Rapid and orthogonal logic gating with a gibberellin-induced dimerization system

Miyamoto, Takafumi; DeRose, Robert; Suarez, Allison; Ueno, Takahiro; Chen, Melinda; Sun, Tai; Wolfgang, Michael J.; Mukherjee, Chandrani; Meyers, David J.; Inoue, Takanari

doi:10.1038/nchembio.922

Cited by 197 publications

(238 citation statements)

References 41 publications

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“…We therefore turned to a genetic approach to directly target RhoA activity in mural cells. hBMSCs were transduced with lentivirus harboring (i) a RhoA(CA) activating mutant, but lacking its endogenous farnesylation site, thus preventing membrane localization and activation; and (ii) a membranetethered anchor, in which the addition of gibberellinanalog (GA 3 -AM) induced coupling of the RhoA construct to the membrane-tethered anchor to rapidly activate the construct (40). hBMSCs expressing these constructs but not yet exposed to gibberellin behaved normally, adhered to the endothelialized channel similar to nontransduced cells, and maintained confluency and barrier function of endothelium (Fig.…”

Section: Inflammatory Cues Drive Mural Cell Detachment and Vascularmentioning

confidence: 99%

Three-dimensional biomimetic vascular model reveals a RhoA, Rac1, and N -cadherin balance in mural cell–endothelial cell-regulated barrier function

Alimperti

Mirabella

Bajaj

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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The integrity of the endothelial barrier between circulating blood and tissue is important for blood vessel function and, ultimately, for organ homeostasis. Here, we developed a vessel-on-a-chip with perfused endothelialized channels lined with human bone marrow stromal cells, which adopt a mural cell-like phenotype that recapitulates barrier function of the vasculature. In this model, barrier function is compromised upon exposure to inflammatory factors such as LPS, thrombin, and TNFα, as has been observed in vivo. Interestingly, we observed a rapid physical withdrawal of mural cells from the endothelium that was accompanied by an inhibition of endogenous Rac1 activity and increase in RhoA activity in the mural cells themselves upon inflammation. Using a system to chemically induce activity in exogenously expressed Rac1 or RhoA within minutes of stimulation, we demonstrated RhoA activation induced loss of mural cell coverage on the endothelium and reduced endothelial barrier function, and this effect was abrogated when Rac1 was simultaneously activated. We further showed that N-cadherin expression in mural cells plays a key role in barrier function, as CRISPRmediated knockout of N-cadherin in the mural cells led to loss of barrier function, and overexpression of N-cadherin in CHO cells promoted barrier function. In summary, this bicellular model demonstrates the continuous and rapid modulation of adhesive interactions between endothelial and mural cells and its impact on vascular barrier function and highlights an in vitro platform to study the biology of perivascular-endothelial interactions.

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Section: Inflammatory Cues Drive Mural Cell Detachment and Vascularmentioning

confidence: 99%

Three-dimensional biomimetic vascular model reveals a RhoA, Rac1, and N -cadherin balance in mural cell–endothelial cell-regulated barrier function

Alimperti

Mirabella

Bajaj

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…The cell expansion occurs partly through effects on downstream signaling and auxin transport (AMBER et al, 2012). On the other hand, excessive gibberellin induces acidifi cation in cell (MIYAMOTO et al, 2012), which infl uences the auxin transport. HENNY & HAMILTON (1992) suggested 500mg L -1 GA 3 as the maximum concentration of anthurium, in agreement with the results obtained in this study.…”

Section: Resultsmentioning

confidence: 99%

Growth and yield of anthurium in response to gibberellic acid

et al. 2014

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“…mammalian-based orthogonal chemically induced dimerization (CID) systems (Liang et al, 2011;Miyamoto et al, 2012). Taken together, target-based screens offer the possibility to identify a small molecule ligand for a specific protein of interest and to further improve on small molecule binding.…”

Section: Target-based Screens and Synthetic Biologymentioning

confidence: 99%