Activation of the NLRP3 Inflammasome Is Associated with Valosin-Containing Protein Myopathy - PubMed (original) (raw)
Activation of the NLRP3 Inflammasome Is Associated with Valosin-Containing Protein Myopathy
Angèle Nalbandian et al. Inflammation. 2017 Feb.
Abstract
Aberrant activation of the NOD-like receptor (NLR) family, pyrin domain-containing protein 3 (NLRP3) inflammasome, triggers a pathogenic inflammatory response in many inherited neurodegenerative disorders. Inflammation has recently been associated with valosin-containing protein (VCP)-associated diseases, caused by missense mutations in the VCP gene. This prompted us to investigate whether NLRP3 inflammasome plays a role in VCP-associated diseases, which classically affects the muscles, bones, and brain. In this report, we demonstrate (i) an elevated activation of the NLRP3 inflammasome in VCP myoblasts, derived from induced pluripotent stem cells (iPSCs) of VCP patients, which was significantly decreased following in vitro treatment with the MCC950, a potent and specific inhibitor of NLRP3 inflammasome; (ii) a significant increase in the expression of NLRP3, caspase 1, IL-1β, and IL-18 in the quadriceps muscles of VCPR155H/+ heterozygote mice, an experimental mouse model that has many clinical features of human VCP-associated myopathy; (iii) a significant increase of number of IL-1β(+)F4/80(+)Ly6C(+) inflammatory macrophages that infiltrate the muscles of VCPR155H/+ mice; (iv) NLRP3 inflammasome activation and accumulation IL-1β(+)F4/80(+)Ly6C(+) macrophages positively correlated with high expression of TDP-43 and p62/SQSTM1 markers of VCP pathology in damaged muscle; and (v) treatment of VCPR155H/+ mice with MCC950 inhibitor suppressed activation of NLRP3 inflammasome, reduced the F4/80(+)Ly6C(+)IL-1β(+) macrophage infiltrates in the muscle, and significantly ameliorated muscle strength. Together, these results suggest that (i) NLRP3 inflammasome and local IL-1β(+)F4/80(+)Ly6C(+) inflammatory macrophages contribute to pathogenesis of VCP-associated myopathy and (ii) identified MCC950 specific inhibitor of the NLRP3 inflammasome with promising therapeutic potential for the treatment of VCP-associated myopathy.
Keywords: NLRP3 inflammasome; macrophage; myopathy; valosin-containing protein.
Conflict of interest statement
Conflict of interest: The authors have declared that no conflict of interest exists.
Figures
Figure 1. NLRP3 inflammasome pathway is active in myoblasts from VCP patients
(A) Immunohistochemical analysis of NLRP3 cascade mediators in iPSC-derived healthy myoblasts (left panel) and VCP disease myoblasts (middle panel, 0 μM) were immunostained with mAbs specific to human TDP-43 (‘classic’ marker of VCP pathology), and NLRP3, IL-18, Caspase 1, and IL-1β (markers of inflammasome activation) and then analyzed by confocal microscopy. White arrows and yellow dotted circles indicate to positive staining in VCP myoblasts. The right panel, 10 μM shows VCP disease myoblasts treated with MCC950 inhibitor at 10 μM and immunostained with TDP-43, NLRP3, IL-18, Caspase 1, and IL-1β mAbs. White arrows and yellow dotted circles point to fading of positive staining in VCP myoblasts following MCC950 inhibitor treatment. DAPI (blue) indicates staining of nuclei, and FITC (green) indicates staining with the various inflammasome markers including NLRP3, IL-18, Caspase 1, and IL-1β in myoblasts from VCP patients. (B) Western blot analysis of TDP-43, NLRP3, IL-18, Caspase 1, and IL-1β proteins from iPSC-derived healthy myoblasts (control) and from VCP disease myoblasts either untreated (VCP untreated) or treated with 10 μM of MCC950 inhibitor (VCP treated). GAPDH was used as a positive loading control. (C) Western blot results from three experiments normalized to GAPDH. Experiments shown are representative of independently performed triplicates. (*) Indicates P < 0.05 and (**) indicates P < 0.01 calculated by Mann-Whitney test and two-tailed _t_-tests when comparing protein expression of each marker in treated vs. untreated human VCP myoblasts.
Figure 2. Level of expression of NLRP3 inflammasome activation markers in myoblasts from VCP patients
(A) FACS analysis of NLRP3 cascade mediators in iPSC-derived myoblasts from VCP patients that are either left untreated (white) or treated with the MCC950 specific inhibitor of NLRP3 inflammasome (black). Treated and untreated VCP myoblasts were stained with mAbs specific to human NLRP3, Caspase 1, IL-1β and IL-18 (markers of inflammasome activation) and then analyzed by flow cytometry. iPSC-derived myoblasts from healthy patients that are either left untreated (gray) or treated with MCC950 specific inhibitor of NLRP3 inflammasome (dotted line) were used as negative controls. (B) FACS analysis of anti-apoptotic markers BIK, BAK, and BAD in iPSC-derived myoblasts from VCP patients and healthy controls. Experiments shown are representative of independently performed triplicates. (*) Indicates P < 0.05 and (**) indicates P < 0.01 calculated by Mann-Whitney test and two-tailed _t_-tests when comparing protein expression of each marker in treated vs. untreated human VCP myoblasts.
Figure 3. Expression of NLRP3 inflammasome and pathology markers in quadriceps femoris muscles of VCPR155H/+ heterozygote mice
(A) Immunohistochemical analysis of NLRP3 cascade mediators in quadriceps muscles of VCPR155H/+ heterozygote mice. Sections of quadriceps femoris muscles from control wild type mice (n = 8, left panel) and 2-year old VCPR155H/+ heterozygote mice (n = 8, middle panel, VCPR155H/+ 0-mg/kg) were immunostained with mAbs specific to mouse TDP-43 („classic‟ marker of VCP pathology), and NLRP3, IL-18, Caspase 1, and IL-1β (inflammasome markers) and then analyzed by fluorescence microscopy. White arrows indicate positive staining of myoblasts; TRITC (red, stained with laminin) identify muscle membrane of muscle fibers. DAPI (blue) indicates staining of nuclei. The right panel shows staining of section of quadriceps muscles from 2-year-old VCPR155H/+ heterozygote mice treated with MCC950 inhibitor at 30 mg/kg (VCPR155H/+, 30-mg/kg) and immunostained with TDP-43, NLRP3, IL-18, Caspase 1, and IL-1β mAbs (FITC, green). White arrows point to decreased positive staining in mouse myoblasts following MCC950 inhibitor treatment. TRITC-stained (red, stained with laminin) identify muscle membrane of muscle fibers. DAPI (blue) indicates staining of nuclei. (B) Western blot analysis of TDP-43, NLRP3, iNOS, Caspase 1 (p20 and p10), IL-1β, IL-18 and α-tubulin proteins from quadriceps muscles of wild type mice (n = 8, WT control) and from myoblasts from 2-year-old VCPR155H/+ heterozygote mice either left untreated (n = 8, VCPR155H/+ untreated) or treated with 30-mg/kg of MCC950 inhibitor (VCPR155H/+ Treated). (C) Western blot results from three experiments normalized to α-tubulin, positive loading control. Experiments shown are representative of independently performed triplicates. (*) Represents the P < 0.05 calculated by two-tailed and Mann-Whitney _t_-tests when comparing protein expression of each marker in quadriceps muscles of treated vs. untreated myoblasts from VCPR155H/+ heterozygote mice.
Figure 4. Histology and FACS analysis of iNOS- and IL-1β-producing inflammatory macrophages infiltrating bone, muscle and brain of VCPR155H/+ heterozygote mice
(A) Two-years-old VCPR155H/+ heterozygote and WT littermate mice were immunostained with CD68, a glycoprotein expressed on monocytes and macrophages (Magnification at 20×) and in parallel stained with Hematoxylin and eosin (Magnification at 20×). H&E sections show cell infiltrates in muscle of 1- and 2-years-old VCPR155H/+ heterozygote and WT littermate mice (Magnifications at 40× and 63×). As depicted, triangular black arrows show fibrosis, blue arrows depict central nucleation indicating regeneration, asterisks show degenerating fibers, and yellow arrows and yellow dotted lines point to persistent neutrophils, macrophages and necrotic myocytes. (B) Cell suspensions were harvested from bones, muscles and brains of 24-month old perfused VCPR155H/+ heterozygote mice (n = 8) and double stained using a mAb specific to mouse macrophages F4/80 surface marker (clone BM8) and a mAb specific to a mouse intracellular IL-1β (clone B122). Cell suspensions from bones, muscles and brains of 24-month old wild type mice (n = 8) used as controls were stained in parallel. Representative dot plot figure showing the percentages of F4/80(+)IL-1β(+) macrophages determined in bones, muscles and brains from VCPR155H/+ heterozygote mice vs. WT mice. The bar graphs represent the means and SD of the percentages (C) and number (D) of IL-1β(+)F4/80(+) macrophages in bones, muscles and brains from a group of 8 VCPR155H/+ heterozygote mice and 8WT mice. (E) Cell suspensions were harvested from bone, muscle and brain of 24-month old perfused VCPR155H/+ heterozygote mice and double stained using a mAb specific to Ly6C activation market of mouse macrophages (clone 1A8) and a mAb specific to a mouse intracellular IL-1β (clone B122). Cell suspensions from bones, muscles and brains of 24-month old wild type mice used as controls were stained in parallel. The percentages of IL-1β(+)Ly6C(+) macrophages were determined in each compartment and compared between VCPR155H/+ heterozygote mice (n = 8) and WT mice (n = 8). (F) The bar graphs represent the means and SD of the percentages of IL-1β(+)Ly6C(+) macrophages in the muscles, brains, and bones from a group of 8 VCPR155H/+ heterozygote mice and 8 WT mice. (G) The bar graphs represent the means and SD of the absolute numbers of IL-1β(+)Ly6C(+)macrophages detected in the muscles, brains, and bones from a group of VCPR155H/+ heterozygote mice and WT mice (n = 8). Experiments shown are representative of two independently experiments. (*) Indicates P < 0.05 when comparing percentages and number of IL-1β(+)F4/80(+)and IL-1β(+)Ly6C(+)macrophages in VCPR155H/+ heterozygote mice and WT mice using the Mann-Whitney test and 2-tails analysis.
Figure 4. Histology and FACS analysis of iNOS- and IL-1β-producing inflammatory macrophages infiltrating bone, muscle and brain of VCPR155H/+ heterozygote mice
(A) Two-years-old VCPR155H/+ heterozygote and WT littermate mice were immunostained with CD68, a glycoprotein expressed on monocytes and macrophages (Magnification at 20×) and in parallel stained with Hematoxylin and eosin (Magnification at 20×). H&E sections show cell infiltrates in muscle of 1- and 2-years-old VCPR155H/+ heterozygote and WT littermate mice (Magnifications at 40× and 63×). As depicted, triangular black arrows show fibrosis, blue arrows depict central nucleation indicating regeneration, asterisks show degenerating fibers, and yellow arrows and yellow dotted lines point to persistent neutrophils, macrophages and necrotic myocytes. (B) Cell suspensions were harvested from bones, muscles and brains of 24-month old perfused VCPR155H/+ heterozygote mice (n = 8) and double stained using a mAb specific to mouse macrophages F4/80 surface marker (clone BM8) and a mAb specific to a mouse intracellular IL-1β (clone B122). Cell suspensions from bones, muscles and brains of 24-month old wild type mice (n = 8) used as controls were stained in parallel. Representative dot plot figure showing the percentages of F4/80(+)IL-1β(+) macrophages determined in bones, muscles and brains from VCPR155H/+ heterozygote mice vs. WT mice. The bar graphs represent the means and SD of the percentages (C) and number (D) of IL-1β(+)F4/80(+) macrophages in bones, muscles and brains from a group of 8 VCPR155H/+ heterozygote mice and 8WT mice. (E) Cell suspensions were harvested from bone, muscle and brain of 24-month old perfused VCPR155H/+ heterozygote mice and double stained using a mAb specific to Ly6C activation market of mouse macrophages (clone 1A8) and a mAb specific to a mouse intracellular IL-1β (clone B122). Cell suspensions from bones, muscles and brains of 24-month old wild type mice used as controls were stained in parallel. The percentages of IL-1β(+)Ly6C(+) macrophages were determined in each compartment and compared between VCPR155H/+ heterozygote mice (n = 8) and WT mice (n = 8). (F) The bar graphs represent the means and SD of the percentages of IL-1β(+)Ly6C(+) macrophages in the muscles, brains, and bones from a group of 8 VCPR155H/+ heterozygote mice and 8 WT mice. (G) The bar graphs represent the means and SD of the absolute numbers of IL-1β(+)Ly6C(+)macrophages detected in the muscles, brains, and bones from a group of VCPR155H/+ heterozygote mice and WT mice (n = 8). Experiments shown are representative of two independently experiments. (*) Indicates P < 0.05 when comparing percentages and number of IL-1β(+)F4/80(+)and IL-1β(+)Ly6C(+)macrophages in VCPR155H/+ heterozygote mice and WT mice using the Mann-Whitney test and 2-tails analysis.
Figure 4. Histology and FACS analysis of iNOS- and IL-1β-producing inflammatory macrophages infiltrating bone, muscle and brain of VCPR155H/+ heterozygote mice
(A) Two-years-old VCPR155H/+ heterozygote and WT littermate mice were immunostained with CD68, a glycoprotein expressed on monocytes and macrophages (Magnification at 20×) and in parallel stained with Hematoxylin and eosin (Magnification at 20×). H&E sections show cell infiltrates in muscle of 1- and 2-years-old VCPR155H/+ heterozygote and WT littermate mice (Magnifications at 40× and 63×). As depicted, triangular black arrows show fibrosis, blue arrows depict central nucleation indicating regeneration, asterisks show degenerating fibers, and yellow arrows and yellow dotted lines point to persistent neutrophils, macrophages and necrotic myocytes. (B) Cell suspensions were harvested from bones, muscles and brains of 24-month old perfused VCPR155H/+ heterozygote mice (n = 8) and double stained using a mAb specific to mouse macrophages F4/80 surface marker (clone BM8) and a mAb specific to a mouse intracellular IL-1β (clone B122). Cell suspensions from bones, muscles and brains of 24-month old wild type mice (n = 8) used as controls were stained in parallel. Representative dot plot figure showing the percentages of F4/80(+)IL-1β(+) macrophages determined in bones, muscles and brains from VCPR155H/+ heterozygote mice vs. WT mice. The bar graphs represent the means and SD of the percentages (C) and number (D) of IL-1β(+)F4/80(+) macrophages in bones, muscles and brains from a group of 8 VCPR155H/+ heterozygote mice and 8WT mice. (E) Cell suspensions were harvested from bone, muscle and brain of 24-month old perfused VCPR155H/+ heterozygote mice and double stained using a mAb specific to Ly6C activation market of mouse macrophages (clone 1A8) and a mAb specific to a mouse intracellular IL-1β (clone B122). Cell suspensions from bones, muscles and brains of 24-month old wild type mice used as controls were stained in parallel. The percentages of IL-1β(+)Ly6C(+) macrophages were determined in each compartment and compared between VCPR155H/+ heterozygote mice (n = 8) and WT mice (n = 8). (F) The bar graphs represent the means and SD of the percentages of IL-1β(+)Ly6C(+) macrophages in the muscles, brains, and bones from a group of 8 VCPR155H/+ heterozygote mice and 8 WT mice. (G) The bar graphs represent the means and SD of the absolute numbers of IL-1β(+)Ly6C(+)macrophages detected in the muscles, brains, and bones from a group of VCPR155H/+ heterozygote mice and WT mice (n = 8). Experiments shown are representative of two independently experiments. (*) Indicates P < 0.05 when comparing percentages and number of IL-1β(+)F4/80(+)and IL-1β(+)Ly6C(+)macrophages in VCPR155H/+ heterozygote mice and WT mice using the Mann-Whitney test and 2-tails analysis.
Figure 5. Activation of NLRP3 inflammasome is reversed in myoblasts from VCPR155H/+ heterozygote mice following in vitro treatment with MCC950 pharmacologic inhibitor
Myoblasts were derived from 12-month old VCPR155H/+ heterozygotes mice (n = 8) and age- and sex-matched healthy mice (n = 8, controls). Mouse myoblasts were left untreated (white line) or treated (black line) for 16 hours with 10 μM MCC950 inhibitor. Treated and untreated myoblasts were stained with mAbs specific to mouse NLRP3, Caspase 1, IL-1β and IL-18 (markers of inflammasome activation) and then analyzed by flow cytometry, as in Fig. 2. Shown are (A) FACS histograms of markers of the NLRP3 inflammasome activation in myoblasts from VCPR155H/+ heterozygote mice (B) and healthy control mice. (C) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) of each marker (NLRP3, Caspase-1, IL-1β, IL-18 and TDP-43) in the myoblasts from a group of 8 treated and 8 untreated VCPR155H/+ heterozygote mice. (D) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) of each marker (NLRP3, Caspase-1, IL-1β, IL-18 and TDP-43) in the myoblasts from a group of 8 treated and 8 untreated WT mice. Experiments shown are representative of two independently experiments. (*) Indicates P < 0.05 and (**) indicates P < 0.01 when comparing treated and untreated mice using Mann-Whitney test and 2-tails analysis.
Figure 5. Activation of NLRP3 inflammasome is reversed in myoblasts from VCPR155H/+ heterozygote mice following in vitro treatment with MCC950 pharmacologic inhibitor
Myoblasts were derived from 12-month old VCPR155H/+ heterozygotes mice (n = 8) and age- and sex-matched healthy mice (n = 8, controls). Mouse myoblasts were left untreated (white line) or treated (black line) for 16 hours with 10 μM MCC950 inhibitor. Treated and untreated myoblasts were stained with mAbs specific to mouse NLRP3, Caspase 1, IL-1β and IL-18 (markers of inflammasome activation) and then analyzed by flow cytometry, as in Fig. 2. Shown are (A) FACS histograms of markers of the NLRP3 inflammasome activation in myoblasts from VCPR155H/+ heterozygote mice (B) and healthy control mice. (C) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) of each marker (NLRP3, Caspase-1, IL-1β, IL-18 and TDP-43) in the myoblasts from a group of 8 treated and 8 untreated VCPR155H/+ heterozygote mice. (D) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) of each marker (NLRP3, Caspase-1, IL-1β, IL-18 and TDP-43) in the myoblasts from a group of 8 treated and 8 untreated WT mice. Experiments shown are representative of two independently experiments. (*) Indicates P < 0.05 and (**) indicates P < 0.01 when comparing treated and untreated mice using Mann-Whitney test and 2-tails analysis.
Figure 6. Treatment of with MCC950 inhibitor suppresses of NLRP3 inflammasome activation and ameliorates muscle strength of VCPR155H/+ heterozygote mice
Two groups of sex-matched 12-month VCPR155H/+ heterozygotes mice (n = 8 mice per group) received oral gavage of 30-mg/kg of MCC950 inhibitor for 4 consecutive weeks. Untreated age- and sex-matched VCPR155H/+ heterozygotes mice were used as controls. (A) The hematoxylin and eosin sections showing cell infiltrates in muscle of treated vs. Untreated VCPR155H/+ heterozygote mice (Magnification at 40× and 63×). As depicted, yellow dotted lines show fibrosis, central nucleation indicating regeneration, and degenerating fibers, and yellow arrows point to persistent neutrophils, macrophages and necrotic myocytes. Quadriceps, brains, and bones were harvested 4 weeks later from untreated and treated mice and stained with mAbs specific to markers of NLRP3 inflammasome activation (NLRP3, Caspase 1, and IL-1β and mAbs specific to markers of pathology TDP-43 and p62_/SQSTM1_) as performed in Figs. 2 and 5. (B) Representative FACS histograms of the levels of markers of inflammasome activation and of pathology (NLRP3, Caspase 1, IL-1β, TDP-43, and P62/SQSTM1) detected by flow cytometry from one treated (black line) and untreated (white line) VCPR155H/+ heterozygote mice. (C) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) each marker (NLRP3, Caspase 1, IL-1β, TDP-43, and P62/SQSTM1) from a group of 8 treated and 8 untreated VCPR155H/+ heterozygotes mice. (*) Indicates P < 0.05 when comparing treated and untreated mice using ANOVA test. (D) Percentages (left graph) and number (right graph) of F4/80(+)IL-1β(+) macrophages determined in the muscle from treated versus untreated VCPR155H/+ heterozygote mice. (E) Percentages (left graph) and numbers (right graph) of IL-1β(+)Ly6C(+) macrophages determined in the muscle from treated versus untreated VCPR155H/+ heterozygote mice. (F and G) Body size/shape of treated versus untreated VCPR155H/+ heterozygote mice, (H) ROC curve showing weight loss (red line = treated; blue line = untreated), and (I and J) muscle mass loss of fore limb quadriceps observed in untreated versus treated VCPR155H/+ heterozygote mice.
Figure 6. Treatment of with MCC950 inhibitor suppresses of NLRP3 inflammasome activation and ameliorates muscle strength of VCPR155H/+ heterozygote mice
Two groups of sex-matched 12-month VCPR155H/+ heterozygotes mice (n = 8 mice per group) received oral gavage of 30-mg/kg of MCC950 inhibitor for 4 consecutive weeks. Untreated age- and sex-matched VCPR155H/+ heterozygotes mice were used as controls. (A) The hematoxylin and eosin sections showing cell infiltrates in muscle of treated vs. Untreated VCPR155H/+ heterozygote mice (Magnification at 40× and 63×). As depicted, yellow dotted lines show fibrosis, central nucleation indicating regeneration, and degenerating fibers, and yellow arrows point to persistent neutrophils, macrophages and necrotic myocytes. Quadriceps, brains, and bones were harvested 4 weeks later from untreated and treated mice and stained with mAbs specific to markers of NLRP3 inflammasome activation (NLRP3, Caspase 1, and IL-1β and mAbs specific to markers of pathology TDP-43 and p62_/SQSTM1_) as performed in Figs. 2 and 5. (B) Representative FACS histograms of the levels of markers of inflammasome activation and of pathology (NLRP3, Caspase 1, IL-1β, TDP-43, and P62/SQSTM1) detected by flow cytometry from one treated (black line) and untreated (white line) VCPR155H/+ heterozygote mice. (C) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) each marker (NLRP3, Caspase 1, IL-1β, TDP-43, and P62/SQSTM1) from a group of 8 treated and 8 untreated VCPR155H/+ heterozygotes mice. (*) Indicates P < 0.05 when comparing treated and untreated mice using ANOVA test. (D) Percentages (left graph) and number (right graph) of F4/80(+)IL-1β(+) macrophages determined in the muscle from treated versus untreated VCPR155H/+ heterozygote mice. (E) Percentages (left graph) and numbers (right graph) of IL-1β(+)Ly6C(+) macrophages determined in the muscle from treated versus untreated VCPR155H/+ heterozygote mice. (F and G) Body size/shape of treated versus untreated VCPR155H/+ heterozygote mice, (H) ROC curve showing weight loss (red line = treated; blue line = untreated), and (I and J) muscle mass loss of fore limb quadriceps observed in untreated versus treated VCPR155H/+ heterozygote mice.
Figure 6. Treatment of with MCC950 inhibitor suppresses of NLRP3 inflammasome activation and ameliorates muscle strength of VCPR155H/+ heterozygote mice
Two groups of sex-matched 12-month VCPR155H/+ heterozygotes mice (n = 8 mice per group) received oral gavage of 30-mg/kg of MCC950 inhibitor for 4 consecutive weeks. Untreated age- and sex-matched VCPR155H/+ heterozygotes mice were used as controls. (A) The hematoxylin and eosin sections showing cell infiltrates in muscle of treated vs. Untreated VCPR155H/+ heterozygote mice (Magnification at 40× and 63×). As depicted, yellow dotted lines show fibrosis, central nucleation indicating regeneration, and degenerating fibers, and yellow arrows point to persistent neutrophils, macrophages and necrotic myocytes. Quadriceps, brains, and bones were harvested 4 weeks later from untreated and treated mice and stained with mAbs specific to markers of NLRP3 inflammasome activation (NLRP3, Caspase 1, and IL-1β and mAbs specific to markers of pathology TDP-43 and p62_/SQSTM1_) as performed in Figs. 2 and 5. (B) Representative FACS histograms of the levels of markers of inflammasome activation and of pathology (NLRP3, Caspase 1, IL-1β, TDP-43, and P62/SQSTM1) detected by flow cytometry from one treated (black line) and untreated (white line) VCPR155H/+ heterozygote mice. (C) The bar graphs represent the means and SD of the mean fluorescent intensity (MFI) each marker (NLRP3, Caspase 1, IL-1β, TDP-43, and P62/SQSTM1) from a group of 8 treated and 8 untreated VCPR155H/+ heterozygotes mice. (*) Indicates P < 0.05 when comparing treated and untreated mice using ANOVA test. (D) Percentages (left graph) and number (right graph) of F4/80(+)IL-1β(+) macrophages determined in the muscle from treated versus untreated VCPR155H/+ heterozygote mice. (E) Percentages (left graph) and numbers (right graph) of IL-1β(+)Ly6C(+) macrophages determined in the muscle from treated versus untreated VCPR155H/+ heterozygote mice. (F and G) Body size/shape of treated versus untreated VCPR155H/+ heterozygote mice, (H) ROC curve showing weight loss (red line = treated; blue line = untreated), and (I and J) muscle mass loss of fore limb quadriceps observed in untreated versus treated VCPR155H/+ heterozygote mice.
References
- Cook GP, Savic S, Wittmann M, McDermott MF. The NLRP3 inflammasome, a target for therapy in diverse disease states. Eur J Immunol. 2010;40:631–634. - PubMed
- Schroder K, Zhou R, Tschopp J. The NLRP3 inflammasome: a sensor for metabolic danger? Science. 2010;327:296–300. - PubMed
- Strowig T, Henao-Mejia J, Elinav E, Flavell R. Inflammasomes in health and disease. Nature. 2012;481:278–286. - PubMed
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