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Reference reagents for Reference reagents for monitoring minimal residual monitoring minimal residual
Concept of minimal residual disease (MRD)Concept of minimal residual disease (MRD)
1012
1010
106
Cell number
Time
Morphological detection limit
PCR detection limit
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New molecular-based New molecular-based therapiestherapies
1.1. Therapeutic breakthroughTherapeutic breakthrough O Brien et al NEJM, 2003; 348; 994-1004O Brien et al NEJM, 2003; 348; 994-1004 GleevecGleevec a tyrosine kinase inhibitor and CML ( a tyrosine kinase inhibitor and CML (BCR ABLBCR ABL))
– Therapeutic follow up on cytogenetic: t(9;21) or Ph Chr.Therapeutic follow up on cytogenetic: t(9;21) or Ph Chr.– -- CCR status (18 months): CCR status (18 months):
• Interferon + ARA C: 14.4%Interferon + ARA C: 14.4%• Gleevec 74% p<0,0001Gleevec 74% p<0,0001
2.2. Biological surveyBiological survey::• Hughes TP et al NEJM 2003; 349: 1399-401Hughes TP et al NEJM 2003; 349: 1399-401
• 3 laboratories3 laboratories• reference value: pool of 30 CML at diagnosisreference value: pool of 30 CML at diagnosis• Responder: reduction of at least 3 logs by 12 months.Responder: reduction of at least 3 logs by 12 months.
Sudden need of reference biological material and Sudden need of reference biological material and Quality ControlsQuality Controls
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•Partners:Partners:• SANCO European CommissionSANCO European Commission
RQ PCR - EAC network- CONCLUSIONS -1. The PCR step was highly reproducible,2. Variations mainly due to the pre -PCR steps (RNA stability, RT),3. Normalisation by a control gene is mandatory,4. The EAC protocol allowed data comparison,5. Proposal: use of the EAC protocol as International reference
E. Beillard et al & Gabert J et al Leukemia, dec 2003
developpment of a biological reference developpment of a biological reference materialmaterial
• RationalRational– Allows data comparison between and within plat-forms / Allows data comparison between and within plat-forms /
laboratorieslaboratories • Needed for research, Needed for research, • Strategic for network programs,Strategic for network programs,• Indispensable for medical useIndispensable for medical use (ex. therapeutic decision (ex. therapeutic decision
made on gene profiling signatures)made on gene profiling signatures)
• Collaboration ERT MEIDIA & NIBSC (UK)Collaboration ERT MEIDIA & NIBSC (UK)– NAT Standards in virology (Dr. J. Saldanha)NAT Standards in virology (Dr. J. Saldanha)– Freeze dried cells?Freeze dried cells?
• can be sent at room temperature worldwidecan be sent at room temperature worldwide– French patent n°French patent n°03/0319903/03199
MRD by gene expression analysis MRD by gene expression analysis (RQ PCR)(RQ PCR)
and Hematological malignanciesand Hematological malignancies
• Towards the definition of new biological Towards the definition of new biological makersmakers
• Potentially available wapon within the Potentially available wapon within the medical arsenal against hematological medical arsenal against hematological malignanciesmalignancies– at diagnosis (Lossos et al NEJM, 2004)at diagnosis (Lossos et al NEJM, 2004)– For therapeutic efficiency monitoring (MRD)For therapeutic efficiency monitoring (MRD)
reference materialsreference materials
• Required forRequired for innovative technologies innovative technologies development :development :– gene expression profiling, CGH array, SNP …gene expression profiling, CGH array, SNP …