Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I

Coleman, Mathew L.; Sahai, Erik; Yeo, Margaret; Bosch, Marta; Dewar, A.; Olson, Michael F.

doi:10.1038/35070009

Cited by 1,138 publications

(1,094 citation statements)

References 30 publications

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“…Thus, U2OS cells were treated with 1530 distinct chemicals from the ICCB Known Bioactives Library or the US-Drug Collection (encompassing most US Food and Drug Administration-approved and some experimental drugs), alone or in combination with MTX, for 18 h, stained with quinacrine and then analyzed by automated, quantitative epifluorescence microscopy ( Figure 2a). This screen led to the identification of several compounds that are capable of preventing the MTX-triggered loss of quinacrine-dependent fluorescence (Figure 2b): monensin, which blocks intracellular trafficking and exocytosis; 38 blebbistatin, which inhibits myosin II, thereby blocking the apoptosis-associated blebbing of the plasma membrane; 39 Y-27632, which inhibits ROCK1, 40 a protein that -upon cleavage by caspase-3 -participates in apoptotic blebbing; 41,42 and mefloquine, which inhibits PANX1 channels. 43 Subsequent validation experiments confirmed that monensin, blebbistatin and several chemically-unrelated inhibitors of ROCK1 (i.e., Y-27632, bearing with a central aminoethyl group, and H-1152, characterized by a central sulfonyl junction) and PANX1 (i.e., the synthetic quinine analogue mefloquine and the disulfonic stilbene derivatives 4,4 0 -diisothiocyano-2,2 0 -stilbenedisulfonic acid (DIDS) and 4-acetamido-4-isothiocyano-stilbene-2,2-disulfonic acid (SITS)) blunt the loss of quinacrine fluorescence as triggered in U2OS cells by MTX (Figure 2c).…”

Section: Resultsmentioning

confidence: 99%

Molecular mechanisms of ATP secretion during immunogenic cell death

et al. 2013

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The immunogenic demise of cancer cells can be induced by various chemotherapeutics, such as anthracyclines and oxaliplatin, and provokes an immune response against tumor-associated antigens. Thus, immunogenic cell death (ICD)-inducing antineoplastic agents stimulate a tumor-specific immune response that determines the long-term success of therapy. The release of ATP from dying cells constitutes one of the three major hallmarks of ICD and occurs independently of the two others, namely, the pre-apoptotic exposure of calreticulin on the cell surface and the postmortem release of high-mobility group box 1 (HMBG1) into the extracellular space. Pre-mortem autophagy is known to be required for the ICD-associated secretion of ATP, implying that autophagy-deficient cancer cells fail to elicit therapy-relevant immune responses in vivo. However, the precise molecular mechanisms whereby ATP is actively secreted in the course of ICD remain elusive. Using a combination of pharmacological screens, silencing experiments and techniques to monitor the subcellular localization of ATP, we show here that, in response to ICD inducers, ATP redistributes from lysosomes to autolysosomes and is secreted by a mechanism that requires the lysosomal protein LAMP1, which translocates to the plasma membrane in a strictly caspase-dependent manner. The secretion of ATP additionally involves the caspase-dependent activation of Rho-associated, coiled-coil containing protein kinase 1 (ROCK1)-mediated, myosin II-dependent cellular blebbing, as well as the opening of pannexin 1 (PANX1) channels, which is also triggered by caspases. Of note, although autophagy and LAMP1 fail to influence PANX1 channel opening, PANX1 is required for the ICD-associated translocation of LAMP1 to the plasma membrane. Altogether, these findings suggest that caspase-and PANX1-dependent lysosomal exocytosis has an essential role in ATP release as triggered by immunogenic chemotherapy.

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Section: Resultsmentioning

confidence: 99%

Molecular mechanisms of ATP secretion during immunogenic cell death

et al. 2013

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“…However, recent evidence suggests that ROCKs exert effects by means of Rho-independent pathways (Coleman et al, 2001;Sebbagh et al, 2001). Overexpression of dominant negative and constitutively active forms of RhoA during heart development causes cardiac dysfunction, resulting in dilation of the ventricles and atria (Sah et al, 1999).…”

Section: Figmentioning

confidence: 99%

Rho‐associated kinases play an essential role in cardiac morphogenesis and cardiomyocyte proliferation

Zhao

Rivkees

2002

Developmental Dynamics

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Rho-associated coiled-coil kinases (ROCKs), initially identified as effectors for Rho GTPases, play a role in cardiac cell physiology and are also expressed in the developing heart. However, their role in cardiac development is not known. To investigate the role of these kinases in cardiac development, we examined cardiac development in cultured murine embryos treated with the ROCK inhibitor Y27632. After inhibition of ROCK activity, we found disturbed cardiac chamber formation and trabeculation. To further examine the mechanisms by which ROCK blockade causes cardiac hypoplasia, we assessed programmed cell death and cell proliferation in the hearts. We found decreased cell proliferation in the Y27632-treated hearts, but no changes in programmed cell death. We further observed that ROCK inhibition decreased cardiac myocyte proliferation, suggesting that ROCK kinases regulate cardiomyocyte division. To identify factors involved in ROCK action in regulation of cardiac cell division, we examined expression of cell cycle proteins by using Western blot analysis. We found that ROCK blockade decreased expression of cell cycle proteins, cyclin D3, CDK6, and p27 KIP1 in the hearts and cardiomyocytes, which are required for initiation of cell cycle and G1/S phase transition. These observations show that ROCK kinases play a role in cardiac development and that ROCK kinases regulate cardiac cell proliferation and cell cycle protein expression. Developmental Dynamics 226:24 -32, 2003.

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“…The small GTPase Rho and its target Rho-associated protein kinases (Rho kinase, p160ROCK) [9,18,19] control cell adhesion [10] and motility [35] through reorganization of the actin cytoskeleton and regulation of actomyosin contractility [3] in a number of cellular processes, including vascular contraction [31], tumor invasion [12], penile erection [2], and apoptosis [4]. Further, several lines of evidence indicated that Rho-ROCK signaling also has been involved in cellular transformation.…”

Section: Introductionmentioning

confidence: 99%

Stimulation of Ectopic Bone Formation in Response to BMP-2 by Rho Kinase Inhibitor: A Pilot Study

Yoshikawa

Yoshioka

Nakase

et al. 2009

Clin Orthop Relat Res

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The small GTPase Rho and Rho-associated protein kinase (Rho kinase, ROCK) signal participates in a variety of biological functions including vascular contraction, tumor invasion, and penile erection. Evidence also suggests Rho-ROCK is involved in signaling for mesenchymal cellular differentiation. However, whether it is involved in osteoblastic differentiation is unknown. We therefore asked whether Rho-ROCK signaling participates in recombinant human bone morphogenetic protein (rhBMP-2)-induced osteogenesis both in vitro and in vivo. Continuous delivery of a specific ROCK inhibitor (Y-27632) enhanced ectopic bone formation induced by rhBMP-2 impregnated into an atelocollagen carrier in mice without affecting systemic bone metabolism. Treatment with Y-27632 also enhanced the osteoblastic differentiation of cultured murine neonatal calvarial cells. These effects were associated with increased expression of BMP-4 gene. Expression of a dominant negative mutant of ROCK in ST2 cells promoted osteoblastic differentiation, while a constitutively active mutant of ROCK attenuated osteoblastic differentiation and the ROCK inhibitor reversed this phenotype. Thus, ROCK inhibits osteogenesis, and a ROCK inhibitor in combination with the local delivery of rhBMP/collagen composite may be clinically applicable for stimulating bone formation.

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Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I

Cited by 1,138 publications

References 30 publications

Molecular mechanisms of ATP secretion during immunogenic cell death

Molecular mechanisms of ATP secretion during immunogenic cell death

Rho‐associated kinases play an essential role in cardiac morphogenesis and cardiomyocyte proliferation

Stimulation of Ectopic Bone Formation in Response to BMP-2 by Rho Kinase Inhibitor: A Pilot Study

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