CIBER de Bioingenieria, Biomateriales y Nanomedicina, CIBER-BBN, Madrid, Spain; Hospital Universitario La Paz-IdiPAZ, Paseo de la Castellana 261, 28046 Madrid, Spain.
CIBER de Bioingenieria, Biomateriales y Nanomedicina, CIBER-BBN, Madrid, Spain; Hospital Universitario La Paz-IdiPAZ, Paseo de la Castellana 261, 28046 Madrid, Spain.
J Photochem Photobiol B. 2023 May;242:112697. doi: 10.1016/j.jphotobiol.2023.112697. Epub 2023 Mar 23.
This study investigated whether noninvasive near-infrared (NIR) energy could be transduced into heat in deep-seated organs in which adenovirus type-5 vectors tend to accumulate, thereby activating heat shock protein (HSP) promoter-mediated transgene expression, without local administration of photothermal agents. NIR irradiation of the subdiaphragmatic and left dorsocranial part of the abdominal cavity of adult immunocompetent C3H/HeNRj mice with an 808-nm laser effectively increased the temperature of the irradiated regions of the liver and spleen, respectively, resulting in the accumulation of the heat-inducible HSP70 protein. Spatial control of transgene expression was achieved in the NIR-irradiated regions of the mice administered an adenoviral vector carrying a firefly luciferase (fLuc) coding sequence controlled by a human HSP70B promoter, as assessed by bioluminescence and immunohistochemistry analyses. Levels of reporter gene expression were modulated by controlling NIR power density. Spatial control of transgene expression through NIR-focused activation of the HSP70B promoter, as well as temporal regulation by administering rapamycin was achieved in the spleens of mice inoculated with an adenoviral vector encoding a rapamycin-dependent transactivator driven by the HSP70B promoter and an adenoviral vector carrying a fLuc coding sequence controlled by the rapamycin-activated transactivator. Mice that were administered rapamycin and exposed to NIR light expressed fLuc activity in the splenic region, whereas no activity was detected in mice that were only administered rapamycin or vehicle or only NIR-irradiated. Thus, in the absence of any exogenously supplied photothermal material, remote control of heat-induced transgene expression can be achieved in the liver and spleen by means of noninvasive NIR irradiation.
本研究旨在探讨非侵入性近红外（NIR）能量是否可以转化为深层器官中的热能，腺病毒 5 型载体往往会在这些器官中积聚，从而激活热休克蛋白（HSP）启动子介导的转基因表达，而无需局部给予光热剂。用 808nm 激光对成年免疫功能正常的 C3H/HeNRj 小鼠的腹腔下和左背侧部分进行 NIR 照射，分别有效地提高了肝脏和脾脏照射区域的温度，导致热诱导 HSP70 蛋白的积累。在给予携带萤火虫荧光素酶（fLuc）编码序列的腺病毒载体的小鼠的 NIR 照射区域中，通过生物发光和免疫组织化学分析，实现了转基因表达的空间控制，该腺病毒载体由人 HSP70B 启动子控制。通过控制 NIR 功率密度来调节报告基因表达水平。在接种了由 HSP70B 启动子驱动的雷帕霉素依赖性转录激活子编码的腺病毒载体和由雷帕霉素激活的转录激活子控制的 fLuc 编码序列的腺病毒载体的小鼠脾脏中，通过 NIR 聚焦激活 HSP70B 启动子实现了转基因表达的空间控制，并且通过给予雷帕霉素进行了时间调节。给予雷帕霉素并暴露于 NIR 光的小鼠在脾区表达 fLuc 活性，而仅给予雷帕霉素或载体或仅 NIR 照射的小鼠则未检测到活性。因此，在没有任何外源供应的光热材料的情况下，可以通过非侵入性的 NIR 照射，在肝脏和脾脏中实现远程控制热诱导的转基因表达。
