Inactivation of RhoA for Hypertension Treatment Through the TRPV4–RhoA–RhoGDI1 Axis
Abstract
BACKGROUND: The RhoA (Ras homolog family member A) signaling pathway is pivotal in regulating vascular smooth muscle cells (VSMCs) function and blood pressure homeostasis. Current inhibitors of the RhoA signaling pathway are limited in hypertension treatment, suffering from poor efficacy, insufficient specificity, and developmental challenges. METHODS: Cryo-electron microscopy (EM), proximity ligation assay (PLA), and site-directed mutagenesis were used to explore the mechanism of RhoA activity regulation. VSMC, hypertensive animal models, Trpv4 -/- and Arhgdia f/f Myh11-CRE ERT2 (smooth muscle–specific RhoGDI1 knockout) mice were used to investigate the role of the TRPV4 (transient receptor potential cation channel subfamily V member 4)–RhoA–RhoGDI1 (Rho GDP dissociation inhibitor 1) axis in hypertension. RESULTS: AH001 (( R )-1-(3-ethylphenyl) ethane-1,2-diol) was identified as a novel inhibitor of the RhoA signaling pathway. It targets the TRPV4–RhoA–RhoGDI1 axis to effectively sequester inactive RhoA–GDP in the plasma membrane and cytoplasm, which is distinct from typical RhoA inhibition modes. The cryo-EM structure of the TRPV4 AH001 –RhoA complex showed that AH001-bound TRPV4 adopts a closed state with RhoA in an inactive GDP-bound state. Functional studies further revealed that AH001 reduced the pool of active RhoA by enhancing TRPV4–RhoA binding and facilitating RhoGDI1–RhoA interaction in VSMC. This inhibition notably decreased both acute and long-term blood pressure and prevented vascular remodeling in Ang II–induced hypertensive mice and spontaneously hypertensive rats. However, these antihypertensive effects were weakened in Trpv4 -/- and Arhgdia f/f Myh11-CRE ERT2 mice. Additionally, AH001 effectively inhibited VSMC contraction via the RhoA/ROCK (Rho-associated protein kinase)/MYPT1 (myosin phosphatase target subunit 1)/MLC (myosin light chain 2) signaling pathway and suppressed VSMC phenotype switching to myofibroblasts through the RhoA/ROCK/LIMK1 (LIM domain kinase)/cofilin/MRTF-A (myocardin-related transcription factor A)/SRF (serum response factor) signaling cascade. TRPV4 and RhoGDI1 knockdown attenuated AH001’s inhibition of VSMC contraction and phenotypic switching to myofibroblasts. CONCLUSIONS: This study revealed a novel mode of RhoA signaling inhibition targeting the TRPV4–RhoA–RhoGDI1 axis, offering new insights for future antihypertensive drug development and proposing innovative strategies for targeting challenging Rho GTPases.
Article Details
Authors (21)
Jiawen Wang
Institute of Functional Nano & Soft Materials (FUNSOM)
Zhen Yuan
Beijing National Laboratory for Molecular Sciences, CAS Key Laboratory of Molecular Recognition and Function, Institute of Chemistry
Na Yu
State Key Laboratory of Quantum Functional Materials, School of Physical Science and Technology
Qian Jiao
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science & Technology, Shanghai, China (J.W., Z.Y., N.Y., Q.J., H.Z., W.L., J.S., S.R., Y.M., L.Q., J.F., Y.X., X.Q., J.Z., Z.Z., S.L., R.W., H.L.).
Honglei Zhou
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science & Technology, Shanghai, China (J.W., Z.Y., N.Y., Q.J., H.Z., W.L., J.S., S.R., Y.M., L.Q., J.F., Y.X., X.Q., J.Z., Z.Z., S.L., R.W., H.L.).
Wenjie Liao
Jiwei Shan
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science & Technology, Shanghai, China (J.W., Z.Y., N.Y., Q.J., H.Z., W.L., J.S., S.R., Y.M., L.Q., J.F., Y.X., X.Q., J.Z., Z.Z., S.L., R.W., H.L.).
Shanshan Ruan
Yi Zhao
State Key Laboratory of Quantum Functional Materials, School of Physical Science and Technology
Ya Mo
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science & Technology, Shanghai, China (J.W., Z.Y., N.Y., Q.J., H.Z., W.L., J.S., S.R., Y.M., L.Q., J.F., Y.X., X.Q., J.Z., Z.Z., S.L., R.W., H.L.).
Luyao Qi
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science & Technology, Shanghai, China (J.W., Z.Y., N.Y., Q.J., H.Z., W.L., J.S., S.R., Y.M., L.Q., J.F., Y.X., X.Q., J.Z., Z.Z., S.L., R.W., H.L.).
Tiejun Li
CAS Key Laboratory of Colloid, Interface and Thermodynamics, Institute of Chemistry, Chinese Academy of Sciences, No. 2 North First Street, Zhongguancun, Beijing 100190, China
Jianjun Fu
Bowen Ke
Yufang Xu
Shandong Provincial Key Laboratory of Development and Regeneration, School of Life Sciences, Shandong University
Xuhong Qian
State Key Laboratory of Bioreactor Engineering, Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, Meilong Road 130, Shanghai 200237, China
Jian Zhang
Zhenjiang Zhao
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology
Shiliang Li
Beijing National Laboratory for Condensed Matter Physics
Rui Wang
Honglin Li
Innovation Center for AI and Drug Discovery, School of Pharmacy, East China Normal University