Infusion of Autologous Retrodifferentiated Stem Cells into Patients with Beta-Thalassemia

Beta-thalassemia is a genetic, red blood cell disorder affecting the beta-globin chain of the adult hemoglobin gene. This results in excess accumulation of unpaired alpha-chain gene products leading to reduced red blood cell life span and the development of severe anemia. Current treatment of this disease involves regular blood transfusion and adjunct chelation therapy to lower blood transfusion–induced iron overload. Fetal hemoglobin switching agents have been proposed to treat genetic blood disorders, such as sickle cell anemia and beta-thalassemia, in an effort to compensate for the dysfunctional form of the beta-globin chain in adult hemoglobin. The rationale behind this approach is to pair the excess normal alpha-globin chain with the alternative fetal gamma-chain to promote red blood cell survival and ameliorate the anemia. Reprogramming of differentiation in intact, mature, adult white blood cells in response to inclusion of monoclonal antibody CR3/43 has been described. This form of retrograde development has been termed “retrodifferentiation”, with the ability to re-express a variety of stem cell markers in a heterogeneous population of white blood cells. This form of reprogramming, or reontogeny, to a more pluripotent stem cell state ought to recapitulate early hematopoiesis and facilitate expression of a fetal and/or adult program of hemoglobin synthesis or regeneration on infusion and subsequent redifferentiation. Herein, the outcome of infusion of autologous retrodifferentiated stem cells (RSC) into 21 patients with beta-thalassemia is described. Over 6 months, Infusion of 3-h autologous RSC subjected to hematopoietic-conducive conditions into patients with beta-thalassemia reduced mean blood transfusion requirement, increased mean fetal hemoglobin synthesis, and significantly lowered mean serum ferritin. This was always accompanied by an increase in mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) in such patients. No adverse side effects in response to the infusion of autologous RSC were noted.This novel clinical procedure may profoundly modify the devastating course of many genetic disorders in an autologous setting, thus paving the way to harnessing pluripotency from differentiated cells to regenerate transiently an otherwise genetically degenerate tissue such as thalassemic blood.

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Association of haplotypes for SNPs in the LTR regions of bovine leukemia virus with hematological indices of cattle

Vavilov Journal of Genetics and Breeding ◽

10.18699/vj19.491 ◽

2019 ◽

Vol 23 (3) ◽

pp. 262-269 ◽

Cited By ~ 2

Author(s):

N. V. Blazhko ◽

S. Kh. Vyshegurov ◽

A. S. Donchenko ◽

K. S. Shatokhin ◽

T. I. Krytsyna ◽

...

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Leukemia Virus ◽

White Blood Cells ◽

Biological Species ◽

Mean Corpuscular Hemoglobin ◽

Corpuscular Hemoglobin ◽

Distribution Width ◽

Hematological Indices ◽

Blood Indices

Molecular typing of BLV samples isolated from Holsteinized Russian Black Pied cattle was carried out, and various cytofluorometric and morphological blood indices were examined. We performed the total count of white blood cells (WBC), lymphocyte (lymf), granulocyte (gran), monocyte (mon), red blood cell (RBC), hemoglobin (HGB), hematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW), platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), and platelet crit count (PCT). The LTR-region of BLV was haplotyped. Only viruses of haplotypes I (0.33±0.03) and III (0.67±0.03) of the eight possible were detected. The ratio of hematologically sick, healthy, and suspected carriers of BLV of haplotypes I and II was comparable with the results of other researchers. The numbers of leukocytes, erythrocytes and platelets in the blood of carriers of haplotype III exceeded the corresponding parameters of cattle affected by the virus of haplotype I. It is interesting to note that the difference in the hemolytic status of animals was manifested not only by the concentration of leukocytes as direct immune agents but also by the count of erythrocytes and platelets, which are not directly involved in the immune response. The number of particles of haplotype III of the BLV circulating in the blood of infected individuals exceeded that of the carriers of haplotype I. In this connection, an assumption was made about the evolutionary advantage of the more virulent haplotype III. However, the results of our own research in conjunction with the data of other scientists indicate that the high virulence of individual virus strains is a consequence of the tendency to implement the maximum possible intensity of the synthesis of virus particles but not of the high damaging effect alone. It is shown that high lethality is evolutionarily disadvantageous for viruses, since the extinction of the carrier as a biological species is fraught with the disappearance of the virus itself.

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Biochemical bases for difference in oxygen affinity of maternal and fetal red blood cells of rattlesnakes

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1993.264.3.r481 ◽

1993 ◽

Vol 264 (3) ◽

pp. R481-R486

Author(s):

F. R. Ragsdale ◽

R. L. Ingermann

Keyword(s):

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Oxygen Affinity ◽

Fetal Hemoglobin ◽

Nucleoside Triphosphate ◽

Biochemical Basis ◽

Crotalus Viridis ◽

Adult Hemoglobin

Pregnancy in Crotalus viridis oreganus is associated with an increase in the nucleoside triphosphate (NTP) concentration and a concomitant decrease in the oxygen affinity of the adult red blood cell. However, although the red blood cells of non-pregnant adults and fetuses have indistinguishable NTP concentrations, they have different oxygen affinities. Therefore, red blood cell NTP concentrations alone cannot account for the oxygen-affinity difference between fetal and maternal red blood cells. Hemoglobins from adult and fetal snakes had similar intrinsic oxygen affinities; however, adult hemoglobin was more responsive to organic phosphate modulation compared with fetal hemoglobin. Structural differences, indicated by native gel electrophoresis and electrophoresis of the globins under denaturing conditions at high pH, corroborated functional differences of hemoglobins from fetus and adult. Therefore, the biochemical basis for the oxygen-affinity difference between maternal and fetal red blood cells in this rattlesnake appears to be unique. It appears to be caused by a functionally distinct fetal hemoglobin and the pregnancy-associated rise in red blood cell NTP levels in the mother.

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The role of membrane skeletal-associated alpha-globin in the pathophysiology of beta-thalassemia

Blood ◽

10.1182/blood.v75.6.1333.bloodjournal7561333 ◽

1990 ◽

Vol 75 (6) ◽

pp. 1333-1336 ◽

Cited By ~ 2

Author(s):

S Sorensen ◽

E Rubin ◽

H Polster ◽

N Mohandas ◽

S Schrier

Keyword(s):

Transgenic Mice ◽

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Beta Thalassemia ◽

Beta Globin ◽

Cell Deformability ◽

Red Blood Cell Deformability ◽

Alpha Globin

The beta-thalassemic mouse provides a useful model for testing hypotheses about the pathophysiology in human beta-thalassemia. The clinical picture of these mice and their red blood cell deformability characteristics are quite similar to those observed in human beta- thalassemia intermedia. The creation of transgenic mice that express human beta-globin (beta s) has provided an opportunity to study the effect of increasing the non-alpha-globin chain production on the thalassemic phenotype. A small increase in beta-globin production produces transgenic mice that are healthier, have almost normal hemoglobin values, and whose red blood cell deformability is increased. We quantified and characterized the membrane skeletal-associated globin in normal, transgenic thal/sickle, and thalassemic mice and showed that only alpha-globin was associated with the membrane skeleton in the pathologic red blood cells, and that the degree of rigidity as measured in the rheoscope correlated directly and closely with the amount of membrane skeletal-associated globin in these abnormal red blood cells.

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Characterization and comparison of the red blood cell membrane damage in severe human alpha- and beta-thalassemia

Blood ◽

10.1182/blood.v79.4.1058.1058 ◽

1992 ◽

Vol 79 (4) ◽

pp. 1058-1063 ◽

Cited By ~ 1

Author(s):

R Advani ◽

S Sorenson ◽

E Shinar ◽

W Lande ◽

E Rachmilewitz ◽

...

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Membrane Damage ◽

Beta Thalassemia ◽

Beta Globin ◽

Cell Membrane Damage ◽

Protein 4.1 ◽

Globin Chains ◽

Alpha Thalassemia ◽

Alpha Globin

Abstract The aim of the present work was to understand the pathophysiology of the severe human thalassemias as represented by beta-thalassemia intermedia and hemoglobin (Hb) H (alpha-thalassemia) disease. We have previously shown that the material properties of the red blood cell (RBC) and its membrane differ in severe alpha- and beta-thalassemia, and we now show that this difference is probably caused by accumulation of alpha-globin chains at the cytoskeleton in beta-thalassemia, whereas beta-globin chains are associated with the cytoskeleton in alpha- thalassemia. In both alpha- and beta-thalassemia, some of these globin chains have become oxidized as evidenced by loss of the free thiols. Furthermore, there is similar evidence of oxidation of protein 4.1 in beta-thalassemia, whereas beta-spectrin appears to be subject to oxidation in alpha-thalassemia. These observations support the idea that the association of partly oxidized globin chains with the cytoskeleton results in oxidation of adjacent skeletal proteins. The abnormality of protein 4.1 in beta-thalassemia is consistent with a prior observation, and is also in accord with the known importance of protein 4.1 in maintenance of membrane stability, a property that is abnormal in beta-thalassemic membranes.

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Characterization and comparison of the red blood cell membrane damage in severe human alpha- and beta-thalassemia

Blood ◽

10.1182/blood.v79.4.1058.bloodjournal7941058 ◽

1992 ◽

Vol 79 (4) ◽

pp. 1058-1063 ◽

Cited By ~ 73

Author(s):

R Advani ◽

S Sorenson ◽

E Shinar ◽

W Lande ◽

E Rachmilewitz ◽

...

Keyword(s):

Blood Cell ◽

Red Blood Cell ◽

Membrane Damage ◽

Beta Thalassemia ◽

Beta Globin ◽

Cell Membrane Damage ◽

Protein 4.1 ◽

Globin Chains ◽

Alpha Thalassemia ◽

Alpha Globin

The aim of the present work was to understand the pathophysiology of the severe human thalassemias as represented by beta-thalassemia intermedia and hemoglobin (Hb) H (alpha-thalassemia) disease. We have previously shown that the material properties of the red blood cell (RBC) and its membrane differ in severe alpha- and beta-thalassemia, and we now show that this difference is probably caused by accumulation of alpha-globin chains at the cytoskeleton in beta-thalassemia, whereas beta-globin chains are associated with the cytoskeleton in alpha- thalassemia. In both alpha- and beta-thalassemia, some of these globin chains have become oxidized as evidenced by loss of the free thiols. Furthermore, there is similar evidence of oxidation of protein 4.1 in beta-thalassemia, whereas beta-spectrin appears to be subject to oxidation in alpha-thalassemia. These observations support the idea that the association of partly oxidized globin chains with the cytoskeleton results in oxidation of adjacent skeletal proteins. The abnormality of protein 4.1 in beta-thalassemia is consistent with a prior observation, and is also in accord with the known importance of protein 4.1 in maintenance of membrane stability, a property that is abnormal in beta-thalassemic membranes.

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The role of membrane skeletal-associated alpha-globin in the pathophysiology of beta-thalassemia

Blood ◽

10.1182/blood.v75.6.1333.1333 ◽

1990 ◽

Vol 75 (6) ◽

pp. 1333-1336 ◽

Cited By ~ 22

Author(s):

S Sorensen ◽

E Rubin ◽

H Polster ◽

N Mohandas ◽

S Schrier

Keyword(s):

Transgenic Mice ◽

Blood Cell ◽

Red Blood Cells ◽

Red Blood Cell ◽

Blood Cells ◽

Beta Thalassemia ◽

Beta Globin ◽

Cell Deformability ◽

Red Blood Cell Deformability ◽

Alpha Globin

Abstract The beta-thalassemic mouse provides a useful model for testing hypotheses about the pathophysiology in human beta-thalassemia. The clinical picture of these mice and their red blood cell deformability characteristics are quite similar to those observed in human beta- thalassemia intermedia. The creation of transgenic mice that express human beta-globin (beta s) has provided an opportunity to study the effect of increasing the non-alpha-globin chain production on the thalassemic phenotype. A small increase in beta-globin production produces transgenic mice that are healthier, have almost normal hemoglobin values, and whose red blood cell deformability is increased. We quantified and characterized the membrane skeletal-associated globin in normal, transgenic thal/sickle, and thalassemic mice and showed that only alpha-globin was associated with the membrane skeleton in the pathologic red blood cells, and that the degree of rigidity as measured in the rheoscope correlated directly and closely with the amount of membrane skeletal-associated globin in these abnormal red blood cells.

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beta-thalassemia intermedia in a Brazilian patient with - 101(C > T) and codon 39 (C > T) mutations

Sao Paulo Medical Journal ◽

10.1590/s1516-31802003000100007 ◽

2003 ◽

Vol 121 (1) ◽

pp. 28-30

Author(s):

Sylvia Morais de Sousa ◽

Letícia Khater ◽

Luís Antônio Peroni ◽

Karine Miranda ◽

Marcelo Jun Murai ◽

...

Keyword(s):

Clinical Course ◽

Globin Gene ◽

Fetal Hemoglobin ◽

Hypochromic Anemia ◽

Beta Thalassemia ◽

Beta Globin ◽

Thalassemia Intermedia ◽

Mean Cell Volume ◽

Molecular Alterations ◽

Brazilian Patient

CONTEXT: We verified molecular alterations in a 72-year-old Brazilian male patient with a clinical course of homozygous beta-thalassemia intermedia, who had undergone splenectomy and was surviving without regular blood transfusions. The blood cell count revealed microcytic and hypochromic anemia (hemoglobin = 6.5 g/dl, mean cell volume = 74 fl, mean cell hemoglobin = 24 pg) and hemoglobin electrophoresis showed fetal hemoglobin = 1.3%, hemoglobin A2 = 6.78% and hemoglobin A = 79.4%. OBJECTIVE: To identify mutations in a patient with the symptoms of beta-thalassemia intermedia. DESIGN: Molecular inquiry into the mutations possibly responsible for the clinical picture described. SETTING: The structural molecular biology and genetic engineering center of the Universidade Estadual de Campinas, Campinas, Brazil. PROCEDURES: DNA extraction was performed on the patient's blood samples. The polymerase chain reaction (PCR) was done using five specific primers that amplified exons and the promoter region of the beta globin gene. The samples were sequenced and then analyzed via computer programs. RESULTS: Two mutations that cause the disease were found: -101 (C > T) and codon 39 (C > T). CONCLUSIONS: This case represents the first description of 101 (C > T) mutation in a Brazilian population and it is associated with a benign clinical course.

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