laboratory of cellular therapy “g. lanzani” scientific
TRANSCRIPT
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LABORATORY OF CELLULAR THERAPY “G. LANZANI”Scientific Director: Dr. M Introna
USC EMATOLOGIA Ospedali Riuniti BergamoDirector: Prof. A Rambaldi
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DL 24/4/2006 n° 219 “Codice comunitario”(attua le 2001/83/CE, 2003/94/CE, 2003/63/CE)
Regola tutto il ciclo di vita dei prodotti farmaceuticidalla produzione alla commercializzazione.
Allegato I. Parte IV: medicinali per terapie avanzate (celluleper terapia somatica e/o terapia genica).
Contiene il discusso articolo 28 sulle eccezioni ospedaliereVedi legge 07/07/2009 n° 88, G.U. 161 del 14/07/2009
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Norme per la Sperimentazione Clinica
DPR 21/09/2001 n° 439 per Fasi I
DL026/04/2002 Documentazione ISS per fasi I
DL 24/06/2003 N° 211 (attua 2001/20/CE) per GCP e successivo DL 21/12/2007 (IMPD)
DL 06/11/2007 n°200 (attua 2005/28/CE) per GCP
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DL 02/03/2004 e successivi per “Banca Dati”
DL 05/12/2006 per “Usi Consolidati”e successivi decreti “proroga”
DL 05/12/2006 per “Usi Compassionevoli”
DM 08/05/2003 per “Usi Compassionevoli”
Altre norme per la Sperimentazione Clinica
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DL 06/11/2007 n°191 (attua 2004/23/CE)
DL 25/01/2010 N° 16 (attua 2006/17/CE e 2006/68/CE)
ISTITUTO DEI TESSUTI/CNT
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AIFA
Area produzione e controllo
Ufficio autorizzazioni officine
Il Laboratorio di Terapia Cellulare “G.Lanzani” è autorizzato aprodurre prodotti per terapia cellulare sottoforma di
preparazioni liquide di piccolo volume per uso parenteralepreparate in asepsi.
Roma, 27.07.2007
N° aM-144/2007
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30/07/2007 Preliminary authorization of the “Lanzani” Cell Factory by AIFA
28/11/2008 Official authorization of the “Lanzani” Cell Factoryby AIFA n° aM 189/2008
27/02/2008 2nd General inspection
JULY 07
FEBRUARY 08
NOVEMBER 08
12/04/2007 Inspection of follow up of corrective actionsAPRIL 07
NOVEMBER 09 3-6/11/09 Inspection of follow up
19/12/2005 1st General InspectionDECEMBER 2005
Chronology of the AIFA inspections up to finalauthorization of the Cell Factory
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Other antigens or naive
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Ruolo soppressorio sulla immunità da parte delle cellule MSC
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Le Blanc et al. Lancet 2004
Clinical course and immunosuppression of the patient
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Le Blanc et al, Lancet 2008
Survival from time of haemopoietic stem –cell transplantation inpatients given mesenchymal stem cells
n = 30n = 25
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Production MSC
Expansion from BM Quality controls
Week0 12345678910
1incubatorMedium,½literPL 2‐3bagswith80‐100x106/batch
o1with100x106+2‐3smallbags/batch
LaboratoryTC2:2incubators
EndotoxinLAL
Sterility
MycoplasmaCulture(EP)
Phenotype,viability
From frozenvial P1
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13
25
10
3
Number of frozenvials at p1
(~2x106cells/vial)
3
2
4
1
Frozen bags(mean:80x106cells/bag)
200.0
147.0
375.2
95.0
hMSC Frozen(x 106)
washout
washout
washout
washout
Origin ofmaterial
BG-MSC12
BG-MSC11
BG-MSC7
BG-MSC2
Code
1
20
5
1
n° vials usedFrozen bags
(mean 1x107cells/bags)Code
1BG-MSC12
32BG-MSC11
10BG-MSC7
3BG-MSC2
hMSC EXPANSIONS
…from “fresh material”
…from “frozen vials bank”
Capelli C. et al, BMT, 2007; Capelli C et al., Cytotherapy, 2009
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24/11/2008 Submission of Phase I MSC/aGVHD IMPD to ISS
24/04/2009 Authorization by ISS
16/07/2009 Authorization by E.C. Monza
NOVEMBER 08
APRIL 09
JULY 09
Time scale of approval of Phase I clinicalstudy with MSC in aGVHD
NOVEMBER 09 Authorisation by E.C. Bergamo
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MSC for the treatment of severe aGVHD
• Dossier n° Eudra CT: 2008/007869/23
• Usage of MSC for steroid resistant grade III-IV aGVHD
• Scientific project shared with Prof. A. Biondi and dott. E. Biagi,Unità di Pediatria, Ospedale San Gerardo, Monza, UniversitàMilano Bicocca.
• Clinical PI: Dott. E. Biagi
Authorization by ISS on 24/04/2009Authorization by E.C. Monza on 16/07/2009
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Pts GVHD Grading at MSC Infusion
No. of MSC
Infusion
Infused MSC (Dose/kg)
Response After MSC Infusion Follow-up
1A Chronic overlap syndrome (skin and mucosae grade III)
1 1 x 106/kg Partial response temporary Skin and mucosae grade I
Alive with chronic skin and mucosae GVHD grade II
2A Chronic overlap syndrome (skin and mucosae grade II)
1 1 x 106/kg Complete response definitive No GVHD
Alive with skin chronic GVHD grade I
2B Chronic mucosae and liver grade III
1 1.4 x 106/kg Partial response temporary Chronic mucosae and liver grade II
3A Acute skin grade I 1 1 x 106/kg Complete response definitive No GVHD
Alive, no GVHD
4A Acute skin grade I 1 0.7 x 106/kg No response Skin GVHD grade I
Alive, no GVHD
4B Chronic skin and liver grade III 4 2.8 x 106/kg
Partial response temporary Skin and liver grade II
5A Acute skin grade II 1 1.2 x 106/kg Complete response definitive No GVHD
Alive, no GVHD
5B Chronic skin GVHD grade I 1 1.2 x 106/kg No response Skin GVHD grade I
6A Chronic overlap (skin and mucosae grade III)
1 1.2 x 106/kg Complete response definitive
Alive, no GVHD
7A Acute skin grade I 2 2.4 x 106/kg Complete response definitive No GVHD
Alive, no GVHD
8A Acute (skin II and gut grade IV) 2 3.2 x 106/kg Partial response temporary Skin and gut grade II
Death for progressive gut GVHD And pulmonary aspergillosis
9A Acute (skin and gut grade IV) 2 2 x 106/kg No response Skin and gut grade IV
Death for sepsis
10A Acute gut grade IV 2 2.4 x 106/kg Partial response temporary Grade III
Alive, no GVHD
11A Acute gut grade IV 1 3.7 x 106/kg No response Deah for gut GVHD and pneumonia
Lucchini G. et al, BBMC, 2010, in press
MSC infusions in pediatric patients
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Richiesta di emendamento sostanziale a ISSper allargamento a 13 centri ematologici italiani
e cambio dosaggio (3 somministrazioni al posto di 2)Aspetti logistici critici
Coordinamento pediatrico: MonzaCoordinamento adulti: Bergamo
In attesa di risposta formale da ISS
A novembre 2010 già arruolati 13 pazienti
MSC per il trattamento della GVHD acuta grave
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CIK culturePBMCsDay 0
INFgamma1000 IU/ml
OKT3 50 ng/mlIL2 500 IU/ml
IL2 500 IU/mlevery 3-4 days
Day 1 Day 21
Protocol for generation of cytokine-induced killer cells
CD3
CD
56
CD
56
CD3
CD
56 NK
CD56
T
CD3
CIK
CD3CD56
Introna M. et al, BMT, 2006Franceschetti M. et al., Exp Hematol, 2009Pievani A et al., Blood in pressPievani A. et al., submitted
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Murine CIK cells GFP transduced and i.v. injected in syngenic animals carrying s.c.lymphoma
Edinger, M. et al, Blood 2003
Homing of cytokine-induced killer cells in vivo
Day 0: early localization of CIK cells to the lungs;Day 1: distribution to other sites, including liver and spleen;Day 3: preferential infiltration of subcutaneous tumor site;Day 12: regression of tumor and minimal signal from remaining CIK cells
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Nishimura, R. et al.Blood 2008;112:2563-2574
Trafficking and survival of CIK cells upon adoptive transfer
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Produzione CIK
Produzione Controlli di qualità
Week0 12345678910
1incubatoreTerreno,OKT3,IFNg,IL‐23‐4sacche/produzione
LaboratorioTC1:3incubatori
NostroLALemycoplasma(Mycoalert)
Sterilità
MycoplasmaColtura(P.E.)
Feno[po,vitalità
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• Dossier n° 64499-PRE.21-848
• Titolo: Impiego linfociti CIK: Fase I
• Approvazione in data 30/05/2005
• Progetto scientifico in condivisione con il Prof. A. Biondi eil dott. E. Biagi, Unità di Pediatria, Ospedale San Gerardo,Monza, Università Milano Bicocca.
Cellule CIK da donatore per le ricaduteleucemiche
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Chiusura studio CIK Fase I
05/06 Arruolamento di 17 pazienti tutti singolarmenteautorizzati dall’Ufficio comma c del ISS dietro specifica
richiesta
06.12.06 Invio manoscritto con dati di 11 pazientiPubblicazione su Hematologica 2007, 92, 07, 948-955
06.10.08 Invio relazione finale chiusura Fase I
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Results
Clinical responseNone 6Stable Disease 1Hematologic Improvement 1Complete response 3
Alive/dead 4/7
Days of Median Follow upfrom transplant 782 (188-1327)
Introna et al, 2007
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RECIPIENT
DONOR
Days posttranplant
27
90
120
132
155
174
377
DLI (+102)
CIK (+132)
CIK (+174)
77%
22%
25%
6%
45%
97%
100%
0%
100%
Response of pt 765 as determined by chimerism analysis of CD3+ cells
Introna et al, 2007
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AIFA
•Progetto scientifico in condivisione con il Prof. A. Biondi e il dott. E. Biagi,Unità di Pediatria, Ospedale San Gerardo, Monza, Università Milano Bicocca,con il Prof. S. Cortelazzo, Ospedale Bolzano.
•PI clinico: Prof. A Rambaldi
Infusione sequenziale di linfociti non manipolati del donatore (DLI) e linfocitikiller indotti da citochine (CIK) dopo il trapianto allogenico di cellule staminali
Protocollo CIK 2
Numero EudraCT: 2008-003185-26
Fase II autorizzata da AIFA il 14.04.09, Prot.n° AIFAII/37533/P
Autorizzazione da CE Bergamo: 30/07/2009
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Therapeutic use
1st DLI: 1x106/kg
2nd DLI: 1x106/kg
10x106/kg5x106/kg5x106/kg4
10x106/kg5x106/kg1x106/Kg3
5x106/kg5x106/kg1x106/Kg2
5x106/kg1x106/Kg1x106/Kg1
3rd CIK cells infusion
2nd CIK cells infusion
1st CIK cells infusion
Combina-tion
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None1,5,101-5-10BGFL.H.VIII
None1,5,101-5-10BZFA.G.VII
None1,51-5-5BZFM.A.VI
None1,51-5-5BZFV.D.V
None1-5-51-5-5BGMP.P.IV
None1-1-51-1-5BGMT.G.III
None1-11-1-5BGFV.A.II
None1-51-1-5BGMM.C.M.I
Toxicity(aGVHD)
TreatmentGroupCityGenderPatient N°
CIK II protocol: enrollment at November 2010
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Laboratorio di Terapia Cellulare “G. Lanzani”Responsabile: M. Introna
1. Processazione dicellule staminali
emopoieticheResponsabile: M.
Introna
Referente Qualità delLaboratorio (RQL):
J. Golay
OperatoriG. BorleriC. CapelliE. Gotti
2. Produzione di prodotti diTerapia Cellulare in GMPQualified person: M. Introna
Assicurazione di QualitàResponsabile: J. Golay
Operatori:G. BorleriC. CapelliA. Pievani
E. GottiL. BolognaA. Amaru
3. Ricerca e SviluppoGroup leaders
M. IntronaJ. Golay
Personale AmministrativoE. Traini
Programma di Trapianto delleCellule Staminali EmopoieticheDirettore: Alessandro Rambaldi
USC Ematologia
OrganigrammaCentro di Raccolta
SIMTResponsabile: Dott.essa A.
Falanga
Unità Clinica
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Laboratorio di Terapia Cellulare“G.Lanzani”
USC Ematologia, Ospedali Riuniti di Bergamo
Associazione ItalianaContro le Leucemie
Linfomi e Mielomi – ONLUSsezione PAOLO BELLI
Alessandro Rambaldi Direttore USC EmatologiaMartino Introna Qualified Person DT e RRPJosée Golay Quality Control e RRCQGianmaria Borleri Operator ProductionChiara Capelli Operator ProductionAlice Pievani Operator ProductionElisa Gotti Operator ProductionLuca Bologna Operator Quality ControlAriel Amaru PhD, Ricerca e sviluppoElisabetta Traini Operatore di supporto gestionale
Microbiologia OORRBGUfficio Tecnico OORRBGUfficio Qualità OORRBG
Uffici Amministrativi OORRBGUfficio Ambientale OORRBG/Impresa pulizie
Ricerca donatori: E. OldaniGestione software: F. Delaini
Gestione GCP: ML Ferrari