Molecular photoacoustic imaging of angiogenesis with integrin-targeted gold nanobeacons

摘要

Photoacoustic tomography (PAT) combines optical and acoustic imaging to generate high-resolution images of microvasculature. Inherent sensitivity to hemoglobin permits PAT to image blood vessels but precludes discriminating neovascular from maturing microvasculature. αvβ3-Gold nanobeacons (αvβ3-GNBs) for neovascular molecular PAT were developed, characterized, and demonstrated in vivo using a mouse Matrigel-plug model of angiogenesis. PAT results were microscopically corroborated with fluorescent αvβ3-GNB localization and supporting immunohistology in Rag1tm1Mom Tg(Tie-2-lacZ)182-Sato mice. αvβ3-GNBs (154 nm) had 10-fold greater contrast than blood on an equivolume basis when imaged at 740 nm to 810 nm in blood. The lowest detectable concentration in buffer was 290 nM at 780 nm. Noninvasive PAT of angiogenesis using a 10-MHz ultrasound receiver with αvβ3-GNBs produced a 600% increase in signal in a Matrigel-plug mouse model relative to the inherent hemoglobin contrast pretreatment. In addition to increasing the contrast of neovessels detected at baseline, αvβ3-GNBs allowed visualization of numerous angiogenic sprouts and bridges that were undetectable before contrast injection. Competitive inhibition of αvβ3-GNBs with αvβ3-NBs (no gold particles) almost completely blocked contrast enhancement to pretreatment levels, similar to the signal from animals receiving saline only. Consistent with other studies, nontargeted GNBs passively accumulated in the tortuous neovascular but provided less than half of the contrast enhancement of the targeted agent. Microscopic studies revealed that the vascular constrained, rhodamine-labeled αvβ3-GNBs homed specifically to immature neovasculature (PECAM+, Tie-2?) along the immediate tumor periphery, but not to nearby mature microvasculature (PECAM+, Tie-2+). The combination of PAT and αvβ3-GNBs offered sensitive and specific discrimination and quantification of angiogenesis in vivo, which may be clinically applicable to a variety of highly prevalent diseases, including cancer and cardiovascular disease.—Pan, D., Pramanik, M., Senpan, A., Allen, J. S., Zhang, H., Wickline, S. A., Wang, L. V., Lanza, G. M. Molecular photoacoustic imaging of angiogenesis with integrin-targeted gold nanobeacons.