Hierarchical - PowerPoint Presentation

Slide1

Hierarchical Cluster analysis of Florescent in Situ Hybridisation in newly diagnosed myeloma patients

Ieuan Walker BSc. (hons) MSc. 4.11.15Slide2

Introduction

Cytogenetics/FISH is a key part of myeloma risk stratification and has recently been included in R-ISS

Clear associations between individual FISH abnormalities and effect on outcome e.g. 17p deletion

Co-associations between individual abnormalities

recognised

but less well defined

Single

centre

, retrospective review of FISH abnormality clustering and outcomeSlide3

MethodsPatients and Sample analysis

Retrospective analysis of all newly diagnosed symptomatic myeloma patients between 2009 and 2014

153 patients with median follow-up 36 months

Heterogeneous 1

st

line treatmentStandard FISH panel (see next slide)Hierarchical cluster analysis for abnormality groupingEffects on overall outcome and relationship with ISS and R-ISS staging systemsHierarchical cluster analysisUsing SPSS we formed a similarity score by making each of the cytogenetic lesions proportional to the ‘square Euclidian distance’We then merged these in turn based on how ‘similar’ they using an algorithm we designed. Survival curves were generated using a standard Kaplan Meier method, and P values by a log-rank testSlide4

Cytogenetic Abnormality

Genes of interest

at Loci

Ch1q21

CKS1B, PMSD4

Ch1p32.3CDKN2C

Ch5p15.2CSF1R

Ch11q13

CCND1

Ch11q22.3

ATM

Ch13q34

LAMP1

ch14q23Re

IgH Heavy chain

t

[4:14]IgH

/FGFR3t

[6:14]

IgH/CCND3

t[11:14}

IgH/CCND1t[14:16]IgH/MAFt[14;20]IgH/MAFBch17p13.TP53CEP9triHyperdiploid ChromosomeCEP12TriHyperdiploid ChromosomeCEP13Monosomy or HyperdiploidCEP15Hyperdiploid Chromosome

1q21 Gain of Copy

Rearranged 11:14

FISH panelSlide5

Hierarchical cluster analysis.

1

Ch1q21 Tri

Ch13 mon CLUSTER 1

IgH Re

Ch5p15.2 Tri CEP9 Tri CEP15 Tri CLUSTER 2 Ch11q22.3 Tri

Dendrogram using Linkage by PatientSlide6

What do the clusters mean in terms of prognosis?

Median overall survival for our total population has not been reached.

Median overall survival for Cluster 1 patients = 42 months

Median overall survival for Cluster 2 Not reached

Median OS for ISS 1: Not Reached

2: 45 Months

3: 40 monthsSlide7

Can we use clusters to provide any additional stratification to the ISS staging criteria?

Patients categorized according to their cluster status (1 or 2) and ISS stage 3

Survival benefit

for

cluster 2

Patients with lesions in both clusters were excluded from the analysisCluster 1 + ISS3 (n=19)Cluster 2 + ISS 3 (n= 23)

ISS 3 (n= 65)

Median OSCluster 1 + ISS 3

= 27mCluster 2 + ISS 3 Not reached

Stage 3 = 40m

*

* p=0.047

Cluster 1

Ch1q21

Tri Ch13 monIgH Re

Cluster 2

Ch5p15.2 TriCEP9 Tri

CEP15 Tri

Ch11q22.3 TriSlide8

Does cluster analysis complement the revised ISS?

109 patients restaged according to R-ISS (n =28, 60, 19 respectively

)

ISS stage 3 patients also

analysed

according to cluster 1 or 2 (black solid and purple)ISS-3+Cluster 1 has worse prognosis than R-ISS3Median OS survival:

ISS3 + Cluster 1 = 20 months

R-ISS3= 23 Months

**

** P<0.005Slide9

Conclusions

FISH can be used as a marker of prognosis but needs to be used in conjunction with biomarkers to identify the poorest prognostic groups.

Specific cytogenetic lesions undoubtedly are important prognosticators for survival such asTP53

loss.

The FISH “signatures” is also important. While poor prognosis markers counteract positive lesions, the absence of any cluster 2 abnormalities appears to convey even higher risk

.Using our unbiased cluster analysis we show that cytogenetics lesions group together.To further validate our algorithm we need to expand the data set… watch this space! Slide10

Thanks to

Guy’s And St Thomas Hospital Haematology

Dr Matt Streetly

Dr George Double

Dr Majid Kazmi

Dr Ines El-NajirGrace Miller King’s College Hospital HaematologyProf Steve ScheyViapath (cytogenetics at Guys Hopsital)Dr Michael Neat