Deciphering the role of ADAM15 vs. ADAM17 in fibroblast function: Mechanobiology vs. Proteolysis

解读 ADAM15 与 ADAM17 在成纤维细胞功能中的作用：机械生物学与蛋白水解

• 批准号: RGPIN-2022-03408
• 负责人: Kassiri, Zamaneh
• 金额: $ 2.84万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=763294
• 关键词: Deciphering; role; ADAM15; vs.; ADAM17

Fibroblasts (FBs) are the main cells for synthesis and homeostasis of the extracellular matrix (ECM) in various tissues. FBs respond to external stimuli and adjust their function (proliferation, migration, activation) accordingly. Excess FB activation leads to overproduction of ECM (scar/fibrosis), while the inverse results in structural instability. Disintegrin and metalloproteinase-15 (ADAM15) and ADAM17 are two highly expressed ADAMs in FBs, and are (incorrectly) assumed to have similar functions, though recent findings suggest otherwise. Our prior research in cardiovascular disease models has revealed the dire need to understand the unique mechanistic function of each ADAM, and the fundamental processes by which they regulate FB function and ECM remodeling. ADAM15 is unique as the only ADAM to cleave an ECM protein, whereas ADAM17 is the most versatile sheddase with >80 substrates. ADAMs have the ability to proteolytically process membrane-bound proteins and to also interact with integrins (through their disintegrin domain), but it is yet to be determined which mechanism takes priority, and if this is specific to an ADAM or to the nature of the stimulus. We hypothesize that in FBs, ADAM15 predominantly functions through its C-terminus-mediated signaling, and by interacting with integrins and activating the associated intracellular signaling; whereas the function of ADAM17 is primarily through proteolytic shedding of membrane-bound proteins (growth factors, cytokines) enhancing their bioavailability and their downstream pathways. Over the next 5 years, we will investigate the comparative function of ADAM15 and ADAM17 in the function of FBs in ECM production and homeostasis in response to pharmacological versus mechanical stimuli. We will use a primary FB culture system, and will knockdown ADAM15 or ADAM17 using siRNA (scrambled siRNA will be used as control in parallel). Subsequently, FBs will be treated with pharmacological stimuli (Angiotensin II or Phenylephrine) or subjected to mechanical stress (rhythmic or static stretching, FlexCell5000 system). FB activity (synthesis of ECM proteins, contractility), proliferation, migration, and survival will be investigated in control, ADAM15- and ADAM17-deficient FBs (±stimulus). Detailed molecular and mechanistic analyses will be performed to compare and contrast the role of these two ADAMs in agonist-induced versus mechanical activation of FBs and the overall ECM remodeling. My lab has extensive experience in studying ECM remodeling and FB functions. We have all the equipment and expertise necessary to conduct the described experiments. These experiments will reveal novel mechanism of action for ADAM15 and -17 in FB function and ECM remodeling, and will create new knowledge in cell physiology and natural sciences by determining the proteolytic and mechano-sensory functions of these ADAMs in FBs, which would be beneficial in the fields of cell- and mechano-biology.

FIBB对外部的反应并记录了其功能（迁移，迁移，激活）。 （错误地）假定具有相似的功能，尽管我们在心血管疾病模型中的先前研究表明，他们了解了每个ADAM L过程的独特机械，它们可以调节FB功能和ECM重塑。裂解曲线蛋白，而ADAM17是具有> 80个底物的最通用的SHEDDASE。 . We HypotheSize That in fbs, Adam15 Predominnants Throoth is the C-Terminus-MediaTed Signaling Nteracting with Integrings and Activating The Associated Intracellular Signaling; Shedding of Membrane-Bound Proteins (Growth Factors, Cytokines) ENHANCING THEIR BIOAVAILABILITY and THEIR DOWNSTREAM PATHWAYS. We Will研究ADAM15在ECM生产中的FBS和机械刺激中的比较功能。受到机械应力（有节奏或静态拉伸，flexcell5000系统），并将研究在对照中，ADAM17缺陷型FBS（±刺激） - 我的ECM重塑。 ，通过确定ESE ADAM在FBS中ESE ADAM的蛋白质和机械感官功能，在细胞生理和自然科学方面的Willedge，这在细胞和机械生物学领域将是有益的。