• DYPSL3
• ROCK2
• SHH
• medulloblastoma
• migration

• rho-Associated Kinases/metabolism
• rho-Associated Kinases/genetics
• Medulloblastoma/pathology
• Medulloblastoma/metabolism
• Medulloblastoma/genetics
• Humans
• Hedgehog Proteins/metabolism
• Hedgehog Proteins/genetics
• Cell Movement/genetics
• Down-Regulation
• Signal Transduction
• Cell Line, Tumor
• Child
• Gene Expression Regulation, Neoplastic
• Cerebellar Neoplasms/pathology
• Cerebellar Neoplasms/genetics
• Cerebellar Neoplasms/metabolism
• rhoA GTP-Binding Protein/metabolism

• CD44
• Chemoresistance
• Hyaluronic acid
• Mechanobiology
• Ovarian cancer
• Tumor microenvironment

• Alzheimer’s disease
• Huntington’s disease
• Parkinson’s disease
• Rho-associated kinases
• amyotrophic lateral sclerosis
• fasudil
• multiple sclerosis
• neurodegenerative diseases

• National Natural Science Foundation of China
• Heilongjiang University of Chinese Medicine

• 3D-QSAR
• MPBs
• ROCK1 inhibitors
• molecular dynamics simulation
• pharmacophore model

• rho-Associated Kinases/antagonists & inhibitors
• rho-Associated Kinases/chemistry
• rho-Associated Kinases/metabolism
• Quantitative Structure-Activity Relationship
• Molecular Docking Simulation
• Protein Kinase Inhibitors/chemistry
• Protein Kinase Inhibitors/pharmacology
• Molecular Dynamics Simulation
• Benzamides/chemistry
• Benzamides/pharmacology
• Humans
• Binding Sites
• Protein Binding
• Drug Discovery/methods
• Hydrogen Bonding

• NF-κB
• ROCK inhibitors
• ROCK1
• ROCK2
• RhoA
• microglia
• neuroinflammation

• Mice
• Animals
• Microglia/metabolism
• Tumor Necrosis Factor-alpha/metabolism
• Neuroinflammatory Diseases
• Interleukin-6/metabolism
• Lipopolysaccharides/pharmacology
• Lipopolysaccharides/metabolism
• Mice, Transgenic

• K01 AG000994 NIA NIH HHS
• AG00000994 NIH HHS

• Animals
• Humans
• Mice
• Cells, Cultured
• Hepatic Stellate Cells/metabolism
• rho-Associated Kinases/metabolism
• Signal Transduction

• Deutsche Forschungsgemeinschaft
• Bundesministerium für Bildung und Forschung
• Hessisches Ministerium für Wissenschaft und Kunst
• Eurostars
• Horizon 2020

The endothelial cells that compose the vascular system in the body display a wide range of mechanotransductive behaviors and responses to biomechanical stimuli, which act in concert to control overall blood vessel structure and function. Such mechanosensitive activities allow blood vessels to constrict, dilate, grow, or remodel as needed during development as well as normal physiological functions, and the same processes can be dysregulated in various disease states. Mechanotransduction represents cellular responses to mechanical forces, translating such factors into chemical or electrical signals which alter the activation of various cell signaling pathways. Understanding how biomechanical forces drive vascular growth in healthy and diseased tissues could create new therapeutic strategies that would either enhance or halt these processes to assist with treatments of different diseases. In the cardiovascular system, new blood vessel formation from preexisting vasculature, in a process known as angiogenesis, is driven by vascular endothelial growth factor (VEGF) binding to VEGF receptor 2 (VEGFR-2) which promotes blood vessel development. However, physical forces such as shear stress, matrix stiffness, and interstitial flow are also major drivers and effectors of angiogenesis, and new research suggests that mechanical forces may regulate VEGFR-2 phosphorylation. In fact, VEGFR-2 activation has been linked to known mechanobiological agents including ERK/MAPK, c-Src, Rho/ROCK, and YAP/TAZ. In vascular disease states, endothelial cells can be subjected to altered mechanical stimuli which affect the pathways that control angiogenesis. Both normalizing and arresting angiogenesis associated with tumor growth have been strategies for anti-cancer treatments. In the field of regenerative medicine, harnessing biomechanical regulation of angiogenesis could enhance vascularization strategies for treating a variety of cardiovascular diseases, including ischemia or permit development of novel tissue engineering scaffolds. This review will focus on the impact of VEGFR-2 mechanosignaling in endothelial cells (ECs) and its interaction with other mechanotransductive pathways, as well as presenting a discussion on the relationship between VEGFR-2 activation and biomechanical forces in the extracellular matrix (ECM) that can help treat diseases with dysfunctional vascular growth.

• VEGFR-2
• angiogenesis
• endothelial cells
• mechanobiology
• mechanoregulation

• 3D model
• Cornea
• Corneal nerve regeneration
• Neurotrophic keratopathy
• Rho kinase inhibitor

Biological therapies have dramatically improved the therapeutic landscape of psoriatic arthritis (PsA); however, 40–50% of patients are primary non-responders with response rates declining significantly with each successive biological therapy. Therefore, there is a pressing need to develop a coherent strategy for effective initial and subsequent selection of biologic agents. We interrogated 40 PsA patients initiating either tumour necrosis factor inhibitors (TNFi) or interleukin-17A inhibitors (17Ai) for active PsA. Patients achieving low disease activity according to the Disease Activity Index for PsA (DAPSA) at 3 months were classified as responders. Baseline and 3-month CD4⁺ transcript profiling were performed, and novel signaling pathways were identified using a multi-omics profiling and integrative computational analysis approach. Using transcriptomic data at initiation of therapy, we identified over 100 differentially expressed genes (DEGs) that differentiated IL-17Ai response from non-response and TNFi response from non-response. Integration of cell-type-specific DEGs with protein–protein interactions and further comprehensive pathway enrichment analysis revealed several pathways. Rho GTPase signaling pathway exhibited a strong signal specific to IL-17Ai response and the genes, RAC1 and ROCKs, are supported by results from prior research. Our detailed network and pathway analyses have identified the rewiring of Rho GTPase pathways as potential markers of response to IL17Ai but not TNFi. These results need further verification.

• Adult
• Amoeba
• Cell Line, Tumor
• Cell Movement
• Glioblastoma
• Humans
• Hyaluronic Acid

• P30 CA016058 NCI NIH HHS

• AR-12286 (Pubchem CID: 66906051)
• AT13148 (Pubchem CID: 24905401)
• Ewing’s sarcoma
• Fasudil (Pubchem CID: 3547)
• H-1152 (Pubchem CID: 448043)
• LX7101 (Pubchem CID: 56962369)
• Osteosarcoma
• Pediatric tumor
• RKI-1447 (Pubchem CID: 60138149)
• ROCK
• Ripasudil (Pubchem CID: 9863672)
• Wilm’s tumor
• Y-27632
• Y-27632 (Pubchem CID: 448042)
• Y-33075 (Pubchem CID: 9810884)
• YM529 (Pubchem CID: 130956)

• HSG
• Rho protein
• glioma
• invasion
• proliferation

The present study aimed to investigate variations in the Ras homolog gene family, member A (RhoA)-Rho-associated protein kinase 2 (ROCK2)-myosin light chain (MYL) pathway in a rat model of alcoholic cardiomyopathy (ACM) and the role of angiotensin-converting enzyme inhibitor drugs. Rat models of ACM were established via alcoholic gavage + free access to alcohol. The structural and functional changes of the heart were analyzed by hematoxylin-eosin staining, Masson's trichrome staining, immunohistochemistry staining, western blotting and fluorescence quantitative polymerase chain reaction. A total of 16 weeks later, a decreased ejection fraction and left ventricular fractional shortening in the alcohol group compared with the control group were demonstrated resulting in an increased left ventricular end diastolic diameter. These adverse effects were ameliorated following treatment with valsartan. In addition, the alcohol group revealed a disorganized arrangement of myocardial filaments, which was improved upon treatment with valsartan. RhoA and ROCK2 protein expression significantly increased in myocardial cells in the alcohol compared with the control group. Following drug intervention with valsartan, expression of RhoA and ROCK2 proteins were inhibited in the alcohol group. Furthermore, significantly elevated RhoA and ROCK2 and decreased MYL protein and mRNA expression in the alcohol group was demonstrated compared with the control group. Administration of valsartan reversed the expression profile of RhoA, ROCK and MYL in ACM. Expression of RhoA and ROCK were elevated with downregulation of MYL resulting in heart failure. However, the angiotensin receptor antagonist diminished the expression of RhoA and ROCK and enhanced the expression of MYL. The results of the present study suggest a curative effect of valsartan in ACM.

• Ras homolog gene family
• Rho-associated protein kinase
• alcoholic cardiomyopathy
• member A
• myosin light chain
• valsartan

Radioresistance remains a challenge to the successful treatment of various tumors. Intrinsic factors like alterations in signaling pathways regulate response to radiation. RhoC, which has been shown to modulate several tumor phenotypes has been investigated in this report for its role in radioresistance. In vitro and clinical sample-based studies have been performed to understand its contribution to radiation response in cervical cancer and this is the first report to establish the role of RhoC and its effector ROCK2 in cervical cancer radiation response.

Biochemical, transcriptomic and immunological approaches including flow cytometry and immunofluorescence were used to understand the role of RhoC and ROCK2. RhoC variants, siRNA and chemical inhibitors were used to alter the function of RhoC and ROCK2. Transcriptomic profiling was performed to understand the gene expression pattern of the cells. Live sorting using an intracellular antigen has been developed to isolate the cells for transcriptomic studies.

Enhanced expression of RhoC conferred radioprotection on the tumor cells while inhibition of RhoC resulted in sensitization of cells to radiation. The RhoC overexpressing cells had a better DNA repair machinery as observed using transcriptomic analysis. Similarly, overexpression of ROCK2, protected tumor cells against radiation while its inhibition increased radiosensitivity in vitro. Further investigations revealed that ROCK2 inhibition abolished the radioresistance phenotype, conferred by RhoC on SiHa cells, confirming that it is a downstream effector of RhoC in this context. Additionally, transcriptional analysis of the live sorted ROCK2 high and ROCK2 low expressing SiHa cells revealed an upregulation of the DNA repair pathway proteins. Consequently, inhibition of ROCK2 resulted in reduced expression of pH2Ax and MRN complex proteins, critical to repair of double strand breaks. Clinical sample-based studies also demonstrated that ROCK2 inhibition sensitizes tumor cells to irradiation.

Our data primarily indicates that RhoC and ROCK2 signaling is important for the radioresistance phenotype in cervical cancer tumor cells and is regulated via association of ROCK2 with the proteins of DNA repair pathway involving pH2Ax, MRE11 and RAD50 proteins, partly offering insights into the mechanism of radioresistance in tumor cells. These findings highlight RhoC-ROCK2 signaling involvement in DNA repair and urge the need for development of these molecules as targets to alleviate the non-responsiveness of cervical cancer tumor cells to irradiation treatment.

The online version of this article (10.1186/s13046-019-1385-7) contains supplementary material, which is available to authorized users.

• Cervical cancer
• DNA repair
• Radioresistance
• RhoC ROCK2

• Osteosarcoma
• Prognosis
• ROCK
• miR-138
• microRNA

• Axon regeneration
• ganglion cells
• snRPN

Ewing's sarcoma (EWS) is a highly aggressive bone cancer that affects children and adolescents. Despite advances in multimodal management, 5-year event-free survival rates for patients presenting with metastases at diagnosis remain at 25%. As key regulators of actin organization, the Rho-associated coiled-coil containing protein kinases, ROCK1 and ROCK2, have been associated with cancer dissemination and poorer prognosis. Recently, in vitro data indicating ROCK2 as a molecular target for the treatment of EWS has been presented. Nonetheless, a deeper exploration of the contribution of this kinase dysregulation in EWS is still necessary. In this regard, the present study aimed to evaluate the expression of ROCK1 and ROCK2 in 23 pediatric tumor samples and to verify the prospect of using their pharmacological inhibition through functional assays. Our results showed positive immunostaining for ROCK1 and ROCK2 in the majority samples (75 and 65%, respectively). A significantly increased risk of incomplete remission in patients with positive immunostaining for ROCK2 was found (P=0.026), though no correlations with other prognostic features (huvos classification, FLI1/EWS status, relapse, metastasis or death) were observed. Associations with survival were merely suggestive. Apparent protein expression of both kinases was also found in EWS cell lines (SK-ES-1 and RD-ES). Treatments with selective ROCK inhibitors did not alter cell viability or migration in vitro. However, a significant increase in invasion was observed after treatment with SR3677 (ROCK2 inhibitor) and hydroxyfasudil (pan-inhibitor). Consequently, even though the majority of EWS samples included in our study showed positivity for ROCK1 and ROCK2, the lack of significant associations with prognosis and absence of appropriate responses to their inhibition in vitro does not support their prospective use as therapeutic targets for the treatment of this metastatic tumor. Larger cohort studies might provide more evidence on whether there is a specific role of ROCK kinases in EWS physiopathology.

• Ewing sarcoma
• ROCK
• expression
• inhibition
• target

Graded induction of regulatory light chain (RLC) activators MLCK and ROCK were used to explore the relationship between RLC phosphorylation and actin-myosin stress fiber viscoelasticity. MLCK controls peripheral stress fiber mechanics by monophosphorylation of RLC, whereas ROCK acts on central stress fibers via diphosphorylation.

The assembly and mechanics of actomyosin stress fibers (SFs) depend on myosin regulatory light chain (RLC) phosphorylation, which is driven by myosin light chain kinase (MLCK) and Rho-associated kinase (ROCK). Although previous work suggests that MLCK and ROCK control distinct pools of cellular SFs, it remains unclear how these kinases differ in their regulation of RLC phosphorylation or how phosphorylation influences individual SF mechanics. Here, we combine genetic approaches with biophysical tools to explore relationships between kinase activity, RLC phosphorylation, SF localization, and SF mechanics. We show that graded MLCK overexpression increases RLC monophosphorylation (p-RLC) in a graded manner and that this p-RLC localizes to peripheral SFs. Conversely, graded ROCK overexpression preferentially increases RLC diphosphorylation (pp-RLC), with pp-RLC localizing to central SFs. Interrogation of single SFs with subcellular laser ablation reveals that MLCK and ROCK quantitatively regulate the viscoelastic properties of peripheral and central SFs, respectively. The effects of MLCK and ROCK on single-SF mechanics may be correspondingly phenocopied by overexpression of mono- and diphosphomimetic RLC mutants. Our results point to a model in which MLCK and ROCK regulate peripheral and central SF viscoelastic properties through mono- and diphosphorylation of RLC, offering new quantitative connections between kinase activity, RLC phosphorylation, and SF viscoelasticity.

The role of cancer-associated fibroblasts (CAFs) as regulators of tumor progression, specifically vascular growth, has only recently been described. CAFs are thought to be more mechanically active but how this trait may alter the tumor microenvironment is poorly understood. We hypothesized that enhanced mechanical activity of CAFs, as regulated by the Rho/ROCK pathway, contributes to increased blood vessel growth. Using a 3D in vitro tissue model of vasculogenesis, we observed increased vascularization in the presence of breast cancer CAFs compared to normal breast fibroblasts. Further studies indicated this phenomenon was not simply a result of enhanced soluble signaling factors, including vascular endothelial growth factor (VEGF), and that CAFs generated significantly larger deformations in 3D gels compared to normal fibroblasts. Inhibition of the mechanotransductive pathways abrogated the ability of CAFs to deform the matrix and suppressed vascularization. Finally, utilizing magnetic microbeads to mechanically stimulate mechanically-inhibited CAFs showed partial rescue of vascularization. Our studies demonstrate enhanced mechanical activity of CAFs may play a crucial and previously unappreciated role in the formation of tumor-associated vasculature which could possibly offer potential novel targets in future anti-cancer therapies.

• Angiosarcoma
• Hemangioendothelioma
• Hemangioma
• Hemangiopericytoma
• Rho kinase
• Rock
• Vascular sarcoma
• shRNA

• HDAC
• Migration
• ROCK
• Rho
• SIRT

• Bone cells
• Differentiation
• Mineralization
• ROCK
• Signal transduction
• Vesicles

Understanding the molecular processes characterizing Ewing sarcoma (EWS) cell migration is crucial to highlight novel therapies for patients with disseminated disease. In this study we analyzed the role of ROCK kinases in the regulation of cell migration, growth and differentiation of EWS cells. Overexpression of ROCK promotes invasion and metastasis in many solid tumors. However, the effect of ROCK in EWS has not been extensively investigated. Expression of ROCK1 and ROCK2 was analyzed by western blotting in a representative panel of human EWS cell lines, in comparison with the parameters of in vitro malignancy. We investigated the effects of a ROCK2 specific inhibitor toward those of a pan-ROCK inhibitor on the growth, migration and differentiation of two EWS cell lines. ROCK2 but not ROCK1 expression was found to be associated with in vitro cell migration and anchorage-independent growth capabilities. Exposure of EWS cells to ROCK inhibitors significantly reduced migration and growth, while favoring morphology changes and neural differentiation. These effects were more striking when cells were specifically deprived of ROCK2 activity. Our findings lead to consider ROCK2, rather than ROCK1, as a possible molecular target for the treatment of EWS.

Nonexudative (dry) age-related macular degeneration (AMD), a leading cause of blindness in the elderly, is associated with the loss of retinal pigmented epithelium (RPE) cells and the development of geographic atrophy, which are areas devoid of RPE cells and photoreceptors. One possible treatment option would be to stimulate RPE attachment and proliferation to replace dying/dysfunctional RPE and bring about wound repair. Clinical trials are underway testing injections of RPE cells derived from pluripotent stem cells to determine their safety and efficacy in treating AMD. However, the factors regulating RPE responses to AMD-associated lesions are not well understood. Here, we use cell culture to investigate the role of RhoA coiled coil kinases (ROCKs) in human embryonic stem cell–derived RPE (hESC-RPE) attachment, proliferation, and wound closure.

H9 hESC were spontaneously differentiated into RPE cells. hESC-RPE cells were treated with a pan ROCK1/2 or a ROCK2 only inhibitor; attachment, and proliferation and cell size within an in vitro scratch assay were examined.

Pharmacological inhibition of ROCKs promoted hESC-RPE attachment and proliferation, and increased the rate of closure of in vitro wounds. ROCK inhibition decreased phosphorylation of cofilin and myosin light chain, suggesting that regulation of the cytoskeleton underlies the mechanism of action of ROCK inhibition.

ROCK inhibition promotes attachment, proliferation, and wound closure in H9 hESC-RPE cells. ROCK isoforms may have different roles in wound healing.

Modulation of the ROCK-cytoskeletal axis has potential in stimulating wound repair in transplanted RPE cells and attachment in cellular therapies.

• ROCK inhibition
• age related macular degeneration
• retinal pigment epithelium

• DRP1
• Glioma
• RHOA
• ROCK1

• ROCK
• Y-27632
• breast cancer metastasis
• microtentacles
• reattachment

• Cancer
• Contractility
• Invadopodia
• Invasion
• ROCK
• Rigidity

• ROCK
• actin cytoskeleton
• actomyosin contractility
• cancer biology
• cell biology
• cell proliferation
• cellular senescence
• mouse
• rho kinase

• Animals
• Carcinogenesis
• Carcinoma, Non-Small-Cell Lung/pathology
• Cell Proliferation
• Gene Knockout Techniques
• Melanoma/pathology
• Mice
• rho-Associated Kinases/genetics
• rho-Associated Kinases/metabolism

• 12905 Cancer Research UK
• C107/A16512 Cancer Research UK
• C107/A1205 Cancer Research UK
• 19258 Cancer Research UK
• C107/A10433 Cancer Research UK
• 16512 Cancer Research UK

Rho-associated coiled-coil kinase (ROCK) is a major downstream effector of the small GTPase RhoA. The ROCK family, consisting of ROCK1 and ROCK2, plays a central role in the organization of the actin cytoskeleton, and is involved in a wide range of fundamental cellular functions such as contraction, adhesion, migration, proliferation, and apoptosis. Since the discovery of effective inhibitors such as fasudil and Y27632, the biological roles of ROCK have been extensively explored in numerous diseases, including cancer. Accumulating evidence supports the concept that ROCK plays important roles in tumor development and progression through regulating many key cellular functions associated with malignancy, including tumorigenicity, tumor growth, metastasis, angiogenesis, tumor cell apoptosis/survival and chemoresistance as well. This review focuses on the new advances of the most recent 5 years from the studies on the roles of ROCK in cancer development and progression; the discussion is mainly focused on the potential value of ROCK inhibitors in cancer therapy.

• Cancer therapy
• ROCK isoform-specific inhibitor
• Rho kinase
• Rho kinase pan-inhibitor

• Glioblastoma
• NMDAR
• fasudil
• glutamate

• Animals
• Cerebrovascular Disorders/drug therapy
• Cerebrovascular Disorders/enzymology
• Cerebrovascular Disorders/metabolism
• Humans
• Protein Kinase Inhibitors/chemistry
• Protein Kinase Inhibitors/pharmacology
• rho-Associated Kinases/antagonists & inhibitors
• rho-Associated Kinases/metabolism

• Z01 HL001786 Intramural NIH HHS
• ZIA HL001786-35 Intramural NIH HHS

Gastric cancer is one of the most frequent and lethal malignancies worldwide because of high frequency of metastasis. Tumor cell motility and invasion play fundamental roles in cancer metastasis. Recent studies have revealed that the Rho/Rho-associated protein kinases (ROCK) pathway plays a critical role in the regulation of cancer cell motility and invasion. In addition, the Rho/ROCK pathway plays important roles in invasion and metastasis on the basis of its predominant function of cell cytoskeletal regulation in gastric cancer. According to the current understanding of tumor motility, there are two modes of tumor cell movement: mesenchymal and amoeboid. In addition, cancer cell movement can be interchangeable between the mesenchymal and amoeboid movements under certain conditions. Control of cell motility through the actin cytoskeleton creates the potential for regulating tumor cell metastasis. In this review we discuss Rho GTPases and ROCK signaling and describe the mechanisms of Rho/ROCK activity with regard to motility and metastasis in gastric cancer. In addition, we provide an insight of the therapeutic potential of targeting the Rho/ROCK pathway.

• RhoGTPases
• Rho-associated protein kinases
• Gastric cancer
• Motility
• Metastasis

• Animals
• Antineoplastic Agents/therapeutic use
• Cell Movement/drug effects
• Drug Design
• Humans
• Molecular Targeted Therapy
• Neoplasm Invasiveness
• Neoplasm Metastasis
• Protein Kinase Inhibitors/therapeutic use
• Signal Transduction/drug effects
• Stomach Neoplasms/drug therapy
• Stomach Neoplasms/enzymology
• Stomach Neoplasms/pathology
• rho GTP-Binding Proteins/antagonists & inhibitors
• rho GTP-Binding Proteins/metabolism
• rho-Associated Kinases/antagonists & inhibitors
• rho-Associated Kinases/metabolism