells, ΔI and ΔV cells failed to exhibit homophilic ligation and deadhesion responses on a substratum of T-cad protein. In the wound assay, migration was increased for ΔII cells but impaired for ΔI and ΔV cells. In endothelial cell-spheroid assay, angiogenic sprouting was augmented for ΔII cells but inhibited for ΔI and ΔV cells. We conclude that EC1 and EC5 domains of T-cad are essential for its proangiogenic effects. ΔI and ΔV constructs may serve as dominant-negative mutants and as potential tools targeting excessive angiogenesis.

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