Dissection of Thrombospondin-4 Domains Involved in Intracellular Adaptive Endoplasmic Reticulum Stress-Responsive Signaling

Brody, Matthew J.; Schips, Tobias G.; Vanhoutte, Davy; Kanisicak, Onur; Karch, Jason; Maliken, Bryan D.; Blair, N. Scott; Sargent, Michelle A.; Prasad, Vikram; Molkentin, Jeffery D.

doi:10.1128/mcb.00607-15

Cited by 23 publications

(57 citation statements)

References 58 publications

(105 reference statements)

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“…The P387-TSP-4 variant is associated with an increased risk of cardiovascular disease and myocardial infarction [15][16][17][18][19][20][21][22] and is more active than WT A387-TSP-4 in mediating cellular effects 24,37,42,49 . The region of TSP-4 molecule harboring P387 SNP appears to be important in regulation of many TSP-4 functions and was found to mediate the intracellular effects of TSP-4 50 and its effects in the nervous system 32 .…”

Section: Introductionmentioning

confidence: 99%

Effects of thrombospondin-4 on pro-inflammatory phenotype differentiation and apoptosis in macrophages

Rahman

Muppala

et al. 2020

Cell Death Dis

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Thrombospondin-4 (TSP-4) attracted renewed attention recently as a result of assignment of new functions to this matricellular protein in cardiovascular, muscular, and nervous systems. We have previously reported that TSP-4 promotes local vascular inflammation in a mouse atherosclerosis model. A common variant of TSP-4, P387-TSP-4, was associated with increased cardiovascular disease risk in human population studies. In a mouse atherosclerosis model, TSP-4 had profound effect on accumulation of macrophages in lesions, which prompted us to examine its effects on macrophages in more detail. We examined the effects of A387-TSP-4 and P387-TSP-4 on mouse macrophages in cell culture and in vivo in the model of LPS-induced peritonitis. In tissues and in cell culture, TSP-4 expression was associated with inflammation: TSP-4 expression was upregulated in peritoneal tissues in LPS-induced peritonitis, and pro-inflammatory signals, INFγ, GM-CSF, and LPS, induced TSP-4 expression in macrophages in vivo and in cell culture. Deficiency in TSP-4 in macrophages from Thbs4 −/− mice reduced the expression of pro-inflammatory macrophage markers, suggesting that TSP-4 facilitates macrophage differentiation into a pro-inflammatory phenotype. Expression of TSP-4, especially more active P387-TSP-4, was associated with higher cellular apoptosis. Cultured macrophages displayed increased adhesion to TSP-4 and reduced migration in presence of TSP-4, and these responses were further increased with P387 variant. We concluded that TSP-4 expression in macrophages increases their accumulation in tissues during the acute inflammatory process and supports macrophage differentiation into a pro-inflammatory phenotype. In a model of acute inflammation, TSP-4 supports pro-inflammatory macrophage apoptosis, a response that is closely related to their pro-inflammatory activity and release of pro-inflammatory signals. P387-TSP-4 was found to be the more active form of TSP-4 in all examined functions.

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Section: Introductionmentioning

confidence: 99%

Effects of thrombospondin-4 on pro-inflammatory phenotype differentiation and apoptosis in macrophages

Rahman

Muppala

et al. 2020

Cell Death Dis

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“…These mutations result in mutant Thbs5 proteins that accumulate in the endoplasmic reticulum (ER) and have delayed secretion, resulting in defective chondrocytes and altered ECM composition (15-18, 22, 23, 26, 27). Similarly, we have shown that mutation of DXDXDG calcium binding sites within the T3R domains of Thbs4 results in a loss of secretion by cultured cardiomyocytes, in contrast to the robustly secreted wild-type Thbs4 protein (19).…”

mentioning

confidence: 72%

“…binding motifs within the T3R domains of the Thbs proteins are critical for their flux through the secretory pathway and deposition briefly outside the cell (15)(16)(17)(18)(19). For example, mutations in calcium binding sites in the T3R domains of the human Thbs5/ COMP gene cause skeletal dysplasia, including pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED) (15,16,(20)(21)(22)(23)(24)(25).…”

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confidence: 99%

Defective Flux of Thrombospondin-4 through the Secretory Pathway Impairs Cardiomyocyte Membrane Stability and Causes Cardiomyopathy

Brody

Vanhoutte

Schips

et al. 2018

Molecular and Cellular Biology

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Thrombospondins are stress-inducible secreted glycoproteins with critical functions in tissue injury and healing. Thrombospondin-4 (Thbs4) is protective in cardiac and skeletal muscle where it activates an adaptive endoplasmic reticulum (ER) stress-response, induces expansion of the ER, and enhances sarcolemmal stability. However, it is unclear if Thbs4 has these protective functions from within the cell, from the extracellular matrix, or from the secretion process itself. Here we generated transgenic mice with cardiac-specific overexpression of a secretion-defective mutant of Thbs4 to evaluate its exclusive intracellular and secretion-dependent functions. Like wild-type Thbs4, the secretion-defective mutant upregulates the adaptive ER stress response and expands the ER and intracellular vesicles in cardiomyocytes. However, only the secretion-defective Thbs4 mutant produces cardiomyopathy with sarcolemmal weakness and rupture that is associated with reduced adhesion-forming glycoproteins in the membrane. Similarly, deletion of in the mouse model of Duchenne muscular dystrophy enhances cardiomyocyte membrane instability and cardiomyopathy. Finally, overexpression of the secretion-defective Thbs4 mutant in , but not wild-type Thbs4, impaired muscle function and sarcomere alignment. These results suggest that transit through the secretory pathway is required for Thbs4 to augment sarcolemmal stability, while ER stress induction and vesicular expansion mediated by Thbs4 is an exclusively intracellular process.

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“…Immunoblot analysis was performed using standard methods with primary antibodies for LRRC10 (28) and ␣-sarcomeric actin (Sigma) as previously described (8,9). For coimmunoprecipitation, cardiac lysates were made in extraction buffer as described for GST pulldown assays from mice 4 wk after sham or TAC surgery and diluted in lysate incubation buffer containing rabbit IgG (Santa Cruz Biotechnology) or anti-LRRC10 (1) coupled to protein G Dynabeads (Invitrogen) and incubated overnight at 4°C.…”

Section: Experiments Used Lrrc10mentioning

confidence: 99%

“…For coimmunoprecipitation, cardiac lysates were made in extraction buffer as described for GST pulldown assays from mice 4 wk after sham or TAC surgery and diluted in lysate incubation buffer containing rabbit IgG (Santa Cruz Biotechnology) or anti-LRRC10 (1) coupled to protein G Dynabeads (Invitrogen) and incubated overnight at 4°C. Protein G beads were then washed three times with lysate incubation buffer, and immunoprecipitated proteins were eluted, resolved by SDS-PAGE, and immunoblotted as described above (9).…”

Section: Experiments Used Lrrc10mentioning

confidence: 99%

LRRC10 is required to maintain cardiac function in response to pressure overload

Brody

Feng

Grimes

et al. 2016

American Journal of Physiology-Heart and Circulatory Physiology

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Brody MJ, Feng L, Grimes AC, Hacker TA, Olson TM, Kamp TJ, Balijepalli RC, Lee Y. LRRC10 is required to maintain cardiac function in response to pressure overload. Am J Physiol Heart Circ Physiol 310: H269 -H278, 2016. First published November 25, 2015 doi:10.1152/ajpheart.00717.2014.-We previously reported that the cardiomyocyte-specific leucine-rich repeat containing protein (LRRC)10 has critical functions in the mammalian heart. In the present study, we tested the role of LRRC10 in the response of the heart to biomechanical stress by performing transverse aortic constriction on Lrrc10-null (Lrrc10 Ϫ/Ϫ ) mice. Mild pressure overload induced severe cardiac dysfunction and ventricular dilation in Lrrc10 Ϫ/Ϫ mice compared with control mice. In addition to dilation and cardiomyopathy, Lrrc10 Ϫ/Ϫ mice showed a pronounced increase in heart weight with pressure overload stimulation and a more dramatic loss of cardiac ventricular performance, collectively suggesting that the absence of LRRC10 renders the heart more disease prone with greater hypertrophy and structural remodeling, although rates of cardiac fibrosis and myocyte dropout were not different from control mice. Lrrc10 Ϫ/Ϫ cardiomyocytes also exhibited reduced contractility in response to ␤-adrenergic stimulation, consistent with loss of cardiac ventricular performance after pressure overload. We have previously shown that LRRC10 interacts with actin in the heart. Here, we show that His 150 of LRRC10 was required for an interaction with actin, and this interaction was reduced after pressure overload, suggesting an integral role for LRRC10 in the response of the heart to mechanical stress. Importantly, these experiments demonstrated that LRRC10 is required to maintain cardiac performance in response to pressure overload and suggest that dysregulated expression or mutation of LRRC10 may greatly sensitize human patients to more severe cardiac disease in conditions such as chronic hypertension or aortic stenosis. CARDIOMYOCYTE HYPERTROPHY in response to extracellular stimuli, such as neurohumoral or growth factor stimulation of membrane-bound receptors (13, 23, 44), or in response to mechanical stretch or strain that is sensed by mechanosensory machinery embedded in the Z-disc cytoskeleton, and sarcolemma (25, 38, 49). At the whole organ level, the heart undergoes hypertrophy as an adaptive mechanism to maintain cardiac output and reduce ventricular wall stress (20,23,38). While initially beneficial, prolonged cardiac hypertrophy becomes maladaptive and progresses to cardiac dilation, decompensation, heart failure, and/or sudden death (20,23,38). Cardiac hypertrophy can be induced by a number of cardiovascular diseases, such as myocardial infarction or chronic hypertension (23, 37). Indeed, high blood pressure is a common precursor to heart disease, particularly in the United States, where chronic hypertension is widespread (18,21,42). Despite increased awareness and treatment of hypertension, recent epidemiological studies have suggested that its prevalence con...

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Dissection of Thrombospondin-4 Domains Involved in Intracellular Adaptive Endoplasmic Reticulum Stress-Responsive Signaling

Cited by 23 publications

References 58 publications

Effects of thrombospondin-4 on pro-inflammatory phenotype differentiation and apoptosis in macrophages

Effects of thrombospondin-4 on pro-inflammatory phenotype differentiation and apoptosis in macrophages

Defective Flux of Thrombospondin-4 through the Secretory Pathway Impairs Cardiomyocyte Membrane Stability and Causes Cardiomyopathy

LRRC10 is required to maintain cardiac function in response to pressure overload

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