Central nervous system tumors carry grave clinical prognoses due to limited effectiveness of surgical resection, radiation, and
chemotherapy. Thus, improved strategies for
brain tumor visualization and targeted treatment are critically needed. We demonstrate that mouse cerebellar
medulloblastoma (MB) can be targeted and illuminated with a fluorescent, engineered
cystine knot (
knottin)
peptide that binds with high affinity to αvβ3, αvβ5, and α5β1
integrin receptors. This
integrin-binding
knottin peptide, denoted EETI 2.5F, was evaluated as a molecular imaging probe in both orthotopic and genetic models of MB. Following tail vein injection, fluorescence arising from
dye-conjugated EETI 2.5F was localized to the
tumor compared with the normal surrounding brain tissue, as measured by optical imaging. The imaging signal intensity correlated with
tumor volume. Due to its unique ability to bind to α5β1
integrin, EETI 2.5F showed superior in vivo and ex vivo
brain tumor imaging contrast compared with other engineered
integrin-binding
knottin peptides and with c(RGDfK), a well-studied
integrin-binding
peptidomimetic. Next, EETI 2.5F was fused to an
antibody fragment crystallizable (Fc) domain (EETI 2.5F-Fc) to determine if a larger
integrin-
binding protein could also target intracranial
brain tumors. EETI 2.5F-Fc, conjugated to a
fluorescent dye, illuminated MB following i.v. injection and was able to distribute throughout the
tumor parenchyma. In contrast,
brain tumor imaging signals were not detected in mice injected with EETI 2.5F
proteins containing a scrambled
integrin-binding sequence, demonstrating the importance of target specificity. These results highlight the potential of using EETI 2.5F and EETI 2.5-Fc as targeted
molecular probes for
brain tumor imaging.