Miller Austin D C, Chowdhury Soham P, Hanson Hadley W, Linderman Sarah K, Ghasemi Hannah I, Miller Wyatt D, Morrissey Meghan A, Richardson Chris D, Gardner Brooke M, Mukherjee Arnab
Biomolecular Science and Engineering Graduate Program, University of California, Santa Barbara, CA, 93106, USA.
Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, 93106, USA.
J Biol Eng. 2024 Apr 22;18(1):30. doi: 10.1186/s13036-024-00424-5.
Aquaporin-1 (Aqp1), a water channel, has garnered significant interest for cell-based medicine and in vivo synthetic biology due to its ability to be genetically encoded to produce magnetic resonance signals by increasing the rate of water diffusion in cells. However, concerns regarding the effects of Aqp1 overexpression and increased membrane diffusivity on cell physiology have limited its widespread use as a deep-tissue reporter. In this study, we present evidence that Aqp1 generates strong diffusion-based magnetic resonance signals without adversely affecting cell viability or morphology in diverse cell lines derived from mice and humans. Our findings indicate that Aqp1 overexpression does not induce ER stress, which is frequently associated with heterologous expression of membrane proteins. Furthermore, we observed that Aqp1 expression had no detrimental effects on native biological activities, such as phagocytosis, immune response, insulin secretion, and tumor cell migration in the analyzed cell lines. These findings should serve to alleviate any lingering safety concerns regarding the utilization of Aqp1 as a genetic reporter and should foster its broader application as a noninvasive reporter for in vivo studies.
水通道蛋白-1（Aqp1）作为一种水通道，因其能够通过增加细胞内水扩散速率进行基因编码以产生磁共振信号，在基于细胞的医学和体内合成生物学领域引起了广泛关注。然而，对Aqp1过表达及其增加的膜扩散性对细胞生理学影响的担忧，限制了其作为深部组织报告基因的广泛应用。在本研究中，我们提供证据表明，Aqp1在源自小鼠和人类的多种细胞系中产生强烈的基于扩散的磁共振信号，而不会对细胞活力或形态产生不利影响。我们的研究结果表明，Aqp1过表达不会诱导内质网应激，内质网应激通常与膜蛋白的异源表达相关。此外，我们观察到Aqp1表达对分析细胞系中的天然生物学活性，如吞噬作用、免疫反应、胰岛素分泌和肿瘤细胞迁移，没有不利影响。这些发现应有助于缓解对将Aqp1用作基因报告基因的任何残留安全担忧，并应促进其作为体内研究的非侵入性报告基因的更广泛应用。
