ARCHITETTURA DELLA RICERCA Lucio Luzzatto, Scientific Director, Istituto Toscano Tumori, Firenze, ITALY “Cosa stai cercando” 17 novembre 2010
Welcome message from author
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
ARCHITETTURA DELLA RICERCA
Lucio Luzzatto,Scientific Director,
Istituto Toscano Tumori,Firenze, ITALY
“Cosa stai cercando”17 novembre 2010
RICERCA BIOMEDICA
•Osservazionale•Sperimentale
MacroMacro--evolutionevolution
Problemipostidallapraticaclinica
Nuoviportatidallaricercadi base
PROGETTO DI RICERCA
• Delineare il progetto: obiettivi e criteri di importanza scientifica
• Elaborazione: letteratura, metodologie, piano sperimentale, considerazioni statistiche
• Preventivo economico• Principi etici e Comitato Etico• Realizzazione del progetto• Rendicontazione scientifica• Rendicontazione economica
HemoglobinuriaHemoglobinuria: : Sometimes Paroxysmal, Sometimes NocturnalSometimes Paroxysmal, Sometimes Nocturnal
WORLDWIDE PNH PATIENTSWORLDWIDE PNH PATIENTS
22
88
77
3838 132132
33
• Intravascular Hemolysis
• Thrombosis
• Cytopenias
Paroxysmal Nocturnal Hemoglobinuria(PNH) is an acquired disorder defined by a clinical triad:
Patient RBCPatient RBCControlControlRBCRBC
dAcdAc dSdS dAcdAc dHidHi pHipHi pAcpAc HH2200
Ham Test in a PNH Patient
QuickTime™ and aGIF decompressor
are needed to see this picture.
CLONAL ORIGIN OF PNH CELLS CLONAL ORIGIN OF PNH CELLS IN A G6PD A/B HETEROZYGOTEIN A G6PD A/B HETEROZYGOTE PATIENTPATIENT
G6PDG6PD--AA G6PDG6PD--BB Whole RBCWhole RBC PNH PNH RBCRBC
ControlsControls PatientPatient
University College University College HospitalHospitalIbadan NigeriaIbadan Nigeria
TThe Complement he Complement CCascaascadede
Cell ActivationCell ActivationLysisLysis
C6 C7 C8 C9C6 C7 C8 C9
Potent AnaphylatoxinPotent AnaphylatoxinChemotaxisChemotaxis
Cell ActivationCell Activation
C5bC5b--99
C3 ConvertaseC3 Convertase
C3C3
C1qC1q Activated C1Activated C1
C4+C2C4+C2
C5 ConvertaseC5 Convertase
C3bC3b
Classical PathwayClassical PathwayActivationActivation
Antibody/Antigen ComplexesAntibody/Antigen Complexes
C3aC3aWeakWeak
AnaphylatoxinAnaphylatoxin
C4b2aC4b2a C4b2a3bC4b2a3b
C5bC5b
C5aC5a
C5C5Immune ComplexImmune Complexand Microbialand MicrobialOpsonizationOpsonization
Factor B+DFactor B+DC3 ConvertaseC3 Convertase C5 ConvertaseC5 ConvertaseC3, C3HC3, C3H22OO C3bC3b
Alternative PathwayAlternative PathwayActivationActivation
Microbiological MembranesMicrobiological MembranesBacterial LPSBacterial LPS
Immune ComplexesImmune ComplexesMammalian Cell MembranesMammalian Cell Membranes
C3bBb3bC3bBb3bC3bBbC3bBb
LectinLectin PathwayPathwayActivationActivation
MBLMBL
adaptedadapted fromfrom: Meri: Meri etet al, al, 19901990
C5bC5b--8844ººCCC9 loosely boundC9 loosely bound(displaceable)(displaceable)
C9C9
C5bC5b--883737ººCCC9 tightly boundC9 tightly bound(not displaceable)(not displaceable)
C9C9
C5bC5b--88
PolyPoly--C9C9
C5bC5b--88
C9C9
C5bC5b--88
CD59CD59C9C9
Mechanism of Action of CD59Mechanism of Action of CD59
with CD59with CD59
without CD59without CD59QuickTime™ and aGIF decompressor
are needed to see this picture.
CD59, CD90, CD109
CD55CD58*CD59CD48CDw52PrPcCD16*
CD24 CD55CD58* CD59 CD48 PrPCCD73
CDw108
CD55CD58*CD59CD109PrPCGP500Gova/b
CD55CD58*CD59PrPCAChEJMH AgDombrochHG Ag
CD55 CD58*CD59 CD14CD16 CD24CD48 CD66bCD66c CD87CD109 CD157LAPNB1 PrPCp50-80 GPI-80ADP-RT
NA1/NA2
CD14 CD55CD58*
CD59 CD48 CDw52
CD87 CD109CD157
Group 8 PrPC GPI-80CD16*
CD55 CD58*CD59 CD48CDw52 CD87 CDw108 PrPcADP-RT CD73CD90
CD109CD16*
Proteins Deficient on PNH Blood Cells
Hematopoietic Stem Cell
Platelets
PMNMonocytes
NK cells
RBC
B cells
T cells
QuickTime™ and aGIF decompressor
are needed to see this picture.
all these proteins are GPIall these proteins are GPI--linkedlinked
[From Kinoshita et al., J Biochem (JPN) 144:287,2008]
THE BIOSYNTHETIC PATHWAY OF THE GPI ANCHOR
INVOLVES AT LEAST TEN DISCRETE STEPS
nucleus
ER
X chr.
PIG-A
+GPIprotein
Etn-P GlcN
Inositol-P
glycan
PIG-A
GPI anchornucleus
ER
X chr.
PIG-A
+GPIprotein
PIG-A
GPI anchor
Etn-P GlcN
Inositol-P
glycan
Cellula Staminale Ematopoietica
Normale NormaleEPN
Emoglobinuria Parossistica Notturna (EPN)Patogenesi
..
4
982 11898497161
1
1452Null mutations
3
AvaIpolymorphism
del 735 bp
Large deletions
***** *
****
***
**
50 bp
**
652
del exons 3-4-5
Non-null mutations
Mutations in the PIG-A Gene
A. Normal, steady state
C3b
C3
+
CD55
CD55 CD59
C3b
C3b
C5 convertase
Classicalpathway
Lectinpathway
Alternativepathway
PhysiologicalC3 tick-over
C6 C7 C8 C9 MAC
CD55
CD55 CD59
C5b
C5
A normal (CD55+, CD59+) red cell can cope with the threat of complement activation
(From Luzzatto, Risitano & Notaro, 2010)
C3b
C3
+ C5b
C5
C3b
C5 convertase
MAC-mediated intravascular hemolysis
Classicalpathway
Lectinpathway
Alternativepathway
C6 C7 C8 C9 MAC
PhysiologicalC3 tick-over
C3bC3b
B. PNH, steady state
An abnormal (CD55-, CD59-) red cell (PNH red cell) will be lysed sooner or later by activated complement
(From Luzzatto, Risitano & Notaro, 2010)
TARGETING THE COMPLEMENT TARGETING THE COMPLEMENT CCASCAASCADEDE
Cell ActivationCell ActivationLysisLysis
C6 C7 C8 C9C6 C7 C8 C9
Potent Potent AnaphylatoxinAnaphylatoxinChemotaxisChemotaxis
Cell ActivationCell Activation
C5bC5b--99
C3 C3 ConvertaseConvertase
C3C3
C1qC1q Activated C1Activated C1
C4+C2C4+C2
C5 C5 ConvertaseConvertase
C3bC3b
Classical PathwayClassical PathwayActivationActivation
Antibody/Antigen ComplexesAntibody/Antigen Complexes
C3aC3aWeakWeak
AnaphylatoxinAnaphylatoxin
C4b2aC4b2a C4b2a3bC4b2a3b
C5bC5b
C5aC5a
C5C5Immune ComplexImmune Complexand Microbialand MicrobialOpsonizationOpsonization
Factor B+DFactor B+DC3 C3 ConvertaseConvertase C5 C5 ConvertaseConvertase
C3, C3HC3, C3H22OO C3bC3b
Alternative PathwayAlternative PathwayActivationActivation
Microbiological MembranesMicrobiological MembranesBacterial LPSBacterial LPS
Immune ComplexesImmune ComplexesMammalian Cell MembranesMammalian Cell Membranes
C3bBb3bC3bBb3bC3bBbC3bBb
LectinLectin PathwayPathwayActivationActivation
MBLMBL
XX
AntiAnti--C5 C5 (ECULIZUMAB)(ECULIZUMAB)
CH1
CH
3C
H2
hinge
EculizumabEculizumab (anti(anti--C5 antibody)C5 antibody)
human IgG4 heavy chainconstant regions 2 and 3
complementarity determining regions(murine origin)
CL
human framework regions
human IgG2 heavy chainconstant region 1 and hinge
Levels of Lactate Dehydrogenase and PNH Type III Erythrocytes during Treatment with Eculizumab
Hillmen P et al. N Engl J Med 2006;355:1233-1243
ECULIZUMAB ARRESTS THE SELECTIVE INTRAVASCULAR
LYSIS OF PNH III RED CELLS
(Hillmen et al., New Eng J Med355:1233,2006)
Kaplan-Meier Curves for the Time to the First Transfusion during Treatment
Hillmen P et al. N Engl J Med 2006;355:1233-1243
ECULIZUMAB CAN ABROGATE THE NEED FOR BLOOD TRANSFUSION
(Hillmen et al., New Eng J Med 355:1233,2006)
Change in Fatigue Scores from Baseline to Week 26
Hillmen P et al. N Engl J Med 2006;355:1233-1243
ECULIZUMAB HAS AN IMPACT ON THE QUALITY OF LIFE
(Hillmen et al., New Eng J Med 355:1233,2006)
3D STRUCTUREOF THE EPIDERMALGROWTH FACTORRECEPTOR
THE IRESSA/LUNG CANCER STORY. I.
• Non-small cell lung cancer (NSCLC) is a major cause of death in many countries
• NSCLC over-express the epidermal growth factor receptor (EGFR) in 40-80% of cases
• Gefitinib (Iressa) is an inhibitor of the TyrKin activity of EGFR
(From Lynch et al., NEJM 350:2129,2004)
Response to Gefitinib in a Patient with Refractory Non–Small-Cell Lung Cancer
(From Lynch et al., NEJM 350:2129,2004)
THE IRESSA/LUNG CANCER STORY. II.
• In a study at Mass General Hospital 25 out of 275 patients had an impressive clinical response to gefitinib
• In 8 out of 9 responders in whom the EGFR gene was sequenced a somatic mutation was found within the kinase domain
• No mutations were found in 7 out of 7 non-responders(From Lynch et al., NEJM 350:2129,2004)
RICERCA NEL SETTORE SALUTE
• Geografia• Storia• Tipologie
– Epidemiologia– Prevenzione– Diagnosi– Cure– Procedure infermieristiche– Qualità di vita– Modalità somministrazione cure– Aspetti economici
PROGETTI ITT
• Ricerca sul CANCRO in TOSCANA– Cancer Research– Clinical Cancer Research– Epidemiology– Supportive care
• Università, Ospedali, CNR• Peer review in due stadi: referees
esterni, ISAB• Progetti approvati
– 2007: 25 su 85– 2009: 33 su 109
ITT INTERNATIONAL SCIENTIFIC ADVISORY BOARD
• Sydney BRENNER (Cambridge), Chair• Dino AMADORI, Forlì• Andrea ARDIZZONI, Parma• Paolo BRUZZI, Genova• Giannino DEL SAL, Trieste• Jean-Claude Horiot, Genolier• Pier Paolo PANDOLFI, Boston• Robert PINEDO, Amsterdam• Paolo VINEIS, London
NOMINA NUMINA(what’s in a name?)
• Grant• Principal investigator
(PI)• Post-doctoral Fellow
(post-doc)• Graduate Student• Resident• Fellow
• Finanziamento• Titolare della ricerca
(capo-cordata)• Borsista; Co.co.co.;
(precario)• Dottorando• Specializzando• Specializzando
CLINICAL QUALITY CONTROLSmade user friendly
MORTALITY AND MORBIDITY REVIEWS•Regular periodicity•Appropriate notice•Mix of flagged and random cases•Collegial•Analysis carried out by an independent
but internal colleague•Frank discussion•No attacks•Experienced sensible chairman essential
ETHICAL and REGULATORY ISSUES IN THE CLINICAL USE
OF STEM CELLS & GENE TRANSFER
• Compliance with Good Manufacturing Practice (GMP) standards
• Compliance with all rules and regulations on clinical trials
• Ethics Committee approval
ETHICAL and REGULATORY ISSUES IN THE CLINICAL USE OF GENE
TRANSFER
An investigator involved in a clinical trial should not have any financial interests in any aspect of the trial itself
American Society of Gene Therapy. 1999
STEPS IN CANCER MANAGEMENT
• Diagnosis• Staging• Radical treatment• Surveillance of remission• Treatment of relapse• Salvage treatment• Management of patient with
advanced cancer
LA COMUNICAZIONE CON I PAZIENTI CON TUMORI
• Dire/non dire• Quanto dire• Quando dire• Comunicazione con
familiari• Comunicazione con
paziente• Comunicazione e sostegno
psicologico
CAPIRE,CURARE.PREVENIRE IL CANCROAL MEGLIOPER TUTTI
www.ittumori.it
Related Documents
