Chevillet John R, Khokhlova Tatiana D, Giraldez Maria D, Schade George R, Starr Frank, Wang Yak-Nam, Gallichotte Emily N, Wang Kai, Hwang Joo Ha, Tewari Muneesh
From the Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Wash (J.R.C., E.N.G., M.D.G., M.T.); Institute for Systems Biology, Seattle, Wash (J.R.C., K.W.); Department of Medicine (T.D.K., J.H.H.), Department of Urology (G.R.S.), and Applied Physics Laboratory (F.S., Y.N.W.), University of Washington, Seattle, Wash; and Departments of Internal Medicine (M.D.G., M.T.) and Biomedical Engineering, Center for Computational Medicine and Bioinformatics, and the Biointerfaces Institute (M.T.), University of Michigan, 109 Zina Pitcher Pl, 1502 BSRB, SPC 2200, Ann Arbor, MI 48109.
Radiology. 2017 Apr;283(1):158-167. doi: 10.1148/radiol.2016160024. Epub 2016 Nov 1.
Purpose To compare the abilities of three pulsed focused ultrasound regimes (that cause tissue liquefaction, permeabilization, or mild heating) to release tumor-derived microRNA into the circulation in vivo and to evaluate release dynamics. Materials and Methods All rat experiments were approved by the University of Washington Institutional Animal Care and Use Committee. Reverse-transcription quantitative polymerase chain reaction array profiling was used to identify candidate microRNA biomarkers in a rat solid tumor cell line. Rats subcutaneously grafted with these cells were randomly assigned among three pulsed focused ultrasound treatment groups: (a) local tissue liquefaction via boiling histotripsy, (b) tissue permeabilization via inertial cavitation, and (c) mild (<10°C) heating of tissue, as well as a sham-treated control group. Blood specimens were drawn immediately prior to treatment and serially over 24 hours afterward. Plasma microRNA was quantified with reverse-transcription quantitative polymerase chain reaction, and statistical significance was determined with one-way analysis of variance (Kruskal-Wallis and Friedman tests), followed by the Dunn multiple-comparisons test. Results After tissue liquefaction and cavitation treatments (but not mild heating), plasma quantities of candidate biomarkers increased significantly (P value range, <.0001 to .04) relative to sham-treated controls. A threefold to 32-fold increase occurred within 15 minutes after initiation of pulsed focused ultrasound tumor treatment, and these increases persisted for 3 hours. Histologic examination confirmed complete liquefaction of the targeted tumor area with boiling histotripsy, in addition to areas of petechial hemorrhage and tissue disruption by means of cavitation-based treatment. Conclusion Mechanical tumor tissue disruption with pulsed focused ultrasound-induced bubble activity significantly increases the plasma abundance of tumor-derived microRNA rapidly after treatment. RSNA, 2016 Online supplemental material is available for this article.
目的 比较三种脉冲聚焦超声模式（导致组织液化、通透化或轻度加热）在体内将肿瘤衍生的微小RNA释放到循环系统中的能力，并评估释放动力学。材料与方法 所有大鼠实验均经华盛顿大学机构动物护理与使用委员会批准。采用逆转录定量聚合酶链反应阵列分析来鉴定大鼠实体瘤细胞系中的候选微小RNA生物标志物。将皮下移植了这些细胞的大鼠随机分为三个脉冲聚焦超声治疗组：(a) 通过沸腾组织粉碎术实现局部组织液化，(b) 通过惯性空化实现组织通透化，(c) 对组织进行轻度（<10°C）加热，以及一个假治疗对照组。在治疗前即刻采集血液样本，并在治疗后的24小时内连续采集。用逆转录定量聚合酶链反应对血浆微小RNA进行定量，并通过单因素方差分析（Kruskal-Wallis和Friedman检验）确定统计学意义，随后进行Dunn多重比较检验。结果 在组织液化和空化治疗后（但不是轻度加热后），相对于假治疗对照组，候选生物标志物的血浆量显著增加（P值范围为<.0001至.04）。在脉冲聚焦超声肿瘤治疗开始后的15分钟内增加了3倍至32倍，并且这些增加持续了3小时。组织学检查证实，通过沸腾组织粉碎术，靶向肿瘤区域完全液化，此外，基于空化的治疗还导致了瘀点状出血和组织破坏区域。结论 脉冲聚焦超声诱导的气泡活动对肿瘤组织进行机械破坏后，可在治疗后迅速显著增加肿瘤衍生微小RNA的血浆丰度。RSNA，2016 本文提供在线补充材料。
