A Study of Genetic A Study of Genetic Susceptibility to Susceptibility to

Hodgkin’s Lymphoma in Hodgkin’s Lymphoma in a Cohort of Familiesa Cohort of Families

Cheshire and Merseyside Cheshire and Merseyside Regional Molecular Genetics Regional Molecular Genetics

LaboratoryLaboratory

Abi Rousseau

LymphomaLymphoma

Cancer of the Cancer of the lymphatic systemlymphatic system

Broadly subdivided Broadly subdivided into non-Hodgkin’s into non-Hodgkin’s and Hodgkin’sand Hodgkin’s

Hodgkin’s defined Hodgkin’s defined by:by: Reed-Sternberg Reed-Sternberg

cellscells Hodgkin cellsHodgkin cells

LymphomaLymphoma

Cancer of the Cancer of the lymphatic systemlymphatic system

Broadly subdivided Broadly subdivided into non-Hodgkin’s into non-Hodgkin’s and Hodgkin’sand Hodgkin’s

Hodgkin’s defined Hodgkin’s defined by:by: Reed-Sternberg Reed-Sternberg

cellscells Hodgkin cellsHodgkin cells

LymphomaLymphoma

Cancer of the Cancer of the lymphatic systemlymphatic system

Broadly subdivided Broadly subdivided into non-Hodgkin’s into non-Hodgkin’s and Hodgkin’sand Hodgkin’s

Hodgkin’s defined Hodgkin’s defined by:by: Reed-Sternberg Reed-Sternberg

cellscells Hodgkin cellsHodgkin cells

Clinical FeaturesClinical Features

LymphodenopathyLymphodenopathy Other symptoms:Other symptoms:

Significant weight lossSignificant weight loss Itchy skinItchy skin Recurrent feversRecurrent fevers Drenching night sweatsDrenching night sweats FatigueFatigue Increased sensitivity to Increased sensitivity to

alcoholalcohol

IncidenceIncidence Rare – accounts for 5% of all cancers Rare – accounts for 5% of all cancers

diagnosed in UKdiagnosed in UK Cancer Research UK figures for 2006:Cancer Research UK figures for 2006:

1611 new cases – incidence 2.7/100,0001611 new cases – incidence 2.7/100,000 Bimodal age distributionBimodal age distribution

CausesCauses EnvironmentalEnvironmental

Developed countries show higher incidence than Developed countries show higher incidence than developing countriesdeveloping countries

Study of incidence trends among Chinese Study of incidence trends among Chinese immigrants to British Columbia supports an immigrants to British Columbia supports an environmental influence (Au environmental influence (Au et alet al, 2004), 2004)

Impact of environmental risk factors such as Impact of environmental risk factors such as smoking and diet – weak and inconsistent evidencesmoking and diet – weak and inconsistent evidence

ClusteringClustering 31 cases connected by common contacts in 31 cases connected by common contacts in

Albany, New York (Vianna Albany, New York (Vianna et alet al, 1971 & 1972) , 1971 & 1972) Lacked control group and results not replicated in Lacked control group and results not replicated in

similar studiessimilar studies

Causes continuedCauses continued ViralViral

Epstein-Barr Virus (EBV) infection in ~50% of casesEpstein-Barr Virus (EBV) infection in ~50% of cases Localised to HRS cellsLocalised to HRS cells HRS cells arise from B cells that have acquired HRS cells arise from B cells that have acquired

disadvantageous mutations – rescued from apoptosis disadvantageous mutations – rescued from apoptosis by EBV infectionby EBV infection

3 viral proteins expressed: EBNA1, LMP1 and 3 viral proteins expressed: EBNA1, LMP1 and LMP2ALMP2A

Causes continuedCauses continued GeneticGenetic

Reports of familial HL (Robertson et al, 1987)Reports of familial HL (Robertson et al, 1987)

Risk of HL higher in individuals with a family history Risk of HL higher in individuals with a family history of the condition (Razis et al, 1959)of the condition (Razis et al, 1959)

Risk of developing HL higher in gender concordant Risk of developing HL higher in gender concordant siblings (Grufferman et al, 1977)siblings (Grufferman et al, 1977)

Increased risk in monozygotic twins (Mack Increased risk in monozygotic twins (Mack et alet al, , 1995)1995)

HL co-occurring with congenital genetic disorders, HL co-occurring with congenital genetic disorders, e.g. LWDe.g. LWD

3 families with multiple 3 families with multiple casescases

MP HD116

PL HD115

Family 2

JP HD105

JC KK746

IC KK746.1

JC KK746.2

Family 3

HL

Family 1

SPHD141

CPHD141.2

EWHD141.1

Aims of this studyAims of this study

Analyse affected members of each Analyse affected members of each family for copy number variation using family for copy number variation using oligo arrayCGHoligo arrayCGH BlueGnome Cytochip Oligo 4x44K and BlueGnome Cytochip Oligo 4x44K and

2x105K 2x105K 4x44K – 350Kb genome wide backbone4x44K – 350Kb genome wide backbone 2x105K – 150Kb genome wide backbone2x105K – 150Kb genome wide backbone

Investigate any shared regions of copy Investigate any shared regions of copy number variation for potential number variation for potential candidate genes or regulatory elements candidate genes or regulatory elements for Hodgkin’s lymphoma susceptibilityfor Hodgkin’s lymphoma susceptibility

Family 1 results – 1p21.2 Family 1 results – 1p21.2 deletiondeletion

Disrupts 3’ end of OLFM3Disrupts 3’ end of OLFM3

Family 1 - DiscussionFamily 1 - Discussion

OLFM3OLFM3 Encodes olfactomedin 3, expressed in Encodes olfactomedin 3, expressed in

ocular tissues, brain, kidney and lungocular tissues, brain, kidney and lung May play a role in pathogenesis of May play a role in pathogenesis of

glaucoma and other ocular disordersglaucoma and other ocular disorders Does an ocular disorder co-Does an ocular disorder co-

segregate with Hodgkin’s in this segregate with Hodgkin’s in this family? family? More clinical information and DNA from More clinical information and DNA from

further family members required to further family members required to investigate significanceinvestigate significance

Family 2 resultsFamily 2 results 4q28.1 duplication4q28.1 duplication

18p11.31 duplication18p11.31 duplication

Family 2 - DiscussionFamily 2 - Discussion 4q28.14q28.1

No genes disruptedNo genes disrupted 2 predicted CTCF binding sites2 predicted CTCF binding sites

18p11.3118p11.31 1 predicted CTCF binding site1 predicted CTCF binding site TGIF – transforming growth interacting TGIF – transforming growth interacting

factorfactor Represses transcription of EBNA1 (essential Represses transcription of EBNA1 (essential

for replication of EBV genome)for replication of EBV genome) EBV- related genetic susceptibility??EBV- related genetic susceptibility??

Further work required to elucidate Further work required to elucidate targets of the CTCF binding sites targets of the CTCF binding sites

Family 3 results – 7q36.3 Family 3 results – 7q36.3 deletiondeletion

Discussion – Family 3Discussion – Family 3

CNPY1CNPY1 Interacts with FGFR1 and ACTRIIInteracts with FGFR1 and ACTRII FGFR1 upregulated in various cancersFGFR1 upregulated in various cancers Reduction of canopy1 would lead to Reduction of canopy1 would lead to

downregulation of FGFR1downregulation of FGFR1 ACTRII loss of function mutations – ACTRII loss of function mutations –

colorectal cancercolorectal cancer More clinical information and DNA More clinical information and DNA

from further family members from further family members required to investigate significancerequired to investigate significance

SummarySummary

HL is rare and familial HL accounts for HL is rare and familial HL accounts for only a small proportion of cases only a small proportion of cases

Familial HL may be genetic, viral, Familial HL may be genetic, viral, environmental or a combinationenvironmental or a combination

Hodgkin’s likely to be heterogeneous Hodgkin’s likely to be heterogeneous In each family disruption of a gene or In each family disruption of a gene or

CTCF binding site has been identifiedCTCF binding site has been identified Findings need to be confirmed by Findings need to be confirmed by

another methodanother method Further studies requiredFurther studies required

AcknowledgmentsAcknowledgments

Liverpool Liverpool Molecular GeneticsMolecular Genetics David GokhaleDavid Gokhale Vicky StintonVicky Stinton Roger MountfordRoger Mountford Kym SpencerKym Spencer Katrina SmithKatrina Smith

Liverpool Liverpool CytogeneticsCytogenetics Anna ToppingAnna Topping Una MayeUna Maye

NGRL, ManchesterNGRL, Manchester William FergusonWilliam Ferguson

Cancer Cancer Immunogenetics Immunogenetics Group, ManchesterGroup, Manchester G Malcolm TaylorG Malcolm Taylor Adiba HussainAdiba Hussain

BlueGnome LtdBlueGnome Ltd David ChrimesDavid Chrimes

Sheffield Sheffield CytogeneticsCytogenetics Simon WebsterSimon Webster

ReferencesReferences Au W.Y. et al. (2004). Hodgkin’s lymphoma in Chinese migrants to British Columbia: a 25 year Au W.Y. et al. (2004). Hodgkin’s lymphoma in Chinese migrants to British Columbia: a 25 year

survey. Annals of Oncology 15: 626-630survey. Annals of Oncology 15: 626-630 Vianna N.J. et al (1972). Hodgkin’s disease: Cases with features of a community outbreak. Vianna N.J. et al (1972). Hodgkin’s disease: Cases with features of a community outbreak.

Annals of Internal Medicine, 77(2): 169-180Annals of Internal Medicine, 77(2): 169-180 Küppers R (2009). The biology of hodgkin’s lymphoma. Nature Reviews Cancer 9(1): 15-27Küppers R (2009). The biology of hodgkin’s lymphoma. Nature Reviews Cancer 9(1): 15-27 Robertson S.J. et al (1987. Familial Hodgkin’s Disease. Cancer 59: 1314-1319Robertson S.J. et al (1987. Familial Hodgkin’s Disease. Cancer 59: 1314-1319 Razis D.V. et al (1959). Familial Hodgkin’s disease: its significance and implications. Annals of Razis D.V. et al (1959). Familial Hodgkin’s disease: its significance and implications. Annals of

Internal Medicine 51: 933-971Internal Medicine 51: 933-971 Grufferman S. et al (1977). Hodgkin’s disease in siblings. NEJM 296: 248-250Grufferman S. et al (1977). Hodgkin’s disease in siblings. NEJM 296: 248-250 Mack T.M. et al (1995). Concordance for Hodgkin’s disease in identical twins suggesting Mack T.M. et al (1995). Concordance for Hodgkin’s disease in identical twins suggesting

genetic susceptibility to the young-adult form of the disease. NEJM 332: 413-418genetic susceptibility to the young-adult form of the disease. NEJM 332: 413-418 Gokhale D.A. et al (1995). Molecular genetic analysis of a family with a history of Hodgkin’s Gokhale D.A. et al (1995). Molecular genetic analysis of a family with a history of Hodgkin’s

disease and dyschrondrosteosis. Leukemia 9: 826-833disease and dyschrondrosteosis. Leukemia 9: 826-833 Bao L. et al (2007). CTCFBSDB: a CTCF-binding site database for characterisation of Bao L. et al (2007). CTCFBSDB: a CTCF-binding site database for characterisation of

vertebrate genomic insulators. Nucleic Acids Research 36: D83-D87vertebrate genomic insulators. Nucleic Acids Research 36: D83-D87 Goldin L.R. et al (2005). A genome screen of families at high risk for Hodgkin lymphoma: Goldin L.R. et al (2005). A genome screen of families at high risk for Hodgkin lymphoma:

evidence for a susceptibility gene on chromosome 4. Journal of Medical Genetics 42: 595-601evidence for a susceptibility gene on chromosome 4. Journal of Medical Genetics 42: 595-601 Joos S. et al (2000). Genomic imbalances including amplification of the tyrosine kinase gene Joos S. et al (2000). Genomic imbalances including amplification of the tyrosine kinase gene

JAK2 in CD30+ Hodgkin cells. Cancer Research 60: 549-552 JAK2 in CD30+ Hodgkin cells. Cancer Research 60: 549-552 Torrado M. et al (2002). Optimedin: a novel olfactomedin-related protein that interacts with Torrado M. et al (2002). Optimedin: a novel olfactomedin-related protein that interacts with

myocilin. Human Molecular Genetics 11: 1291-1301myocilin. Human Molecular Genetics 11: 1291-1301 Liang C.L. et al (2000). Transcription of Epstein-Barr virus-encoded nuclear antigen 1 Liang C.L. et al (2000). Transcription of Epstein-Barr virus-encoded nuclear antigen 1

promoter Qp is repressed by transforming growth factor-beta via Smad4 binding element in promoter Qp is repressed by transforming growth factor-beta via Smad4 binding element in human BL cells. Virology 277(1): 184-192human BL cells. Virology 277(1): 184-192

Hirate Y. et al (2006). Canopy1, a novel regulator of FGF signalling around the midbrain-Hirate Y. et al (2006). Canopy1, a novel regulator of FGF signalling around the midbrain-hindbrain boundary in zebrafish. Current Biology 16: 421-427hindbrain boundary in zebrafish. Current Biology 16: 421-427

Olaru A. et al (2003). Loss of heterozygosity and mutational analyses of the ACTRII gene locus Olaru A. et al (2003). Loss of heterozygosity and mutational analyses of the ACTRII gene locus in human colorectal tumors. Laboratory Investigation 83(12): 1867-1871in human colorectal tumors. Laboratory Investigation 83(12): 1867-1871