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Laura Breda,
Carla Casu,
Sara Gardenghi,
Nicoletta Bianchi,
Luca Cartegni,
Mohandas Narla,
Karina Yazdanbakhsh,
Marco Musso,
Deepa Manwani,
Jane Little,
Lawrence B Gardner, Dorothy A Kleinert,
Eugenia Prus,
Eitan Fibach,
Robert W Grady,
Patricia J Giardina,
Roberto Gambari,
Stefano Rivella
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ABSTRACT: Preclinical and clinical studies demonstrate the feasibility of treating β-thalassemia and Sickle Cell Disease (SCD) by lentiviral-mediated transfer of the human β-globin gene. However, previous studies have not addressed whether the ability of lentiviral vectors to increase hemoglobin synthesis might vary in different patients.We generated lentiviral vectors carrying the human β-globin gene with and without an ankyrin insulator and compared their ability to induce hemoglobin synthesis in vitro and in thalassemic mice. We found that insertion of an ankyrin insulator leads to higher, potentially therapeutic levels of human β-globin through a novel mechanism that links the rate of transcription of the transgenic β-globin mRNA during erythroid differentiation with polysomal binding and efficient translation, as reported here for the first time. We also established a preclinical assay to test the ability of this novel vector to synthesize adult hemoglobin in erythroid precursors and in CD34(+) cells isolated from patients affected by β-thalassemia and SCD. Among the thalassemic patients, we identified a subset of specimens in which hemoglobin production can be achieved using fewer copies of the vector integrated than in others. In SCD specimens the treatment with AnkT9W ameliorates erythropoiesis by increasing adult hemoglobin (Hb A) and concurrently reducing the sickling tetramer (Hb S).Our results suggest two major findings. First, we discovered that for the purpose of expressing the β-globin gene the ankyrin element is particularly suitable. Second, our analysis of a large group of specimens from β-thalassemic and SCD patients indicates that clinical trials could benefit from a simple test to predict the relationship between the number of vector copies integrated and the total amount of hemoglobin produced in the erythroid cells of prospective patients. This approach would provide vital information to select the best candidates for these clinical trials, before patients undergo myeloablation and bone marrow transplant.
PLoS ONE 01/2012; 7(3):e32345. · 4.09 Impact Factor
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Lauren Mednick,
Shuli Yu,
Felicia Trachtenberg,
Yan Xu, Dorothy A Kleinert,
Patricia J Giardina,
Janet L Kwiatkowski,
Dru Foote,
Vivekanandan Thayalasuthan,
John B Porter,
Alexis A Thompson,
Leann Schilling,
Charles T Quinn,
Ellis J Neufeld,
Robert Yamashita
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ABSTRACT: Thalassemia is an inherited blood disorder that requires lifelong adherence to a complicated and burdensome medical regimen which could potentially impact emotional functioning of patients. The importance of understanding and promoting healthy emotional functioning is crucial not only to psychological well-being, but also to physical health as it has been shown to impact adherence to medical regimens [1-4]. The current study aimed to [1] determine the prevalence of depressive and anxiety symptoms in adolescent and adult patients with thalassemia; and [2] explore possible demographic, medical, and psychosocial correlates of these symptoms in 276 patients (14-58 years old, M age = 27.83; 52% female). Overall, most patients did not report experiencing significant symptoms of anxiety and depression (33% of participants indicated experiencing symptoms of anxiety and 11% symptoms of depression). Females and older patients were more likely to experience these symptoms than males and younger patients. Symptoms of anxiety and depression were positively associated with self-report of difficulty with adherence and negatively associated with quality of life. Given these findings, regular screening for anxiety and depression symptoms could help to identify at-risk individuals to provide them with appropriate psychological support with the goal of improving both emotional and physical health.
American Journal of Hematology 10/2010; 85(10):802-5. · 4.67 Impact Factor
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ABSTRACT: Lentiviral-mediated beta-globin gene transfer successfully treated beta-thalassemic mice. Based on this result, clinical trials were initiated. To date, however, no study has investigated the efficacy of gene therapy in relation to the nature of the different beta-globin mutations found in patients. Most mutations can be classified as beta(0) or beta(+), based on the amount of beta-globin protein produced. Therefore, we propose that a screening in vitro is necessary to verify the efficacy of gene transfer prior to treatment of individual patients. We used a two-phase liquid culture system to expand and differentiate erythroid progenitor cells (ErPCs) transduced with lentiviral vectors. We propose the use of this system to test the efficiency of lentiviral vectors carrying the human beta-globin gene, to correct the phenotype of ErPCs from patients preparing for gene therapy. This new approach might have profound implications for designing gene therapy and for understanding the genotype/phenotype variability observed in Cooley's anemia patients.
Annals of the New York Academy of Sciences 08/2010; 1202:134-40. · 3.15 Impact Factor
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Felicia Trachtenberg,
Dru Foote,
Marie Martin,
Susan Carson,
Thomas Coates,
Owen Beams,
Olivia Vega,
Manuela Merelles-Pulcini,
Patricia J Giardina, Dorothy A Kleinert,
Janet Kwiatkowski,
Alexis A Thompson,
Ellis J Neufeld,
Leann Schilling,
Vivek Thayalasuthan,
Zahra Pakbaz,
Robert Yamashita
American Journal of Hematology 02/2010; 85(5):367-70. · 4.67 Impact Factor
