Kumar Gaurav, Goldberg S Nahum, Gourevitch Svetlana, Levchenko Tatyana, Torchilin Vladimir, Galun Eithan, Ahmed Muneeb
From the Laboratory for Minimally Invasive Tumor Therapies, Department of Radiology, Beth Israel Deaconess Medical Center, Harvard Medical School, 1 Deaconess Rd, WCC 308-B, Boston, MA 02215 (G.K., S.N.G., M.A.); Division of Image-guided Therapy and Interventional Oncology, Department of Radiology (S.N.G.), and Goldyne Savad Institute of Gene Therapy (S.G., E.G.), Hadassah Hebrew University Hospital, Jerusalem, Israel; and Department of Pharmaceutical Sciences, Northeastern University, Boston, Mass (T.L., V.T.).
Radiology. 2018 Feb;286(2):524-536. doi: 10.1148/radiol.2017162943. Epub 2017 Sep 6.
Purpose To (a) identify key expressed genes in the periablational rim after radiofrequency ablation (RFA) and their role in driving the stimulation of distant tumor growth and (b) use adjuvant drug therapies to block key identified mediator(s) to suppress off-target tumorigenic effects of hepatic RFA. Materials and Methods This institutional animal care and use committee-approved study was performed in C57BL6 mice (n = 20) and F344 rats (n = 124). First, gene expression analysis was performed in mice after hepatic RFA or sham procedure; mice were sacrificed 24 hours to 7 days after treatment. Data were analyzed for differentially expressed genes (greater than twofold change) and their functional annotations. Next, animals were allocated to hepatic RFA or sham treatment with or without STAT3 (signal transducer and activator of transcription 3) inhibitor S3I-201 for periablational phosphorylated STAT3 immunohistochemistry analysis at 24 hours. Finally, animals with subcutaneous R3230 adenocarcinoma tumors were allocated to RFA or sham treatment with or without a STAT3 inhibitor (S3I-201 or micellar curcumin, eight arms). Outcomes included distant tumor growth, proliferation (Ki-67 percentage), and microvascular density. Results At 24 hours, 217 genes had altered expression (107 upregulated and 110 downregulated), decreasing to 55 genes (27 upregulated and 28 downregulated) and 18 genes (four upregulated, 14 downregulated) at 72 hours and 7 days, respectively. At 24 hours, STAT3 occurred in four of seven activated pathways associated with pro-oncogenic genes at network analysis. Immunohistochemistry analysis confirmed elevated periablational phosphorylated STAT3 24 hours after RFA, which was suppressed with S3I-201 (percentage of positive cells per field: 31.7% ± 3.4 vs 3.8% ± 1.7; P < .001). Combined RFA plus S3I-201 reduced systemic distant tumor growth at 7 days (end diameter: 11.8 mm ± 0.5 with RFA plus S3I-201, 19.8 mm ± 0.7 with RFA alone, and 15 mm ± 0.7 with sham procedure; P < .001). STAT3 inhibition with micellar curcumin also suppressed postablation stimulation of distant tumor growth, proliferation, and microvascular density (P < .01). Conclusion Gene expression analysis identified multiple pathways upregulated in the periablational rim after hepatic RFA, of which STAT3 was active in four of seven. Postablation STAT3 activation is linked to increased distant tumor stimulation and can be suppressed with adjuvant STAT3 inhibitors. RSNA, 2017.
目的 （a） 识别射频消融（RFA）后消融边缘的关键表达基因及其在促进远处肿瘤生长中的作用，（b） 使用辅助药物疗法阻断关键的已识别介质，以抑制肝脏RFA的脱靶致瘤效应。材料与方法 本研究经机构动物护理和使用委员会批准，在C57BL6小鼠（n = 20）和F344大鼠（n = 124）中进行。首先，对接受肝脏RFA或假手术的小鼠进行基因表达分析；在治疗后24小时至7天处死小鼠。分析数据以找出差异表达基因（变化超过两倍）及其功能注释。接下来，将动物分配接受肝脏RFA或假手术，同时或不同时使用STAT3（信号转导和转录激活因子3）抑制剂S3I-201，于24小时进行消融边缘磷酸化STAT3免疫组织化学分析。最后，将患有皮下R3230腺癌肿瘤的动物分配接受RFA或假手术，同时或不同时使用STAT3抑制剂（S3I-201或胶束姜黄素，共八个组）。观察指标包括远处肿瘤生长、增殖（Ki-67百分比）和微血管密度。结果 在24小时时，有217个基因表达发生改变（107个上调，110个下调），在72小时和7天时分别降至55个基因（27个上调，28个下调）和18个基因（4个上调，14个下调）。在24小时时，网络分析显示STAT3出现在与促癌基因相关的七个激活通路中的四个中。免疫组织化学分析证实RFA后24小时消融边缘磷酸化STAT3升高，而S3I-201可抑制这种升高（每视野阳性细胞百分比：31.7% ± 3.4对3.8% ± 1.7；P <.001）。联合RFA加S3I-201可在7天时减少全身远处肿瘤生长（最终直径：RFA加S3I-201组为11.8 mm ± 0.5，单纯RFA组为19.8 mm ± 0.7，假手术组为15 mm ± 0.7；P <.001）。用胶束姜黄素抑制STAT3也可抑制消融后对远处肿瘤生长、增殖和微血管密度的刺激（P <.01）。结论 基因表达分析确定了肝脏RFA后消融边缘中多个上调的通路，其中STAT3在七个通路中的四个中处于激活状态。消融后STAT3激活与远处肿瘤刺激增加有关，可被辅助性STAT3抑制剂抑制。RSNA，2017年。
