GRK6 deficiency in mice causes autoimmune disease due to impaired apoptotic cell clearance

Nakaya, Michio; Tajima, Mitsuru; Kosako, Hidetaka; Nakaya, Takeo; Hashimoto, Akiko; Watari, Kenji; Nishihara, H. K.; Ohba, Mina; Komiya, Shiori; Tani, Naoki; Nishida, Motohiro; Taniguchi, Hiroyuki; Sato, Yoji; Matsumoto, Machiko; Tsuda, Makoto; Kuroda, Masahiko; Inoue, Kazuhide; Kurose, Hitoshi

doi:10.1038/ncomms2540

Cited by 52 publications

(43 citation statements)

References 48 publications

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“…(A) *P < 0.05; ***P < 0.001, 1-way ANOVA followed by Newman-Keuls analysis. (C and E) ***P < 0.001; ## jci.org Volume 127 Number 1 January 2017 engulfed, they appear yellow as an overlay of green and red (24)(25)(26). Confocal microscopy showed yellow signal inside αSMA-positive cells at the infarct and border areas of infarcted hearts, suggesting that cardiac myofibroblasts engulfed apoptotic cells in these areas (Figure 2A).…”

Section: Cardiac Myofibroblasts Efficiently Engulf Apoptotic Cells Inmentioning

confidence: 99%

“…After digestion, cells were stained with eFluor 780 viability dye for 30 minutes. The following antibodies were then added, and the reaction was allowed for 30 minutes: FITC antiLy-6C ( For the phagocytosis assay, thymocytes were collected from female C57BL/6 mice, labeled with 5-chloromethylfluorescein diacetate (CMFDA), pHrodo Red succinimidyl ester (P36600; Thermo Fisher Scientific), or CellTrace Far Red dye (C34564; Thermo Fisher Scientific), and treated with 10 μM dexamethasone at 37°C for 4.5 hours to induce apoptosis (26). CMFDA-labeled apoptotic thymocytes were cocultured with cardiac macrophages or cardiac myofibroblasts at 37°C, after which the slides were washed vigorously with PBS to remove the attached apoptotic thymocytes.…”

Section: Methodsmentioning

confidence: 99%

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Cardiac myofibroblast engulfment of dead cells facilitates recovery after myocardial infarction

Nakaya

Watari

Tajima

et al. 2016

Journal of Clinical Investigation

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114

109

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digested by collagenase/trypsin, and the digested cardiac cells were allowed to attach to the plates overnight. The attached cells included macrophages and myofibroblasts (positive for α smooth muscle actin [αSMA]) as well as other cardiac cells (Supplemental Figure 1B). Notably, cardiac myofibroblasts seemed to be more difficult than cardiac macrophages to collect using our isolation method from infarcted hearts because, as revealed by our immunohistochemical analysis, the number of cardiac myofibroblasts was the same as that of cardiac macrophages in the infarcted area (Supplemental Figure 1C). When the overnight-attached cells were cultured in 10% FBS/DMEM for more than 6 days, almost all of the cells on the plates were positive for αSMA and SM22α, 2 myofibroblast marker proteins (18, 19) (>97.9% and >93.8%, respectively) (Supplemental Figure 1, D and E), indicating that the cells were primarily composed of cardiac myofibroblasts. This is probably because only myofibroblasts were able to grow rapidly in the culture medium.Isolated cardiac macrophages and myofibroblasts were allowed to engulf fluorescently labeled apoptotic cells, and we assessed the fluorescence taken up by cardiac macrophages and administration promoted the restoration of cardiac function and morphology after MI, suggesting that MFG-E8 is a new therapeutic target for the treatment of MI.

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Section: Cardiac Myofibroblasts Efficiently Engulf Apoptotic Cells Inmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Cardiac myofibroblast engulfment of dead cells facilitates recovery after myocardial infarction

Nakaya

Watari

Tajima

et al. 2016

Journal of Clinical Investigation

Self Cite

114

109

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“…Loss of GRK6 in mice reduces and enhances chemotaxis in lymphocytes and PMN, respectively, suggesting a cell-specific role of GRK6. Acute inflammation response was also increased in GRK6 KO mice (124), and GRK6 deficiency in mice has been implicated with autoimmune disease related to impaired apoptotic cell clearance (165). Nevertheless, GRK6-mediated immune re-sponse and inflammation impact on the heart still remain to be fully elucidated.…”

Section: B Grk4 and Grk6 In Hypertensionmentioning

confidence: 99%

The Evolving Impact of G Protein-Coupled Receptor Kinases in Cardiac Health and Disease

Sato

Chuprun

Schwartz

et al. 2015

Physiological Reviews

126

152

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G protein-coupled receptors (GPCRs) are important regulators of various cellular functions via activation of intracellular signaling events. Active GPCR signaling is shut down by GPCR kinases (GRKs) and subsequent β-arrestin-mediated mechanisms including phosphorylation, internalization, and either receptor degradation or resensitization. The seven-member GRK family varies in their structural composition, cellular localization, function, and mechanism of action (see sect. II). Here, we focus our attention on GRKs in particular canonical and novel roles of the GRKs found in the cardiovascular system (see sects. III and IV). Paramount to overall cardiac function is GPCR-mediated signaling provided by the adrenergic system. Overstimulation of the adrenergic system has been highly implicated in various etiologies of cardiovascular disease including hypertension and heart failure. GRKs acting downstream of heightened adrenergic signaling appear to be key players in cardiac homeostasis and disease progression, and herein we review the current data on GRKs related to cardiac disease and discuss their potential in the development of novel therapeutic strategies in cardiac diseases including heart failure.

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“…-/ -mature hematopoietic cells, abnormal signaling and in vitro migration were thus predicted and observed [17,18], with functional consequences for immune function in vivo [19,20]. In agreement with the paradigm of need-adapted shuttling between cell surface and intracellular location, chemokine receptors have been reported to be predominantly located intracellularly [21,22].…”

Section: Introductionmentioning

confidence: 62%

Functional Consequences of Perturbed CXCL12 Signal Processing: Analyses of Immature Hematopoiesis in GRK6-Deficient Mice

Chudziak

Spohn

Karpova

et al. 2015

Stem Cells and Development

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Hematopoietic stem and progenitor cells (HSPCs) reside in bone marrow (BM) in an environment rich in CXCL12, the ligand for CXCR4, which is constitutively expressed on all immature hematopoietic cells in BM. This ligand-receptor pair critically controls HSPC retention and (relative) quiescence in BM. Interestingly, in a chemokine-abundant environment, CXCR4 surface expression and CXCL12 sensitivity of BM-residing HSPCs are continuously maintained. The mechanisms underlying this peculiar pattern of G-protein signal integration by BM-HSPCs are unknown. G-protein receptor kinases (GRKs) control receptor function by phosphorylating the intracellular domains upon ligand-induced activation, which results in receptor internalization and transient refractoriness. Using, therefore, a GRK6-deficient (GRK6 -/ -) mouse, we sought to address how perturbed ligand-induced CXCR4 (in)activation affects HSPC behavior in vitro and in vivo. In vitro, GRK6-/ -HSPCs were characterized by hyper-responsiveness to CXCL12, as expected. In vivo, GRK6-/ -immature hematopoiesis was characterized by a marked expansion of immature hematopoiesis in spleens and a modest repopulation defect in serial competitive transplantation. Enforced mobilization with granulocyte colonystimulating factor (G-CSF) and AMD3100 was normal, as was hematopoietic regeneration after noncompetitive transplantation or pharmacological myelosuppression. These observations illustrate that GRK-mediated restriction of CXCR4 signal input after ligand engagement is largely dispensable for BM-resident HSPCs, which may explain how continuous CXCL12 responsiveness of BM-HSPCs can be maintained.

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GRK6 deficiency in mice causes autoimmune disease due to impaired apoptotic cell clearance

Cited by 52 publications

References 48 publications

Cardiac myofibroblast engulfment of dead cells facilitates recovery after myocardial infarction

Cardiac myofibroblast engulfment of dead cells facilitates recovery after myocardial infarction

The Evolving Impact of G Protein-Coupled Receptor Kinases in Cardiac Health and Disease

Functional Consequences of Perturbed CXCL12 Signal Processing: Analyses of Immature Hematopoiesis in GRK6-Deficient Mice

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