A RhoA-FRET Biosensor Mouse for Intravital Imaging in Normal Tissue Homeostasis and Disease Contexts

: Nobis M, Herrmann D, Warren SC, Kadir S, Leung W, Killen M, Magenau A, Stevenson D, Lucas MC, Reischmann N, Vennin C, Conway JRW, Boulghourjian A, Zaratzian A, Law AM, Ormandy CJ, Gallego-Ortega D, Grey ST, Walters SN, Chtanova T, Bailey J, Baldock PA, Quinn JMW, Schwarz JP, Croucher PI, Zhang L, Mrowinska A, Masedunskas A, Herzog H, Gunning PW, Hardeman EC, Samuel MS, Pajic M, Del Monte-Nieto G, Harvey RP, Sansom OJ, Welch HCE, McGhee EJ, Johnsson AE, Anderson KI, Timpson P, Morton JP, Strathdee D

: 2017

: Cell Reports

: Cell reports

: Cell Rep

: 21

: 1

: 274

: 288

: 15

: 2211-1247

DOI: https://doi.org/10.1016/j.celrep.2017.09.022

The small GTPase RhoA is involved in a variety of fundamental processes in normal tissue. Spatiotemporal control of RhoA is thought to govern mechanosensing, growth, and motility of cells, while its deregulation is associated with disease development. Here, we describe the generation of a RhoA-fluorescence resonance energy transfer (FRET) biosensor mouse and its utility for monitoring real-time activity of RhoA in a variety of native tissues in vivo. We assess changes in RhoA activity during mechanosensing of osteocytes within the bone and during neutrophil migration. We also demonstrate spatiotemporal order of RhoA activity within crypt cells of the small intestine and during different stages of mammary gestation. Subsequently, we reveal co-option of RhoA activity in both invasive breast and pancreatic cancers, and we assess drug targeting in these disease settings, illustrating the potential for utilizing this mouse to study RhoA activity in vivo in real time.