Leptin-deficient obesity prolongs survival in a murine model of myelodysplastic syndrome

Obesity enhances the risk of developing myelodysplastic syndromes. However, the effect of obesity on survival is unclear. Obese people present with monocytosis due to inflammatory signals emanating from obese adipose tissue. We hypothesized that obesity-induced myelopoiesis would promote the transition of myelodysplastic syndrome to acute myeloid leukemia and accelerate mortality in obesity. Obese Ob/Ob mice or their lean littermate controls received a bone marrow transplant from NUP98-HOXD13 transgenic mice, a model of myelodysplastic syndrome. The metabolic parameters of the mice were examined throughout the course of the study, as were blood leukocytes. Myeloid cells were analyzed in the bone, spleen, liver and adipose tissue by flow cytometry halfway through the disease progression and at the endpoint. Survival curves were also calculated. Contrary to our hypothesis, transplantation of NUP98-HOXD13 bone marrow into obese recipient mice significantly increased survival time compared with lean recipient controls. While monocyte skewing was exacerbated in obese mice receiving NUP98-HOXD13 bone marrow, transformation to acute myeloid leukemia was not enhanced. Increased survival of obese mice was associated with a preservation of fat mass as well as increased myeloid cell deposition within the adipose tissue, and a concomitant reduction in detrimental myeloid cell accumulation within other organs. The study herein revealed that obesity increases survival in animals with myelodysplastic syndrome. This may be due to the greater fat mass of Ob/Ob mice, which acts as a sink for myeloid cells, preventing their accumulation in other key organs, such as the liver

Exosomes containing HIV protein Nef reorganize lipid rafts potentiating inflammatory response in bystander cells

HIV infection has a profound effect on "bystander" cells causing metabolic co-morbidities. This may be mediated by exosomes secreted by HIV-infected cells and containing viral factors. Here we show that exosomes containing HIV-1 protein Nef (exNef) are rapidly taken up by macrophages releasing Nef into the cell interior. This caused down-regulation of ABCA1, reduction of cholesterol efflux and sharp elevation of the abundance of lipid rafts through reduced activation of small GTPase Cdc42 and decreased actin polymerization. Changes in rafts led to re-localization of TLR4 and TREM-1 to rafts, phosphorylation of ERK1/2, activation of NLRP3 inflammasome, and increased secretion of pro-inflammatory cytokines. The effects of exNef on lipid rafts and on inflammation were reversed by overexpression of a constitutively active mutant of Cdc42. Similar effects were observed in macrophages treated with exosomes produced by HIV-infected cells or isolated from plasma of HIV-infected subjects, but not with exosomes from cells and subjects infected with ΔNef-HIV or uninfected subjects. Mice injected with exNef exhibited monocytosis, reduced ABCA1 in macrophages, increased raft abundance in monocytes and augmented inflammation. Thus, Nef-containing exosomes potentiated pro-inflammatory response by inducing changes in cholesterol metabolism and reorganizing lipid rafts. These mechanisms may contribute to HIV-associated metabolic co-morbidities

Defective AMPK regulation of cholesterol metabolism accelerates atherosclerosis by promoting HSPC mobilization and myelopoiesis

Objectives: Dysregulation of cholesterol metabolism in the liver and hematopoietic stem and progenitor cells (HSPCs) promotes atherosclerosis development. Previously, it has been shown that HMG-CoA-Reductase (HMGCR), the rate-limiting enzyme in the mevalonate pathway, can be phosphorylated and inactivated by the metabolic stress sensor AMP-activated protein kinase (AMPK). However, the physiological significance of AMPK regulation of HMGCR to atherogenesis has yet to be elucidated. The aim of this study was to determine the role of AMPK/HMGCR axis in the development of atherosclerosis. Methods: We have generated a novel atherosclerotic-prone mouse model with defects in the AMPK regulation of HMGCR (Apoe−/−/Hmgcr KI mice). Atherosclerotic lesion size, plaque composition, immune cell and lipid profiles were assessed in Apoe−/− and Apoe−/−/Hmgcr KI mice. Results: In this study, we showed that both male and female atherosclerotic-prone mice with a disruption of HMGCR regulation by AMPK (Apoe−/−/Hmgcr KI mice) display increased aortic lesion size concomitant with an increase in plaque-associated macrophages and lipid accumulation. Consistent with this, Apoe−/−/Hmgcr KI mice exhibited an increase in total circulating cholesterol and atherogenic monocytes, Ly6-Chi subset. Mechanistically, increased circulating atherogenic monocytes in Apoe−/−/Hmgcr KI mice was associated with enhanced egress of bone marrow HSPCs and extramedullary myelopoiesis, driven by a combination of elevated circulating 27-hydroxycholesterol and intracellular cholesterol in HSPCs. Conclusions: Our results uncovered a novel signalling pathway involving AMPK-HMGCR axis in the regulation of cholesterol homeostasis in HSPCs, and that inhibition of this regulatory mechanism accelerates the development and progression of atherosclerosis. These findings provide a molecular basis to support the use of AMPK activators that currently undergoing Phase II clinical trial such as O–3O4 and PXL 770 for reducing atherosclerotic cardiovascular disease risks

Apoptotic ablation of platelets reduces atherosclerosis in mice with diabetes

Objective: People with diabetes are at a significantly higher risk of cardiovascular disease, in part, due to accelerated atherosclerosis. Diabetic subjects have increased number of platelets that are activated, more reactive, and respond suboptimally to antiplatelet therapies. We hypothesized that reducing platelet numbers by inducing their premature apoptotic death would decrease atherosclerosis. Approach and Results: This was achieved by targeting the antiapoptotic protein Bcl-x(L) (B-cell lymphoma-extra large; which is essential for platelet viability) via distinct genetic and pharmacological approaches. In the former, we transplanted bone marrow from mice carrying the Tyr15 to Cys loss of function allele of Bcl-x (known as Bcl-x(Plt20)) or wild-type littermate controls into atherosclerotic-prone Ldlr(+/−) mice made diabetic with streptozotocin and fed a Western diet. Reduced Bcl-x(L) function in hematopoietic cells significantly decreased platelet numbers, exclusive of other hematologic changes. This led to a significant reduction in atherosclerotic lesion formation in Bcl-x(Plt20) bone marrow transplanted Ldlr(+/−) mice. To assess the potential therapeutic relevance of reducing platelets in atherosclerosis, we next targeted Bcl-x(L) with a pharmacological strategy. This was achieved by low-dose administration of the BH3 (B-cell lymphoma-2 homology domain 3) mimetic, ABT-737 triweekly, in diabetic Apoe(−/−) mice for the final 6 weeks of a 12-week study. ABT-737 normalized platelet numbers along with platelet and leukocyte activation to that of nondiabetic controls, significantly reducing atherosclerosis while promoting a more stable plaque phenotype. Conclusions: These studies suggest that selectively reducing circulating platelets, by targeting Bcl-x(L) to promote platelet apoptosis, can reduce atherosclerosis and lower cardiovascular disease risk in diabetes.Man K.S. Lee, Michael J. Kraakman, Dragana Dragoljevic, Nordin M.J. Hanssen, Michelle C. Flynn, Annas Al-Sharea, Gopalkrishna Sreejit, Camilla Bertuzzo-Veiga, Olivia D. Cooney, Fatima Baig, Elizabeth Morriss, Mark E. Cooper, Emma C. Josefsson, Benjamin T. Kile, Prabhakara R. Nagareddy, Andrew J. Murph

IL-18 production from the NLRP1 inflammasome prevents obesity and metabolic syndrome

Interleukin-18 (IL-18) is activated by Caspase-1 in inflammasome complexes and has anti-obesity effects; however, it is not known which inflammasome regulates this process. We found that mice lacking the NLRP1 inflammasome phenocopy mice lacking IL-18, with spontaneous obesity due to intrinsic lipid accumulation. This is exacerbated when the mice are fed a high-fat diet (HFD) or a high-protein diet, but not when mice are fed a HFD with low energy density (high fiber). Furthermore, mice with an activating mutation in NLRP1, and hence increased IL-18, have decreased adiposity and are resistant to diet-induced metabolic dysfunction. Feeding these mice a HFD further increased plasma IL-18 concentrations and strikingly resulted in loss of adipose tissue mass and fatal cachexia, which could be prevented by genetic deletion of IL-18. Thus, NLRP1 is an innate immune sensor that functions in the context of metabolic stress to produce IL-18, preventing obesity and metabolic syndrome.Andrew J. Murphy, Michael J. Kraakman, Helene L. Kammoun, Dragana Dragoljevic, Man K.S. Lee ... Benjamin T. Kile ... et al

ABC Transporters, Cholesterol Efflux, and Implications for Cardiovascular Diseases

Most types of cells in the body have no or very limited capacity of catabolizing cholesterol, so cholesterol efflux is essential for cholesterol homeostasis. There are multiple mechanisms responsible for cellular cholesterol efflux. Among them, the active efflux pathways are mediated primarily by the ATP-binding cassette (ABC) transporters ABCA1 and ABCG1. ABCA1 is essential for cholesterol and phospholipid efflux to apolipoprotein A-I and high density lipoprotein (HDL) biogenesis. ABCG1 promotes cholesterol efflux primarily to HDL particles. Atherosclerotic cardiovascular disease is a chronic inflammatory disease characterized by marked macrophage foam cell accumulation in atherosclerotic plaques and associated pro-inflammatory responses in lesional cells. Findings from both animal and human studies indicate a critical role of disturbed cholesterol homeostasis in pro-inflammatory responses in these cells, particularly in lesional macrophages. ABCA1 and ABCG1 are highly expressed in macrophages, particularly in response to cholesterol accumulation, and are essential in maintenance of cholesterol homeostasis. Functional deficiency of ABCA1 and ABCG1 in macrophage markedly increases atherogenesis, with exacerbated inflammatory responses. ABCA1 and ABCG1 also play a critical role in control of hematopoietic stem and progenitor cell (HSPC) proliferation and extramedullary hematopoiesis. Hematopoietic ABCA1 and ABCG1 deficiencies cause marked HSPC expansion and extramedullary hematopoiesis, particularly in hypercholesterolemia, and lead to marked monocytosis and neutrophilia with exacerbated pro-inflammatory responses. All these contribute to atherosclerosis. In this chapter, we describe these findings and discuss the current understanding of the underlying mechanisms. We also discuss other ABC transporters such as ABCG4, which also promotes cholesterol efflux to HDL and controls megakaryocyte proliferation and platelet biogenesis. By this pathway, ABCG4 also modulates atherogenesis. Therapeutic approaches targeting the pathways and mechanisms described also are discussed.</p