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  • Endothelial Cell Interactions and Inflammation in Sickle Cell Disease

    FERNANDO F.COSTA

    University of Campinas -

    UNICAMP, Brazil

    http://www.fapesp.br/

  • Hemoglobina

  • HEMOGLOBINOPHATIES

    World :every year 300 000 infants are born

    with major hemoglobin disorders

    Brazil :25000-35000 SCA patients

    Bahia:1 out of every 500 births

    US :70000 -100000 SCA patients

    1 out of every 500 African

    Americans births

  • Costa: Hematologia, Fundamentos e Prtica 2001

    The Pathophysiology of Sickle Cell Anemia

  • Vaso-occlusion in Sickle Cell Disease

    Vascular

    Occlusion

    Tissue Infarction Hemolytic Anemia

    Pain episodes

    Splenic Hypofunction

    Infections

    Acute chest syndrome Pulmonary hypertension

    Stroke Multi-organ damage

    HbS

  • Vaso-occlusion in Sickle Cell Disease

    Vascular

    Occlusion

    Tissue Infarction Hemolytic Anemia

    Pain episodes

    Splenic Hypofunction

    Infections

    Acute chest syndrome Pulmonary hypertension

    Stroke Multi-organ damage

    HbS

    HU therapy

  • Vaso-occlusion in Sickle Cell Disease

    Vascular

    Occlusion

    Tissue Infarction Hemolytic Anemia

    Pain episodes

    Splenic Hypofunction

    Infections

    Acute chest syndrome Pulmonary hypertension

    Stroke Multi-organ damage

    HbS

    HU therapy

  • NO

    Hypoxia

    ROS

    ET-1

    Vaso-occlusion pathophysiology: Role of endothelial

    activation, vascular inflammation and cell adhesion

  • NO

    Hypoxia

    ROS

    ET-1

    Vaso-occlusion pathophysiology: Role of endothelial

    activation, vascular inflammation and cell adhesion

    Adhesion molecule

    expression

  • NO

    Hypoxia

    ROS

    ET-1 IL-8

    IL-1

    TNF

    Vaso-occlusion pathophysiology: Role of endothelial

    activation, vascular inflammation and cell adhesion

    Adhesion molecule

    expression

  • NO

    Hypoxia

    ROS

    ET-1 IL-8

    IL-1

    TNF

    TF

    PAF

    vWF

    Vaso-occlusion pathophysiology: Role of endothelial

    activation, vascular inflammation and cell adhesion

    Adhesion molecule

    expression

  • Vaso-occlusion pathophysiology: Role of endothelial

    activation, vascular inflammation and cell adhesion

  • Vaso-occlusion pathophysiology: Role of endothelial

    activation, vascular inflammation and cell adhesion

  • Reduce Hemolysis

    Reduce Inflammation

    Reduce Endothelial Activation

    Reduce Red Cell Adhesion

    Reduce Neutrophil Adhesion

    Increase NO Bioavailability

    Reduce Oxidative Stress

    Targeting Vaso-occlusion:

  • Reduce Hemolysis

    Reduce Inflammation

    Reduce Endothelial Activation

    Reduce Red Cell Adhesion

    Reduce Neutrophil Adhesion

    Increase NO Bioavailability

    Reduce Oxidative Stress

    Targeting Vaso-occlusion:

  • Plasma Inflammatory Proteins in SCD

    Lanaro et al., 2009

    Targeting Vascular Inflammation:

    Control SCA SCAHU0.0

    2.5

    5.0

    7.5

    P=0.006

    P=0.006

    TN

    F-

    (p

    g/m

    l)

    TNF-

  • IL-8

    Plasma Inflammatory Proteins in SCD

    Lanaro et al., 2009; Garrido, in submisso

    Targeting Vascular Inflammation:

    Control SCA SCAHU0

    10

    20

    30

    40

    50

    60

    P

  • IL-8

    Plasma Inflammatory Proteins in SCD

    Lanaro et al., 2009; Garrido, in submisso

    Targeting Vascular Inflammation:

    Control SCA SCAHU0

    10

    20

    30

    40

    50

    60

    P

  • Plasma Inflammatory Proteins in SCD

    Fertrin, unpublished

    Targeting Vascular Inflammation:

    IL-6 (pg/mL)

    CTR

    LSS

    SSH

    U

    0

    5

    10

    15

    pla

    sm

    a I

    L-6

  • Pro-coagulant factors in SCD

    Garrido, in submission

    Targeting Vascular Inflammation:

    Tissue Factor

    Control SCA SCAHU0

    100

    200

    300

    400

    500

    P

  • Endothelial Adhesion Molecules

    Targeting Vascular Inflammation:

    Control SCA SCAHU0

    100

    200

    300

    400

    P

  • TNF- SS RBC Contact

    Heme

    Hypoxia

    ROS

    NO

    The Endothelium and Vascular Inflammation

  • NFB

    eNOS function

    XO BH4

    TNF- SS RBC Contact

    Heme

    Hypoxia

    ROS

    NO

    The Endothelium and Vascular Inflammation

    ET-1

    ET-3

    Vasoconstriction

  • NFB

    eNOS function

    XO BH4

    Cell Adhesion

    Adhesion Molecule Expression

    TNF-

    The Endothelium and Vascular Inflammation

    ET-1

    ET-3

    Vasoconstriction

  • NFB

    eNOS function

    XO BH4

    Cell Adhesion

    Adhesion Molecule Expression

    IL-1, IL-8,

    GM-CSF,

    IL-6

    Inflammation

    TNF-

    Leukocyte Activation

    CRP

    The Endothelium and Vascular Inflammation

    ET-1

    ET-3

    Vasoconstriction

  • NFB

    eNOS function

    XO BH4

    Cell Adhesion

    Adhesion Molecule Expression

    IL-1, IL-8,

    GM-CSF,

    IL-6

    vWF, TXA2, PAF,

    Tissue Factor

    Inflammation

    Coagulation

    TNF-

    Leukocyte Activation

    CRP

    The Endothelium and Vascular Inflammation

    ET-1

    ET-3

    Vasoconstriction

  • NFB

    eNOS function

    XO BH4

    Cell Adhesion

    Adhesion Molecule Expression

    IL-1, IL-8,

    GM-CSF,

    IL-6

    vWF, TXA2, PAF,

    Tissue Factor

    Inflammation

    Coagulation

    TNF-

    Platelets CD40L

    LIGHT

    Leukocyte Activation

    CRP

    Thrombin

    The Endothelium and Vascular Inflammation

    ET-1

    ET-3

    Vasoconstriction

  • NFB

    eNOS function

    XO BH4

    Cell Adhesion

    Adhesion Molecule Expression

    IL-1, IL-8,

    GM-CSF,

    IL-6

    vWF, TXA2, PAF,

    Tissue Factor

    Inflammation

    Coagulation

    TNF-

    Platelets CD40L

    LIGHT

    Leukocyte Activation

    CRP

    Thrombin

    ET-1

    ET-3

    Vasoconstriction

    The Endothelium and Vascular Inflammation

  • NFB

    eNOS function

    XO BH4

    Cell Adhesion

    Adhesion Molecule Expression

    IL-1, IL-8,

    GM-CSF,

    IL-6

    vWF, TXA2, PAF,

    Tissue Factor

    Inflammation

    Coagulation

    TNF-

    Platelets CD40L

    LIGHT

    Leukocyte Activation

    CRP

    Thrombin

    ET-1

    ET-3

    Vasoconstriction

    Hydroxyurea Therapy and Vascular Inflammation

  • Reduce Hemolysis

    Reduce Inflammation

    Reduce Endothelial Activation

    Reduce Red Cell Adhesion

    Reduce Neutrophil Adhesion

    Increase NO Bioavailability and cGMP signalling

    Reduce Oxidative Stress

    Targeting Vaso-occlusion:

  • Control SCD SCDHU0

    3

    6

    9

    12

    15

    18

    ***

    #

    % A

    dh

    esio

    n t

    o F

    N

    Control SCD SCDHU0

    5

    10

    15

    20

    25*

    #

    % A

    dh

    esio

    n t

    o IC

    AM

    -1

    Basal 10nM TNF- Basal 10nM TNF-0

    10

    20

    30

    40

    50

    *

    *

    Control SCD

    #

    % n

    eu

    tro

    ph

    il a

    dh

    es

    ion

    to

    HU

    VE

    C

    SCA Neutrophil adhesion properties are increased in vitro

    FN

    ICAM-1

    HUVEC

    Canalli et al., 2008; 2011

  • Reduce Hemolysis

    Reduce Inflammation

    Reduce Endothelial Activation

    Reduce Red Cell Adhesion

    Reduce Neutrophil Adhesion

    Increase NO Bioavailability and cGMP signalling

    Reduce Oxidative Stress

    Targeting Vaso-occlusion:

  • NO-cGMP pathway in SCD

    PKG

    NO

    Soluble guanylate

    cyclase (sGC)

    GTP cGMP

    -globin

    HbF

    Cell adhesion

    Almeida et al., 2008; Cokic et

    al., 2003; Ikuta et al., 2001

    RED

    CELLS

    WHITE

    CELLS

  • BASAL

    DEA

    NO

    BASAL

    DEA

    NO

    0

    5

    10

    15

    Control SCD

    ***

    ##

    % A

    dh

    esio

    n t

    o F

    N

    Bas

    al

    DEANO

    Bas

    al

    DEANO

    0

    5

    10

    15

    20

    25

    Control SCD

    **

    ##

    % A

    dh

    es

    ion

    to

    IC

    AM

    -1

    NO donors reduce SCD neutrophil adhesive properties in vitro

    Canalli et al., 2008

  • Hydroxyurea

    PKG

    NO

    Soluble guanylate cyclase

    (sGC)

    GTP cGMP

    Erythrocytic lineage cells

    -globin

    HbF

    Leukocytes Cell adhesion

    The cGMP-dependent pathway as a potential therapeutic target for SCA

    vaso-occlusion

    King, 2004

    http://upload.wikimedia.org/wikipedia/commons/c/c9/Hydroxyurea.png

  • Hydroxyurea

    PKG

    NO

    Soluble guanylate cyclase

    (sGC)

    GTP cGMP PDE 9