Radiation enhancement using focussed ultrasound-stimulated microbubbles for head and neck cancer: A phase 1 clinical trial.

BACKGROUND AND PURPOSE: Preclinical research demonstrated that the exposure of microbubbles (intravascular gas microspheres) to focussed ultrasound within the targeted tumour upregulates pro-apoptotic pathways and enhances radiation-induced tumour cell death. This study aimed to assess the safety and efficacy of magnetic resonance (MR)-guided focussed ultrasound-stimulated microbubbles (MRgFUS-MB) for head and neck cancers (HN). MATERIALS AND METHODS: This prospective phase 1 clinical trial included patients with newly diagnosed or recurrent HN cancer (except nasopharynx malignancies) for whom locoregional radiotherapy with radical- or palliative-intent as deemed appropriate. Patients with contraindications for microbubble administration or contrast-enhanced MR were excluded. MR-coupled focussed ultrasound sonicated intravenously administered microbubbles within the MR-guided target volume. Patients receiving 5-10 and 33-35 radiation fractions were planned for 2 and 3 MRgFUS-MB treatments, respectively. Primary endpoint was toxicity per CTCAEv5.0. Secondary endpoint was tumour response at 3 months per RECIST 1.1 criteria. RESULTS: Twelve patients were enrolled between Jun/2020 and Nov/2023, but 1 withdrew consent. Eleven patients were included in safety analysis. Median follow-up was 7 months (range, 0.3-38). Most patients had oropharyngeal cancer (55 %) and received 20-30 Gy/5-10 fractions (63 %). No systemic toxicity or MRgFUS-MB-related adverse events occurred. The most severe acute adverse events were radiation-related grade 3 toxicities in 6 patients (55 %; dermatitis in 3, mucositis in 1, dysphagia in 6). No radiation necrosis or grade 4/5 toxicities were reported. 8 patients were included in the 3-month tumour response assessment: 4 had partial response (50 %), 3 had complete response (37.5 %), and 1 had progressive disease (12.5 %). CONCLUSIONS: MRgFUS-MB treatment was safe and associated with high rates of tumour response at 3 months.

R2* Impact on Hepatic Fat Quantification With a Commercial Single Voxel Technique at 1.5 and 3.0 T.

Rationale and Objectives: Fat quantification accuracy using a commercial single-voxel high speed T2-corrected multi-echo (HISTO) technique and its robustness to R2* variations at 3.0 T, such as those introduced by iron in liver, has not been fully established. This study evaluated HISTO at 3.0 T and sought to reproduce results at 1.5 T. Methods: Phantoms were prepared with a range of fat content and R2*. Data were acquired at 1.5 T and 3.0 T, using HISTO and a Dixon technique. Fat quantification accuracy was evaluated as a function of R2*. The patient study included 239 consecutive patients. Data were acquired at 1.5 T or 3.0 T, using HISTO and Dixon techniques. The techniques were compared using Bland-Altman plots. Bias significance was evaluated using a one-sample t-test. Results: In phantoms, HISTO was accurate within 10% up to a R2* of 100 s-1 at both field strengths, while Dixon was accurate within 10% where R2* was accurately quantified (up to 350 s-1 at 1.5 T, and 550 s-1 at 3.0 T). In patients, where R2* was <100 s-1, fat quantification from both techniques agreed at 1.5 T (P = .71), but not at 3.0 T (P = .007), with a bias <1%. Conclusion: Results suggest that HISTO is reliable when R2* is <100 s-1, corresponding to patients with at most mild liver iron overload, and that it should be used with caution when R2* is >100 s-1. Dixon should be preferred for hepatic fat quantification due to its robustness to R2* variations.

Radiation enhancement using focussed ultrasound-stimulated microbubbles for breast cancer: A Phase 1 clinical trial.

BACKGROUND: Preclinical studies have demonstrated that tumour cell death can be enhanced 10- to 40-fold when radiotherapy is combined with focussed ultrasound-stimulated microbubble (FUS-MB) treatment. The acoustic exposure of microbubbles (intravascular gas microspheres) within the target volume causes bubble cavitation, which induces perturbation of tumour vasculature and activates endothelial cell apoptotic pathways responsible for the ablative effect of stereotactic body radiotherapy. Subsequent irradiation of a microbubble-sensitised tumour causes rapid increased tumour death. The study here presents the mature safety and efficacy outcomes of magnetic resonance (MR)-guided FUS-MB (MRgFUS-MB) treatment, a radioenhancement therapy for breast cancer. METHODS AND FINDINGS: This prospective, single-center, single-arm Phase 1 clinical trial included patients with stages I-IV breast cancer with in situ tumours for whom breast or chest wall radiotherapy was deemed adequate by a multidisciplinary team (clinicaltrials.gov identifier: NCT04431674). Patients were excluded if they had contraindications for contrast-enhanced MR or microbubble administration. Patients underwent 2 to 3 MRgFUS-MB treatments throughout radiotherapy. An MR-coupled focussed ultrasound device operating at 800 kHz and 570 kPa peak negative pressure was used to sonicate intravenously administrated microbubbles within the MR-guided target volume. The primary outcome was acute toxicity per Common Terminology Criteria for Adverse Events (CTCAE) v5.0. Secondary outcomes were tumour response at 3 months and local control (LC). A total of 21 female patients presenting with 23 primary breast tumours were enrolled and allocated to intervention between August/2020 and November/2022. Three patients subsequently withdrew consent and, therefore, 18 patients with 20 tumours were included in the safety and LC analyses. Two patients died due to progressive metastatic disease before 3 months following treatment completion and were excluded from the tumour response analysis. The prescribed radiation doses were 20 Gy/5 fractions (40%, n = 8/20), 30 to 35 Gy/5 fractions (35%, n = 7/20), 30 to 40 Gy/10 fractions (15%, n = 3/20), and 66 Gy/33 fractions (10%, n = 2/20). The median follow-up was 9 months (range, 0.3 to 29). Radiation dermatitis was the most common acute toxicity (Grade 1 in 16/20, Grade 2 in 1/20, and Grade 3 in 2/20). One patient developed grade 1 allergic reaction possibly related to microbubbles administration. At 3 months, 18 tumours were evaluated for response: 9 exhibited complete response (50%, n = 9/18), 6 partial response (33%, n = 6/18), 2 stable disease (11%, n = 2/18), and 1 progressive disease (6%, n = 1/18). Further follow-up of responses indicated that the 6-, 12-, and 24-month LC rates were 94% (95% confidence interval [CI] [84%, 100%]), 88% (95% CI [75%, 100%]), and 76% (95% CI [54%, 100%]), respectively. The study's limitations include variable tumour sizes and dose fractionation regimens and the anticipated small sample size typical for a Phase 1 clinical trial. CONCLUSIONS: MRgFUS-MB is an innovative radioenhancement therapy associated with a safe profile, potentially promising responses, and durable LC. These results warrant validation in Phase 2 clinical trials. TRIAL REGISTRATION: clinicaltrials.gov, identifier NCT04431674.

Novel MRI-guided focussed ultrasound stimulated microbubble radiation enhancement treatment for breast cancer.

Preclinical studies have demonstrated focused ultrasound (FUS) stimulated microbubble (MB) rupture leads to the activation of acid sphingomyelinase-ceramide pathway in the endothelial cells. When radiotherapy (RT) is delivered concurrently with FUS-MB, apoptotic pathway leads to increased cell death resulting in potent radiosensitization. Here we report the first human trial of using magnetic resonance imaging (MRI) guided FUS-MB treatment in the treatment of breast malignancies. In the phase 1 prospective interventional study, patients with breast cancer were treated with fractionated RT (5 or 10 fractions) to the disease involving breast or chest wall. FUS-MB treatment was delivered before 1st and 5th fractions of RT (within 1 h). Eight patients with 9 tumours were treated. All 7 evaluable patients with at least 3 months follow-up treated for 8 tumours had a complete response in the treated site. The maximum acute toxicity observed was grade 2 dermatitis in 1 site, and grade 1 in 8 treated sites, at one month post RT, which recovered at 3 months. No RT-related late effect or FUS-MB related toxicity was noted. This study demonstrated safety of combined FUS-MB and RT treatment. Promising response rates suggest potential strong radiosensitization effects of the investigational modality.Trial registration: clinicaltrials.gov, identifier NCT04431674.

Ultrasound-stimulated microbubbles enhancement of fractionated radiation for tumor treatment.

BACKGROUND: Radiation therapy (XRT) causes numerous biological changes in tumor microenvironment. Radiation vascular response, due to endothelial disruption, can influence treatment outcomes in a dose-dependent manner. Ultrasound-stimulated microbubbles (USMB) have also been demonstrated to create a vascular response in the tumor microenvironment and enhance tumor response when used in combination with XRT. Single doses of 8-10 Gy are known to induce activation of acid sphingomyelinase (ASMase)-induced ceramide production, causing vascular damage. Destruction of vasculature results in endothelial apoptosis followed by tumor cell death. The effect of tumor response is known to be synergistic by 10-fold higher cell kill observed when USMB is combined with radiation. METHODS: In this study, we used an USMB approach in combination with conventional low dose fractionated radiation to enhance endothelial cell responses to XRT in human PC3 prostate cancer xenograft model. Mice were divided into untreated, USMB therapy, fractionated XRT, and combined USMB therapy followed by XRT (USMB + XRT) groups. USMB therapy was delivered twice per week in the USMB-alone and combined USMB + XRT treatment groups over four weeks. Radiation treatments were delivered in fractions of 2 Gy/day (total 40 Gy in 20 fractions, BED10 = 48 Gy) in the XRT-alone and combined USMB + XRT groups. The treatment outcome was evaluated using histopathology, power Doppler, and immunohistochemistry assays. RESULTS: Tumor growth assessment showed that sizes of tumors increased in the control and the single treatment groups over a treatment period of four weeks, but significantly decreased with the combined treatments of USMB + XRT. Immunohistochemical analysis indicated a statistically significant vascular disruption in mice that received treatment involving a full 4-week schedule of combined (USMB + XRT) treatments. A statistically significant increase in vascular disruption was demonstrated through CD68 and trichrome fibrosis staining. Changes in local perfusion assessed using high-frequency power Doppler imaging demonstrated attenuated blood flow in the combined group. DISCUSSION AND CONCLUSIONS: This work demonstrates the efficacy of using USMB as a radiation sensitizer in a mouse model of human PC3 tumor xenograft. This radiation treatment enhancement modality has the advantage of targeting tumor vasculature with ultrasound stimulation that can be implemented prior to radiation treatment.

MR-guided ultrasound-stimulated microbubble therapy enhances radiation-induced tumor response.

High intensity focused ultrasound (HIFU) systems have been approved for therapeutic ultrasound delivery to cause tissue ablation or induced hyperthermia. Microbubble agents have also been used in combination with sonication exposures. These require temperature feedback and monitoring to prevent unstable cavitation and prevent excess tissue heating. Previous work has utilized lower power and pressure to oscillate microbubbles and transfer energy to endothelial cells in the absence of thermally induced damage that can radiosensitize tumors. This work investigated whether reduced acoustic power and pressure on a commercial available MR-integrated HIFU system could result in enhanced radiation-induced tumor response after exposure to ultrasound-stimulated microbubbles (USMB) therapy. A commercially available MR-integrated HIFU system was used with a hyperthermia system calibration provided by the manufacturer. The ultrasound transducer was calibrated to reach a peak negative pressure of - 750 kPa. Thirty male New Zealand white rabbits bearing human derived PC3 tumors were grouped to receive no treatment, 14 min of USMB, 8 Gy of radiation in a separate irradiation cabinet, or combined treatments. In vivo temperature changes were collected using MR thermometry at the tumor center and far-field muscle region. Tissues specimens were collected 24 h post radiation therapy. Tumor cell death was measured and compared to untreated controls through hematoxylin and eosin staining and immunohistochemical analysis. The desired peak negative pressure of - 750 kPa used for previous USMB occurred at approximately an input power of 5 W. Temperature changes were limited to under 4 °C in ten of twelve rabbits monitored. The median temperature in the far-field muscle region of the leg was 2.50 °C for groups receiving USMB alone or in combination with radiation. Finally, statistically significant tumor cell death was demonstrated using immunohistochemical analysis in the combined therapy group compared to untreated controls. A commercial MR-guided therapy HIFU system was able to effectively treat PC3 tumors in a rabbit model using USMB therapy in combination with radiation exposures. Future work could find the use of reduced power and pressure levels in a commercial MR-guided therapy system to mechanically stimulate microbubbles and damage endothelial cells without requiring high thermal doses to elicit an antitumor response.

Ultrasound-stimulated microbubble radiation enhancement of tumors: Single-dose and fractionated treatment evaluation.

The use of ultrasound-stimulated microbubble therapy has successfully been used to target tumor vasculature and enhance the effects of radiation therapy in tumor xenografts in mice. Here, we further investigate this treatment using larger, more clinically relevant tumor models. New Zealand white rabbits bearing prostate tumor (PC3) xenografts received a single treatment of either ultrasound-stimulated microbubbles (USMB), ionizing radiation (XRT; 8Gy), or a combination of both treatments (USMB+XRT). Treatment outcome was evaluated 24 hours after treatment using histopathology, immunolabeling, 3D Doppler ultrasound and photoacoustic imaging. A second cohort of rabbits received multiple treatments over a period of three weeks, where USMB treatments were delivered twice weekly with daily XRT treatments to deliver a fractionated 2Gy dose five days per week. A significant decrease in vascular function, observed through immunolabeling of vascular endothelial cells, was observed in tumors receiving the combined treatment (USMB+XRT) compared to control and single treatment groups. This was associated with an increase in cell death as observed through in situ end labeling (ISEL), a decrease in vascular index measured by Power Doppler imaging, and a decrease in oxygen saturation. In rabbits undergoing the long-term fractionated combined treatment, a significant growth delay was observed after 1 week and a significant reduction in tumor size was observed after 3 weeks with combined therapy. Results demonstrated an enhancement of radiation effect and superior anti-tumor effect of the combination of USMB+XRT compared to the single treatments alone. Tumor growth was maximally inhibited with fractionated radiotherapy combined with the ultrasound-stimulated microbubble-based therapy.

Differentiating platinum coated brachytherapy seeds and gold fiducial markers with varying off-resonant frequency offsets.

PURPOSE: To develop an off-resonant frequency filtered method to selectively differentiate between implanted gold fiducial markers and platinum coated brachytherapy seeds. MATERIALS AND METHODS: The magnetic susceptibilities for gold fiducial markers and brachytherapy seeds differ in magnitude and also in their signs, resulting in B0-field inhomogeneity patterns with opposite main lobes. A pulse sequence used to localize brachytherapy seeds with positive contrast, centre-out radial sampling with off-resonance reception (co-RASOR), was used to reconstruct images with a range of off-resonant frequency offsets. The proposed method utilizes two frequency filters to selectively reconstruct maximum intensity projections through band-pass regions where each seed has its maximal localized hyperintensity. Seeds were simulated and then placed in gel and tissue phantoms to validate the technique using orthogonal 2D slices with seeds both parallel and perpendicular to the B0-field. RESULTS: Dual-plane 2D co-RASOR sequences were reconstructed off-resonance with applied frequency filters to create two projections displaying each seed, which were then colour-coded to negative and positive frequencies. Phantom validation showed that each seed contains its maximal CNR in opposing frequency regions as predicted. Local maxima can also appear in both negative and positive frequency regions. The relative difference between the signal of each seed and these local maxima ranged from 1.19 to 3.73, and an image threshold was determined in all cases. Tissue validation showed the technique differentiates seeds correctly and is limited by the hyperintensity patterns observed in the co-RASOR method. CONCLUSIONS: Dual-plane co-RASOR offers sub-millimetre positive contrast from implanted seeds that contain unique off-resonant frequency maxima, which frequency filters can selectively differentiate.

Automatic ROI Placement in the Upper Trapezius Muscle in B-mode Ultrasound Images.

Research involving B-mode ultrasound imaging often requires user defined regions of interest (ROIs) for analysis, traditionally drawn/selected by a trained operator. This manual process is incredibly time consuming and subjective. Here, we propose a fast and simple method of detecting the average location of aponeurosis layers in ultrasound images of the upper trapezius to place a rectangular ROI for quantitative image analysis. A total of 56 B-mode ultrasound images were analyzed, where rectangular ROIs were manually placed in the skeletal muscle by two trained operators. Interoperator agreement was determined between the ROI border locations using intercorrelation coefficient (ICC). Next, our automatic algorithm was applied (image thresholding, binary masking, and pixel intensity peak detection), estimating the mean position of both aponeurosis layers for rectangular ROI placement. The automatic estimation method was compared with the manual (visual) method by various statistics ( t test, linear correlation, Bland-Altman plot). The performance was also evaluated under additive noise conditions (Speckle). Finally, agreement of the overlapping ROI area between the manual and automatic methods was also computed. Performance of the automatic method compared with manual placement was excellent for both the superficial and deep ROI borders, performing consistently even with additive noise (error <0.674 ± 1.69 mm). Manual measurements indicated excellent consensus (ICC = 0.902) between operators. The overlapping area between the manual and automatic measurements demonstrated good agreement (90.65 ± 11.3%). With constraints, our method is robust even under large levels of noise addition making the automatic algorithm an acceptable replacement for manual ROI placement in the upper trapezius.

Resolution and registration in dual-plane co-RASOR MR.

Magnetic resonance imaging (MRI) has superior soft tissue contrast and lower interobserver variability compared to computed tomography and advances in equipment and pseudo-CT estimation have allowed for MR-only radiation therapy planning. Dedicated MR sequences have been used to localize paramagnetic structures with positive contrast, and most implanted seeds are gold fiducial markers (GFMs). We used a fast, dual-plane co-RASOR sequence to localize implanted GFMs with positive contrast in phantom and tissue to assess their resolution and registration accuracy of registration to CT. Off-resonant reconstructions of co-RASOR images were able to resolve GFMs down to 5 mm apart at 12 cm FOV. No systematic biases were observed by comparing registration of co-RASOR and bSSFP to CT images in an MR-compatible Lego phantom with a set of highly visible known points. The standard deviations of the MR to CT distance errors were <0.5 mm in all directions. We separated the component due to registration by comparing the two MR sequences, which had a maximum standard deviation of 0.36 mm in the SI-direction. Registration using the positive contrast points in a porcine sample phantom showed increased errors, but co-RASOR still performs acceptably with a target registration error of <0.75 mm. The dual-plane co-RASOR sequence could then be used for both registration and image tracking when performing MR-only radiation therapy planning.

Localizing implanted fiducial markers using undersampled co-RASOR MR imaging.

The goal of this work was to use an undersampled, dual-plane centre-out radial sampling acquisition pulse sequence, with off-resonance reception, to localize fiducial markers with reduced acquisition time. Two iterative reconstruction techniques, conjugate gradient CG-SENSE and the variational penalty Total Generalized Variation (TGV), were investigated to minimize the undersampling artifacts in off-resonant radial imaging. Simulations of a field perturber were performed at sub-millimeter resolution and reconstructed to display signal pileups that can be radially compressed towards the geometric centre of the perturber for high contrast visualization, but contrast is non-recoverable as the echo time increases. A cylindrical platinum fiducial marker, placed in a phantom parallel and perpendicular to the B0-field was imaged with a short-TE half-echo readout. Contrast-to-Noise (CNR) between the signal of the fiducial its adjacent surrounding shell and half-maximum area were used to compare reconstruction methods and undersampling factors. For single slice acquisitions centred about the fiducial, each slice can be performed in as little as 2.8s. The total acquisition time to localize the fiducial marker in a phantom was reduced to 73s by undersampling (R=8) 37 axial and 15 coronal slices, effectively encoding 1.4s/slice. The noise present in undersampled images, for both scan planes and fiducial orientations, decreased significantly using TGV and CG-SENSE reconstructions, with TGV displaying better spatial resolution from reduced half-maximum area.