Identification of Multiple Oxidative Post-Translational Modifications of Plasma Albumin in Patients with Pulmonary Hypertension of Sickle Cell Anemia

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4814-4814
Author(s):  
Adam Odhiambo ◽  
David Perlman ◽  
Martin Steinberg ◽  
Catherine E. Costello ◽  
Mark E. McComb ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Pulmonary Hypertension ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Plasma Proteins ◽  
Maldi Mass Spectrometry ◽  
Disease Pathogenesis ◽  
Post Translational Modifications ◽  
Redox Biology ◽  
Extensive Evaluation

Abstract Background: Pulmonary hypertension (PH) in sickle cell anemia (SCA) is characterized by decreased nitric oxide bioavailability; which may, in part, be related to increased oxidative stress. It is possible that via protein post-translational modifications, oxidants are able to affect both protein structure and function. We hypothesized that, in patients with SCA and PH, oxidative post-translational modifications (PTMs) occur on plasma proteins and are important in disease pathogenesis. We previously reported the identification of one PTM, a malondialdehyde adduct on peptide 146–159 of albumin and sought to do a more extensive evaluation of this protein to determine the presence of other abnormalities. Methods: Plasma was obtained from subjects with: SCA and PH (n=5); SCA steady-state without PH (n=4); Pulmonary Arterial Hypertension (PAH) (n=4); no evidence of cardiopulmonary disease (n=4). Platelet-poor plasma was separated into albumin-enriched and albumin-depleted fractions. The albumin-enriched fraction was subjected to proteolytic digestion by trypsin and studied by matrix-assisted-laser desorption/ionization (MALDI) mass spectrometry (MS) and liquid chromatography (LC)-MS/MS tandem mass spectrometry. Proteomic analyses were performed on all samples and post-translational modifications characterized by MS/MS. Results: We have characterized several additional peptides of albumin from patients with PH of SCA that bear lipid peroxidation and glycation adducts. Our comprehensive LC-MS/MS results have allowed us to identify 4-hydroxynonenal (HNE), hexose (HEX) and malonyl adduction at numerous, distinct cysteine, histidine, lysine, and threonine albumin residues. Conclusion: Increased oxidant burden and altered redox biology is characteristic of PH in SCA. Plasma proteins, such as albumin, are a target for oxidants and changes in their structure may play a role in disease pathogenesis.

Identification of Oxidative Post-Translational Modifications on Plasma Albumin in Patients with Pulmonary Hypertension of Sickle Cell Anemia.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1215-1215
Author(s):  
Elizabeth S. Klings ◽  
Adam Odhiambo ◽  
David Perlman ◽  
Hua Huang ◽  
Catherine Costello ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pulmonary Hypertension ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Post Translational Modification ◽  
Dot Blot ◽  
Plasma Albumin ◽  
Post Translational Modifications ◽  
And Function ◽  
Dot Blot Analysis

Abstract Pulmonary hypertension (PH) in sickle cell anemia (SCA) is characterized by decreased nitric oxide bioavailability; which may, in part, be related to increased oxidative stress. It is possible that via protein post-translational modifications, oxidants are able to affect both protein structure and function. We hypothesized that, in patients with SCA and PH, oxidative post-translational modifications occur on plasma proteins and are important in disease pathogenesis. To examine this hypothesis, we chose to determine if oxidative post-translational modifications occur in a large-abundance protein, albumin, as a reflection of the presence of more widespread protein oxidative damage. Plasma was obtained from subjects with:SCA and PH (n=5);SCA steady-state without PH (n=4);Pulmonary Arterial Hypertension (PAH) (n=4);no evidence of cardiopulmonary disease (n=4). Platelet-poor plasma was separated into albumin-enriched and albumin-depleted fractions. The albumin-enriched fraction was subjected to proteolytic digestion by trypsin and studied by matrix assisted laser desorption ionization (MALDI) mass spectroscopy (MS) and liquid chromatography (LC)-MS/MS tandem mass spectrometry. Proteomics analyses were performed on all samples and post-translational modifications characterized by MS/MS. Results were confirmed by Western and dot-blot analysis using commercially available antibodies. In the albumin fraction, we identified peaks of differential post-translational modification on albumin peptide 146–159 only in PAH and SCA and PH samples; this PTM was identified to be a malondialdehyde adduct. The presence of a malondialdehyde adduct on albumin was confirmed by LC-MS/MS and Western analysis. We have identified a malondialdehyde adduct in plasma albumin that appears to be a common link between PH related to SCA and PAH. This finding supports the notion that oxidative stress modulates the pathogenesis of PH of SCA and suggests that this and other post-translational modifications may be important biomarkers of disease.

scholarly journals Prevalence of Pulmonary Hypertension in Sickle Cell Anemiai Patient in KSA

2021 ◽  
pp. 214-220
Author(s):  
Eman AbdulAziz Balbaid ◽  
Manal abdulaziz Murad ◽  
Hoda Jehad Abousada ◽  
Abdurrahman Yousuf Banjar ◽  
Mashael Abdulghani Taj ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Nervous System ◽  
Saudi Arabia ◽  
Risk Factor ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Cardiac Disease ◽  
Kingdom Of Saudi Arabia ◽  
Cross Sectional ◽  
Increased Risk

Introduction: Pulmonary hypertension (PH) is a relatively common and severe complication of SCI and an independent risk factor for mortality. Sickle cell disease is considered one of the most common diseases in the Kingdom of Saudi Arabia. When a healthy disease related to cardiovascular health is highlighted, sickle cell anemia may be the most common and related disease in high pulmonary pressure. In this study, we aimed to determine prevalence of PHTN in SCA patient, and associated risk factors with it.   Methodology: This is an analytical cross-sectional study conducted in kingdom of Saudi Arabia (General population, SCA patient and CVD patient), from 29/7/2020 till 15/11/2020. The study was depending on online self-reported questionnaire that included assessing the demographic factors as gender, nationality besides, disease-related information:  SCA patient , CVD patient and DM patient. Results: we received 794 responses to our questionnaire where 93.5% of them were Saudi Arabian.  The prevalence of sickle cell anemia is 8.8%. Male represented 29.8% of patients while female represented 52.2% of patients. In SCA patients, the prevalence of PHTN was 31.8%. Moreover, it was found that having cardiac disease is considered a risk factor for developing PHTN where 37.7% of patients having cardiac disease had PHTN compared with 6.2% of health patients (OD: 9.16, 95% CI: 5.5479 to 15.13, P=0.000) while diabetes mellitus increase risk for developing PHTN by more than seven fold (OD: 7.6, 95% CI; 4.7175 to 12.4, P=0.000) and disorder of nervous system by 12 folds (OD: 12.7; 95% CI: 7.6658 to 21.09, P=0.000). Conclusion: we had found that the prevalence if SCA among Saudi Arabia is 8.8% with a higher prevalence in female than male. Moreover, the prevalence of PHTN in SCA patients was high about 31.8% which is much higher than its prevalence in normal individuals. Moreover, it was found that having cardiac disease is considered a risk factor for developing PHTN besides, having diabetic condition and disorder of nervous system which increased risk for developing PHTN in SCA by nine, seven and 12-fold respectively.

Severity of Sickle Cell Disease: Modeling Interrelationships among Hemolysis, Pulmonary Hypertension and Risk of Death.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 786-786
Author(s):  
Paola Sebastiani ◽  
Vikki G. Nolan ◽  
Clinton T. Baldwin ◽  
Maria M. Abad-Grau ◽  
Ling Wang ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Sickle Cell Disease ◽  
Disease Severity ◽  
Hemolytic Anemia ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Clinical Severity ◽  
Acute Chest Syndrome ◽  
Cell Disease ◽  
Risk Of Death

Abstract A single point mutation in the β hemoglobin gene causes sickle cell disease (SCD), but patients have extremely variable phenotypes. Hemolysis-related complications include pulmonary hypertension (PHT), priapism, stroke and leg ulceration; blood viscosity and sickle vasoocclusion are associated with painful episodes, acute chest syndrome and osteonecrosis. Predicting who is at highest risk of death would be useful therapeutically and prognostically. Applying Bayesian network modeling that describes complex interactions among many variables by factorizing their joint probability distribution into modules, to data from 3380 SCD patients, we constructed a disease severity score (DSS: 0, least severe; 1, most severe), defining severity as risk of death within 5 years. A network of 24 variables described complex associations among clinical and laboratory complications of SCD. The analysis was validated in 140 patients whose SCD severity was assessed by expert clinicians and 210 adults where severity was also assessed by the echocardiographic diagnosis of PHT and death. Information about PHT allowed a comparison of the DSS with the tricuspid regurgitant jet velocity (TRJV), an objective marker of PHT and an independent risk factor for death. DSS and three indices of clinical severity (severity ranking of individuals by expert clinicians; objective measurement of the presence and severity of PHT; risk of prospective death) were correlated. Among living subjects, the median score was 0.57 in 135 patients without PHT, 0.64 in 40 patients with mild PHT and 0.86 in 15 patients with severe PHT. The difference in average score between living patients with and without PHT is significant. The same increasing trend was noticeable in the subjects who died during follow-up: 0.60 in subjects without PHT; 0.68 in subjects with mild PHT; 0.79 in subjects with severe PHT. The utility of the DSS is also supported by the ability to assign a score to subjects for whom the TRJV cannot be measured. Surprisingly, besides known risk factors like renal insufficiency and leukocytosis, we identified the intensity of hemolytic anemia and clinical events associated with hemolytic anemia as contributing to risk for death. Priapism, an excellent reflection of the hemolytic anemia-related complications of SCD, is associated with PHT and its association with death was unexpected. Laboratory variables predictive of disease severity included LDH and reticulocytes that reflect the intensity of hemolytic anemia. Elevated systolic blood pressure increased the odds of death by 3.4, consistent with hypertension as a marker of early death in SCD. Subjects with sickle cell anemia are at greatest risk compared with subjects with sickle cell anemia-α thalassemia and with subjects with HbSC disease. Our model suggests that the intensity of hemolytic anemia, estimated by LDH, reticulocyte count and AST, and shown previously to be associated with PHT, priapism, leg ulceration and possibly stroke, is an important contributor to death. This model can be used to compute a personalized measure of disease severity that might be useful for guiding therapeutic decisions and designing clinical trials.

Pulmonary Hypertension as a Risk Factor for Death in Patients with Sickle Cell Anemia

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4819-4819
Author(s):  
Rodolfo D Cancado ◽  
Maria Cristina A Olivato ◽  
Newton Nunes Lima Filho ◽  
Orlando Campos ◽  
Carlos Chiattone
Keyword(s):  
Pulmonary Hypertension ◽  
Sickle Cell Disease ◽  
Pulmonary Artery ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Systolic Pressure ◽  
Pulmonary Artery Systolic Pressure ◽  
Cell Disease ◽  
Risk Of Death ◽  
Hydroxyurea Therapy

Abstract Pulmonary hypertension develops in most forms of hereditary and chronic hemolytic anemia, including sickle cell disease, thalassemia, hereditary spherocytosis, and paroxysmal nocturnal hemoglobinuria, suggesting that there is a clinical syndrome of hemolysis-associated pulmonary hypertension. Retrospective studies from tertiary care referral centers suggest a prevalence of pulmonary hypertension in adults with sickle cell disease ranging from 20 to 40%. Despite the fact the elevations in pulmonary artery pressures are slight, morbidity and mortality are high. In adult sickle cell anemia patients, pulmonary hypertension is emerging as a major risk factor for death. We performed Doppler echocardiographic assessments of pulmonary-artery systolic pressure in 80 consecutive patients (20 men and 60 women; mean [±SD] age, 30 ± 10.8 years) between 1/20/2006 and 1/20/2008. The genotype on the basis of hematologic and hemoglobin characteristics was hemoglobin SS in all patients. Pulmonary hypertension was prospectively defined as a tricuspid regurgitant Jet velocity (TFJV) of at least 2.5 m per second. Patients were followed for a mean of 18 months (6–24 months), and data were censored at the time of death or loss to follow-up. Doppler-defined pulmonary hypertension occurred in 37.5 percent of patients (30/80). Multiple logistic-regression analysis, with the use of the dichotomous variable of a tricuspid regurgitant jet velocity of less than 2.5 m per second or 2.5 m per second or more, identified age, female sex, deferasirox therapy, left ventricular mass index, pulmonary artery systolic pressure, reticulocytes, white-cell count, platelet count, lactate dehydrogenase (a marker of hemolysis), blood urea nitrogen, creatinine, uric acid and self-reported history of cardiovascular complication, billiary stones, retinopathy and acute chest syndrome, as significant independent correlates of pulmonary hypertension. The hemoglobin level, fetal hemoglobin level, hydroxyurea therapy and serum ferritin level were unrelated to pulmonary hypertension. Hazard rate for death according to the TFJV of at least 2.5 m per second, as compared with a velocity of less than 2.5 m per second, was associated with an increased risk of death (0.00 versus 2.54; P=0.998). Mortality rate in 24 months was 6.7% (2/30) for patients with TRJ velocity ≥ 2.5 m/sec versus 0.0% (0/50) for patients without pulmonary hypertension. Pulmonary hypertension, diagnosed by Doppler echocardiography, is common in adults with sickle cell disease. It appears to be a complication of chronic hemolysis, is resistant to hydroxyurea therapy, and confers a high risk of death. Large trials evaluating the effects of treatment for pulmonary hypertension in the sickle cell anemia population are indicated.

scholarly journals Proteomics Pathways of Sickle Cell Anemia (P2SCA): A Comprehensive Analysis By Liquid Chromatography Mass Spectrometry of Erythrocyte Membrane Proteins Characterized from the Muhimbili Sickle Cell Programme, Tanzania

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3653-3653
Author(s):  
Haddy KS Fye ◽  
Paul Mrosso ◽  
Frédéric B Piel ◽  
Simon Davis ◽  
Roman Fischer ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Erythrocyte Membrane ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Liquid Chromatography Mass Spectrometry ◽  
Potential Implication ◽  
Chromatography Mass Spectrometry ◽  
Hydroxyurea Treatment ◽  
Significant Difference

Abstract Annually, there are 312 000 births with Sickle Cell Anemia (SCA), which has been recognized as one of the most common inherited conditions in Africa and is currently emerging as a condition of prominence in much of the developed world due to migration patterns. Despite its growing importance, there has been a significant lag in the application of emerging methodologies to its research, in particular to support the discovery of disease-associated markers of potential implication in therapeutics and informing a more comprehensive understanding of the condition. To bring SCA in line with cutting-edge 'omics research, we conducted a study applying advanced mass spectrometry methods for the comprehensive characterization of the protein profiles of erythrocyte membranes. One hundred and twenty participants of confirmed Hemoglobin (Hb) phenotypes HbAA (18), HbAS (21) and HbSS (81) were enrolled from the Muhimbili Sickle Cell Programme in Tanzania. All consented individuals were confirmed to not be on hydroxyurea treatment and not having received a blood transfusion in the preceding 3 months. Whole blood was collected and mixed in 10 ml EDTA vials sent within 72 hours to the Target Discovery Institute (TDI), University of Oxford, UK. Once received, packed erythrocyte fractions were isolated and underwent a series of wash steps followed by erythrocyte lysis and the isolation of the ghost membranes by ultracentrifugation. The ghost cells were then prepared for liquid chromatography mass spectrometry (LCMS) analysis using a novel published method. Proteomics analysis on a Thermo Scientific Q-Exactive High Field MS identified 2,288 membrane or membrane associated proteins of which 1,605 showed significant difference between at least two of the Hb phenotypes with fold changes of up to 22. Specific comparisons identified 1,286 proteins showing significant (p value cut-off, 0.05) changes in expression between the HbAA and HbSS groups; 1,248 between the HbAS and HbSS groups and 278 between the HbAA and HbAS groups. A summary of 30 markers have been presented including a range of proteins of potential therapeutic impact. These were selected based on their ranked significance following statistical analysis, the overall distinction seen between the controls and the HbSS group and the robustness of their identification. The results obtained show significant changes in the presence or absence as well as levels of numerous proteins in the HbSS population compared to controls. This research is a significant contribution to the field of SCA research and forms the foundation to direct further study on the sickle erythrocyte membrane. The large number of proteins identified to be of association to individuals' Hb genotypes represents a significant breakthrough and could contribute to a better understanding of the pathophysiological mechanisms involved in the disease. Figure. Figure. Disclosures No relevant conflicts of interest to declare.

The Hyperhemolysis Phenotype in Sickle Cell Anemia: Increased Risk of Leg Ulcers, Priapism, Pulmonary Hypertension and Death with Decreased Risk of Vasoocclusive Events.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 787-787 ◽  
Author(s):  
James G. Taylor ◽  
Vikki G. Nolan ◽  
Gregory J. Kato ◽  
Mark Gladwin ◽  
Martin H. Steinberg
Keyword(s):  
Pulmonary Hypertension ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Phenotypic Expression ◽  
Vascular Complications ◽  
Clinical Variable ◽  
Leg Ulcers ◽  
Risk Of Death ◽  
Arginase 1 ◽  
Increased Risk

Abstract Polymerization of hemoglobin in sickle cell anemia (HbSS) results in red blood cell damage, vasoocclusive disease and hemolytic anemia. Plasma hemoglobin and arginase released during hemolysis produce endothelial dysfunction and lead to vascular complications such as leg ulceration, priapism, pulmonary hypertension and increased risk of death. Examination of 2 independent patient groups from the NIH and CSSCD revealed considerable heterogeneity in the rates of hemolysis as estimated by LDH. We therefore hypothesized that inter-individual variation in hemolytic rate influences the phenotypic expression of HbSS. To test this hypothesis, we have defined a hemolysis phenotype as a clinical variable in our 2 HbSS groups. At NIH, LDH was analyzed as a quantitative trait in 166 adults to define a subgroup above the 75th percentile suggestive of exaggerated hemolysis (n= 42, mean LDH 645). When compared with 42 cases in the lowest LDH quartile (mean 230), the hyperhemolysis group had significantly lower hemoglobin and higher AST, bilirubin, and absolute reticulocyte levels supporting this phenotypic classification. HbF, arginine:ornithine ratios and arginase 1 activity were also significantly different. Clinically, the hyperhemolysis group had an increased prevalence of leg ulcers (OR 5.10; P=0.007), priapism (OR 3.67; P=0.16), and pulmonary hypertension (TRV > 3.0 m/s; OR 12.00; P<0.001). In addition, these patients have fewer severe vasoocclusive pain crises (3.1 ER visits/yr. with high LDH vs. 9.1/yr. with low LDH; P=0.004), supported by a lower prescription rate for hydroxyurea (37% with high LDH vs. 61% with low LDH; P=0.06). To validate these findings, we repeated the LDH phenotypic assignment analysis using 451 comparable CSSCD adults greater than 30 years of age. An identical pattern of laboratory results was seen when comparing high and low LDH quartiles. Hyperhemolysis was defined by anemia, elevated total bilirubin and AST and lower HbF. The most striking observations for this group were the significantly higher proportion of males (55% vs. 27% with low LDH; P<0.001), occurrence of leg ulcers (OR 3.44; P<0.001), and frequent death (33% vs. 16% with low LDH; P=0.003). There were also trends towards more prevalent priapism and stroke in the hemolysis group. After adjustment for HbF and gender, the associations between hyperhemolysis and risk of leg ulceration and risk of death remained significant in the CSSCD. Together, these findings suggest that excessive hemolysis results in a vascular HbSS phenotype characterized by leg ulcers, priapism, pulmonary hypertension, possibly stroke, and death. The lack of an association between excess hemolysis and pain/ACS suggests this phenotype is distinct from vasoocclusion/hyperviscosity. The consistent association of hyperhemolysis with this spectrum of symptoms, even after HbF correction, indicates the existence of unknown determinants for hemolysis. Future environmental and genetic studies of hyperhemolysis may provide novel mechanistic insights into the pathogenesis and treatment of HbSS vascular complications. From a clinical standpoint, these data further support the use of LDH as a simple biomarker for identifying a sub-set of HbSS patients at high risk for hemolysis-driven complications and death, who may otherwise be overlooked by clinicians because of infrequent vasoocclusive pain events.

Plasma Hemoglobin-Binding Capacity in Sickle Cell Disease

Blood ◽  
1959 ◽  
Vol 14 (9) ◽  
pp. 1047-1056 ◽  
Author(s):  
WILLOUGHBY LATHEM ◽  
WALLACE N. JENSEN
Keyword(s):  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Plasma Proteins ◽  
Sickle Cell Trait ◽  
Binding Capacity ◽  
Cell Disease ◽  
Plasma Hemoglobin ◽  
Hemoglobin C ◽  
Hemoglobin Binding ◽  
Made In

Abstract Studies of the capacity of plasma proteins to bind hemoglobin were made in patients with sickle cell anemia (SS), sickle cell trait (SA), hemoglobin C disease and in patients with hemolytic anemias. Hemoglobin binding was quantitatively normal in sickle cell trait, but was greatly reduced or absent in sickle cell anemia, hemoglobin C disease and in other hemolytic disorders. These alterations have been attributed to a reduction in the level of hemoglobin-binding proteins in circulating plasma. The mechanism of this reduction was not established, but the observed changes were correlated with the presence of increased hemolytic activity. The binding of hemoglobin C and hemoglobin S by normal plasma was quantitatively normal.

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