Publications

Photoacoustic tomography (PAT), as a novel biomedical imaging technique, is able to capture temporal, spatial and spectral tomographic information from organisms. Organ-level multi-parametric analysis of continuous PAT images are of interest since it enables the quantification of organ specific morphological and functional parameters in small animals. Accurate organ delineation is imperative for organ-level image analysis, yet the low contrast and blurred organ boundaries in PAT images pose challenge for their precise segmentation. Fortunately, shared structural information among continuous images in the time-space-spectrum domain may be used to enhance segmentation. In this paper, we introduce a structure fusion enhanced graph convolutional network (SFE-GCN), which aims at automatically segmenting major organs including the body, liver, kidneys, spleen, vessel and spine of abdominal PAT image of mice. SFE-GCN enhances the structural feature of organs by fusing information in continuous image sequence captured at time, space and spectrum domains. As validated on large-scale datasets across different imaging scenarios, our method not only preserves fine structural details but also ensures anatomically aligned organ contours. Most importantly, this study explores the application of SFE-GCN in multi-dimensional organ image analysis, including organ-based dynamic morphological analysis, organ-wise light fluence correction and segmentation-enhanced spectral un-mixing. Code will be released at https://github.com/lzc-smu/SFEGCN.git.

Triple-negative breast cancer (TNBC) is recognized as a particularly aggressive subtype of breast cancer that is devoid of effective therapeutic targets. Immune checkpoint inhibitors (ICIs) have demonstrated promising results in TNBC treatment. Nonetheless, most patients either develop resistance to ICIs or fail to respond to them initially. Owing to its spatio-temporal precision and non-invasive nature, photoimmunotherapy offers a targeted therapeutic strategy for TNBC. Herein, we report hyaluronic acid (HA)-functionalized indocyanine green-based supramolecular nanoparticles (HGI NPs), with biodegradable characteristics, for high-performance photoacoustic imaging and targeted phototherapy for TNBC. Notably, HGI NPs can significantly gather in TNBC tissues because of the enhanced permeability and retention effect of the tumor, and the tumor-targeting properties of HA. The strong amplification of HGI nanoparticles triggers a significant immunogenic cell death (ICD) response when exposed to 808 nm light, thus shifting the immunosuppressive tumor microenvironment (iTME) into a tumor attack mode and ‘hot’ state. Antitumor experiments demonstrate the high efficiency of the supramolecular photosensitizers HGI NPs for TNBC elimination and good biosafety. This synergistic strategy reshapes the iTME and amplifies the antitumor immune response, providing a theoretical foundation for combining phototherapy and ICIs as potential treatments for TNBC.

Pancreatic cancer is known for its high invasiveness and metastasis, making rapid visualization and precise treatment critical for improving patient outcomes. Current diagnostic tools lack abilities to provide rapid and accurate tumor localization, particularly for real-time intraoperative guidance. To address this gap, the study has developed a novel Förster Resonance Energy Transfer (FRET)-mediated dual-ratiometric near-infrared fluorescence (NIRF)/photoacoustic (PA) bimodal probe, SiRho-SHD-NTR, specifically designed for the fast and accurate navigation of pancreatic tumor resection. The probe, due to its excellent binding affinity with nitroreductase (NTR), can rapidly reach response saturation. Cellular experiments demonstrate that the probe rapidly and efficiently penetrates cancer cells, enhancing the effectiveness of PA imaging for preliminary diagnosis and tumor localization, while also enabling the rapid visualization of pancreatic tumors through NIRF imaging. Leveraging the rapid response characteristics of the probe to NTR, the study achieves precise tumor imaging in orthotopic pancreatic cancer mice by spraying the probe, within ≈5 min. More importantly, the probe even allows for the fast visualization of metastatic tumors and fluorescence-guided surgical resection. It is believed that SiRho-SHD-NTR will offer a promising method in the rapid visualization of pancreatic cancer and provide a powerful tool for imaging-guided tumor surgery, targeting both primary and metastatic tumors.

Cellular senescence is considered an important tumour suppression mechanism in response to damage and oncogenic stress in early lesions. However, when senescent cells are not immune-cleared and persist in the tumour microenvironment, they can drive a variety of tumour-promoting activities, including cancer initiation, progression, and metastasis. Additionally, there is compelling evidence demonstrating a direct connection between chemo(radio)therapy-induced senescence and the development of drug resistance and cancer recurrence. Therefore, detection of senescent cells in tissues holds great promise for predicting cancer occurrence earlier, assessing tumour progression, aiding patient stratification and prognosis, and informing about the efficacy of potential senotherapies. However, effective detection of senescent cells is limited by lack of biomarkers and readout strategies suitable for in vivo clinical imaging. To this end, a nanoprobe composed of biocompatible polydopamine (PDA) nanoparticle doped with FDA-approved indocyanine green (ICG) dye, namely PDA-ICG, was designed as a contrast agent for senescence detection using photoacoustic imaging (PAI). In an in vitro model of chemotherapy-induced senescence, PDA-ICG nanoprobe showed an elevated uptake in senescent cells relative to cancer cells. In addition to its improved photostability, 2.5-fold enhancement in photoacoustic signal relative to ICG was observed. Collectively, the results indicate that the PDA-ICG nanoprobe has the potential to be used as a contrast agent for senescence detection of chemotherapy-induced senescence using PAI.

Purpose: Preclinical models of cutaneous wound healing can be useful for improving clinical wound care. Oxygen Enhanced Photoacoustic imaging (OE PAI) can measure oxygenation, and Dynamic Contrast Enhanced (DCE) PAI can measure vascular perfusion. We investigated how a combined OE-DCE PAI protocol can measure vascular oxygenation and perfusion to a cutaneous healing model.
Procedures: We developed a cutaneous “punch” wound model and photographed the wounds to track healing for 9 days. We performed OE-DCE PAI on Day 0, 3, 6, and 9. OE PAI was performed with 21% O2 and 100% O2 breathing gases to measure oxyhemoglobin (HbO2), deoxyhemoglobin (Hb), total hemoglobin (HbT), and oxygen saturation (%sO2), along with changes in these parameters caused by a change in breathing gas (ΔHb, ΔHbO2, ΔHbT, ΔsO2). To perform DCE PAI, indocyanine green (ICG) was administered intravenously while monitoring the PAI signal for 10 min as the agent washed through the wound area, which was used to evaluate the wash-out rate.
Results: The average wound size was significantly smaller only by Day 6. For comparison, OE PAI measured a significant increase in HbO2, Hb, HbT, and %sO2 immediately after creating the wound, which significantly decreased by Day 3 and continued to decrease towards values for normal tissue by Day 9. The vascular wash-out rate significantly increased by Day 3, and continued to increase during the healing process. Notably, the wash-out rate can be assessed at a single PAI absorbance wavelength and by simply comparing signal amplitudes without advanced analysis, which may facilitate clinical translation.
Conclusions: OE-DCE PAI can monitor significant changes in vascular perfusion and oxygenation prior to significant changes in cutaneous wound size. These results establish OE-DCE PAI as a tool for future pre-clinical wound healing studies and eventual clinical translation.

Hydrogen persulfide (H2S2) plays a significant role in redox biology and signal transduction; therefore, quantitative visualization of H2S2 in the deep tissue of living organisms is essential for obtaining reliable information about relevant pathophysiological processes directly. However, currently reported H2S2 probes are unsuitable for this purpose because of their poor selectivity for many polysulfide species or their short wavelength, which hinders precise imaging in deep tissues. Herein, for the first time, we report a unique H2S2-mediated dithiole formation reaction. Based on this reaction, we construct the first NIR-II fluorescence (FL) and photoacoustic (PA) dual-ratiometric probe (NIR-II-H2S2) for quantitatively visualizing H2S2 in vivo. This probe shows dual-ratiometric NIR-II fluorescence (I840/I1000, 107-fold) and photoacoustic (PA800/PA900, 6.5-fold) responses towards Na2S2 species with high specificity, excellent sensitivity (1.8 nM), improved water solubility, and deep-tissue penetration. More importantly, using NIR-II dual-ratiometric FL/PA imaging, we successfully demonstrated that the probe could be used to accurately quantify the fluctuating H2S2 levels in the liver-injury mouse models induced by lipopolysaccharides or metformin drugs. Overall, this study not only presents a promising tool for H2S2-related pathological research, but also provides a unique recognition site that may be generalized for designing more useful H2S2 imaging agents in the future.

Imaging guided cancer therapy is a comprehensive strategy that combines the diagnosis and treatment to eradicate tumors. Ferroptosis is a distinct programmed cell death and holds great potential in cancer therapy. In this study, a hydrogen sulfide (H2S)-activated PEGylated Pd@Cu2O core-shell nanocomposite (termed PCO) that in situ transformed into Pd@Cu2-xS (termed PCS) at colorectal tumor tissues is developed for colorectal cancer photoacoustic (PA) imaging and photothermal-enhanced ferroptosis therapy in NIR-II window. The Cu+ on the surface of PCS can catalyze the Fenton-like reaction with overexpressed H2O2 in the colon tumor tissues, yielding hydroxyl radicals (·OH) and Cu2+. Moreover, the PCS accelerates the Fenton-like reaction to generate more ·OH. The PCS displays dual peroxidase- and glutathione oxidase-mimic enzymatic activity in weakly acidic tumor microenvironment (TME). Additionally, the glutathione depletion by Cu2+ results in the production of Cu+ and glutathione disulfide as well as the down-regulation of glutathione peroxidase 4. The interaction of polyunsaturated fatty acids with ·OH induces the up-regulation of lipid peroxides on cellular membrane, thereby causing ferroptosis. Hence, this study has developed the H2S-activated PCO that in situ transforms into PCS, as a novel colon cancer diagnosis-treatment nanoprobe, for PA imaging guided precise diagnosis and efficient therapy of colon cancer.

Multispectral photoacoustic tomography (PAT) is an imaging modality that utilizes the photoacoustic effect to achieve non-invasive and high-contrast imaging of internal tissues but also molecular functional information derived from multi-spectral measurements. However, the hardware cost and computational demand of a multispectral PAT system consisting of up to thousands of detectors are huge. To address this challenge, we propose an ultra-sparse spiral sampling strategy for multispectral PAT, which we named U3S-PAT. Our strategy employs a sparse ring-shaped transducer that, when switching excitation wavelengths, simultaneously rotates and translates. This creates a spiral scanning pattern with multispectral angle-interlaced sampling. To solve the highly ill-conditioned image reconstruction problem, we propose a self-supervised learning method that is able to introduce structural information shared during spiral scanning. We simulate the proposed U3S-PAT method on a commercial PAT system and conduct in vivo animal experiments to verify its performance. The results show that even with a sparse sampling rate as low as 1/30, our U3S-PAT strategy achieves similar reconstruction and spectral unmixing accuracy as non-spiral dense sampling. Given its ability to dramatically reduce the time required for three-dimensional multispectral scanning, our U3S-PAT strategy has the potential to perform volumetric molecular imaging of dynamic biological activities.

Microvascular endothelial dysfunction may provide insights into systemic diseases, such as carotid artery disease. Raster-scan optoacoustic mesoscopy (RSOM) can produce images of skin microvasculature during endothelial dysfunction challenges via numerous microvascular features. Herein, RSOM was employed to image the microvasculature of 26 subjects (13 patients with single carotid artery disease, 13 healthy participants) to assess the dynamics of 18 microvascular features at three scales of detail, i.e., the micro- (<100 μm), meso- (≈100-1000 μm) and macroscale (<1000 μm), during post-occlusive reactive hyperemia challenges. The proposed analysis identified a subgroup of 9 features as the most relevant to carotid artery disease because they achieved the most efficient classification (AUC of 0.93) between the two groups in the first minute of hyperemia (sensitivity/specificity: 0.92/0.85). This approach provides a non-invasive solution to microvasculature quantification in carotid artery disease, a main form of cardiovascular disease, and further highlights the possible link between systemic disease and microvascular dysfunction.

The analysis of vascular morphology and functionality enables the assessment of disease activity and therapeutic effects in various pathologies. Raster-scanning optoacoustic mesoscopy (RSOM) is an imaging modality that enables the visualization of superficial vascular networks in vivo. In murine models of colitis, deep vascular networks in the colon wall can be visualized by transrectal absorber guide raster-scanning optoacoustic mesoscopy (TAG-RSOM). In order to accelerate the implementation of this technology in translational studies of inflammatory bowel disease, an image-processing pipeline for TAG-RSOM data has been developed. Using optoacoustic data from a murine model of chemically-induced colitis, different image segmentation methods are compared for visualization and quantification of deep vascular patterns in terms of vascular network length and complexity, blood volume, and vessel diameter. The presented image-processing pipeline for TAG-RSOM enables label-free in vivo assessment of changes in the vascular network in murine colitis with broad applications for inflammatory bowel disease research.