Key result
Thrombospondin-1 type 1 repeats block neovascularization through two distinct antiangiogenic subdomains.
Why the study?
The specific regions and sequences within thrombospondin-1 responsible for its antiangiogenic activity and mechanisms of inhibiting neovascularization were not fully defined.
The type 1 repeats of thrombospondin-1 contain two independent subdomains that inhibit FGF-2 and VEGF-driven angiogenesis.
May guide targeted antiangiogenic peptide design; leaves open translation from animal models to clinical use.
BACKGROUND: Suppression of tumor growth by thrombospondin-1 (TSP-1) has been associated with its ability to inhibit neovascularization. The antiangiogenic activity of TSP-1, as defined by cornea pocket assays, was previously mapped to the amino-terminal portion of the protein within the procollagen region and the type 1 repeats. METHODS AND RESULTS: We evaluated the specificity and efficacy of different regions of TSP-1 using recombinant fragments of the protein on chorioallantoic membrane (CAM) angiogenesis and endothelial cell proliferation assays. In both assays, fragments containing the second and third type 1 repeats but not the procollagen region inhibited angiogenesis and endothelial cell proliferation. To further define the sequences responsible for the angiostatic effect of TSP-1, we used synthetic peptides. The CAM assay defined 2 sequences that independently suppressed angiogenesis. The amino-terminal end of the type 1 repeats showed higher potency for inhibiting angiogenesis driven by basic fibroblast growth factor (FGF-2), whereas the second region equally blocked angiogenesis driven by either FGF-2 or vascular endothelial growth factor (VEGF). Modifications of the active peptides revealed the specific amino acids required for the inhibitory response. One sequence included the conserved tryptophan residues in the amino-terminal end of the second and third type 1 repeats, and the other involved the amino acids that follow the CSVTCG sequence in the carboxy-terminus of these repeats. Both inhibition in the CAM assay and inhibition of breast tumor xenograft growth in nude mice were independent of the TGF-beta-activating sequence located in the second type 1 repeat. CONCLUSIONS: These results indicate that the type 1 repeats of TSP-1 contain 2 subdomains that may independently inhibit neovascularization. They also identify 2 independent pathways by which TSP-1 can block FGF-2 and VEGF angiogenic signals on endothelial cells.
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Iruela‐Arispe et al. (1999) studied Angiogenesis and tumor growth. Thrombospondin-1 (TSP-1) recombinant fragments and synthetic peptides was evaluated on Angiogenesis and endothelial cell proliferation. The type 1 repeats of thrombospondin-1 contain two independent subdomains that inhibit neovascularization and block FGF-2 and VEGF angiogenic signals on endothelial cells.