Heme-induced cell adhesion in the pathogenesis of sickle-cell disease and inflammation

doi:10.1016/S0165-6147(00)01609-6

Trends in Pharmacological Sciences

Volume 22, Issue 2, 1 February 2001, Pages 52-54

https://doi.org/10.1016/S0165-6147(00)01609-6 Get rights and content

Abstract

Vascular occlusion is a major cause of the morbidity associated with sickle-cell disease. Although such obstruction is generally attributed to the clumping of sickle-shaped erythrocytes within vessels, other factors might be involved. Heme, which is released into the circulation during the active phase of sickle-cell disease, causes increased expression of endothelial adhesion molecules and adherence of leukocytes and reticulocytes to endothelial cells. Counteracting heme-mediated adhesion could form the basis for novel therapies to prevent the complications associated with this disabling disease.

Section snippets

Heme-induced adhesion and sickle-cell disease

The mutated hemoglobin protein HbS gives rise to fragile erythrocytes that are prone to hemolysis, resulting in large concentrations of free intravascular hemoglobin and heme, both of which can interact with the vascular endothelium 7., 8., 9.. Heme is the functional group of several important proteins (including hemoglobin in erythrocytes) and is therefore essential for various biological processes. However, free heme can have detrimental effects because it catalyzes the formation of reactive

Protective mechanisms against heme insults

The first line of defense against heme- and/or hemoglobin-induced insults within the vasculature are the scavengers of heme and hemoglobin, hemopexin and haptoglobin, respectively. However, in patients suffering from sickle-cell disease or thalassemia, hemopexin and haptoglobin concentrations are usually reduced 7., 21., which results in a loss of protection against heme-induced adhesion. Moreover, such scavengers might be unable to reach the locally elevated concentrations of heme that are

Novel treatment modalities in sickle-cell disease

Current treatment of sickle-cell disease is mainly based on erythrocyte transfusions (HbA) and induction of fetal hemoglobin (HbF) expression by administration of hydroxyurea ¹. Because HbS exhibits significantly increased auto-oxidation compared with HbA ⁸ and is less stable than HbA or HbF, lower HbS:HbA or HbS:HbF ratios will result in reduced hemolysis, lower concentrations of free vascular heme and ultimately alleviation of heme-induced complications.

Thus, it appears that prevention of the

Acknowledgments

This study was supported by the Vanderes Foundation. Owing to space constraints, only limited references are cited. We apologize to those authors whose work is not cited.

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VWF–ADAMTS13 axis dysfunction in children with sickle cell disease treated with hydroxycarbamide vs blood transfusion
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Previous studies have reported elevated von Willebrand factor (VWF) levels in patients with sickle cell disease (SCD) and demonstrated a key role for the VWF-ADAMTS13 axis in the pathobiology of SCD vaso-occlusion. Although blood transfusion is the gold standard for stroke prevention in SCD, the biological mechanisms underpinning its improved efficacy compared with hydroxycarbamide are not fully understood. We hypothesized that the improved efficacy of blood transfusion might relate to differences in VWF–ADAMTS13 axis dysfunction. In total, 180 children with a confirmed diagnosis of SCD (hemoglobin SS) on hydroxycarbamide (n = 96) or blood transfusion (n = 84) were included. Despite disease-modifying treatment, plasma VWF and VWF propeptide were elevated in a significant proportion of children with SCD (33% and 47%, respectively). Crucially, all VWF parameters were significantly higher in the hydroxycarbamide compared with the blood transfusion cohort (P < .05). Additionally, increased levels of other Weibel-Palade body–stored proteins, including factor VIII (FVIII), angiopoietin-2, and osteoprotegerin were observed, indicated ongoing endothelial cell activation. Children treated with hydroxycarbamide also had higher FVIII activity and enhanced thrombin generation compared with those in the blood transfusion cohort (P < .001). Finally, hemolysis markers strongly correlated with VWF levels (P < .001) and were significantly reduced in the blood transfusion cohort (P < .001). Cumulatively, to our knowledge, our findings demonstrate for the first time that despite treatment, ongoing dysfunction of the VWF–ADAMTS13 axis is present in a significant subgroup of pediatric patients with SCD, especially those treated with hydroxycarbamide.
Oxidative effects of cyanogenic glycosides residuals in cassava products on human haemoglobin
2021, Food Bioscience
Citation Excerpt :
Excess heme released during oxidation of oxy-Hb in the kidney as well as other organs and tissues is implicated in some other damages in the body such as denaturation of DNA and increased damage to proteins, depletion of cytosolic enzymes including glucose-6-phosphate dehydrogenase and glutathione reductase, activation of cell-damaging enzymes such as caspases and cathepsins, cytoskeleton damage and disruption of mitochondria function shown as inhibition of ATP synthesis (Leavesley et al., 2008; Nagy et al., 2010; Nath et al., 1998; Tracz et al., 2007). Free heme and excess free radicals released during the oxidation of Hb have been implicated in a number of disease states such as inflammation, sickle cell anaemia, malaria, cardiovascular disease and a lot of other disease state, more often as an exacerbating factor (Ferreira et al., 2008; Nagy et al., 2010; Wagener et al., 2001). In vivo, the reduction in oxy-Hb concentration is synonymous with the accumulation of met-Hb (Wallace et al., 1978).
Cyanogenic glycosides are phytotoxins in certain foods such as cassava (Manihot esculenta Crantz), making the food harmful for human consumption if inadequately processed. Cassava root tubers are used throughout much of Africa for their carbohydrate content, processed in various forms as a staple food. The quantity of cyanoglycosides in some cassava products can be a serious threat to life. To maximize production and profit, producers of cassava products frequently adopt shorter and quicker processing techniques, leading to potentially higher levels of residual cyanoglycosides in those products. Studies have suggested that human Hb may be oxidized by the consumption of such inadequately processed cassava products. However, no scientific investigation has been carried out to confirm this hypothesis. Consequently, this study was done to determine the effects of the residual poisons in cassava products on Hb oxidation. Hb oxidation was analysed using UV–visible spectrophotometry using 13 groups: a standard control, a positive control and test materials. The study showed that apart from garri roasted for 15–25 min and 6 days fermented fufu and abacha, other categories of processed cassava products led to complete oxidation of oxy-Hb. Therefore, roasting garri for 15–25 min, fermentation for 6 days during fufu processing and sun drying of abacha could help reduce the oxidation of Hb by residual toxins in cassava products.
Formation of novel N-acetylcysteine-hemin adducts abrogates hemin-induced cytotoxicity and suppresses the NRF2-driven stress response in human pro-erythroid K562 cells
2020, European Journal of Pharmacology
Heme (iron protoporphyrin IX), as the prosthetic group in hemoproteins, regulates vital cellular functions in human tissues. However, free heme released during hemolysis events promotes severe complications to millions of people worldwide. Over the years, thiols like glutathione (GSH) were known to antagonize heme toxicity. In this study, we have uncovered the underlying molecular mechanism by which N-acetylcysteine (NAC), a well-known thiol prevents hemin-induced cytotoxicity (HIC). Hemin-responsive human pro-erythroid K562 cells were employed to assess hemin intracellular accumulation and cytotoxicity at concentrations ≥50 μΜ, in cultures exposed only to hemin and/or both hemin and NAC. NAC inhibited the intracellular accumulation of hemin and prevented hemin-induced cell growth inhibition, cell death, oxidative stress, and accumulation of ubiquitinated proteins. Meanwhile, the activation of the NF-E2-related factor-2 (NRF2)-driven stress gene activation, a key element involved in HIC, was suppressed by NAC. A refined mechanism of the chemical reaction between NAC and hemin leading to adduct formation via a nucleophilic attack on hemin was uncovered for the first time by tandem mass spectrometry analysis (LC-MS/MS). Such thiol-hemin adducts acted as intermediates to mitigate HIC and to suppress hemin-induced NRF2-driven gene activation. Our findings support the concept that NAC-hemin adduct formation is the major novel molecular mechanism rather than the reactive oxygen species-scavenging capacity of thiols to protect cells from HIC. Our results imply that thiols and their derivatives can be of potential therapeutic value in hemolytic disorders.
Mn porphyrins as a novel treatment targeting sickle cell NOXs to reverse and prevent acute vaso-occlusion in vivo
2020, Blood Advances
Citation Excerpt :
Furthermore, the substantial levels of oxidative stress in SSRBCs enhance HbS autoxidation, which could contribute to cell membrane damage, premature erythrocyte aging, and hemolysis.16 Extracellular hemoglobin and heme in plasma or microparticles62 then promote severe oxidative stress, not only to blood cells but also to blood vessels, leading to vascular dysfunction and vaso-occlusion in SCD.63,64 Increased NOX-dependent ROS generation can signal NF-кB activation in the endothelium, which in turn triggers EC adhesion molecule expression (eg, VCAM-1, ICAM-1, P-selectin [the latter of which can be directly upregulated by ROS]),10,63 promoting leukocyte recruitment, SSRBC/leukocyte adhesion, transient vaso-occlusion, and ischemia/reperfusion.
In sickle cell disease (SCD), adhesion of sickle red blood cells (SSRBCs) and activated leukocytes in inflamed venules affects blood rheology, causing vaso-occlusive manifestations and vital reduction in microvascular blood flow. Recently, we found that NADPH oxidases (NOXs) create a vicious feedback loop within SSRBCs. This positive feedback loop mediates SSRBC adhesion to the endothelium. We show for the first time the therapeutic effectiveness of the redox-active manganese (Mn) porphyrins MnTnBuOE-2-PyP⁵⁺ (MnBuOE; BMX-001) and MnTE-2-PyP⁵⁺ (MnE; BMX-010, AEOL10113) to treat established vaso-occlusion in a humanized sickle mouse model of an acute vaso-occlusive crisis using intravital microscopy. These Mn porphyrins can suppress SSRBC NOX activity. Subcutaneous administration of only 1 dose of MnBuOE or MnE at 0.1 to 2 mg/kg after the inflammatory trigger of vaso-occlusion, or simultaneously, reversed and reduced leukocyte and SSRBC adhesion, diminished leukocyte rolling, restored blood flow, and increased survival rate. Furthermore, MnBuOE and MnE administered to sickle mice subcutaneously at 0.1 to 1 mg/kg for 28 days (except on weekends) did not exacerbate anemia, which seemed to be due to downregulation of both SSRBC reactive oxygen species production and exposure of the eryptotic marker phosphatidylserine. In addition, Mn porphyrins ameliorated leukocytosis, venous blood gases, endothelial activation, and organ oxidative damage. Our data suggest that Mn porphyrins, likely by repressing NOX-mediated adhesive function of SSRBCs and activated leukocytes, could represent a novel, safe therapeutic intervention to treat or prevent the establishment of acute pain crises. These NOX-targeted antioxidants merit further assessment in SCD clinical trials.
Pulmonary hypertension in children with hemolytic disorders
2020, Progress in Pediatric Cardiology
Pulmonary hypertension (PH) is an insidious and potentially devastating complication in adults with hemolytic disorders, with pathophysiologic changes starting in childhood. Hemolysis predisposes to PH through nitric oxide depletion from extracellular release of hemoglobin and arginase, and through red blood cell membrane alterations and formation of microparticles, all promoting oxidative damage, vasoconstriction, inflammation, vascular remodeling, and thromboembolic state. PH is best diagnosed by right heart catheterization, though screening often utilizes Doppler echocardiographic measurement of tricuspid regurgitant velocity, and functional assessment by six minute walk test. Understanding the pathophysiology and clinical presentation of PH in sickle cell disease, thalassemias, paroxysmal nocturnal hemoglobinuria, and other hemolytic disorders can improve recognition, evaluation, and management of this condition.
Patrolling monocytes scavenge endothelial-adherent sickle RBCs: A novel mechanism of inhibition of vaso-occlusion in SCD
2019, Blood
Citation Excerpt :
Attachment of sickle RBCs activates the underlying endothelium and, together with other blood components, leads to vaso-occlusion.9-12 Intravascular hemolysis, also a hallmark of SCD, further activates the vascular endothelium, increasing the attachment of sickle RBCs to the vessel wall.13-16 Haptoglobin and hemopexin, which normally remove free hemoglobin and heme, respectively, from the circulation, are depleted in patients with SCD.17-19
Painful vaso-occlusive crisis (VOC) is the most common complication of sickle cell disease (SCD). Increasing evidence suggests that vaso-occlusion is initiated by increased adherence of sickle red blood cells (RBCs) to the vascular endothelium. Thus, the mechanisms that remove endothelial-attached sickle RBCs from the microvasculature are expected to be critical for optimal blood flow and prevention of VOC in SCD. We hypothesized that patrolling monocytes (PMos), which protect against vascular damage by scavenging cellular debris, could remove endothelial-adherent sickle RBCs and ameliorate VOC in SCD. We detected RBC (GPA⁺)-engulfed material in circulating PMos of patients with SCD, and their frequency was further increased during acute crisis. RBC uptake by PMos was specific to endothelial-attached sickle, but not control, RBCs and occurred mostly through ICAM-1, CD11a, and CD18. Heme oxygenase 1 induction, by counteracting the cytotoxic effects of engulfed RBC breakdown products, increased PMo viability. In addition, transfusions, by lowering sickle RBC uptake, improved PMo survival. Selective depletion of PMos in Townes sickle mice exacerbated vascular stasis and tissue damage, whereas treatment with muramyl dipeptide (NOD2 ligand), which increases PMo mass, reduced stasis and SCD associated organ damage. Altogether, these data demonstrate a novel mechanism for removal of endothelial attached sickle RBCs mediated by PMos that can protect against VOC pathogenesis, further supporting PMos as a promising therapeutic target in SCD VOC.

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Trends in Pharmacological Sciences

Research UpdateHeme-induced cell adhesion in the pathogenesis of sickle-cell disease and inflammation

Abstract

Section snippets

Heme-induced adhesion and sickle-cell disease

Protective mechanisms against heme insults

Novel treatment modalities in sickle-cell disease

Acknowledgments

Blood

Blood

Blood

Blood

Blood

J. Lipid Res.

Biochim. Biophys. Acta

J. Biol. Chem.

Pathogenesis and treatment of sickle cell disease

New Engl. J. Med.

Adhesive interactions of sickle erythrocytes with endothelium

J. Clin. Invest.

Molecular basis of sickle cell–endothelial cell interactions

Curr. Opin. Hematol.

In vivo demonstration of red cell–endothelial cell interaction, sickling and altered microvascular response to oxygen in the sickle transgenic mouse

J. Clin. Invest.

Endothelial-cell heme uptake from heme proteins: induction of sensitization and desensitization to oxidant damage

Proc. Natl. Acad. Sci. USA

Research Update
Heme-induced cell adhesion in the pathogenesis of sickle-cell disease and inflammation