Abstract:This article contains additional data related to the original research article entitled “KRIT1 loss-of-function induces a chronic Nrf2-mediated adaptive homeostasis that sensitizes cells to oxidative stress: implication for Cerebral Cavernous Malformation disease” (Antognelli et al., 2017) [1].Data were obtained by si-RNA-mediated gene silencing, qRT-PCR, immunoblotting, and immunohistochemistry studies, and enzymatic activity and apoptosis assays. Overall, they support, complement and extend original findings… Show more
“…These results were confirmed in KRIT1-silenced hBMEC cells treated with Tiron (see Fig. 3 a-c in [49] ), demonstrating that KRIT1 downregulation causes a redox-sensitive upregulation of Glo1 and depletion of AP adducts in distinct cellular models of CCM disease. Fig.…”
Section: Resultssupporting
confidence: 58%
“…GR252439-11, Abcam). In addition, specific IHC analyses were also performed to detect Glo1, phospho-JNK and KLF4 protein levels (see Experimental Design, Materials and Methods in [49] ). ICH labeling was then developed by a 5- to 10-min incubation with 3,3′ diaminobenzidine (DAB) + H 2 O 2 substrate chromogen, which results in a brown-colored precipitate at antigen site.…”
Section: Methodsmentioning
confidence: 99%
“…In addition, further measurements of cellular H 2 O 2 levels were performed with the Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit (A22188, Invitrogen) (see Experimental Design, Materials and Methods, and Fig. 1 in [49] ). However, as it is still not possible to exclude the involvement of other oxidative species not yet analyzed in addition to O 2 •− and H 2 O 2 , we resort to the use of the generic term ROS to refer to O 2 •− and H 2 O 2 as well as to secondary oxidative products that might be implicated without certainty, according to the recommendations of FRBM´s Editors [53] .…”
KRIT1 (CCM1) is a disease gene responsible for Cerebral Cavernous Malformations (CCM), a major cerebrovascular disease of proven genetic origin affecting 0.3–0.5% of the population.Previously, we demonstrated that KRIT1 loss-of-function is associated with altered redox homeostasis and abnormal activation of the redox-sensitive transcription factor c-Jun, which collectively result in pro-oxidative, pro-inflammatory and pro-angiogenic effects, suggesting a novel pathogenic mechanism for CCM disease and raising the possibility that KRIT1 loss-of-function exerts pleiotropic effects on multiple redox-sensitive mechanisms.To address this possibility, we investigated major redox-sensitive pathways and enzymatic systems that play critical roles in fundamental cytoprotective mechanisms of adaptive responses to oxidative stress, including the master Nrf2 antioxidant defense pathway and its downstream target Glyoxalase 1 (Glo1), a pivotal stress-responsive defense enzyme involved in cellular protection against glycative and oxidative stress through the metabolism of methylglyoxal (MG). This is a potent post-translational protein modifier that may either contribute to increased oxidative molecular damage and cellular susceptibility to apoptosis, or enhance the activity of major apoptosis-protective proteins, including heat shock proteins (Hsps), promoting cell survival.Experimental outcomes showed that KRIT1 loss-of-function induces a redox-sensitive sustained upregulation of Nrf2 and Glo1, and a drop in intracellular levels of MG-modified Hsp70 and Hsp27 proteins, leading to a chronic adaptive redox homeostasis that counteracts intrinsic oxidative stress but increases susceptibility to oxidative DNA damage and apoptosis, sensitizing cells to further oxidative challenges. While supporting and extending the pleiotropic functions of KRIT1, these findings shed new light on the mechanistic relationship between KRIT1 loss-of-function and enhanced cell predisposition to oxidative damage, thus providing valuable new insights into CCM pathogenesis and novel options for the development of preventive and therapeutic strategies.
“…These results were confirmed in KRIT1-silenced hBMEC cells treated with Tiron (see Fig. 3 a-c in [49] ), demonstrating that KRIT1 downregulation causes a redox-sensitive upregulation of Glo1 and depletion of AP adducts in distinct cellular models of CCM disease. Fig.…”
Section: Resultssupporting
confidence: 58%
“…GR252439-11, Abcam). In addition, specific IHC analyses were also performed to detect Glo1, phospho-JNK and KLF4 protein levels (see Experimental Design, Materials and Methods in [49] ). ICH labeling was then developed by a 5- to 10-min incubation with 3,3′ diaminobenzidine (DAB) + H 2 O 2 substrate chromogen, which results in a brown-colored precipitate at antigen site.…”
Section: Methodsmentioning
confidence: 99%
“…In addition, further measurements of cellular H 2 O 2 levels were performed with the Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit (A22188, Invitrogen) (see Experimental Design, Materials and Methods, and Fig. 1 in [49] ). However, as it is still not possible to exclude the involvement of other oxidative species not yet analyzed in addition to O 2 •− and H 2 O 2 , we resort to the use of the generic term ROS to refer to O 2 •− and H 2 O 2 as well as to secondary oxidative products that might be implicated without certainty, according to the recommendations of FRBM´s Editors [53] .…”
KRIT1 (CCM1) is a disease gene responsible for Cerebral Cavernous Malformations (CCM), a major cerebrovascular disease of proven genetic origin affecting 0.3–0.5% of the population.Previously, we demonstrated that KRIT1 loss-of-function is associated with altered redox homeostasis and abnormal activation of the redox-sensitive transcription factor c-Jun, which collectively result in pro-oxidative, pro-inflammatory and pro-angiogenic effects, suggesting a novel pathogenic mechanism for CCM disease and raising the possibility that KRIT1 loss-of-function exerts pleiotropic effects on multiple redox-sensitive mechanisms.To address this possibility, we investigated major redox-sensitive pathways and enzymatic systems that play critical roles in fundamental cytoprotective mechanisms of adaptive responses to oxidative stress, including the master Nrf2 antioxidant defense pathway and its downstream target Glyoxalase 1 (Glo1), a pivotal stress-responsive defense enzyme involved in cellular protection against glycative and oxidative stress through the metabolism of methylglyoxal (MG). This is a potent post-translational protein modifier that may either contribute to increased oxidative molecular damage and cellular susceptibility to apoptosis, or enhance the activity of major apoptosis-protective proteins, including heat shock proteins (Hsps), promoting cell survival.Experimental outcomes showed that KRIT1 loss-of-function induces a redox-sensitive sustained upregulation of Nrf2 and Glo1, and a drop in intracellular levels of MG-modified Hsp70 and Hsp27 proteins, leading to a chronic adaptive redox homeostasis that counteracts intrinsic oxidative stress but increases susceptibility to oxidative DNA damage and apoptosis, sensitizing cells to further oxidative challenges. While supporting and extending the pleiotropic functions of KRIT1, these findings shed new light on the mechanistic relationship between KRIT1 loss-of-function and enhanced cell predisposition to oxidative damage, thus providing valuable new insights into CCM pathogenesis and novel options for the development of preventive and therapeutic strategies.
“…Genetic studies in CCM patient cohorts have led to the identification of three disease genes, CCM1/KRIT1, CCM2, and CCM3, which have been implicated in all major mechanisms of vascular integrity maintenance and endothelial barrier function, including the coordination of redox signaling and autophagy governing cell homeostasis and stress responses [43,[171][172][173][174][175][176][177][178][179][180]. Consistent with such pleiotropic functions, accumulated evidence from animal models and patient cohorts has demonstrated that loss-of-function mutations of CCM genes only predispose to the development of CCM disease.…”
Section: Vitamin D Deficiency and Ccm Diseasementioning
Vitamin D deficiency has been clearly linked to major chronic diseases associated with oxidative stress, inflammation, and aging, including cardiovascular and neurodegenerative diseases, diabetes, and cancer. In particular, the cardiovascular system appears to be highly sensitive to vitamin D deficiency, as this may result in endothelial dysfunction and vascular defects via multiple mechanisms. Accordingly, recent research developments have led to the proposal that pharmacological interventions targeting either vitamin D deficiency or its key downstream effects, including defective autophagy and abnormal pro-oxidant and pro-inflammatory responses, may be able to limit the onset and severity of major cerebrovascular diseases, such as stroke and cerebrovascular malformations. Here we review the available evidence supporting the role of vitamin D in preventing or limiting the development of these cerebrovascular diseases, which are leading causes of disability and death all over the world.
“…Accumulated evidence demonstrates that loss-of-function mutations of CCM genes have pleiotropic effects on several redox-sensitive molecules and mechanisms that control cellular homeostasis and defenses against oxidative stress and inflammation, thereby sensitizing cells to local oxidative stress and inflammatory events [ 84 , 86 , 88 – 95 ]. In particular, KRIT1 loss-of-function has been shown to affect major antioxidant pathways and mechanisms, including the FOXO1-SOD2 axis and the Nrf2 antioxidant pathway [ 89 , 94 ], and the autophagic degradation of dysfunctional, ROS-generating mitochondria [ 89 , 91 ].…”
Section: Avenanthramides As Potential Therapeutics For Cerebral Camentioning
Oat (Avena sativa) is a cereal known since antiquity as a useful grain with abundant nutritional and health benefits. It contains distinct molecular components with high antioxidant activity, such as tocopherols, tocotrienols, and flavanoids. In addition, it is a unique source of avenanthramides, phenolic amides containing anthranilic acid and hydroxycinnamic acid moieties, and endowed with major beneficial health properties because of their antioxidant, anti-inflammatory, and antiproliferative effects. In this review, we report on the biological activities of avenanthramides and their derivatives, including analogs produced in recombinant yeast, with a major focus on the therapeutic potential of these secondary metabolites in the treatment of aging-related human diseases. Moreover, we also present recent advances pointing to avenanthramides as interesting therapeutic candidates for the treatment of cerebral cavernous malformation (CCM) disease, a major cerebrovascular disorder affecting up to 0.5% of the human population. Finally, we highlight the potential of foodomics and redox proteomics approaches in outlining distinctive molecular pathways and redox protein modifications associated with avenanthramide bioactivities in promoting human health and contrasting the onset and progression of various pathologies.
The paper is dedicated to the memory of Adelia Frison
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