Biology subjects

Mallidi, S.

Publications and source records attributed to Mallidi, S..

8 recordsLinked to original sources

bioRxiv2024

In vivo Online label-free monitoring of heterogenous oxygen utilization during phototherapy with real-time ultrasound guided photoacoustic imaging

Understanding the tumor microenvironment, particularly the vascular density and the availability of oxygen, is key in individualizing treatment approaches and determining their efficacy. While there are many therapies including radiotherapy that are ineffective in hypoxic tumor microenvironments, here we demonstrate the heterogeneous oxygen consumption during photodynamic therapy (PDT), a non-invasive treatment method using localized light to activate a photosensitive drug in the presence of oxygen that has shown high effectiveness in the treatment of various types of tumors, including those presented in head and neck cancer (HNC) patients. While our previous work has demonstrated that blood oxygen saturation (StO2) mapped before and after treatment with ultrasound-guided photoacoustic imaging (US-PAI) can be used as a surrogate marker for the regionalized long-term efficacy of PDT, real-time monitoring of StO2 during PDT could provide additional insights on oxygen consumption and inform dose design for "on the spot" treatment decisions. Specifically, in this work, we integrated the US-PAI transducer probe with PDT light delivery fibers. We tested the setup on murine tumor models intravenously injected with liposomal benzoporphyrin derivative (BPD) photosensitizer at 0.5 mg/kg dose and photodynamic illumination at 100 and 400 mW/cm2 fluence rate. As expected, we observed with our US-PAI StO2 images that the rate of oxygen utilization increases when using a high fluence rate (HFR) light dose. Particularly in the higher fluence rate group, we observed StO2 reaching a minimum mid-light dose, followed by some degree of re-oxygenation. US-PAI added the advantage of spatial information to StO2 monitoring, which allowed us to match regions of re-oxygenation during therapy to retained vascular function with immunohistochemistry. Overall, our results have demonstrated the potential of US-PAI for applications in online dosimetry for cancer therapies such as PDT, using oxygen changes to detect regionalized physiological vascular response in the tumor microenvironment.

bioengineering↗

bioRxiv2024

Temporal dynamics of fluorescence and photoacoustic signals of a Cetuximab-IRDye800 conjugate in EGFR-overexpressing tumors

Molecular fluorescence-guided surgery has shown promise for tumor margin delineation but is limited by its depth profiling capability. Interestingly, most fluorophores, either clinically approved or in clinical trials, can also be used as photoacoustic contrast agents, yet their use is limited due to the low light fluence permitted for clinical use and the limited sensitivity of current photoacoustic imaging systems. There is therefore an urgent unmet need to establish methods for enhancing contrast in molecular targeted PA imaging which could potentially complement and overcome limitations in molecular fluorescence guided therapies. In this study, we compare the photoacoustic (PA) and fluorescence imaging capabilities of a cetuximab-IRDye800 conjugate in a subcutaneous tumor xenograft model. We demonstrate that while the fluorescence signal increases steadily over time after administration of cetuximab-IRDye800, PA signal peaks early ([~]2 fold higher at 6-hour as compared to pre-injection controls) and then decreases ([~]1.3 fold higher at 24-hour as compared to pre-injection controls). This pattern aligns with previous findings using other antibody-conjugated PA contrast agents. Mechanistically, we demonstrate that the formation of H-aggregates upon antibody conjugation enhances PA contrast of the IRDye800. The disruption of these H-aggregates, as the antibody-dye conjugate is degraded post receptor-mediated endocytosis, decreases PA signal intensity. The timeframe of maximum PA signal and decrease thereafter is consistent with the time frame of receptor-mediated endocytosis of cetuximab-IRDye800. Our data suggests that tumor cell surface binding results in peak PA signal while lysosomal localization and degradation results in a significant drop in PA signal. Our study sheds light on the distinct temporal dynamics of PA and fluorescence signals of Cetuximab-IRDye800 conjugate and we propose that optimizing IRDye800 conjugation to antibodies can further enhance PA signal intensity when timed to precisely to capture IRDye800 in an H-aggregate form.

cancer biology↗

bioRxiv2024

Vascular regional analysis unveils differential responses to anti-angiogenic therapy in pancreatic xenografts through macroscopic photoacoustic imaging

Pancreatic cancer (PC) is a highly lethal malignancy and the third leading cause of cancer deaths in the U.S. Despite major innovations in imaging technologies, there are limited surrogate radiographic indicators to aid in therapy planning and monitoring. Amongst the various imaging techniques Ultrasound-guided photoacoustic imaging (US-PAI) is a promising modality based on endogenous blood (hemoglobin) and blood oxygen saturation (StO2) contrast to monitor response to anti-angiogenic therapies. Adaptation of US-PAI to the clinical realm requires macroscopic configurations for adequate depth visualization, illuminating the need for surrogate radiographic markers, including the tumoral microvessel density (MVD). In this work, subcutaneous xenografts with PC cell lines AsPC-1 and MIA-PaCa-2 were used to investigate the effects of receptor tyrosine kinase inhibitor (sunitinib) treatment on MVD and StO2. Through histological correlation, we have shown that regions of high and low vascular density (HVD and LVD) can be identified through frequency domain filtering of macroscopic PA images which could not be garnered from purely global analysis. We utilized vascular regional analysis (VRA) of treatment-induced StO2 and total hemoglobin (HbT) changes. VRA as a tool to monitor treatment response allowed us to identify potential timepoints of vascular remodeling, highlighting its ability to provide insights into the TME not only for sunitinib treatment but also other anti-angiogenic therapies.

bioengineering↗

bioRxiv2024

Oxygen-releasing nanodroplets relieve intratumoral hypoxia in head and neck cancer spheroids.

AO_SCPLOWBSTRACTC_SCPLOWHypoxia in solid tumors, including head and neck cancer (HNC), contributes to treatment resistance, aggressive phenotypes, and poor clinical outcomes. Perfluorocarbon nanodroplets have emerged as promising oxygen carriers to alleviate tumor hypoxia. However, a thorough characterization of the hypoxia alleviation effects in terms of sustenance of oxygenated environments have not been thoroughly studied. In this study, we developed and characterized perfluoropentane nanodroplets (PFP NDs) for co-delivery of oxygen and the photoactivatable drug or photosensitizer benzoporphyrin derivative (BPD) to hypoxic HNC spheroids. The PFP NDs exhibited excellent stability, efficient oxygen loading/release, and biocompatibility. Using 3D multicellular tumor spheroids of FaDu and SCC9 HNC cells, we demonstrated the ability of oxygenated PFP NDs to penetrate the hypoxic core and alleviate hypoxia, as evidenced by reduced fluorescence of a hypoxia-sensing reagent and downregulation of hypoxia-inducible factors HIF-1 and HIF-2. BPD-loaded PFP NDs successfully delivered the photosensitizer into the spheroid core in a time-dependent manner. These findings highlight the potential of PFP NDs as a co-delivery platform to overcome hypoxia-mediated treatment resistance and improve therapy outcomes in HNC.

bioengineering↗

bioRxiv2024

Multi-spectral photoacoustic imaging combined with acoustic radiation force impulse imaging for applications in tissue engineering

Tissue engineering is a dynamic field focusing on the creation of advanced scaffolds for tissue and organ regeneration. These scaffolds are customized to their specific applications and are often designed to be complex, large structures to mimic tissues and organs. This study addresses the critical challenge of effectively characterizing these thick, optically opaque scaffolds that traditional imaging methods fail to fully image due to their optical limitations. We introduce a novel multi-modal imaging approach combining ultrasound, photoacoustic, and acoustic radiation force impulse imaging. This combination leverages its acoustic-based detection to overcome the limitations posed by optical imaging techniques. Ultrasound imaging is employed to monitor the scaffold structure, photoacoustic imaging is employed to monitor cell proliferation, and acoustic radiation force impulse imaging is employed to evaluate the homogeneity of scaffold stiffness. We applied this integrated imaging system to analyze melanoma cell growth within silk fibroin protein scaffolds with varying pore sizes and therefore stiffness over different cell incubation periods. Among various materials, silk fibroin was chosen for its unique combination of features including biocompatibility, tunable mechanical properties, and structural porosity which supports extensive cell proliferation. The results provide a detailed mesoscale view of the scaffolds internal structure, including cell penetration depth and biomechanical properties. Our findings demonstrate that the developed multimodal imaging technique offers comprehensive insights into the physical and biological dynamics of tissue-engineered scaffolds. As the field of tissue engineering continues to advance, the importance of non-ionizing and non-invasive imaging systems becomes increasingly evident, and by facilitating a deeper understanding and better characterization of scaffold architectures, such imaging systems are pivotal in driving the success of future tissue-engineering solutions.

bioengineering↗

bioRxiv2023

Impact of ectopic calcification on vascular oxygenation in placental dysfunction models: Simultaneous in vivo assessment with multi-parametric ultrasound and photoacoustic imaging

Preeclampsia (PE) is a prevalent gestational disorder occurring in 2-10% of all pregnancies. It is associated with placental dysfunction which can result in maternal mortality and preterm birth. Despite scientific advances, the underlying mechanisms of PE and the progression of placental dysfunction remain poorly understood without preventative or curative treatment. Other than blood tests and postdelivery placental diagnostics, there are limited detection methods for monitoring placental health. A major roadblock is the lack of imaging modalities available to monitor placental hemodynamics, as the commonly used clinical imaging systems are either harmful due to ionizing sources, too expensive and inaccessible, or provide insufficient data. We propose the combination of quantitative ultrasound and photoacoustic imaging (QUSPAI) to characterize the material and structural changes of the placenta during pregnancy in normal and disease states. The Slc20a2 murine model of placental dysfunction has previously been characterized by increased ectopic placental vascular calcification, reduced fetal growth, and decreased postnatal bone mineral density. In this study we utilized the Slc20a2 model and imaged knockout (KO) and wildtype (WT) placentae on embryonic day (E)18.5. The dual-wavelength images and radio frequency data were collected in vivo and ex vivo to provide quantitative ultrasound spectral (QUS) parameters and blood oxygen saturation. The photoacoustic images indicated that the WT tissue had statistically higher blood oxygen saturation (StO2) values than the KO tissues while QUS data revealed strain-specific calcification effects. Building upon the E18.5 gestation data collected here, future work will involve monitoring longitudinal changes in these parameters, including early calcification, to support translating this method for early diagnosis of placental vascular diseases.

bioengineering↗

bioRxiv2023

Ultrasound-guided Photoacoustic image Annotation Toolkit in MATLAB (PHANTOM) for preclinical applications

Depth-dependent fluence-compensation in photoacoustic (PA) imaging is paramount for accurate quantification of chromophores from deep tissues. Here we present a user-friendly toolkit named PHANTOM (PHotoacoustic ANnotation TOolkit for MATLAB) that includes a graphical interface and assists in the segmentation of ultrasound-guided PA images. We modelled the light source configuration with Monte Carlo eXtreme and utilized 3D segmented tissues from ultrasound to generate fluence maps to depth compensate PA images. The methodology was used to analyze PA images of phantoms with varying blood oxygenation and results were validated with oxygen electrode measurements. Two preclinical models, a subcutaneous tumor and a calcified placenta, were imaged and fluence-compensated using the PHANTOM toolkit and the results were verified with immunohistochemistry. The PHANTOM toolkit provides scripts and auxiliary functions to enable biomedical researchers not specialized in optical imaging to apply fluence correction to PA images, enhancing accessibility of quantitative PAI for researchers in various fields.

bioengineering↗

bioRxiv2023

A Dual Function Antibody Conjugate Enabled Photoimmunotherapy Complements Fluorescence and Photoacoustic Imaging of Head and Neck Cancer Spheroids

Several molecular-targeted imaging and therapeutic agents are in clinical trials for image-guided surgery and photoimmunotherapy (PIT) of head and neck cancers. In this context, we have previously reported the development, characterization, and specificity of a dual function antibody conjugate (DFAC) for multi-modal imaging and photoimmunotherapy (PIT) of EGFR over-expressing cancer cells. The DFAC reported previously and used in the present study, comprises of an EGFR targeted antibody - Cetuximab conjugated to Benzoporphyrin derivative (BPD) for fluorescence imaging and PIT, and a Si-centered naphthalocyanine dye for photoacoustic imaging. We report here the evaluation and performance of DFAC in detecting microscopic cancer spheroids by fluorescence and photoacoustic imaging along with their treatment by PIT. We demonstrate that while fluorescence imaging can detect spheroids with volumes greater than 0.049 mm3, photoacoustic imaging-based detection was possible even for the smallest spheroids (0.01 mm3), developed in the study. When subjected to PIT, the spheroids showed a dose-dependent response with smaller spheroids (0.01 and 0.018 mm3) showing a complete response with no recurrence when treated with 100 J/cm2. Together our results demonstrate the complementary imaging and treatment capacity of DFAC. This potentially enables fluorescence imaging to assess tumor presence on a macroscopic scale followed by photoacoustic imaging for delineating tumor margins guiding surgical resection and elimination of any residual microscopic disease by PIT, in a single intra-operative setting.

cancer biology↗