TMPRSS6 gene mutations in six Saudi families with iron refractory iron deficiency anemia.

Iron-refractory iron deficiency anemia (IRIDA) is considered an autosomal recessive iron deficiency anemia due to mutations in the transmembrane protease serine 6 (TMPRSS6) gene. Variations in iron parameters and a higher risk of iron deficiency have been linked to the TMPRSS6 mutations. Furthermore, human genome-wide association studies (GWAS) identified a common mutation (rs855791) linked to abnormal hematological parameters, highlighting the importance of the TMPRSS6 gene in the regulation of iron homeostasis. This is the first study to investigate TMPRSS6 gene mutation in six Saudi families of probands with iron deficiency anemia unresponsive to oral iron and partially responsive to parenteral iron administration. Each participant provided a vacutainer tube with three blood samples (2.5 ml each) and analyzed based on hematological, biochemical iron profiles, and followed by genotyping by PCR. The TMPRSS6 gene was amplified, sequenced, and analyzed in all probands and family members. Statistical analysis was done using SPSS and SHEsis software. Few functional mutations in these families were suggested (p.W73X, p.E523K and p.V736A). The proband of family 6 presented numerous hematological abnormalities upon initial consultation, including normocytic anemia accompanied by low Hb, normal MCV, low serum iron, low serum ferritin, and normal TIBC. While the p.W73X variant was only found in 2 families, the p.V736A variant was found in all examined Saudi families with IRIDA. Given the evidence outlined for these six cases, future genotype-phenotype correlation studies in a large number of IRIDA patients in Saudi Arabia may be very informative for patient management, in addition to increasing knowledge of TMPRSS6 function during development as well as factors in the regulation of TMPRSS6 and its effect on iron levels in the body.

Genetic analysis of TMPRSS6 gene in Saudi female patients with iron deficiency anemia.

OBJECTIVE/BACKGROUND: Mutations in transmembrane protease serine 6 (TMPRSS6) gene induce high hepcidin level, which causes iron-refractory iron deficiency anemia (IRIDA) by preventing duodenal iron absorption. This study aims to identify the common genetic variations of the TMPRSS6 gene that affect iron levels among Saudi female patients with iron deficiency anemia (IDA). METHODS: All study participants were Saudi females (12-49 years old): 32 patients with IDA, 32 patients with IRIDA, and 34 healthy individuals comprising the control group. Hematological investigations, iron profile, serum hepcidin level, and TMPRSS6 gene transcription were determined. The TMPRSS6 gene was amplified, sequenced, and analyzed among all study participants. RESULTS: The mean hepcidin and TMPRSS6 RNA transcription levels in IDA and IRIDA groups were significantly lower than those in the control group. TMPRSS6 gene sequence analysis detected 41 variants: two in the 5' untranslated region (5'UTR), 17 in introns, and 22 in exons. Thirty-three variants were previously reported in the Single Nucleotide Polymorphism Database, and eight variants were novel; one novel variant was in 5'UTR (g.-2 T > G); five novel variants were detected in exons (p.W73X, p.D479N, p.E523K, p.L674L, and p.I799I). At the time of the sequence analysis of our samples, two variants-p.D479N and p.674L-were novel. However, these variants are present at a very low allele frequency in other populations (L674L, 0.00007761 and D479N, 0.000003980). CONCLUSION: This is the first study to investigate the genetic variants of TMPRSS6 gene in Saudi female patients with IDA. The generated data will serve as a reference for future studies on IDA in the Arab population.

Role of non-invasive assessment in prediction of preclinical cardiac affection in multi-transfused thalassaemia major patients.

BACKGROUND: The principal cause of mortality and morbidity in ß-thalassemia major (ß-TM) is the iron overload as these patients receive about 20 times the normal intake of iron, which leads to iron accumulation and damage in the liver, heart, and endocrine organs. Chronically transfused patients used to die from cardiac iron overload in their teens and twenties. Monitoring of iron status through cardiac magnetic resonance imaging (CMRI) has replaced the conventional methods of assessment, yet this modality is not readily available in centers where the disease distribution is maximal. Objectives The aim of this work is to study some simple non-invasive tools and their abilities to predict preclinical cardiac affection reflecting cardiac iron deposition (CID) in multi-transfused ß-TM patients taking the T2* CMRI as a gold standard reference test. METHODS: One hundred consecutive multi-transfused, clinically stable ß-TM patients with age ranging from 16 to 30 years (mean ± SD, 21.1 ± 3.9) were included in this study. Assessment of serum ferritin, serum hepcidin, and high-sensitivity C-reactive protein as well as cardiac assessment by echo-doppler and 24-hour Holter were used to predict CID, and consequently predict preclinical cardiac affection, in reference to CMRI results as the standard method of cardiac iron assessment. RESULTS: According to CMRI results, patients were subdivided into a group of 42 patients with detectable myocardial iron (T*≤ 20 ms) and a group of 58 patients with no detectable myocardial iron (T* > 20 ms). No differences in age, gender, or distribution of splenectomized patients were observed between both groups. Patients with detectable myocardial iron received significantly higher number of transfusions per year than those with no detectable myocardial iron (mean ± SD, 14.6 ± 1.7 vs. 12.5 ± 1.7; P < 0.001) yet comparable levels of serum ferritin, serum hepcidin, and hepcidin/ferritin ratio (P > 0.05) were noted. Cardiac iron detection was associated with significantly lower heart rate (mean ± SD, 75 ± 6.1 vs. 80 ± 6.9; P < 0.001), lower left ventricular ejection fraction (LVEF) (mean ± SD 60.1 ± 3.2 vs. 70.1 ± 2.8; P < 0.001), and higher total number of premature ventricular contractions (PVCs) (median 78 vs. 14; P < 0.001). The group with CID comprised significantly more patients with left ventricular diastolic dysfunction (15/42, 35.7% vs. 3/58, 5.2%; P < 0.001); PVCs ≥10/hour (13/42, 31% vs. 2/58, 3.4%; P < 0.001); episodes of sinus pauses (6/42, 14.3% vs. 1/58, 1.7%; P < 0.05); episodes of high-grade atrio-ventricular block (5/42, 11.9% vs. 1/58, 1.7%; P < 0.05) compared to the group with no (CID). In presence of normal LVEF, detection of 10 or more PVCs per hour was the most predictive of cardiac iron loading with a positive predictive value of 86.7% and specificity of 96.6%, and the highest likelihood ratio (9.09). Detection of more than 22 PVCs/24 hours had the best sensitivity (81%) and the best negative predictive value (84%). The positive likelihood ratio of the studied parameters was highest in case of presence of PVCs ≥10/hour and lowest in case of average heart rate with a cut-off level of ≤77.5 bpm (9.09 and 1.46, respectively). CONCLUSION: Our results support our hypothesis that monitoring ß-TM patients with echo and Holter electrocardiogram can help in the detection of preclinical cardiac affection in centers lacking cardiac MRI; however, due to relatively low sensitivity they can not fully replace CMRI. Further work is needed to identify additional simple parameters that can form a diagnostic model with adequate sensitivity.

Plasma PTX3 levels in sickle cell disease patients, during vaso occlusion and acute chest syndrome (data from Saudi population).

BACKGROUND: Sickle cell disease (SCD) is a chronic, incurable hereditary disease. The vaso-occlusive crisis (VOC) is the most frequently occurring acute complication in sickle cell patients and accounts for the majority of SCD-related hospital admissions. Another major complication is the potentially fatal acute chest syndrome (ACS). The prototypic long pentraxin-3 (PTX3), an acute phase protein and a key component of innate immunity, is linked to ischemia-induced inflammation, a condition incriminated in SCD complications. AIM: To investigate the expression of PTX3 in stable SCD and VOC patients and to assess its relation to the development and progression of ACS. SUBJECTS AND METHODS: We conducted this study on 160 patients with confirmed SCD (20 stable SCD and 140 in VOC), and 10 healthy age- and sex-matched controls. Patients were diagnosed as SCD by high-performance liquid chromatography. PTX3 levels were assessed using enzyme-linked immunosorbant assay. RESULTS: In the stable state, all 20 SCD patients had PTX3 levels (range = 0.9-2.1 ng/ml; median = 1.1) comparable to those of healthy controls (range = 0.8-2.0 ng/ml; median = 1.0) (P > 0.05). During the VOC, plasma PTX3 significantly increased (range = 8.7-37.2 ng/ml; median = 22.3) (P < 0.01). Out of 140 VOC patients, 15 (10.7%) developed ACS and four required mechanical ventilation, of which two died. The median plasma level of PTX3 (22.3 ng/ml) was set as a cut-off value to stratify patients into low- and high-PTX3 expressers. Of the 140 VOC patients, 43 (30.7%) had PTX3 levels >22.3 ng/ml, of these, 13 patients developed ACS (13/43; 30.2%); of the remaining 97 patients who had PTX3 ≤22.3 ng/ml, only two patients (2/97; 2.1%) progressed to ACS, with a further increment in PTX3 in all of them. PTX3 levels were correlated with length of hospital stay in VOC patients and markers of lung injury in ACS patients. CONCLUSION: PTX3 levels were higher in SCD patients in VOC, being associated with longer hospital stay. Higher initial PTX3 concentrations were related to the development of ACS with a further increase in PTX3 levels observed upon progression to ACS. Thus, PTX3 could be used as a subjective method to predict occurrence and severity of SCD acute complications.

Changing therapy from Glivecto a "copy" imatinib results in a worsening of chronic myeloid leukemia disease status: two case reports.

INTRODUCTION: Imatinib mesylate (Glivec((R))/Gleevec((R))) is the standard first-line therapy for the treatment of chronic myeloid leukemia due to its high hematologic, cytogenetic, and molecular response rates and favorable long-term safety profile. A copy version of imatinib is currently available in several countries. We report on two cases of CML who were originally treated with Glivec in Egypt and subsequently switched to the copy drug CASE PRESENTATION: Case one was a 35-year old female with chronic myeloid leukemia in blast crisis who began treatment with combination chemotherapy and Glivec. The patient achieved and maintained a complete hematologic response and continued on Glivec 400 mg/day. In March 2007, she was switched to the copy drug In September 2007, the patient presented in hematologic relapse. At this time, treatment with chemotherapy in combination with Glivec 400 mg/day was resumed. The patient quickly achieved, and maintained, complete hematologic response on Glivec 400 mg/day. The second patient was a 64-year old male with chronic myeloid leukemia in blast crisis who began treatment with truncated chemotherapy in combination with Glivec 400 mg/day. After 6 months, the patient achieved a partial hematologic response and continued on alternating cycles of chemotherapy with continuous administration of Glivec 400 mg/day. The patient received Glivec from January 2006 to February 2007, after which time he was switched to the copy drug. In November 2007, he presented with upper gastrointestinal bleeding and multiple gastric erosions and died the same day. CONCLUSION: The safety and efficacy of the copy drug has not been established in randomized clinical trials. It is unknown whether patients, who respond to Glivec and then switch to copy versions of imatinib, will tolerate the copy drug and maintain their response.