Short-duration-focused ultrasound stimulation of Hsp70 expression in vivo. Phys Med Biol

Department of Biomedical Engineering, University of California, Davis, CA 95616, USA.
Physics in Medicine and Biology (Impact Factor: 2.76). 08/2008; 53(13):3641-60. DOI: 10.1088/0031-9155/53/13/017
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The development of transgenic reporter mice and advances in in vivo optical imaging have created unique opportunities to assess and analyze biological responses to thermal therapy directly in living tissues. Reporter mice incorporating the regulatory regions from the genes encoding the 70 kDa heat-shock proteins (Hsp70) and firefly luciferase (luc) as reporter genes can be used to non-invasively reveal gene activation in living tissues in response to thermal stress. High-intensity-focused ultrasound (HIFU) can deliver measured doses of acoustic energy to highly localized regions of tissue at intensities that are sufficient to stimulate Hsp70 expression. We report activation of Hsp70-luc expression using 1 s duration HIFU heating to stimulate gene expression in the skin of the transgenic reporter mouse. Hsp70 expression was tracked for 96 h following the application of 1.5 MHz continuous-wave ultrasound with spatial peak intensities ranging from 53 W cm(-2) up to 352 W cm(-2). The results indicated that peak Hsp70 expression is observed 6-48 h post-heating, with significant activity remaining at 96 h. Exposure durations were simulated using a finite-element model, and the predicted temperatures were found to be consistent with the observed Hsp70 expression patterns. Histological evaluation revealed that the thermal damage starts at the stratum corneum and extends deeper with increasing intensity. These results indicated that short-duration HIFU may be useful for inducing heat-shock expression, and that the period between treatments needs to be greater than 96 h due to the protective properties of Hsp70.

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    • "Previous studies demonstrated that hsp is not only induced by heat, but can also be triggered by mechanical stress [33,34]. Kruse et al. demonstrated that hsp70 overexpression is sometimes observed within a similar pressure range (intensity = 53 W/cm2) and a short duration (1s) of ultrasound exposure without temperature elevation [35], similar to induction caused by heat from HIFU exposure [22]. We therefore evaluated hsp60 expression and found that hsp60 indeed increased in regions of FUS exposure compared to control tumor regions, suggesting that induction of hsp overexpression may be involved in triggering of the immune response by MB-FUS (Additional file 1: Figure S1). "
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    ABSTRACT: Background High-intensity focused-ultrasound (HIFU) has been successfully employed for thermal ablation of tumors in clinical settings. Continuous- or pulsed-mode HIFU may also induce a host antitumor immune response, mainly through expansion of antigen-presenting cells in response to increased cellular debris and through increased macrophage activation/infiltration. Here we demonstrated that another form of focused ultrasound delivery, using low-pressure, pulsed-mode exposure in the presence of microbubbles (MBs), may also trigger an antitumor immunological response and inhibit tumor growth. Methods A total of 280 tumor-bearing animals were subjected to sonographically-guided FUS. Implanted tumors were exposed to low-pressure FUS (0.6 to 1.4 MPa) with MBs to increase the permeability of tumor microvasculature. Results Tumor progression was suppressed by both 0.6 and 1.4-MPa MB-enhanced FUS exposures. We observed a transient increase in infiltration of non-T regulatory (non-Treg) tumor infiltrating lymphocytes (TILs) and continual infiltration of CD8+ cytotoxic T-lymphocytes (CTL). The ratio of CD8+/Treg increased significantly and tumor growth was inhibited. Conclusions Our findings suggest that low-pressure FUS exposure with MBs may constitute a useful tool for triggering an anticancer immune response, for potential cancer immunotherapy.
    Journal of Translational Medicine 11/2012; 10(1):221. DOI:10.1186/1479-5876-10-221 · 3.93 Impact Factor
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    ABSTRACT: Dissertation Ultrasound can be used for a variety of therapeutic purposes. High-intensity focused ultrasound (HIFU) has progressed over the past decade to become a viable therapeutic method and is valuable as a non-invasive alternative to many surgical procedures. Ultrasonic thermal therapies can also be used to release thermally sensitive liposomes encapsulating chemotherapeutic drugs. In the brain, the permeability of the blood-brain barrier to drugs, antibodies, and gene transfer can be increased with a mechanical mechanism using ultrasound and contrast agent. The work presented in this dissertation tests the hypothesis that a diagnostic system can be used for combined imaging and therapeutic applications. In order to evaluate the effectiveness of a diagnostic system for use in therapeutic applications, a set of non-destructive tests is developed that can predict the potential for high acoustic output. A rigorous, nondestructive testing regimen for standard, diagnostic transducers to evaluate their potential for therapeutic use is formulated. Based on this work, transducer heating is identified as the largest challenge. The design and evaluation of several custom diagnostic transducers with various modifications to reduce internal heating are described. These transducers are compared with diagnostic controls using image contrast, face heating, hydrophone, and ARFI displacement measurements. From these results, we conclude that the most promising design is a passively and actively cooled, PZT-4 multilayer composite transducer, while the acoustically lossless lens and capactive micro-machined transducers evaluated herein are determined to be ineffective. Three therapeutic applications are evaluated for the combined system. Image-guided spot ablations, such as in the treatment of early stage liver cancers, could not be successfully performed; however, the additional acoustic output requirements are determined to be on the order of 2.4 times those that can be currently produced without transducer damage in a clinically relevant amount of time (10-20 seconds per spot). The potential of a diagnostic system for a hyperthermia application is shown by producing temperatures for the duration necessary to release chemotherapeutic agents from thermally-activated liposomes without damage to the transducer. Finally, a mechanically-based therapeutic method for opening the BBB with ultrasonic contrast agent and specialized sonication regimes under ultrasonic B-mode guidance is demonstrated. These studies indicate that a diagnostic system is capable of both moderate thermal and mechanical therapeutic applications under co-registered image-guidance.
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    ABSTRACT: Acknowledgments The authors acknowledge the funding of NIH CA 103828 and a grant from the US Department of Defense entitled the Medical Free Electron Laser Program, administered by the Air Force Office of Sponsored Research, grant number FA9550-04-1-0045.
    Physics in Medicine and Biology 11/2008; 53(22). DOI:10.1088/0031-9155/53/22/C01 · 2.76 Impact Factor
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