Genome wide association study of fetal hemoglobin in sickle cell anemia in Tanzania.

BACKGROUND: Fetal hemoglobin (HbF) is an important modulator of sickle cell disease (SCD). HbF has previously been shown to be affected by variants at three loci on chromosomes 2, 6 and 11, but it is likely that additional loci remain to be discovered. METHODS AND FINDINGS: We conducted a genome-wide association study (GWAS) in 1,213 SCA (HbSS/HbSβ0) patients in Tanzania. Genotyping was done with Illumina Omni2.5 array and imputation using 1000 Genomes Phase I release data. Association with HbF was analysed using a linear mixed model to control for complex population structure within our study. We successfully replicated known associations for HbF near BCL11A and the HBS1L-MYB intergenic polymorphisms (HMIP), including multiple independent effects near BCL11A, consistent with previous reports. We observed eight additional associations with P<10(-6). These associations could not be replicated in a SCA population in the UK. CONCLUSIONS: This is the largest GWAS study in SCA in Africa. We have confirmed known associations and identified new genetic associations with HbF that require further replication in SCA populations in Africa

Effect of Lowering Tube Potential and Increase Iodine Concentration of Contrast Medium on Radiation Dose and Image Quality in Computed Tomography Pulmonary Angiography Procedure: A Phantom Study

The aim of the study was to examine the effect of lowering tube potential and increase iodine concentration on image quality and radiation dose in computed tomography pulmonary angiography procedure. The pulmonary arteries were simulated by three syringes. The syringes were filled with 1:10 diluted solutions of 300 mg, 350 mg and 370 mg of iodine per millilitre concentration in three water-filled phantoms simulating thin, intermediate and thick patients. The phantoms were scanned at 80 kVp, 110 kVp and 130 kVp and 0.6 second rotation time using a 16 slice computed tomography (CT) scanner. The tube current was either fixed at 80, 100, 200, 250 and 300 mA or automatically adjusted with quality reference tube current-time product (mAsQR). In comparison with 130 kVp, images acquired at 80 kVp and 110 kVp, respectively, showed 76.2% to 99% and 19% to 26% enhancement in CT attenuation of iodinated contrast material. A volume CT dose index (CTDIvol) reduction by 35.3% was attained in small phantom with the use of 80 kVp, while in the medium phantom, a CTDIvol reduction by 29.9% was attained with the use of 110 kVp instead of 130 kVp. In light of the above, lowering tube potential and increase iodinated CM could substantially reduce the dose to small-sized adults and children

Genome wide association study of fetal hemoglobin in sickle cell anemia in Tanzania.

BACKGROUND: Fetal hemoglobin (HbF) is an important modulator of sickle cell disease (SCD). HbF has previously been shown to be affected by variants at three loci on chromosomes 2, 6 and 11, but it is likely that additional loci remain to be discovered. METHODS AND FINDINGS: We conducted a genome-wide association study (GWAS) in 1,213 SCA (HbSS/HbSβ0) patients in Tanzania. Genotyping was done with Illumina Omni2.5 array and imputation using 1000 Genomes Phase I release data. Association with HbF was analysed using a linear mixed model to control for complex population structure within our study. We successfully replicated known associations for HbF near BCL11A and the HBS1L-MYB intergenic polymorphisms (HMIP), including multiple independent effects near BCL11A, consistent with previous reports. We observed eight additional associations with P<10(-6). These associations could not be replicated in a SCA population in the UK. CONCLUSIONS: This is the largest GWAS study in SCA in Africa. We have confirmed known associations and identified new genetic associations with HbF that require further replication in SCA populations in Africa

Clinical epidemiology of individuals with Sickle cell anemia using Hydroxyurea at Muhimbili National Hospital, Dar Es Salaam, Tanzania

Background: The pathophysiology of sickle cell disease (SCD) is complex and involves nitric oxide depletion, increased inflammation/adhesion molecules and vaso-occlusion in addition to the chronic hemolytic anemia. This pathophysiology results in systemic clinical complications including recurrent episodes of severe pain, stroke, acute chest syndrome (ACS) and an increased susceptibility to infection. SCD severity varies among individuals and fetal hemoglobin (HbF) is known as a major modulator of the disease. To date, hydroxyurea (HU) is a known intervention that acts by increasing HbF in individuals with SCD. The increase in HbF reduces the risk of ‘sickling’ events and improves clinical outcomes. This is the first study on the use of HU in individuals with SCA in Tanzania.Methods: A case-control study to determine the proportion, indications, clinical and laboratory outcomes of SCD patients with HU use was conducted at Muhimbili National Hospital in Dar Es Salaam, Tanzania.Results: Forty-two patients with Sickle cell anemia (SCA) on HU treatment and 32 patients with SCA not on HU treatment were enrolled. The proportion of HU use by individuals with SCA at Muhimbili National Hospital was 10 per 1000. The mean HbF % was 9.8 ± 2.4 vs 6.2 ±1.4 for controls (P &lt;0.001). Thirty (71.4%) were enrolled for HU treatment due to central nervous system (CNS) events, frequent painful crises 11(26.2%) and recurrent anemia 1(2.4%). Thirty-two SCA patients (76.2%) reported improvements after being on HU for at least six months. Of these, 91% reported no history of severe pain that required hospitalizations since they started HU. Twenty patients (66.7%) out of those with CNS events reported not to have experienced convulsions after HU initiation.Conclusions: HbF was higher in patients who were on HU and had positive correlation with clinical outcomes. Further clinical trials are required to evaluate more effects of HU use among SCA individuals in Tanzania. Keywords: Sickle cell anemia, HU, Fetal hemoglobin, Tanzania

Association between malaria (Blank = Negative, Filled = Positive) by SCD and Foetal hemoglobin levels for individuals in all ages (A) and for ≥5 years (B).

<p>Sample size of each group is shown on the x-axis and p-values (p) in the graph were derived from t-tests (HbAA and HbSS) and Wilcox sign rank test (HbAS). The box show median (central line) and inter quartile range (IQR). The top whisker corresponds to the largest observation that is less than or equal to the 75th percentile + 1.5 x IQR. The bottom whisker corresponds to the smallest observation that is greater than or equal to the 25th percentile—1.5 x IQR, while open circles are the outliers.</p

Negative Epistasis between Sickle and Foetal Haemoglobin Suggests a Reduction in Protection against Malaria.

BACKGROUND: Haemoglobin variants, Sickle (HbS) and foetal (HbF) have been associated with malaria protection. This study explores epistatic interactions between HbS and HbF on malaria infection. METHODS: The study was conducted between March 2004 and December 2013 within the sickle cell disease (SCD) programme at Muhimbili National Hospital, Tanzania. SCD status was categorized into HbAA, HbAS and HbSS using hemoglobin electrophoresis and High Performance Liquid Chromatography (HPLC). HbF levels were determined by HPLC. Malaria was diagnosed using rapid diagnostic test and/or blood film. Logistic regression and generalized estimating equations models were used to evaluate associations between SCD status, HbF and malaria. FINDINGS: 2,049 individuals with age range 0-70 years, HbAA 311(15.2%), HbAS 241(11.8%) and HbSS 1,497(73.1%) were analysed. At enrolment, malaria prevalence was significantly higher in HbAA 13.2% compared to HbAS 1.24% and HbSS 1.34% (p<0.001). Mean HbF was lower in those with malaria compared to those without malaria in HbAA (0.43% vs 0.82%) but was the reverse in HbSS (8.10% vs 5.59%). An increase in HbF was associated with a decrease in risk of malaria OR=0.50 (95%CI: 0.28, 0.90; p=0.021) in HbAA, whereas for HbSS the risk of malaria increased OR=2.94 (1.44, 5.98; p=0.003). A similar pattern was seen during multiple visits; HbAA OR=0.52 (0.34, 0.80; p=0.003) vs HbSS OR=2.01 (1.27, 3.23; p=0.003). CONCLUSION: Higher prevalence of malaria in HbAA compared to HbAS and HbSS confirmed the protective effect of HbS. Lower prevalence of malaria in HbAA with high HbF supports a protective effect of HbF. However, in HbSS, the higher prevalence of malaria with high levels of HbF suggests loss of malaria protection. This is the first epidemiological study to suggest a negative epistasis between HbF and HbS on malaria

Coefficient estimates from multivariate logistic (single visit at the time of HbF measurement) and GEE (multiple visits) regression models of malaria in relation to SCD status, HbF levels and age for individuals of all ages.

<p>^§Transformed by square root</p><p>*Seven individuals with missing age excluded</p><p>**20 visits with missing age excluded</p><p>Coefficient estimates from multivariate logistic (single visit at the time of HbF measurement) and GEE (multiple visits) regression models of malaria in relation to SCD status, HbF levels and age for individuals of all ages.</p

Participants in the malaria study in the Sickle Cell Disease programme at Muhimbili National Hospital.

<p>Participants in the malaria study in the Sickle Cell Disease programme at Muhimbili National Hospital.</p

A flow chart diagram showing the flow of samples at different stages of the study.

<p>The genetic quality control measures involved removal of individuals with abnormal heterozygosity, gender mismatch, missingness of data, removal of duplicate samples and low quality/rare variants.</p

Regional association plot of the chromosome 19 region with our suggestive novel SNP (<i>rs28567737</i>).

<p>The left y axis represents the negative log10 P values of the associations of SNPs in this region. Each SNP is represented by a dot, with grey dots indicating low LD with <i>rs28567737</i>. The right y axis represents the recombination rate (in centimorgans [cM] per megabase [Mb]). Genes found in the region are shown in relative position under the plot; arrows indicate the direction of transcription.</p