Matrix metalloproteinase-9 (MMP-9) plays major roles in extracellular matrix (ECM) remodeling and membrane protein cleavage, suggesting a high correlation with cancer cell invasion and tumor metastasis. Here, we present a contrast agent based on a DNA aptamer that can selectively target human MMP-9 in the tumor microenvironment (TME) with high affinity and sensitivity. Surface modification of plasmonic gold nanospheres with the MMP-9 aptamer and its complementary sequences allows the nanospheres to aggregate in the presence of human MMP-9 through DNA displacement and hybridization. Aggregation of gold nanospheres enhances the optical absorption in the first near-infrared window (NIR-I) due to the plasmon coupling effect, thereby allowing us to detect the aggregated gold nanospheres within the TME
via
ultrasound-guided photoacoustic (US/PA) imaging. Selective and sensitive detection of human MMP-9
via
US/PA imaging is demonstrated in solution of nanosensors with the pre-treatment of human MMP-9,
in vitro
in cell culture, and
in vivo
in a xenograft murine model of human breast cancer.
Despite significant research regarding metastasis, there has been limited success in preventing it. However, gold nanoparticle (AuNP) technology has shown the potential to inhibit metastasis. Our earlier studies of gold nanorodassisted plasmonic photothermal therapy (AuNRs-PPTT), where gold nanorods (AuNRs) were irradiated with near-infrared (NIR) light to induce heat, were utilized in slowing cancer cell migration in vitro. Herein, we have expanded the in vitro studies of the AuNRs-PPTT to xenograft mice to inhibit metastasis of mammary gland tumors. The study duration was 32 days from 4T1 cancer cell injections in four treatment groups: control (PBS), NIR Only, AuNRs, and AuNRs + NIR. Multiple AuNRs-PPTT treatment sessions with intratumoral AuNRs injections were conducted every 7 days on average on the mice. Photoacoustic spectroscopy has been utilized to study the distribution and aggregation of AuNRs within the tumors and the drainage of particles to the sentinel right subiliac lymph node. The photoacoustic results revealed that the AuNRs' shapes are still stable regardless of their heterogeneous distributions inside the mammalian tumor and lymph nodes. Bioluminescence imaging was used to monitor metastasis using luciferin labeling techniques and has shown that AuNRs-PPTT inhibited metastasis completely within the first 21 days. Moreover, proteomics was run to determine the most pivotal inhibitory pathways: NETosis, cell growth, cell proliferation, inflammation, and extracellular matrix (ECM) degradation. These five mechanisms are interdependent within related networks, which synergistically explains the molecular mechanism of metastasis inhibition by AuNRs-PPTT. The current in vivo data ensures the viability of PPTT applications in inhibiting metastasis in humans.
.
Significance
To effectively study preclinical animal models, medical imaging technology must be developed with a high enough resolution and sensitivity to perform anatomical, functional, and molecular assessments. Photoacoustic (PA) tomography provides high resolution and specificity, and fluorescence (FL) molecular tomography provides high sensitivity; the combination of these imaging modes will enable a wide range of research applications to be studied in small animals.
Aim
We introduce and characterize a dual-modality PA and FL imaging platform using
in vivo
and phantom experiments.
Approach
The imaging platform’s detection limits were characterized through phantom studies that determined the PA spatial resolution, PA sensitivity, optical spatial resolution, and FL sensitivity.
Results
The system characterization yielded a PA spatial resolution of
in the transverse plane and
in the longitudinal axis, a PA sensitivity detection limit not less than that of a sample with absorption coefficient
, an optical spatial resolution of
in the vertical axis and
in the horizontal axis, and a FL sensitivity detection limit not
concentration of IR-800. The scanned animals displayed in three-dimensional renders showed high-resolution anatomical detail of organs.
Conclusions
The combined PA and FL imaging system has been characterized and has demonstrated its ability to image mice
in vivo
, proving its suitability for biomedical imaging research applications.
Methods to noninvasively increase and assess T cell infiltration are critical for success of adoptive cell therapy (ACT) of solid tumors where only a small fraction of adoptive T cells typically accumulates at the target. We developed an approach based on photoacoustic (PA) and ultrasound (US) imaging to visualize magnetic field driven accumulation of T cells tagged with photo-magnetic nanoparticles (PMNPs). Specifically, Au@Fe3O4 shell-core PMNPs (200 nm diameter), absorbing at 1064 nm wavelength, were synthesized and characterized using a UV-Vis-NIR spectrophotometry, dynamic light scattering (DLS), and transmission electron microscopy (TEM). Ovalbumin (OVA)-targeted PMNP-tagged OT1 murine primary T cells were injected intravenously into OVA-expressing tumor-bearing C57BL/6 mice. US/PA imaging (20 MHz, 1064 nm and 680–970 nm, Vevo LAZR, Visualsonics Inc.) of the primary tumor and regional lymph nodes showed accumulation of PMNP-tagged T cells. Our results indicate feasibility of the T cell tagging with PMNPs and magnetic delivery of adoptive PMNP-tagged T cells using US/PA imaging thus providing critical imaging feedback to improve the adoptive T cell therapy of solid tumors. Overall, our studies show that a synergistic combination of US/PA imaging and magnetic delivery of nanoparticle (NP)-tagged adoptive T cells can expedite development, translation, and expansion of ACT.
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