Identification and the role of stress-activated molecular pathways in cell adaptation during**microbubble-coupled sonoporation

**微泡耦合声孔过程中应激激活分子途径的识别及其在细胞适应中的作用

• 批准号: 529449-2018
• 负责人: Botelho, Roberto
• 金额: $ 1.82万
• 依托单位: Ryerson University
• 依托单位国家: 加拿大
• 项目类别: Engage Grants Program
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=645860
• 关键词: Identification; role; stress; activated; molecular

MD Precision Inc. is a Canadian company that develops and manufactures ultrasound equipment for clinical**use, including medical imaging. MD Precision Inc. is looking to develop new devices and/or strategies that**couple ultrasound with microbubble-based sonoporation to kill solid tumours. In this process, microbubbles,**which are inert gas-filled lipid shells, are injected into the bloodstream and then subjected to a local ultrasound**field, which makes microbubbles in that field rapidly vibrate. These vibrations generate fluid "micro-jets" that**puncture the plasma membrane of nearby cells. Depending on the size and number of pores, cells either die,**internalize larger doses of drugs, or survive the assault. Overall, microbubble-coupled sonoporation imposes a**strong physical stress on cells. However, cells have evolved multiple molecular pathways that sense various**stresses like membrane damage. Ultimately, the goal of these stress-activated pathways is to alter gene**expression and/or protein function to adapt surviving cells to stress and improve survivability during**subsequent assaults. It is not known if and which stress-sensing pathways are engaged during the physical**stress imposed by microbubble-induced sonoporation and if surviving cells become stress-adapted. In this**project, my laboratory will use Western blotting, flow cytometry and ELISA assays to identify which**stress-activated pathways are engaged during microbubble-sonoporation in normal and transformed cell lines.**We will then use cell proliferation assays to examine if surviving cells become stress-adapted and better**survive subsequent assaults such as exposure to DNA-damaging agents. Finally, we will employ cell**proliferation assays to investigate if co-application of inhibitors of stress-activated pathways during**sonoporation neutralize stress adaption. This new fundamental knowledge of how cells respond and adapt to**physical stress imposed by microbubble-coupled sonoporation will help MD Precision Inc. to innovate**strategies that use microbubble-coupled sonoporation to more effectively destroy tumours. If successful, this**would position MD Precision Inc. to expand its market share, increase its labour force and benefit Canadians.

MD Precision Inc.是一家加拿大公司，开发和制造用于临床**用途的超声波设备，包括医学成像。MD Precision Inc.正在寻求开发新的设备和/或策略，**将超声波与基于微泡的声波手术相结合，以杀死实体肿瘤。在这个过程中，充满惰性气体的脂质壳**微泡被注入血液，然后受到局部超声波**场的影响，这会使该场中的微泡快速振动。这些振动产生液体“微喷流”，穿透附近细胞的质膜。根据毛孔的大小和数量，细胞要么死亡，要么内化更大剂量的药物，要么在攻击中幸存下来。总体而言，微泡耦合的声波作用对细胞施加了**强烈的物理压力。然而，细胞进化出了多条分子通路，可以感知各种**压力，比如膜损伤。最终，这些应激激活途径的目标是改变基因**的表达和/或蛋白质的功能，以使存活的细胞适应压力，并在**随后的攻击中提高生存能力。目前尚不清楚在微泡诱导的声学治疗施加的物理**压力过程中，是否以及哪些压力感应通路被参与，以及存活的细胞是否变得适应压力。在这个**项目中，我的实验室将使用Western blotting、流式细胞仪和ELISA法来确定正常和转化的细胞系在微泡超声作用过程中参与了哪些**应激激活途径。**然后我们将使用细胞增殖分析来检查存活的细胞是否变得适应了压力，**是否能够更好地**经受后续的攻击，如暴露于DNA损伤剂。最后，我们将使用细胞增殖分析来研究在声学处理过程中联合应用应激激活通路的抑制剂是否能中和应激适应。这些关于细胞如何响应和适应微泡耦合声波手术施加的**物理压力的新基础知识将帮助MD Precision Inc.创新**使用微泡耦合超声波手术更有效地摧毁肿瘤的**策略。如果成功，这将使MD Precision Inc.扩大其市场份额，增加劳动力，并使加拿大人受益。