Maria Francesca Manchinu
Technologist
Area of interest:
Many years of expertise and experience in molecular and cellular biology, developmental biology, gene regulation and expression and in the use and maintenance of murine models.
My current interests revolve around the identification of molecular and cellular origins of both immunodeficiency and autoimmunity diseases and understand the biological mechanisms that are implicated in immunosenescence by the analysis of knock-out and transgenic mice as models of autoimmunity.
I’m also interested in understanding the regulation of human globin genes expression as well as hemoglobin switching, and transcription factors connected to them through the study in vitro and in vivo.
Alternative therapeutic approaches to improve the diagnostic and treatment of Beta Thalassemia and sickle-cell anemia.
In addition to research, training of undergraduate and post graduate students.
Most significant publications:
2020
Manchinu, M. F.; Simbula, M.; Caria, C. A.; Musu, E.; Perseu, L.; Porcu, S.; Steri, M.; Poddie, D.; Frau, J.; Cocco, E.; Manunza, L.; Barella, S.; Ristaldi, M. S.
Delta-Globin Gene Expression Is Enhanced in vivo by Interferon Type I Journal Article
In: Front Med (Lausanne), 7 , pp. 163, 2020.
@article{pmid32528964,
title = {Delta-Globin Gene Expression Is Enhanced in vivo by Interferon Type I},
author = {Manchinu, M. F. and Simbula, M. and Caria, C. A. and Musu, E. and Perseu, L. and Porcu, S. and Steri, M. and Poddie, D. and Frau, J. and Cocco, E. and Manunza, L. and Barella, S. and Ristaldi, M. S.},
year = {2020},
date = {2020-01-01},
journal = {Front Med (Lausanne)},
volume = {7},
pages = {163},
abstract = {Beta hemoglobinopathies are widely spread monogenic lethal diseases. Delta-globin gene activation has been proposed as a possible approach for curing these pathologies. The therapeutic potential of delta-globin, the non-alpha component of Hemoglobin A2 (α2δ2; HbA2), has been demonstrated in a mouse model of beta thalassemia, while its anti-sickling effect, comparable to that of gamma globin, was established some time ago. Here we show that the delta-globin mRNA level is considerably increased in a Deoxyribonuclease II-alpha knockout mouse model in which type 1 interferon (interferon beta, IFNb) is activated. IFNb activation in the fetal liver improves the delta-globin mRNA level, while the beta-globin mRNA level is significantly reduced. In addition, we show that HbA2 is significantly increased in patients with multiple sclerosis under type 1 interferon treatment. Our results represent a proof of principle that delta-globin expression can be enhanced through the use of molecules. This observation is potentially interesting in view of a pharmacological approach able to increase the HbA2 level.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Beta hemoglobinopathies are widely spread monogenic lethal diseases. Delta-globin gene activation has been proposed as a possible approach for curing these pathologies. The therapeutic potential of delta-globin, the non-alpha component of Hemoglobin A2 (α2δ2; HbA2), has been demonstrated in a mouse model of beta thalassemia, while its anti-sickling effect, comparable to that of gamma globin, was established some time ago. Here we show that the delta-globin mRNA level is considerably increased in a Deoxyribonuclease II-alpha knockout mouse model in which type 1 interferon (interferon beta, IFNb) is activated. IFNb activation in the fetal liver improves the delta-globin mRNA level, while the beta-globin mRNA level is significantly reduced. In addition, we show that HbA2 is significantly increased in patients with multiple sclerosis under type 1 interferon treatment. Our results represent a proof of principle that delta-globin expression can be enhanced through the use of molecules. This observation is potentially interesting in view of a pharmacological approach able to increase the HbA2 level.
2018
Manchinu, M. F.; Brancia, C.; Caria, C. A.; Musu, E.; Porcu, S.; Simbula, M.; Asunis, I.; Perseu, L.; Ristaldi, M. S.
Deficiency in interferon type 1 receptor improves definitive erythropoiesis in Klf1 null mice Journal Article
In: Cell Death Differ, 25 (3), pp. 589–599, 2018.
@article{pmid29230002,
title = {Deficiency in interferon type 1 receptor improves definitive erythropoiesis in Klf1 null mice},
author = {Manchinu, M. F. and Brancia, C. and Caria, C. A. and Musu, E. and Porcu, S. and Simbula, M. and Asunis, I. and Perseu, L. and Ristaldi, M. S.},
year = {2018},
date = {2018-01-01},
journal = {Cell Death Differ},
volume = {25},
number = {3},
pages = {589--599},
abstract = {A key regulatory gene in definitive erythropoiesis is the transcription factor Krüppel-like factor 1 (Klf1). Klf1 null mice die in utero by day 15.5 (E15.5) due to impaired definitive erythropoiesis and severe anemia. Definitive erythropoiesis takes place in erythroblastic islands in mammals. Erythroblastic islands are formed by a central macrophage (Central Macrophage of Erythroblastic Island, CMEI) surrounded by maturating erythroblasts. Interferon-β (IFN-β) is activated in the fetal liver's CMEI of Klf1 null mice. The inhibitory effect of IFN-β on erythropoiesis is known and, therefore, we speculated that IFN-β could have contributed to the impairment of definitive erythropoiesis in Klf1 knockout (KO) mice fetal liver. To validate this hypothesis, in this work we determined whether the inactivation of type I interferon receptor (Ifnar1) would ameliorate the phenotype of Klf1 KO mice by improving the lethal anemia. Our results show a prolonged survival of Klf1/Ifnar1 double KO embryos, with an improvement of the definitive erythropoiesis and erythroblast enucleation, together with a longer lifespan of CMEI in the fetal liver and also a restoration of the apoptotic program. Our data indicate that the cytotoxic effect of IFN-β activation in CMEI contribute to the impairment of definitive erythropoiesis associated with Klf1 deprivation.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A key regulatory gene in definitive erythropoiesis is the transcription factor Krüppel-like factor 1 (Klf1). Klf1 null mice die in utero by day 15.5 (E15.5) due to impaired definitive erythropoiesis and severe anemia. Definitive erythropoiesis takes place in erythroblastic islands in mammals. Erythroblastic islands are formed by a central macrophage (Central Macrophage of Erythroblastic Island, CMEI) surrounded by maturating erythroblasts. Interferon-β (IFN-β) is activated in the fetal liver's CMEI of Klf1 null mice. The inhibitory effect of IFN-β on erythropoiesis is known and, therefore, we speculated that IFN-β could have contributed to the impairment of definitive erythropoiesis in Klf1 knockout (KO) mice fetal liver. To validate this hypothesis, in this work we determined whether the inactivation of type I interferon receptor (Ifnar1) would ameliorate the phenotype of Klf1 KO mice by improving the lethal anemia. Our results show a prolonged survival of Klf1/Ifnar1 double KO embryos, with an improvement of the definitive erythropoiesis and erythroblast enucleation, together with a longer lifespan of CMEI in the fetal liver and also a restoration of the apoptotic program. Our data indicate that the cytotoxic effect of IFN-β activation in CMEI contribute to the impairment of definitive erythropoiesis associated with Klf1 deprivation.
2014
Manchinu, M. F.; Marongiu, M. F.; Poddie, D.; Casu, C.; Latini, V.; Simbula, M.; Galanello, R.; Moi, P.; Cao, A.; Porcu, S.; Ristaldi, M. S.
In vivo activation of the human δ-globin gene: the therapeutic potential in β-thalassemic mice Journal Article
In: Haematologica, 99 (1), pp. 76–84, 2014.
@article{pmid23872310,
title = {In vivo activation of the human δ-globin gene: the therapeutic potential in β-thalassemic mice},
author = {Manchinu, M. F. and Marongiu, M. F. and Poddie, D. and Casu, C. and Latini, V. and Simbula, M. and Galanello, R. and Moi, P. and Cao, A. and Porcu, S. and Ristaldi, M. S.},
year = {2014},
date = {2014-01-01},
journal = {Haematologica},
volume = {99},
number = {1},
pages = {76--84},
abstract = {β-thalassemia and sickle cell disease are widespread fatal genetic diseases. None of the existing clinical treatments provides a solution for all patients. Two main strategies for treatment are currently being investigated: (i) gene transfer of a normal β-globin gene; (ii) reactivation of the endogenous γ-globin gene. To date, neither approach has led to a satisfactory, commonly accepted standard of care. The δ-globin gene produces the δ-globin of hemoglobin A2. Although expressed at a low level, hemoglobin A2 is fully functional and could be a valid substitute of hemoglobin A in β-thalassemia, as well as an anti-sickling agent in sickle cell disease. Previous in vitro results suggested the feasibility of transcriptional activation of the human δ-globin gene promoter by inserting a Kruppel-like factor 1 binding site. We evaluated the activation of the Kruppel-like factor 1 containing δ-globin gene in vivo in transgenic mice. To evaluate the therapeutic potential we crossed the transgenic mice carrying a single copy activated δ-globin gene with a mouse model of β-thalassemia intermedia. We show that the human δ-globin gene can be activated in vivo in a stage- and tissue-specific fashion simply by the insertion of a Kruppel-like factor 1 binding site into the promoter. In addition the activated δ-globin gene gives rise to a robust increase of the hemoglobin level in β-thalassemic mice, effectively improving the thalassemia phenotype. These results demonstrate, for the first time, the therapeutic potential of the δ-globin gene for treating severe hemoglobin disorders which could lead to novel approaches, not involving gene addition or reactivation, to the cure of β-hemoglobinopathies.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
β-thalassemia and sickle cell disease are widespread fatal genetic diseases. None of the existing clinical treatments provides a solution for all patients. Two main strategies for treatment are currently being investigated: (i) gene transfer of a normal β-globin gene; (ii) reactivation of the endogenous γ-globin gene. To date, neither approach has led to a satisfactory, commonly accepted standard of care. The δ-globin gene produces the δ-globin of hemoglobin A2. Although expressed at a low level, hemoglobin A2 is fully functional and could be a valid substitute of hemoglobin A in β-thalassemia, as well as an anti-sickling agent in sickle cell disease. Previous in vitro results suggested the feasibility of transcriptional activation of the human δ-globin gene promoter by inserting a Kruppel-like factor 1 binding site. We evaluated the activation of the Kruppel-like factor 1 containing δ-globin gene in vivo in transgenic mice. To evaluate the therapeutic potential we crossed the transgenic mice carrying a single copy activated δ-globin gene with a mouse model of β-thalassemia intermedia. We show that the human δ-globin gene can be activated in vivo in a stage- and tissue-specific fashion simply by the insertion of a Kruppel-like factor 1 binding site into the promoter. In addition the activated δ-globin gene gives rise to a robust increase of the hemoglobin level in β-thalassemic mice, effectively improving the thalassemia phenotype. These results demonstrate, for the first time, the therapeutic potential of the δ-globin gene for treating severe hemoglobin disorders which could lead to novel approaches, not involving gene addition or reactivation, to the cure of β-hemoglobinopathies.
2011
Porcu, S.; Manchinu, M. F.; Marongiu, M. F.; Sogos, V.; Poddie, D.; Asunis, I.; Porcu, L.; Marini, M. G.; Moi, P.; Cao, A.; Grosveld, F.; Ristaldi, M. S.
Klf1 affects DNase II-alpha expression in the central macrophage of a fetal liver erythroblastic island: a non-cell-autonomous role in definitive erythropoiesis Journal Article
In: Mol Cell Biol, 31 (19), pp. 4144–4154, 2011.
@article{pmid21807894,
title = {Klf1 affects DNase II-alpha expression in the central macrophage of a fetal liver erythroblastic island: a non-cell-autonomous role in definitive erythropoiesis},
author = {Porcu, S. and Manchinu, M. F. and Marongiu, M. F. and Sogos, V. and Poddie, D. and Asunis, I. and Porcu, L. and Marini, M. G. and Moi, P. and Cao, A. and Grosveld, F. and Ristaldi, M. S.},
year = {2011},
date = {2011-01-01},
journal = {Mol Cell Biol},
volume = {31},
number = {19},
pages = {4144--4154},
abstract = {A key regulatory gene in definitive erythropoiesis is the erythroid Kruppel-like factor (Eklf or Klf1). Klf1 knockout (KO) mice die in utero due to severe anemia, while residual circulating red blood cells retain their nuclei. Dnase2a is another critical gene in definitive erythropoiesis. Dnase2a KO mice are also affected by severe anemia and die in utero. DNase II-alpha is expressed in the central macrophage of erythroblastic islands (CMEIs) of murine fetal liver. Its main role is to digest the DNA of the extruded nuclei of red blood cells during maturation. Circulating erythrocytes retain their nuclei in Dnase2a KO mice. Here, we show that Klf1 is expressed in CMEIs and that it binds and activates the promoter of Dnase2a. We further show that Dnase2a is severely downregulated in the Klf1 KO fetal liver. We propose that this downregulation of Dnase2a in the CMEI contributes to the Klf1 KO phenotype by a non-cell-autonomous mechanism.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A key regulatory gene in definitive erythropoiesis is the erythroid Kruppel-like factor (Eklf or Klf1). Klf1 knockout (KO) mice die in utero due to severe anemia, while residual circulating red blood cells retain their nuclei. Dnase2a is another critical gene in definitive erythropoiesis. Dnase2a KO mice are also affected by severe anemia and die in utero. DNase II-alpha is expressed in the central macrophage of erythroblastic islands (CMEIs) of murine fetal liver. Its main role is to digest the DNA of the extruded nuclei of red blood cells during maturation. Circulating erythrocytes retain their nuclei in Dnase2a KO mice. Here, we show that Klf1 is expressed in CMEIs and that it binds and activates the promoter of Dnase2a. We further show that Dnase2a is severely downregulated in the Klf1 KO fetal liver. We propose that this downregulation of Dnase2a in the CMEI contributes to the Klf1 KO phenotype by a non-cell-autonomous mechanism.
- Monserrato
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