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Achille Iolascon - One of the best experts on this subject based on the ideXlab platform.
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rap 011 rescues the disease phenotype in a cellular model of Congenital Dyserythropoietic Anemia type ii by inhibiting the smad2 3 pathway
International Journal of Molecular Sciences, 2020Co-Authors: Gianluca De Rosa, Achille Iolascon, Immacolata Andolfo, Francesco Manna, Roberta Marra, Barbara Eleni Rosato, Roberta RussoAbstract:Congenital Dyserythropoietic Anemia type II (CDA II) is a hypo-productive Anemia defined by ineffective erythropoiesis through maturation arrest of erythroid precursors. CDA II is an autosomal recessive disorder due to loss-of-function mutations in SEC23B. Currently, management of patients with CDA II is based on transfusions, splenectomy, or hematopoietic stem-cell transplantation. Several studies have highlighted benefits of ACE-011 (sotatercept) treatment of ineffective erythropoiesis, which acts as a ligand trap against growth differentiation factor (GDF)11. Herein, we show that GDF11 levels are increased in CDA II, which suggests sotatercept as a targeted therapy for treatment of these patients. Treatment of stable clones of SEC23B-silenced erythroleukemia K562 cells with the iron-containing porphyrin hemin plus GDF11 increased expression of pSMAD2 and reduced nuclear localization of the transcription factor GATA1, with subsequent reduced gene expression of erythroid differentiation markers. We demonstrate that treatment of these SEC23B-silenced K562 cells with RAP-011, a “murinized” ortholog of sotatercept, rescues the disease phenotype by restoring gene expression of erythroid markers through inhibition of the phosphorylated SMAD2 pathway. Our data also demonstrate the effect of RAP-011 treatment in reducing the expression of erythroferrone in vitro, thus suggesting a possible beneficial role of the use of sotatercept in the management of iron overload in patients with CDA II.
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codysan a telemedicine tool to improve awareness and diagnosis for patients with Congenital Dyserythropoietic Anemia
Frontiers in Physiology, 2019Co-Authors: Cristian Tornador, Roberta Russo, Achille Iolascon, Immacolata Andolfo, Edgar Sanchezprados, Beatriz Cadenas, Veronica Venturi, Ines Hernandezrodriguez, Mayka SanchezAbstract:Congenital Dyserythropoietic Anemia (CDA) is a heterogeneous group of hematological disorders characterized by chronic hyporegenerative Anemia and distinct morphological abnormalities of erythroid precursors in the bone marrow. In many cases, a final diagnosis is not achieved due to different levels of awareness for the diagnosis of CDAs and lack of use of advanced diagnostic procedures. Researchers have identified five major types of CDA: types I, II, III, IV, and X-linked Dyserythropoietic Anemia and thrombocytopenia (XLDAT). Proper management in CDA is still unsatisfactory, as the different subtypes of CDA have different genetic causes and different but overlapping patterns of signs and symptoms. For this reason, we developed a new telemedicine tool that will help doctors to achieve a faster diagnostic for this disease. Using open access code, we have created a responsive webpage named CoDysAn (Congenital Dyserythropoietic Anemia) that includes practical information for CDA awareness and a step-by-step diagnostic tool based on a CDA algorithm. The site is currently available in four languages (Catalan, Spanish, Italian, and English). This telemedicine webpage is available at http://www.codysan.eu.
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gata1 erythroid specific regulation of sec23b expression and its implication in the pathogenesis of Congenital Dyserythropoietic Anemia type ii
Haematologica, 2017Co-Authors: Roberta Russo, Immacolata Andolfo, Francesco Manna, Gianluca De Rosa, Antonella Gambale, Alessandra Arillo, Farooq Wandroo, Maria Grazia Bisconte, Achille IolasconAbstract:Biallelic mutations in SEC23B gene cause Congenital Dyserythropoietic Anemia type II (CDAII), an autosomal recessive disorder characterized by ineffective erythropoiesis, hemolytic Anemia, and splenomegaly.[1][1] Approximately 80 SEC23B mutations have been described so far.[1][1]–[3][2] However,
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erfe encoding fam132b in Congenital Dyserythropoietic Anemia type ii
Blood, 2015Co-Authors: Roberta Russo, Immacolata Andolfo, Francesco Manna, Gianluca De Rosa, Luigia De Falco, Antonella Gambale, Mariasole Bruno, Domenico Girelli, L De Franceschi, Achille IolasconAbstract:Recessive mutations in SEC23B gene cause Congenital Dyserythropoietic Anemia type II (CDAII), a rare hereditary disorder hallmarked by ineffective erythropoiesis, iron overload, and reduced expression of hepatic hormone hepcidin (Iolascon, 2013). The most recently described hepcidin regulator is the erythroblast-derived hormone erythroferrone (ERFE), a member of TNF-α superfamily that specifically inhibits hepcidin production in experimental models (Kautz, 2014). However, the function of ERFE in humans remains to be investigated. To determine whether dysregulation of ERFE expression is associated with ineffective erythropoiesis and iron-loading in CDAII, we studied the ERFE-encoding FAM132B gene expression in 48 SEC23B -related CDAII patients and 29 age and gender matched healthy controls (HCs). Twelve new cases and four novel SEC23B mutations were described. Samples were obtained after informed consent, according to the Declaration of Helsinki. Genomic DNA, mutational screening, RNA isolation, cDNA preparation, and qRT-PCR were performed as previously described (Russo, 2013). All patients were young adults (17.0±2.5 years at diagnosis), with increased serum ferritin (395.4±67.6 ng/mL) and transferrin saturation (71.9±5.4 %). We observed a statistically significant overexpression of FAM132B gene in peripheral blood mononuclear cells from CDAII patients (9.09±0.08) compared to HCs (8.32±0.12, p FAM132B expression in reticulocytes from a subset of patients and HCs. Of note, a statistically significant correlation between peripheral blood and reticulocyte FAM132B expression from the same patients was observed (Spearman ρ= 0.78, p=0.02). Although the role of ERFE in peripheral blood is still unknown, our observations suggested that the evaluation of FAM132B mRNA in peripheral blood is a reliable and easy-to-measure marker of ERFE levels. When we divided CDAII patients into two sub-groups accordingly to FAM132B gene expression, we observed a statistically significant reduction in hemoglobin (Hb) level in the high-FAM132B subset (8.6±0.4 g/dL) respect to low-FAM132B one (10.1±0.5 g/dL, p=0.02). Of note, the expression level of FAM132B did not correlate with the transfusion regimen. The higher amount of ERFE reflects the increased iron demand for Hb production as well as the expanding abnormal erythropoiesis, as attested by the increased RDW and sTfR (although not significant) in high-FAM132B patients. This in turn leads to reduced hepcidin in high-FAM132B group (4.2±1.8 nM) compared to low-FAM132B one (5.9±1.8 nM, p=0.05), resulting in augmented iron delivery to the erythron. Although the iron balance data do not differ significantly between the two groups, a tendency to decreased hepcidin/ferritin ratio and increased transferrin saturation was observed in high-FAM132B patients. Thus, FAM132B overexpression seems to contribute to the inappropriate suppression of hepcidin with subsequent hemosiderosis observed in CDAII. Consistent with our previous studies, we observed a reduced SEC23B expression in our patients compared to HC. Indeed, FAM132B and SEC23B gene expression exhibited an inverse correlation (Spearman ρ=-0.36, p=0.01). We confirmed the ex vivo data about inverse correlation between FAM132B and SEC23B expression observed in our patients by establishing K562 SEC23B-silenced cells. To knockdown SEC23B gene expression in K562 cells two different pGIPZ Lentiviral shRNAmir for SEC23B (shSEC23B-70/-74) were used. We observed a higher expression of FAM132B at 5 days of erythroid differentiation in K562 SEC23B-silenced cell compared to not-silenced ones. Conversely, SEC23B expression was lower in both shSEC23B compared to sh-CTR at 2 and 5 days of differentiation. Although the mechanisms of hemin-induced differentiation are quite different from EPO-induced ones, we can hypothesize that FAM132B over-expression is related to the maturative arrest and the subsequent increased number of erythroid precursors. This study provides the first analysis on ERFE regulation in humans. Our data suggest that ERFE over-expression in CDAII patients is the result of both physiological and pathological mechanisms leading to hepcidin suppression in condition of dyserythropoiesis. Nevertheless, it seems that ERFE cannot be the main erythroid regulator of hepcidin suppression, at least in CDAII patients. Disclosures No relevant conflicts of interest to declare.
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retrospective cohort study of 205 cases with Congenital Dyserythropoietic Anemia type ii definition of clinical and molecular spectrum and identification of new diagnostic scores
American Journal of Hematology, 2014Co-Authors: Roberta Russo, Immacolata Andolfo, Antonella Gambale, Concetta Langella, Sule Unal, Achille IolasconAbstract:Congenital Dyserythropoietic Anemia II (CDA II) is a rare hyporegenerative Anemia of variable degree, whose causative gene is SEC23B. More than 60 causative mutations in 142 independent pedigrees have been described so far. However, the prevalence of the CDA II is probably underestimated, since its clinical spectrum was not yet well-defined and thus it is often misdiagnosed with more frequent clinically-related Anemias. This study represents the first meta-analysis on clinical and molecular spectrum of CDA II from the largest cohort of cases ever described. We characterized 41 new cases and 18 mutations not yet associated to CDA II, thus expanding the global series to 205 cases (172 unrelated) and the total number of causative variants to 84. The 68.3% of patients are included in our International Registry of CDA II (Napoli, Italy). A genotype-phenotype correlation in three genotypic groups of patients was assessed. To quantify the degree of severity in each patient, a method based on ranking score was performed. We introduced a clinical index to easily discriminate patients with a well-compensated hemolytic Anemia from those with ineffective erythropoiesis. Finally, the worldwide geographical distribution of SEC23B alleles highlighted the presence of multiple founder effects in different areas of the world.
Hannah Tamary - One of the best experts on this subject based on the ideXlab platform.
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characterization of the interactions between codanin 1 and c15orf41 two proteins implicated in Congenital Dyserythropoietic Anemia type i disease
BMC Molecular and Cell Biology, 2020Co-Authors: Grace Swickley, Hannah Tamary, Sharon Noylotan, Yehoshua Bloch, Lidor Malka, Adi Meiri, Amiel Yanai, Benny MotroAbstract:Congenital Dyserythropoietic Anemia type I (CDA I), is an autosomal recessive disease with macrocytic Anemia in which erythroid precursors in the bone marrow exhibit pathognomonic abnormalities including spongy heterochromatin and chromatin bridges. We have shown previously that the gene mutated in CDA I encodes Codanin-1, a ubiquitously expressed and evolutionarily conserved large protein. Recently, an additional etiologic factor for CDA I was reported, C15Orf41, a predicted nuclease. Mutations in both CDAN1 and C15Orf41 genes results in very similar erythroid phenotype. However, the possible relationships between these two etiologic factors is not clear. We demonstrate here that Codanin-1 and C15Orf41 bind to each other, and that Codanin-1 stabilizes C15Orf41. C15Orf41 protein is mainly nuclear and Codanin-1 overexpression shifts it to the cytoplasm. Phylogenetic analyses demonstrated that even though Codanin-1 is an essential protein in mammals, it was lost from several diverse and unrelated animal taxa. Interestingly, C15Orf41 was eliminated in the exact same animal taxa. This is an extreme case of the Phylogenetic Profiling phenomenon, which strongly suggests common pathways for these two proteins. Lastly, as the 3D structure is more conserved through evolution than the protein sequence, we have used the Phyre2 alignment program to find structurally homologous proteins. We found that Codanin-1 is highly similar to CNOT1, a conserved protein which serves as a scaffold for proteins involved in mRNA stability and transcriptional control. The physical interaction and the stabilization of C15Orf41 by Codanin-1, combined with the phylogenetic co-existence and co-loss of these two proteins during evolution, suggest that the major function of the presumptive scaffold protein, Codanin-1, is to regulate C15Orf41 activities. The similarity between Codanin-1 and CNOT1 suggest that Codanin-1 is involved in RNA metabolism and activity, and opens up a new avenue for the study of the molecular pathways affected in CDAI.
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morphological features of Congenital Dyserythropoietic Anemia type i the role of electron microscopy in diagnosis
European Journal of Haematology, 2017Co-Authors: Hanna Shalev, Joseph Kapelushnik, Dina Shaft, Orly Dgany, Tanya Krasnov, Peretz Resnitzky, Hannah TamaryAbstract:Introduction Congenital Dyserythropoietic Anemias are rare blood disorders characterized by Congenital Anemia and a wide range of morphological and functional abnormalities of erythroid precursors. Objectives To analyze the relative frequency of both light microscopic (LM) and electron microscopic (EM) morphological features of erythroblasts in a large group of patients with molecular proven Congenital Dyserythropoietic Anemia type I (CDAI). Methods We retrospectively evaluated the LM and EM of bone marrow (BM) erythroblasts in 35 patients with CDAI. Thirty-four patients carried the CDAN1 Arg1042Trp founder mutation and one the p.Pro1130Leu mutation. BM slides of 24 patients were available for LM examination. EM studies were performed in all 35 patients. Results On LM, marked erythroid hyperplasia, binuclear erythroblasts and various non-specific Dyserythropoietic features were documented in every case; internuclear chromatin bridges were detected in 19 patients (79%). In all EM of erythroblasts revealed a spongy appearance of heterochromatin, a widening of nuclear pores and invagination of cytoplasm into the nuclear region. Conclusions EM studies revealed high morphological frequency of specific ultrastructural changes in erythroblasts which facilitate prompt diagnosis of CDAI. Due to low specificity of BM LM findings, when BM EM is unavailable diagnostic approach should also include other inherited Anemias. This article is protected by copyright. All rights reserved.
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distal limb anomalies in patients with Congenital Dyserythropoietic Anemia
American Journal of Medical Genetics Part A, 2017Co-Authors: Achiya Amir, Gadi Horev, Joanne Yacobovich, Michael Bennett, Hannah TamaryAbstract:The Congenital Dyserythropoietic Anemias (CDAs) are a group of rare genetic disorders characterized by ineffective erythropoiesis and the development of secondary hemochromatosis. Distal limb anomalies are a well-documented though rare feature of Congenital Dyserythropoietic Anemia type I, that have not been reported so far in other types. We describe a patient with Congenital Dyserythropoietic Anemia type II and four members of a family with clinical features of Congenital Dyserythropoietic Anemia type III with distal limb anomalies. The patient with Congenital Dyserythropoietic Anemia type II presented with bilateral complete osseous syndactyly of the hands, and bilateral complete cutaneous syndactyly of feet. Three of the four affected family members with Congenital Dyserythropoietic Anemia type III had partial absence of fingers, small or absent nails, overlapping toes, and short metatarsals. We suggest that similar to Congenital Dyserythropoietic Anemia type I, distal anomalies may appear in some patients with Congenital Dyserythropoietic Anemia types II and III. Patients presenting with Anemia and distal limb anomalies should be further investigated for the presence of Congenital Dyserythropoietic Anemia. © 2016 Wiley Periodicals, Inc.
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morbidity and mortality of adult patients with Congenital Dyserythropoietic Anemia type i
European Journal of Haematology, 2017Co-Authors: Hanna Shalev, Itai Levi, Kaid Alathamen, Aviva Levitas, Hannah TamaryAbstract:Congenital Dyserythropoietic Anemia type I (CDAI) is a rare autosomal recessive disease characterized by macrocytic Anemia, ineffective erythropoiesis, and secondary hemochromatosis. To better define the natural history of the disease among adult patients, we studied 32 Bedouin patients (median age 34 yr; range 21-60) all carrying the same CDAN1 founder mutation. Follow-up studies included complete blood count, blood chemistry, abdominal ultrasound, echocardiography, and T2*MRI. Main complications were due to Anemia and ineffective erythropoiesis [osteoporosis (8/9, 89%), cholelithiasis (21/30, 70%), pulmonary arterial hypertension (PAH) (6/25, 24%)] and iron overload [hypothyroidism (9/24, 38%), and diabetes mellitus (6/32, 19%)]. T2* MRI revealed increased liver iron but no cardiac iron (13/13). Anemia improved in the majority of patients who underwent splenectomy (5/6). Three patients died (9%) at the age of 46-56 due to PAH (1) and sepsis (2). All previously underwent splenectomy. Analyzing both our patients and the 21 patients previously described by Heimpel et al. (Blood 107:334, 2006), we conclude that adults with CDA I suffer significant morbidity and mortality. Careful monitoring of iron overload and prompt iron chelation therapy is mandatory. Due to possible complications and inconsistent response to splenectomy α-interferon, transfusion therapy or stem cell transplantation should be considered as alternatives to this procedure in severely affected patients.
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codanin 1 binds to key erythroid genes and its knockdown coupled with ectopic mutant expression recapitulates the Congenital Dyserythropoietic Anemia type i cda i phenotype
Blood, 2014Co-Authors: Linette Bosques, Hannah Tamary, Patrick G Gallagher, Mitchell J Weiss, Caroline Tang, Diane S Krause, Sherman M Weissman, Gary M KupferAbstract:Abstract Congenital Dyserythropoietic Anemia type I (CDA I) is an autosomal recessive disorder marked by specific morphological abnormalities of erythroblasts in the bone marrow, resulting in ineffective erythropoiesis and Anemia. CDA I principally affects the erythroid lineage, while other non-erythroid hematopoietic lineages are unaffected. Light microscopy of bone marrow shows erythroid hyperplasia and binucleated erythroblasts connected by internuclear bridges. Electron microscopy of bone marrow demonstrates erythroblasts with spongy heterochromatin and invagination of the nuclear membrane. The gene responsible for causing CDA I, CDAN1, encodes for a ubiquitous protein, codanin-1, with unknown function. The goal of our study is to understand how codanin-1 regulates erythropoiesis and how codanin-1 defects result in CDA I. To determine codanin-1 expression pattern during erythroid differentiation, we analyzed codanin-1 RNA and protein levels in erythroid primary cells and cell lines. RNA-seq conducted in erythroid-induced human primary CD34+ cells revealed codanin-1 RNA expression is maintained in erythroid lineage. Similarly, immunoblotting shows codanin-1 protein is expressed at high levels in human erythroleukemia cell line stimulated down the erythroid lineage. Codanin-1 has been identified as a chromatin-binding protein. Thus, we conducted ChIP-seq analysis to determine the location of DNA binding sites for codanin-1 in K562 cells. The ChIP-seq results identified 2534 binding sites for codanin-1. Interestingly, many of these sites belong to key erythroid genes. Furthermore, 28% of these DNA binding sites overlap with GATA-1 bound sites. Because putative loss of function mutations in CDAN1 result in CDA I, we studied the effects of codanin-1 gene silencing on erythroid differentiation. shRNA-mediated knockdown reduced codanin-1 mRNA by 50% and protein expression levels by 90%. To assess the effects of codanin-1 perturbation on erythroid differentiation, hemoglobin synthesis was quantified using Drabkin’s reagent in which the absorbance measured at 540 nm is proportional to hemoglobin content in the sample. Erythroid-induced knockdown cells displayed decreased hemoglobin content when compared to non-targeting control shRNA (0.20 versus 0.48, measured in 3 separate experiments; p To establish a robust CDA I cell system, the first well-characterized CDAN1 mutation (R1042W) was studied because codanin-1 deletion mutants are embryonic lethal in mice and do not occur in humans. Endogenous codanin-1 knockdown cells were transduced with mutant or wild type codanin-1 and induced down the erythroid lineage. Because CDA I is characterized by abnormal heterochromatin appearance, we conducted electron microscopy studies to examine the ultrastructural features. Studies show that knockdown of endogenous codanin-1 coupled with ectopic expression of mutant codanin-1 results in recapitulation of the CDA I phenotype in the K562 cell line model. Because CDA I is almost exclusively restricted to the erythroid lineage, understanding its pathophysiology has the potential to identify critical pathways in erythropoiesis. Here we show that the full phenotype of CDA I is recapitulated upon not only the loss of function of the codanin-1 protein but also on the expression of the mutant form of codanin-1. Our analysis demonstrates that codanin-1 binds to important erythroid loci, suggesting that codanin-1 affects the expression of erythroid genes. Further work will focus on studying codanin-1 function in human primary erythroid cells and in animal models. Disclosures No relevant conflicts of interest to declare.
Silverio Perrotta - One of the best experts on this subject based on the ideXlab platform.
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Congenital Dyserythropoietic Anemia type ii molecular analysis and expression of the sec23b gene
Orphanet Journal of Rare Diseases, 2011Co-Authors: Francesca Punzo, Maria Domenica Cappellini, Aida M Bertoliavella, Saverio Scianguetta, Fulvio Della Ragione, Maddalena Casale, Luisa Ronzoni, Gian Luca Forni, Ben A Oostra, Silverio PerrottaAbstract:Background Congenital Dyserythropoietic Anemia type II (CDAII), the most common form of CDA, is an autosomal recessive condition. CDAII diagnosis is based on invasive, expensive, and time consuming tests that are available only in specialized laboratories. The recent identification of SEC23B mutations as the cause of CDAII opens new possibilities for the molecular diagnosis of the disease. The aim of this study was to characterize molecular genomic SEC23B defects in 16 unrelated patients affected by CDAII and correlate the identified genetic alterations with SEC23B transcript and protein levels in erythroid precursors.
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mutational spectrum in Congenital Dyserythropoietic Anemia type ii identification of 19 novel variants in sec23b gene
American Journal of Hematology, 2010Co-Authors: Roberta Russo, Maria Rosaria Esposito, Silverio Perrotta, Antonella Gambale, Roberta Asci, J Delaunay, Gian Luca Forni, Ugo Ramenghi, Vedat Uygun, Achille IolasconAbstract:SEC23B gene encodes an essential component of the coat protein complex II (COPII)-coated vesicles. Mutations in this gene cause the vast majority the Congenital Dyserythropoietic Anemia Type II (CDA II), a rare disorder resulting from impaired erythropoiesis. Here, we investigated 28 CDA II patients from 21 unrelated families enrolled in the CDA II International Registry. Overall, we found 19 novel variants [c.2270 A>C p.H757P; c.2149−2 A>G; c.1109+1 G>A; c.387(delG) p.L129LfsX26; c.1858 A>G p.M620V; c.1832 G>C p.R611P; c.1735 T>A p.Y579N; c.1254 T>G p.I418M; c.1015 C>T p.R339X; c.1603 C>T p.R535X; c.1654 C>T p.L552F; c.1307 C>T p.S436L; c.279+3 A>G; c. 2150(delC) p.A717VfsX7; c.1733 T>C p.L578P; c.1109+5 G>A; c.221+31 A>G; c.367 C>T p.R123X; c.1857_1859delCAT; p.I619del] in the homozygous or the compound heterozygous state. Homozygosity or compound heterozygosity for two nonsense mutations was never found. In four cases the sequencing analysis has failed to find two mutations. To discuss the putative functional consequences of missense mutations, computational analysis and sequence alignment were performed. Our data underscore the high allelic heterogeneity of CDA II, as the most of SEC23B variations are inherited as private mutations. In this mutation update, we also provided a tool to improve and facilitate the molecular diagnosis of CDA II by defining the frequency of mutations in each exon. Am. J. Hematol., 2010. © 2010 Wiley-Liss, Inc.
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molecular analysis of 42 patients with Congenital Dyserythropoietic Anemia type ii new mutations in the sec23b gene and a search for a genotype phenotype relationship
Haematologica, 2010Co-Authors: Achille Iolascon, Roberta Russo, Maria Rosaria Esposito, Silverio Perrotta, Roberta Asci, Carmelo Piscopo, Madeleine Feneantthibault, Loic Garcon, Jean DelaunayAbstract:Background The most frequent form of Congenital Dyserythropoietic Anemia is the type II form. Recently it was shown that the vast majority of patients with Congenital Dyserythropoietic Anemia type II carry mutations in the SEC23B gene. Here we established the molecular basis of 42 cases of Congenital Dyserythropoietic Anemia type II and attempted to define a genotype-phenotype relationship. Design and Methods SEC23B gene sequencing analysis was performed to assess the diversity and incidence of each mutation in 42 patients with Congenital Dyserythropoietic Anemia type II (25 described exclusively in this work), from the Italian and the French Registries, and the relationship of these mutations with the clinical presentation. To this purpose, we divided the patients into two groups: (i) patients with two missense mutations and (ii) patients with one nonsense and one missense mutation. Results We found 22 mutations of uneven frequency, including seven novel mutations. Compound heterozygosity for a missense and a nonsense mutation tended to produce a more severe clinical presentation, a lower reticulocyte count, a higher serum ferritin level, and, in some cases, more pronounced transfusion needs, than homozygosity or compound heterozygosity for two missense mutations. Homozygosity or compound heterozygosity for two nonsense mutations was never found. Conclusions This study allowed us to determine the most frequent mutations in patients with Congenital Dyserythropoietic Anemia type II. Correlations between the mutations and various biological parameters suggested that the association of one missense mutation and one nonsense mutation was significantly more deleterious that the association of two missense mutations. However, there was an overlap between the two categories.
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abnormalities of erythrocyte glycoconjugates are identical in two families with Congenital Dyserythropoietic Anemia type ii with different chromosomal localizations of the disease gene
Haematologica, 2007Co-Authors: Ewa Zdebska, Achille Iolascon, Silverio Perrotta, Justyna Spychalska, Carmen Lanzara, Gabriela Smolenskasym, Jerzy KoscielakAbstract:We analyzed erythrocyte glycoconjugates in two families with Congenital Dyserythropoietic Anemia type II (CDA-II): family 2 with the typical localization of the disease gene to chromosome 20q11.2 and family 1 in which this localization was excluded. Despite the different genetics, the erythrocyte glycoconjugate abnormalities in the two families were identical suggesting a complex inheritance of CDA-II. We also found that erythrocyte anion exchanger 1 protein is decreased in CDA-II homozygotes and obligate carriers alike.
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Congenital Dyserythropoietic Anemia type ii exclusion of seven candidate genes
Blood Cells Molecules and Diseases, 2003Co-Authors: Carmela Lanzara, Silverio Perrotta, Paolo Gasparini, A Iolascon, Romina Ficarella, A Totaro, Xin Chen, Roberta Roberto, Carla Lasalandra, Massimo CarellaAbstract:Congenital Dyserythropoietic Anemias (CDA) are genetic disorders characterized by Anemia and ineffective erythropoiesis. Three main types of CDA have been distinguished: CDA I, CDAII and CDA III, whose loci have been already mapped. After the identification of the locus for CDA II, also known as HEMPAS (hereditary erythroblast multinuclearity with positive acidified serum test), on the long arm of chromosome 20 (20q11.2) we have analyzed by a mutational search seven candidate genes in a large series of CDA II patients. In particular, the following genes have been investigated: integrin beta 4 binding protein, ribophorin II, ubiquitin protein ligase ITCH, mannosil-oligosaccharide alpha-1,2-mannosidase like protein, erythrocyte protein band 4.1 like protein, zinc finger protein PLAGL2, and finally novel zinc finger protein. None of them resulted as the causative gene but several protein variants and DNA polymorphisms have been identified. These data exclude the role of the above mentioned genes in causing CDA II and add further information in the process of cloning the CDA II gene.
Roberta Russo - One of the best experts on this subject based on the ideXlab platform.
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rap 011 rescues the disease phenotype in a cellular model of Congenital Dyserythropoietic Anemia type ii by inhibiting the smad2 3 pathway
International Journal of Molecular Sciences, 2020Co-Authors: Gianluca De Rosa, Achille Iolascon, Immacolata Andolfo, Francesco Manna, Roberta Marra, Barbara Eleni Rosato, Roberta RussoAbstract:Congenital Dyserythropoietic Anemia type II (CDA II) is a hypo-productive Anemia defined by ineffective erythropoiesis through maturation arrest of erythroid precursors. CDA II is an autosomal recessive disorder due to loss-of-function mutations in SEC23B. Currently, management of patients with CDA II is based on transfusions, splenectomy, or hematopoietic stem-cell transplantation. Several studies have highlighted benefits of ACE-011 (sotatercept) treatment of ineffective erythropoiesis, which acts as a ligand trap against growth differentiation factor (GDF)11. Herein, we show that GDF11 levels are increased in CDA II, which suggests sotatercept as a targeted therapy for treatment of these patients. Treatment of stable clones of SEC23B-silenced erythroleukemia K562 cells with the iron-containing porphyrin hemin plus GDF11 increased expression of pSMAD2 and reduced nuclear localization of the transcription factor GATA1, with subsequent reduced gene expression of erythroid differentiation markers. We demonstrate that treatment of these SEC23B-silenced K562 cells with RAP-011, a “murinized” ortholog of sotatercept, rescues the disease phenotype by restoring gene expression of erythroid markers through inhibition of the phosphorylated SMAD2 pathway. Our data also demonstrate the effect of RAP-011 treatment in reducing the expression of erythroferrone in vitro, thus suggesting a possible beneficial role of the use of sotatercept in the management of iron overload in patients with CDA II.
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codysan a telemedicine tool to improve awareness and diagnosis for patients with Congenital Dyserythropoietic Anemia
Frontiers in Physiology, 2019Co-Authors: Cristian Tornador, Roberta Russo, Achille Iolascon, Immacolata Andolfo, Edgar Sanchezprados, Beatriz Cadenas, Veronica Venturi, Ines Hernandezrodriguez, Mayka SanchezAbstract:Congenital Dyserythropoietic Anemia (CDA) is a heterogeneous group of hematological disorders characterized by chronic hyporegenerative Anemia and distinct morphological abnormalities of erythroid precursors in the bone marrow. In many cases, a final diagnosis is not achieved due to different levels of awareness for the diagnosis of CDAs and lack of use of advanced diagnostic procedures. Researchers have identified five major types of CDA: types I, II, III, IV, and X-linked Dyserythropoietic Anemia and thrombocytopenia (XLDAT). Proper management in CDA is still unsatisfactory, as the different subtypes of CDA have different genetic causes and different but overlapping patterns of signs and symptoms. For this reason, we developed a new telemedicine tool that will help doctors to achieve a faster diagnostic for this disease. Using open access code, we have created a responsive webpage named CoDysAn (Congenital Dyserythropoietic Anemia) that includes practical information for CDA awareness and a step-by-step diagnostic tool based on a CDA algorithm. The site is currently available in four languages (Catalan, Spanish, Italian, and English). This telemedicine webpage is available at http://www.codysan.eu.
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gata1 erythroid specific regulation of sec23b expression and its implication in the pathogenesis of Congenital Dyserythropoietic Anemia type ii
Haematologica, 2017Co-Authors: Roberta Russo, Immacolata Andolfo, Francesco Manna, Gianluca De Rosa, Antonella Gambale, Alessandra Arillo, Farooq Wandroo, Maria Grazia Bisconte, Achille IolasconAbstract:Biallelic mutations in SEC23B gene cause Congenital Dyserythropoietic Anemia type II (CDAII), an autosomal recessive disorder characterized by ineffective erythropoiesis, hemolytic Anemia, and splenomegaly.[1][1] Approximately 80 SEC23B mutations have been described so far.[1][1]–[3][2] However,
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increased levels of erfe encoding fam132b in patients with Congenital Dyserythropoietic Anemia type ii
Blood, 2016Co-Authors: Roberta Russo, Immacolata Andolfo, Francesco Manna, Gianluca De Rosa, Luigia De Falco, Antonella Gambale, Mariasole Bruno, Alessandro Matte, Paolo Ricchi, Domenico GirelliAbstract:To the editor: Recessive mutations in the SEC23B gene cause Congenital Dyserythropoietic Anemia type II (CDAII),[1][1] a rare hereditary disorder hallmarked by ineffective erythropoiesis, iron overload, and reduced expression of hepatic hormone hepcidin.[2][2],[3][3] Some erythroid regulators have
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erfe encoding fam132b in Congenital Dyserythropoietic Anemia type ii
Blood, 2015Co-Authors: Roberta Russo, Immacolata Andolfo, Francesco Manna, Gianluca De Rosa, Luigia De Falco, Antonella Gambale, Mariasole Bruno, Domenico Girelli, L De Franceschi, Achille IolasconAbstract:Recessive mutations in SEC23B gene cause Congenital Dyserythropoietic Anemia type II (CDAII), a rare hereditary disorder hallmarked by ineffective erythropoiesis, iron overload, and reduced expression of hepatic hormone hepcidin (Iolascon, 2013). The most recently described hepcidin regulator is the erythroblast-derived hormone erythroferrone (ERFE), a member of TNF-α superfamily that specifically inhibits hepcidin production in experimental models (Kautz, 2014). However, the function of ERFE in humans remains to be investigated. To determine whether dysregulation of ERFE expression is associated with ineffective erythropoiesis and iron-loading in CDAII, we studied the ERFE-encoding FAM132B gene expression in 48 SEC23B -related CDAII patients and 29 age and gender matched healthy controls (HCs). Twelve new cases and four novel SEC23B mutations were described. Samples were obtained after informed consent, according to the Declaration of Helsinki. Genomic DNA, mutational screening, RNA isolation, cDNA preparation, and qRT-PCR were performed as previously described (Russo, 2013). All patients were young adults (17.0±2.5 years at diagnosis), with increased serum ferritin (395.4±67.6 ng/mL) and transferrin saturation (71.9±5.4 %). We observed a statistically significant overexpression of FAM132B gene in peripheral blood mononuclear cells from CDAII patients (9.09±0.08) compared to HCs (8.32±0.12, p FAM132B expression in reticulocytes from a subset of patients and HCs. Of note, a statistically significant correlation between peripheral blood and reticulocyte FAM132B expression from the same patients was observed (Spearman ρ= 0.78, p=0.02). Although the role of ERFE in peripheral blood is still unknown, our observations suggested that the evaluation of FAM132B mRNA in peripheral blood is a reliable and easy-to-measure marker of ERFE levels. When we divided CDAII patients into two sub-groups accordingly to FAM132B gene expression, we observed a statistically significant reduction in hemoglobin (Hb) level in the high-FAM132B subset (8.6±0.4 g/dL) respect to low-FAM132B one (10.1±0.5 g/dL, p=0.02). Of note, the expression level of FAM132B did not correlate with the transfusion regimen. The higher amount of ERFE reflects the increased iron demand for Hb production as well as the expanding abnormal erythropoiesis, as attested by the increased RDW and sTfR (although not significant) in high-FAM132B patients. This in turn leads to reduced hepcidin in high-FAM132B group (4.2±1.8 nM) compared to low-FAM132B one (5.9±1.8 nM, p=0.05), resulting in augmented iron delivery to the erythron. Although the iron balance data do not differ significantly between the two groups, a tendency to decreased hepcidin/ferritin ratio and increased transferrin saturation was observed in high-FAM132B patients. Thus, FAM132B overexpression seems to contribute to the inappropriate suppression of hepcidin with subsequent hemosiderosis observed in CDAII. Consistent with our previous studies, we observed a reduced SEC23B expression in our patients compared to HC. Indeed, FAM132B and SEC23B gene expression exhibited an inverse correlation (Spearman ρ=-0.36, p=0.01). We confirmed the ex vivo data about inverse correlation between FAM132B and SEC23B expression observed in our patients by establishing K562 SEC23B-silenced cells. To knockdown SEC23B gene expression in K562 cells two different pGIPZ Lentiviral shRNAmir for SEC23B (shSEC23B-70/-74) were used. We observed a higher expression of FAM132B at 5 days of erythroid differentiation in K562 SEC23B-silenced cell compared to not-silenced ones. Conversely, SEC23B expression was lower in both shSEC23B compared to sh-CTR at 2 and 5 days of differentiation. Although the mechanisms of hemin-induced differentiation are quite different from EPO-induced ones, we can hypothesize that FAM132B over-expression is related to the maturative arrest and the subsequent increased number of erythroid precursors. This study provides the first analysis on ERFE regulation in humans. Our data suggest that ERFE over-expression in CDAII patients is the result of both physiological and pathological mechanisms leading to hepcidin suppression in condition of dyserythropoiesis. Nevertheless, it seems that ERFE cannot be the main erythroid regulator of hepcidin suppression, at least in CDAII patients. Disclosures No relevant conflicts of interest to declare.
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prevalence of left ventricular hypertrabeculation noncompaction among patients with Congenital Dyserythropoietic Anemia type 1 cda1
International Journal of Cardiology, 2020Co-Authors: Hadar Abramovichyoffe, Hanna Shalev, Aryeh Shalev, Orit Barrett, Aviva LevitasAbstract:Abstract Background Congenital Dyserythropoietic Anemia type 1 (CDA1) is a rare autosomal recessive disease characterized by macrocytic Anemia, ineffective erythropoiesis, and secondary hemochromatosis. Left-ventricular noncompaction (LVNC) is a cardiomyopathy that is commonly attributed to intrauterine arrest of normal compaction during the endomyocardial morphogenesis. LV hypertrabeculation/noncompaction (LVHT/NC) morphology, however, might exist in various hemoglobinopathies. Our primary objective was to determine whether the pattern of LVHT/NC is more prevalent among patients with CDA1, in comparison to subjects without CDA1, and to find potential risk factors for LVHT/NC among these patients. Our secondary objective was to evaluate the clinical implication of LVHT/NC. Methods We retrospectively assessed 32 CDA1 patients (median age 17.5, range 6–61) that underwent routine assessment of iron overload by cardiac magnetic resonance. Number and distribution of noncompacted LV segments were assessed in CDA1 patients and compared to 64 age- and gender-matched patients without CDA1. The ratio of noncompacted to compacted myocardium (NC/C ratio) in end-diastole was calculated for each of the three long-axis views. NC/C ratio > 2.3 was considered diagnostic for LVHT/NC. Results In multivariate analysis, the presence of CDA1 was independently associated with NC/C ratio > 2.3, a feature of LVHT/NC (adjusted OR = 11.46, 95%CI = 2.6–50.68, p = .001). CDA1 was strongly associated with increased number of myocardial segments exhibiting LVHT/NC pattern. Cardiac volumes and ejection fraction were preserved without clinical adverse events in long term follow-up. Conclusions CDA1 patients have a higher prevalence of LVHT/NC than normal individuals, independent of myocardial iron overload and without effect on ejection fraction or clinical outcome.
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morphological features of Congenital Dyserythropoietic Anemia type i the role of electron microscopy in diagnosis
European Journal of Haematology, 2017Co-Authors: Hanna Shalev, Joseph Kapelushnik, Dina Shaft, Orly Dgany, Tanya Krasnov, Peretz Resnitzky, Hannah TamaryAbstract:Introduction Congenital Dyserythropoietic Anemias are rare blood disorders characterized by Congenital Anemia and a wide range of morphological and functional abnormalities of erythroid precursors. Objectives To analyze the relative frequency of both light microscopic (LM) and electron microscopic (EM) morphological features of erythroblasts in a large group of patients with molecular proven Congenital Dyserythropoietic Anemia type I (CDAI). Methods We retrospectively evaluated the LM and EM of bone marrow (BM) erythroblasts in 35 patients with CDAI. Thirty-four patients carried the CDAN1 Arg1042Trp founder mutation and one the p.Pro1130Leu mutation. BM slides of 24 patients were available for LM examination. EM studies were performed in all 35 patients. Results On LM, marked erythroid hyperplasia, binuclear erythroblasts and various non-specific Dyserythropoietic features were documented in every case; internuclear chromatin bridges were detected in 19 patients (79%). In all EM of erythroblasts revealed a spongy appearance of heterochromatin, a widening of nuclear pores and invagination of cytoplasm into the nuclear region. Conclusions EM studies revealed high morphological frequency of specific ultrastructural changes in erythroblasts which facilitate prompt diagnosis of CDAI. Due to low specificity of BM LM findings, when BM EM is unavailable diagnostic approach should also include other inherited Anemias. This article is protected by copyright. All rights reserved.
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morbidity and mortality of adult patients with Congenital Dyserythropoietic Anemia type i
European Journal of Haematology, 2017Co-Authors: Hanna Shalev, Itai Levi, Kaid Alathamen, Aviva Levitas, Hannah TamaryAbstract:Congenital Dyserythropoietic Anemia type I (CDAI) is a rare autosomal recessive disease characterized by macrocytic Anemia, ineffective erythropoiesis, and secondary hemochromatosis. To better define the natural history of the disease among adult patients, we studied 32 Bedouin patients (median age 34 yr; range 21-60) all carrying the same CDAN1 founder mutation. Follow-up studies included complete blood count, blood chemistry, abdominal ultrasound, echocardiography, and T2*MRI. Main complications were due to Anemia and ineffective erythropoiesis [osteoporosis (8/9, 89%), cholelithiasis (21/30, 70%), pulmonary arterial hypertension (PAH) (6/25, 24%)] and iron overload [hypothyroidism (9/24, 38%), and diabetes mellitus (6/32, 19%)]. T2* MRI revealed increased liver iron but no cardiac iron (13/13). Anemia improved in the majority of patients who underwent splenectomy (5/6). Three patients died (9%) at the age of 46-56 due to PAH (1) and sepsis (2). All previously underwent splenectomy. Analyzing both our patients and the 21 patients previously described by Heimpel et al. (Blood 107:334, 2006), we conclude that adults with CDA I suffer significant morbidity and mortality. Careful monitoring of iron overload and prompt iron chelation therapy is mandatory. Due to possible complications and inconsistent response to splenectomy α-interferon, transfusion therapy or stem cell transplantation should be considered as alternatives to this procedure in severely affected patients.
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High levels of soluble serum hemojuvelin in patients with Congenital Dyserythropoietic Anemia type I
European Journal of Haematology, 2012Co-Authors: Hanna Shalev, Carole Brasse-lagnel, Galit Perez-avraham, Joseph Kapelushnik, Itai Levi, Anat Rabinovich, Dorine W. Swinkels, Hannah TamaryAbstract:OBJECTIVE: Congenital Dyserythropoietic Anemia (CDA) is a rare group of red blood cell disorders with ineffective erythropoiesis and secondary hemochromatosis. Inappropriate suppression of hepcidin and high levels of growth differentiation factor 15 (GDF15) have been described in CDA I and II patients, probably contributing to secondary hemochromatosis. Hemojuvelin (HJV) is an important regulator of serum hepcidin, while soluble form of HJV (s-HJV) competitively down-regulates hepcidin. METHODS: We determined the soluble hemojuvelin (s-HJV) levels in 17 patients with CDA I and in 17 healthy volunteers (HV) and looked for correlations with other parameters of iron overload and erythropoiesis. RESULTS: Significantly higher levels of s-HJV were found in patients (2.32 +/- 1.40 mg/L) compared with healthy volunteers (0. 69 +/- 0.44 mg/L) (P = 0.001). Western blot analysis confirmed the presence of high levels of s-HJV in CDA I patients. s-HJV positively correlated with serum ferritin, erythropoietin, soluble transferrin receptor, and GDF15 and negatively correlated with hepcidin to ferritin ratios. CONCLUSIONS: We for the first time documented high levels of serum s-HJV in CDA I patients, suggesting that it may contribute to iron loading pathology in CDA I and eventually in other Anemias with ineffective erythropoiesis.
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High Levels of Soluble Serum Hemojuvelin in Patients with Congenital Dyserythropoietic Anemia Type I
Blood, 2011Co-Authors: Hannah Tamary, Carole Brasse-lagnel, Galit Perez-avraham, Joseph Kapelushnik, Itai Levi, Hanna ShalevAbstract:Abstract 3179 Ineffective erythropoiesis in Congenital Dyserythropoietic Anemia type I (CDA I) is characterized by increased iron absorption mediated by down-regulation of hepcidin. It has been suggested that growth differentiation factor 15 (GDF15) contributes to the inappropriate suppression of hepcidin. Hemojuvelin (HJV), an important regulator of hepcidin production, is highly expressed in skeletal muscle and the liver. Mutations in the gene encoding HJV lead to severe hepcidin deficiency and juvenile hemochromatosis. Membrane-bound HJV (m-HJV) acts as a co-receptor for bone morphogenetic proteins (BMPs). HJV also exists in a soluble form (s-HJV) which competitively down-regulates hepcidin by interfering with BMP signaling. At least two proteases, furin and matriptase-2, cleave m-HJV into s-HJV. Recently, Brasse-Lagnel et al. ( Heamtologica 95:2031–7, 2010 ) reported a new assay for measuring both the furin- and the matriptase-2-cleaved forms of s-HJV. The aim of the present study was to evaluate the possible role of s-HJV in the development of iron overload in CDA I. The study group included eight (8) Israeli Bedouins with CDA I who were homozygous for the founder mutation, c.3124C>T. All were young adults of mean age 27 years. One patient had undergone splenectomy and is currently transfusion-dependent; 3 female patients required occasional blood transfusion during intercurrent infection, pregnancy, and delivery. Nine healthy volunteers served as the comparative group. Laboratory data, including levels of serum hepcidin, ferritin, erythropoietin, soluble transferrin receptor (sTfR), and GDF 15, were previously collected in both groups (Tamary et al. Blood 112:5241–4, 2008 ). Serum levels of s-HJV were measured in stored frozen samples using a competitive ELISA assay based on an anti-HJV polyclonal antibody (Brasse-Lagnel et al. Heamtologica 95:2031–7, 2010 ). Compared to the healthy volunteers, the patient group was characterized by significantly higher levels of s-HJV (1.99±0.98mg/L vs 0.36±0.25mg/L; p=0.001), significantly lower level of hemoglobin, and significantly higher levels of serum ferritin, erythropoietin, GDF 15, and sTfR. Serum hepcidin levels were similar in the CDA I patients and the healthy volunteers, but the hepcidin-to-ferritin ratio was significantly lower in the patient group, suggesting that serum hepcidin concentrations were inappropriately low. Levels of s-HJV negatively correlated with hemoglobin levels (R= −0.69 p=0.002) and positively correlated with the iron loading parameter, serum ferritin (R=0.73 p=0.000), and parameters of erythropoiesis, such as serum erythropoietin (R=0.63 p= 0.006) and sTfR (R=0.66 p= 0.008). Additionally, GDF 15 level was positively correlated with s-HJV level (R= 0.64 p=0.006), whereas the hepcidin-to-ferritin ratio (R=−0.797 p=0.000) was negatively correlated with s-HJV level. In summary, our study suggests that s-HJV is overexpressed in patients with CDA I and probably contributes to the down-regulation of hepcidin and the iron-loading pathology. To our knowledge, this is the first study to analyze s-HJV levels in a disease characterized by ineffective erythropoiesis. The main limitation of our study is the small number of patients. Larger studies including patients with CDA and thalassemia are indicated. The source of serum s-HJV and the trigger for its overexpression in CDA I still need to be established. Disclosures: No relevant conflicts of interest to declare.