Molecular Genetics Oncology Service Benedict J' Milner, Dept' of Medical Genetics, NHS Grampian

1
Molecular Genetics Oncology ServiceBenedict J.
Milner, Dept. of Medical Genetics, NHS Grampian
2
MGOS - Services Offered
SUSPECT HAEMATOLOGICAL MALIGNANCY
Bone Marrow
Blood
Lymph Node
Skin Lesion
PML-RARA analysis
inv(16) analysis
AML
t(821) analysis
DNA Extraction
DNA Extraction
RNA Extraction
Flt3 analysis
cDNA synthesis
BCR-ABL RQ-PCR analysis
CML
BCR-ABL analysis
t(411) analysis
Ig / TCR Gene Re-arrangement Analysis
PowerPlex Analysis
ALL
t(1221) analysis
t(119) analysis
Post-PBSC Transplant Monitoring
MPD
JAK2 analysis
Lymphoma
3
History

• Initial 5 year development grant funded by ARH
  Endowments Childhood Cancer Fund - October 1998
  to September 2003
• Need for molecular genetics service identified by
  Haematology Pathology to support Cytogenetics
  Oncology Service
• From October 2003 - Extended on annual basis
  while permanent funding sought
• Permanent NHS-G funding from April 2007
• NSD funding from April 2008

4
Chronic Myeloid Leukaemia (CML) Testing

• Diagnosis RT-PCR
• t(922) translocation BCR/ABL fusion gene
• Upregulation of ABL tyrosine kinase
• First-line treatment with Imatinib (Glivec)
• Tyrosine Kinase Inhibitor

5
Chronic Myeloid Leukaemia (CML) Testing

• Treatment Follow-up
• RQ-PCR 3 monthly following onset of Imatinib
• FusionQuant p210 kit (Ipsogen) using
  Corbett RotorGene 6000
• (also FusionQuant p190 kit)
• CML patients resistant to Glivec chemotherapy
• BCR/ABL KD mutation screening using WAVE dHPLC /
  sequencing

6
Acute Myeloid Leukaemia (AML) Testing

• Good Prognostic Indicators
• AML (M2) t(821) AML1 / ETO
• AML (M3) t(1517) PML / RARA
• AML (M4Eo) inv(16) CBFb / MYH11
• Poor Prognosis Indicator
• AML Flt3 ITD

7
Acute Lymphoblastic Leukaemia (ALL) Testing

• Good Prognostic Indicator
• Childhood ALL t(1221) TEL / AML1
• Poor Prognosis Indicators
• ALL t(922) BCR / ABL p190
• B-ALL t(411) MLL / AF4
• Pre-B cALL t(119) E2A / PBX

8
Myeloproliferative Disease (MPD) Testing

• Definitive diagnosis of MPD
• JAK2 V617F mutation detection using ARMS PCR
• 95 of cases of PCV
• 50 of cases of both ET and PMF
• Drugs currently being developed to target the
  increased activity of the Janus Kinase protein

9
Lymphoma Diagnosis

• In a subset (5-15) of suspect lymphoproliferation
  s, traditional histopathology and
  immunophenotyping will not discriminate between
  malignancy and reactive disease
• Biomed -2 tests available
• IGH gene re-arrangement
• IGK IGL gene re-arrangement
• TCRB TCRG gene re-arrangement
• t(1114) detection - Mantle Cell Lymphoma
• t(1418) detection - Follicular Cell Lymphoma

10
Detection of Clonal Gene Re-arrangements
11
Multiplex PCR analysis of TCRB gene
re-arrangements
12
TCRB tube A Vb-Jb PCR GeneScan Analysis
13
Post-Peripheral Blood Stem Cell Transplant
MonitoringPromega PowerPlex 16 DNA
Fingerprinting Kit
14
Promega PowerPlex 16 DNA Fingerprinting Kit
15
Post-PBSCT Monitoring
Figure
16
MGOS Sample Intake
(projected)
17
Molecular Genetics Oncology ServiceBenedict J.
Milner, Dept. of Medical Genetics, NHS Grampian
E-mail b.j.milner_at_abdn.ac.uk