Focused ultrasound-mediated blood-brain barrier opening is safe and feasible with moderately hypofractionated radiotherapy for brainstem diffuse midline glioma.

BACKGROUND: Diffuse midline glioma (DMG) is a pediatric tumor with dismal prognosis. Systemic strategies have been unsuccessful and radiotherapy (RT) remains the standard-of-care. A central impediment to treatment is the blood-brain barrier (BBB), which precludes drug delivery to the central nervous system (CNS). Focused ultrasound (FUS) with microbubbles can transiently and non-invasively disrupt the BBB to enhance drug delivery. This study aimed to determine the feasibility of brainstem FUS in combination with clinical doses of RT. We hypothesized that FUS-mediated BBB-opening (BBBO) is safe and feasible with 39 Gy RT. METHODS: To establish a safety timeline, we administered FUS to the brainstem of non-tumor bearing mice concurrent with or adjuvant to RT; our findings were validated in a syngeneic brainstem murine model of DMG receiving repeated sonication concurrent with RT. The brainstems of male B6 (Cg)-Tyrc-2J/J albino mice were intracranially injected with mouse DMG cells (PDGFB+, H3.3K27M, p53-/-). A clinical RT dose of 39 Gy in 13 fractions (39 Gy/13fx) was delivered using the Small Animal Radiation Research Platform (SARRP) or XRAD-320 irradiator. FUS was administered via a 0.5 MHz transducer, with BBBO and tumor volume monitored by magnetic resonance imaging (MRI). RESULTS: FUS-mediated BBBO did not affect cardiorespiratory rate, motor function, or tissue integrity in non-tumor bearing mice receiving RT. Tumor-bearing mice tolerated repeated brainstem BBBO concurrent with RT. 39 Gy/13fx offered local control, though disease progression occurred 3-4 weeks post-RT. CONCLUSION: Repeated FUS-mediated BBBO is safe and feasible concurrent with RT. In our syngeneic DMG murine model, progression occurs, serving as an ideal model for future combination testing with RT and FUS-mediated drug delivery.

Modulation of cardio-respiratory activity in mice via transcranial focused ultrasound.

OBJECTIVE: The objective of this study was to investigate the effect of FUS on autonomic nervous system activity, including heart and respiratory rates, and to separate the thermal modulation from combined thermal and mechanical FUS effects. METHODS: The thalamus and hypothalamus of wild-type mice were sonicated with a continuous-wave, 2 MHz FUS transducer at pressures of 425 and 850 kPa for 60 seconds. Cardiac and respiratory rates were monitored as signs of autonomic nervous activity. FUS-induced changes in autonomic activity were compared to FUS targeted to a spatially-distant motor region and to laser-induced heating. RESULTS: FUS delivered to the primary target over the thalamus and hypothalamus at 850 kPa reversibly increased the respiratory rate by 6.5±3.2 breaths per minute and decreased the heart rate by 3.2±1.8 beats per minute. No significant changes occurred in this region at 425 kPa or when targeting the motor regions at 850 kPa. Laser heating with the same temperature rise profile produced by 850 kPa sonication resulted in cardiorespiratory modulation similar to that of FUS. CONCLUSIONS: FUS is capable of reversibly and non-invasively modulating cardiorespiratory activity in mice. Localized changes in temperature may constitute the main cause for this activity, though further investigation is warranted into the distinct and complementary mechanisms of mechanically- and thermally-induced FUS neuromodulation. Close monitoring of vital signs during FUS neuromodulation may be warranted to monitor systemic responses to stimulation.

ROE (Radiotherapy Outcomes Estimator): An open-source tool for optimizing radiotherapy prescriptions.

BACKGROUND AND OBJECTIVES: Radiotherapy prescriptions currently derive from population-wide guidelines established through large clinical trials. We provide an open-source software tool for patient-specific prescription determination using personalized dose-response curves. METHODS: We developed ROE, a plugin to the Computational Environment for Radiotherapy Research to visualize predicted tumor control and normal tissue complication simultaneously, as a function of prescription dose. ROE can be used natively with MATLAB and is additionally made accessible in GNU Octave and Python, eliminating the need for commercial licenses. It provides a curated library of published and validated predictive models and incorporates clinical restrictions on normal tissue outcomes. ROE additionally provides batch-mode tools to evaluate and select among different fractionation schemes and analyze radiotherapy outcomes across patient cohorts. CONCLUSION: ROE is an open-source, GPL-copyrighted tool for interactive exploration of the dose-response relationship to aid in radiotherapy planning. We demonstrate its potential clinical relevance in (1) improving patient awareness by quantifying the risks and benefits of a given treatment protocol (2) assessing the potential for dose escalation across patient cohorts and (3) estimating accrual rates of new protocols.

Focused ultrasound mediated blood-brain barrier opening is safe and feasible in a murine pontine glioma model.

Drug delivery in diffuse intrinsic pontine glioma is significantly limited by the blood-brain barrier (BBB). Focused ultrasound (FUS), when combined with the administration of microbubbles can effectively open the BBB permitting the entry of drugs across the cerebrovasculature into the brainstem. Given that the utility of FUS in brainstem malignancies remains unknown, the purpose of our study was to determine the safety and feasibility of this technique in a murine pontine glioma model. A syngeneic orthotopic model was developed by stereotactic injection of PDGF-B+PTEN-/-p53-/- murine glioma cells into the pons of B6 mice. A single-element, spherical-segment 1.5 MHz ultrasound transducer driven by a function generator through a power amplifier was used with concurrent intravenous microbubble injection for tumor sonication. Mice were randomly assigned to control, FUS and double-FUS groups. Pulse and respiratory rates were continuously monitored during treatment. BBB opening was confirmed with gadolinium-enhanced MRI and Evans blue. Kondziela inverted screen testing and sequential weight lifting measured motor function before and after sonication. A subset of animals were treated with etoposide following ultrasound. Mice were either sacrificed for tissue analysis or serially monitored for survival with daily weights. FUS successfully caused BBB opening while preserving normal cardiorespiratory and motor function. Furthermore, the degree of intra-tumoral hemorrhage and inflammation on H&E in control and treated mice was similar. There was also no difference in weight loss and survival between the groups (p > 0.05). Lastly, FUS increased intra-tumoral etoposide concentration by more than fivefold. FUS is a safe and feasible technique for repeated BBB opening and etoposide delivery in a preclinical pontine glioma model.

Distinguishing metastatic triple-negative breast cancer from nonmetastatic breast cancer using second harmonic generation imaging and resonance Raman spectroscopy.

Triple-negative breast cancer (TNBC) is an aggressive subset of breast cancer that is more common in African-American and Hispanic women. Early detection followed by intensive treatment is critical to improving poor survival rates. The current standard to diagnose TNBC from histopathology of biopsy samples is invasive and time-consuming. Imaging methods such as mammography and magnetic resonance (MR) imaging, while covering the entire breast, lack the spatial resolution and specificity to capture the molecular features that identify TNBC. Two nonlinear optical modalities of second harmonic generation (SHG) imaging of collagen, and resonance Raman spectroscopy (RRS) potentially offer novel rapid, label-free detection of molecular and morphological features that characterize cancerous breast tissue at subcellular resolution. In this study, we first applied MR methods to measure the whole-tumor characteristics of metastatic TNBC (4T1) and nonmetastatic estrogen receptor positive breast cancer (67NR) models, including tumor lactate concentration and vascularity. Subsequently, we employed for the first time in vivo SHG imaging of collagen and ex vivo RRS of biomolecules to detect different microenvironmental features of these two tumor models. We achieved high sensitivity and accuracy for discrimination between these two cancer types by quantitative morphometric analysis and nonnegative matrix factorization along with support vector machine. Our study proposes a new method to combine SHG and RRS together as a promising novel photonic and optical method for early detection of TNBC.

Iterative Curve Fitting of the Bioheat Transfer Equation for Thermocouple-Based Temperature Estimation In Vitro and In Vivo.

Temperature measurements with thin thermocouples embedded in ultrasound fields are strongly subjected to a viscous heating artifact (VHA). The artifact contribution decays over time; therefore, it can be minimized at late temperature readings. However, previous studies have failed to demonstrate a rigorous method for determining the optimal time point at which the artifact contribution is negligible. In this study, we present an iterative processing method based on successive curve fittings using an artifact-independent model. The fitting starting point moves at each iteration until the maximum R2 indicates where the viscous heating is minimum. A solution of the bioheat transfer equation is used to account for blood perfusion, thus enabling in vivo measurements. Three T-type thermocouples with different diameters and sensitivities were assessed in an excised canine liver and in the mouse brain in vivo. We found that the artifact constitutes up to 81% ± 5% of wire thermocouple readings. The best-fit time varied in the liver samples ( n = 3 ) from 0 to 3.335 ± 0.979 s and in the mouse brain ( n = 5 ) from 0 to 0.498 ± 0.457 s at variable experimental conditions, which clearly demonstrates the need of the method for finding the appropriate starting time point of the fit. This study introduces a statistical method to determine the best time to fit a curve that can back-estimate temperature in tissues under ultrasound exposure using thermocouples. This method allows temperature evaluation in vivo and in vitro during a validation and safety assessment of a wide range of therapeutic and diagnostic ultrasound modalities.

Compact OAM microscope for edge enhancement of biomedical and object samples.

The production of orbital angular momentum (OAM) by using a q-plate, which functions as an electrically tunable spatial frequency filter, provides a simple and efficient method of edge contrast in biological and medical sample imaging for histological evaluation of tissue, smears, and PAP smears. An instrument producing OAM, such as a q-plate, situated at the Fourier plane of a 4f lens system, similar to the use of a high-pass spatial filter, allows the passage of high spatial frequencies and enables the production of an image with highly illuminated edges contrasted against a dark background for both opaque and transparent objects. Compared with ordinary spiral phase plates and spatial light modulators, the q-plate has the added advantage of electric control and tunability.

Hybrid generation and analysis of vector vortex beams.

A method is described for generating optical vector vortex beams carrying superpositions of orbital angular momentum states by using a tandem application of a spatial light modulator with a vortex retarder. The vortex component has a spatially inhomogeneous phase front that can carry orbital angular momentum, and the vector nature is a spatially inhomogeneous state of polarization in the laser beam profile. The vector vortex beams are characterized experimentally by imaging the beams at points across the focal plane in an astigmatic system using a tilted lens. Mathematical analysis of the Gouy phase shows good agreement with the phase structure obtained in the experimental images. The polarization structure of the vector beam and the orbital angular momentum of the vortex beam are shown to be preserved.