Cooley Anemia Publications (11121)


Cooley Anemia Publications

Cytotherapy 2017 Feb;19(2):311-326
Molecular Genetics Thalassaemia Department, The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus; Cyprus School of Molecular Medicine, Nicosia, Cyprus.

Primary hematopoietic stem and progenitor cells (HSPCs) are key components of cell-based therapies for blood disorders and are thus the authentic substrate for related research. We propose that ubiquitous small-volume diagnostic samples represent a readily available and as yet untapped resource of primary patient-derived cells for cell- and gene-therapy studies.
In the present study we compare isolation and storage methods for HSPCs from normal and thalassemic small-volume blood samples, considering genotype, density-gradient versus lysis-based cell isolation and cryostorage media with different serum contents. Read More

Downstream analyses include viability, recovery, differentiation in semi-solid media and performance in liquid cultures and viral transductions.
We demonstrate that HSPCs isolated either by ammonium-chloride potassium (ACK)-based lysis or by gradient isolation are suitable for functional analyses in clonogenic assays, high-level HSPC expansion and efficient lentiviral transduction. For cryostorage of cells, gradient isolation is superior to ACK lysis, and cryostorage in freezing media containing 50% fetal bovine serum demonstrated good results across all tested criteria. For assays on freshly isolated cells, ACK lysis performed similar to, and for thalassemic samples better than, gradient isolation, at a fraction of the cost and hands-on time. All isolation and storage methods show considerable variation within sample groups, but this is particularly acute for density gradient isolation of thalassemic samples.
This study demonstrates the suitability of small-volume blood samples for storage and preclinical studies, opening up the research field of HSPC and gene therapy to any blood diagnostic laboratory with corresponding bioethics approval for experimental use of surplus material.

J. Pediatr. Hematol. Oncol.
J Pediatr Hematol Oncol 2017 Jan 12. Epub 2017 Jan 12.
*Thalassemia Center †Molecular Genetics Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt.

The klf10 gene could indirectly modify γ-globin chain production and hence the level of fetal hemoglobin (HbF) ameliorating the phenotype of β-hemoglobinopathies and the response to hydroxycarbamide (hydroxyurea [HU]) therapy. In this study, we aimed to evaluate the frequency of different genotypes for the klf10 gene in β-thalassemia major (B-TM), β-thalassemia intermedia (B-TI), and sickle cell disease (SCD) patients by polymerase chain reaction and to assess its relation to disease phenotypes and HU response.
This cross-sectional study included 75 patients: 50 B-TM, 12 SCD, and 13 B-TI patients (on stable HU dose). Read More

The relation of the klf10 gene polymorphism (TIEG, TIEG1, EGRα) (rs3191333: c*0.141C>T) to phenotype was studied through baseline mean corpuscular volume, HbF, and transfusion history, whereas evaluation of response to HU therapy was carried out clinically and laboratory.
The frequency of the mutant klf10 genotype (TT) and that of the mutant allele (T) was significantly higher among B-TM patients compared with those with B-TI and SCD patients. Only homozygous SCD patients for the wild-type allele within the klf10 gene had a significantly lower transfusion frequency. The percentage of HU responders and nonresponders between different klf10 polymorphic genotypes among B-TI or SCD patients was comparable.
Although the klf10 gene does not play a standalone role as an HbF modifier, our data support its importance in ameliorating phenotype among β-hemoglobinopathies.

Hematology 2017 Jan 13:1-2. Epub 2017 Jan 13.
a Chair of Cardiology, Second University of Naples - Monaldi Hospital , P.zzale E. Ruggeri, 80131 Naples , Italy.
Curr Gene Ther
Curr Gene Ther 2017 Jan 9. Epub 2017 Jan 9.
Division of Hematopoietic Innovative Therapies, CIEMAT/ CIBERER-ISC-III. Madrid, Spain.

Fanconi anemia (FA) is a rare genetic syndrome characterized by progressive marrow failure. Gene therapy by infusion of FA-corrected autologous hematopoietic stem cells (HSCs) may offer a potential cure since it is a monogenetic disease with mutations in the FANC genes, coding for DNA repair enzymes (See review[1]). However, the collection of hCD34 +-cells in FA patients implies particular challenges because of the reduced numbers of progenitor cells present in their bone marrow (BM)[2] or mobilized peripheral blood[3-5]. Read More

In addition, the FA genetic defect fragilizes the HSCs[6]. These particular features might explain why the first clinical trials using murine leukemia virus derived retroviral vectors conducted for FA failed to show engraftment of corrected cells. The gene therapy field is now moving towards the use of lentiviral vectors (LVs) evidenced by recent succesful clinical trials for treatment of patients suffering from adrenoleukodystrophy (ALD)[7], β-thalassemia[8], metachromatic leukodystrophy[9] and Wiskott-Aldrich syndrome[10]. LV trials for X-linked severe combined immunodificiency and Fanconi anemia (FA) defects were recently initiated[11, 12]. Fifteen years of preclinical studies using different FA mouse models and in vitro research allowed us to find the weak points in the in vitro culture and transduction conditions, which most probably led to the initial failure of FA HSC gene therapy. In this review we will focus on the different obstacles, unique to FA gene therapy, and how they been overcome through the development of optimized protocols for FA HSC culture and transduction and the engineering of new gene transfer tools for FA HSCs. These combined advances in the field hopefully will allow the correction of the FA hematological defect in the near future.

Ann Palliat Med
Ann Palliat Med 2016 Dec 9. Epub 2016 Dec 9.
Department of Radiotherapy and Oncology, Kasturba Medical College and Hospital, Manipal University, Manipal, Karnataka 576104, India.
J. Pediatr. Hematol. Oncol.
J Pediatr Hematol Oncol 2017 Jan 5. Epub 2017 Jan 5.
*Department of Pediatrics, Faculty of Medicine, Kuwait University, Safat †Pediatric Hematology Unit, Mubarak Al-Kabeer Hospital, Jabriya ‡Advanced Technology Company, Salmiya, Kuwait.
Int Ophthalmol
Int Ophthalmol 2017 Jan 4. Epub 2017 Jan 4.
Department of Ophthalmology, Kayseri Education and Research Hospital, Sanayi Mah. Atatürk Boulevard Hastane Street No: 78, 38010, Kayseri, Turkey.
Malays J Pathol
Malays J Pathol 2016 Dec;38(3):235-239
Universiti Teknologi MARA (UiTM), Faculty of Medicine, Sg Buloh Campus, Malaysia.

Thalassemia and hemoglobinopathies are inherited red blood cell disorders found worldwide. Hemoglobin (Hb) E disorder is one of the hemoglobinopathies known to have the high prevalence in South East Asia. Most of transfusion-dependent thalassemias were genotypically compound heterozygous Hb E/ β-thalassemia. Read More

In Malaysia, the national screening program for thalassemia was implemented for early pregnancy or secondary school girls; however many participants do not turn-up and missed the screening test. Screening for thalassemia using samples from cord blood is an alternative choice as it is a readily available source of blood and hence early detection of the disease. The purpose of this study was to determine the potential use of cord blood for the screening of HbE hemoglobinopathy by using capillary electrophoresis (CE).
Cord blood samples were collected from 300 newborns of healthy mothers. Hematological parameters were determined and hemoglobin quantitation for all cord blood samples were performed using capillary electrophoresis system (CES) and high performance liquid chromatography (HPLC).
Majority of cord blood samples (63%) revealed Hb AF followed by Hb AFA2 (20%). Hb AFE was detected in 10.7% with the mean value of Hb E ranging from 2.3%-11.1%.
Hemoglobin E was detected in cord blood using capillary electrophoresis system. It can be recommended in areas where Hb E/β is prevalent. Implementation of a screening strategy using CE on cord blood sampling will identify the disease early. With regular follow-up on these patients, the status of their disease can be determined earlier and appropriate management implemented.

Zhongguo Shi Yan Xue Ye Xue Za Zhi
Zhongguo Shi Yan Xue Ye Xue Za Zhi 2016 Dec;24(6):1828-1832
Medical Genetics Center, Key Laboratory of Metabolic and Genetic Diseases of Women and Children,Guangdong Women and Children Hospital, Guangzhou 511442, Guangdong Province, China. E-mail: