In Memory of In Memory of Brian Alan FrithBrian Alan Frith

Utililzation of GeneticsUtililzation of Genetics in in Screening for Fanconi Screening for Fanconi

Anemia and Subsequent Anemia and Subsequent Implications for TherapyImplications for Therapy

Cheryl Vanderford, PA-SCheryl Vanderford, PA-S

Bettye Hollins, PhD, AdvisorBettye Hollins, PhD, Advisor

Why am I interested in this Why am I interested in this topic?topic?

At the age of seventeen months, At the age of seventeen months, my brother was diagnosed with my brother was diagnosed with megakaryoblastic leukemia. He megakaryoblastic leukemia. He fought a short four month battle.fought a short four month battle.

In 2003, doctors informed us In 2003, doctors informed us that Brian’s cell-line (CHRF-288) that Brian’s cell-line (CHRF-288) was being used in research for was being used in research for Fanconi Anemia.Fanconi Anemia.

http://www.affinityonline.net/local/1999-05/mcd-002.asp

OverviewOverview What is Fanconi Anemia?What is Fanconi Anemia? Why are FA genes important?Why are FA genes important? What tests are used to screen for What tests are used to screen for

Fanconi Anemia?Fanconi Anemia? How do you diagnose Fanconi How do you diagnose Fanconi

Anemia?Anemia? How can you treat Fanconi Anemia?How can you treat Fanconi Anemia? ImplicationsImplications

What is Fanconi Anemia?What is Fanconi Anemia?

Fanconi Anemia is an autosomal Fanconi Anemia is an autosomal recessive disorder.recessive disorder.

We all have FA genes. We all have FA genes. Known mutations involved: FANCA, Known mutations involved: FANCA,

FANCB, FANCC, FANCD1, FANCD2, FANCB, FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCE, FANCF, FANCG, FANCI, FANCJ, FANCL, and FANCM FANCJ, FANCL, and FANCM (Kennedy and (Kennedy and

D’Andrea, 2006).D’Andrea, 2006).

http://www.wellspring.in/images/centers_of_excellence/common.jpg

What is Fanconi Anemia?What is Fanconi Anemia?

Fanconi Anemia predisposes an Fanconi Anemia predisposes an individual to numerous types of individual to numerous types of cancer, including acute myeloid cancer, including acute myeloid leukemia, breast cancer, squamous leukemia, breast cancer, squamous cell carcinoma of the head and neck, cell carcinoma of the head and neck, and cancers of the gynecological and cancers of the gynecological system, skin, esophagus, liver, and system, skin, esophagus, liver, and kidney kidney (Kennedy and D’Andrea, 2006).(Kennedy and D’Andrea, 2006).

Function of FA genesFunction of FA genes DNA repair DNA repair Cell cycle controlCell cycle control Oxygen sensitivityOxygen sensitivity Apoptosis and telomere maintenanceApoptosis and telomere maintenance Haemopoiesis Haemopoiesis

Tischkowitz, M., Hodgson, S. Fanconi anaemia. J. Med. Genet. 2003; 40:1-10.

Fanconi Anemia Pathway: DNA Fanconi Anemia Pathway: DNA RepairRepair

Mirchandani, K., D’Andrea, A. The fanconi anemia/BRCA pathway: a coordinator of cross-link repair. Experimental Cell Research. 2006; 312: 2647-2653.

FA Role in Cell Cycle FA Role in Cell Cycle RegulationRegulation

Checkpoint exists at Checkpoint exists at G2/M stage of mitosis to G2/M stage of mitosis to prevent progression of prevent progression of damaged DNA. damaged DNA.

Mutated FA genes result Mutated FA genes result in a prolonged G2/M in a prolonged G2/M checkpoint and reduced checkpoint and reduced efficacy of the DNA efficacy of the DNA repair mechanism.repair mechanism.

http://en.wikipedia.org/wiki/Mitosis

Tischkowitz, M., Hodgson, S. Fanconi anaemia. J. Med. Genet. 2003; 40:1-10.

G0G1

FA Role in HaemopoiesisFA Role in Haemopoiesis Common presentations of patients with Common presentations of patients with

Fanconi Anemia:Fanconi Anemia: AnemiaAnemia PancytopeniaPancytopenia Bone marrow hypoplasiaBone marrow hypoplasia ThrombocytopeniaThrombocytopenia

Gene therapy to target this role of FA Gene therapy to target this role of FA genes in individuals with FA gene genes in individuals with FA gene mutations. mutations.

S/SxS/Sx Low Birth WeightLow Birth Weight

http://www.successby6ottawa.ca/lbwfpn/images/pink.jpghttp://www.successby6ottawa.ca/lbwfpn/images/pink.jpg

S/SxS/Sx

This is a three year old boy with Fanconi This is a three year old boy with Fanconi Anemia.Anemia.

http://www.emedicine.com/ped/images/19911991PED3022-01A.JPG

Short stature

S/SxS/Sx Absence of or malformity in hands and Absence of or malformity in hands and

arms, for example the absence of a thumb arms, for example the absence of a thumb or the presence of polydactyly or the presence of polydactyly

http://www.fanconi.org/aboutfa/http://www.fanconi.org/aboutfa/Diagnosis.htmDiagnosis.htm

Tischkowitz, M., Hodgson, S. Fanconi

anaemia. J. Med. Genet. 2003; 40:1-10.

S/SxS/Sx Presence of only one kidney or of a horseshoe Presence of only one kidney or of a horseshoe

kidneykidney

Horseshoe KidneyHorseshoe Kidneyhttp://www.md.huji.ac.il/mirror/webpath/RENAL004.jpg

S/SxS/Sx Café-au-lait spots Café-au-lait spots

http://www.dental.mu.edu/oralpath/lesions/cafe/cafe1.JPGhttp://www.dental.mu.edu/oralpath/lesions/cafe/cafe1.JPG

ScreeningScreening DEB testDEB test MMC testMMC test Prenatal screeningPrenatal screening Carriers of Fanconi AnemiaCarriers of Fanconi Anemia

Blood testBlood test Rapid means of screening population at Rapid means of screening population at

largelarge Immunoblotting and immunofluorescence Immunoblotting and immunofluorescence

SubtypingSubtyping Retroviral vectorsRetroviral vectors

DiagnosisDiagnosis DEB and MMC testsDEB and MMC tests Diagnosis typically occurs before the age Diagnosis typically occurs before the age

of twelve of twelve (Fanconi Anemia Research Fund, Inc., 2006). (Fanconi Anemia Research Fund, Inc., 2006).

MMC test is used to diagnose Fanconi MMC test is used to diagnose Fanconi Anemia at the University of Kentucky.Anemia at the University of Kentucky.

Subtyping via use of retroviruses needs to Subtyping via use of retroviruses needs to be incorporated into standard protocol be incorporated into standard protocol when diagnosing a patient with Fanconi when diagnosing a patient with Fanconi Anemia.Anemia.

TreatmentTreatment

Retrovirus Retrovirus mediated gene mediated gene transfertransfer

Lentivirus Lentivirus mediated gene mediated gene transfertransfer http://www.kfdunn.com/vector/gene-therapy.gif

Risks of Gene TherapyRisks of Gene Therapy Retrovirus potential to stimulate Retrovirus potential to stimulate

oncogenes.oncogenes. Lentivirus association with arthritis and Lentivirus association with arthritis and

encephalitis in goats, leukemia in cattle, encephalitis in goats, leukemia in cattle, anemia in horses, and immunodeficiency anemia in horses, and immunodeficiency in cats, cattle, primates, and humans.in cats, cattle, primates, and humans.

Anson, D. The use of retroviral vectors for gene therapy-what are the risks? A review of retroviral pathogenesis and its relevance to retroviral vector-mediated gene delivery. Genetic Vaccines and Therapy. 2004; 2(1): 9. (Published online).

Somatic MosaicismSomatic Mosaicism Reversal of mutated FA genes has been Reversal of mutated FA genes has been

successfully accomplished by nature successfully accomplished by nature alone.alone. How? How?

Better understanding of nature’s own Better understanding of nature’s own gene therapy for Fanconi Anemia may gene therapy for Fanconi Anemia may allow us to someday mimic the same allow us to someday mimic the same mechanism to eliminate Fanconi mechanism to eliminate Fanconi Anemia.Anemia.

Sensitivity to TreatmentSensitivity to Treatment

Tischkowitz, M., Ameziane, N., Waisfisz, Q., De Winter, J., Harris, R., Taniguchi, T., D’Andrea, A., Hodgson, S., Matthew, C., Joenje, H. Bi-allelic silencing of the fanconi anaemia gene FANCF in acute myeloid leukaemia. Br J Haematol. 2003; 123: 469-471.

CHRF-288 is Brian’s cell CHRF-288 is Brian’s cell line.line. Hypermethylation of Hypermethylation of

FANCF is hypothesized FANCF is hypothesized to “be an early step in to “be an early step in sporadic carcinogenesis”sporadic carcinogenesis”

Recent studies have Recent studies have shown that the degree of shown that the degree of methylation to FANCF methylation to FANCF can indicate the can indicate the responsiveness of the responsiveness of the individual to a specific individual to a specific treatment. treatment.

Genetic CounselingGenetic Counseling Provide knowledge about:Provide knowledge about:

What Fanconi Anemia isWhat Fanconi Anemia is What symptoms are What symptoms are

associatedassociated Who can be affectedWho can be affected What the disorder means for What the disorder means for

the future of the individualthe future of the individual What treatments are What treatments are

available and their risks and available and their risks and benefitsbenefits

Importance of screening Importance of screening other family members at other family members at riskrisk

http://www.childbirthsolutions.com/articles/preconception/geneticcounseling/Gen-1.jpg

Is this ethical?Is this ethical? A genetic disorder that predisposes an A genetic disorder that predisposes an

individual to cancer—why not screen the individual to cancer—why not screen the population at large?population at large?

Cost/benefitCost/benefit Wait until Fanconi Anemia shows Wait until Fanconi Anemia shows

symptoms before finding out you carry symptoms before finding out you carry mutated FA genes?mutated FA genes? Early interventionEarly intervention

CarriersCarriers Guilt if not aware of the risk of possibly passing Guilt if not aware of the risk of possibly passing

mutated FA genes to your child.mutated FA genes to your child.

SummarySummary Fanconi Anemia is an autosomal recessive Fanconi Anemia is an autosomal recessive

disorder that predisposes individuals to a disorder that predisposes individuals to a variety of cancers.variety of cancers.

Screening techniques exist, such as the Screening techniques exist, such as the DEB and MMC tests, that should be used to DEB and MMC tests, that should be used to screen the population at large.screen the population at large.

Subtyping of which FA gene(s) is (are) Subtyping of which FA gene(s) is (are) mutated should become standard protocol mutated should become standard protocol with diagnosis.with diagnosis.

Gene therapy may someday eliminate Gene therapy may someday eliminate Fanconi Anemia. Fanconi Anemia.

ReferencesReferences Anson, D. The use of retroviral vectors for gene therapy-what are Anson, D. The use of retroviral vectors for gene therapy-what are

the risks? A review of retroviral pathogenesis and its relevance to the risks? A review of retroviral pathogenesis and its relevance to retroviral vector-mediated gene delivery. retroviral vector-mediated gene delivery. Genetic Vaccines and Genetic Vaccines and Therapy. Therapy. 2004; 2(1): 9. (Published online). 2004; 2(1): 9. (Published online).

Fanconi Anemia Research Fund, Inc., Diagnosis. Fanconi Anemia Research Fund, Inc., Diagnosis. 2006, Available 2006, Available at: at: http://www.fanconi.org/aboutfa/Diagnosis.htmhttp://www.fanconi.org/aboutfa/Diagnosis.htm, Accessed on , Accessed on 2006 Nov 26.2006 Nov 26.

Kennedy, R., D’Andrea, A. DNA repair pathways in clinical Kennedy, R., D’Andrea, A. DNA repair pathways in clinical practice: lessons from pediatric cancer susceptibility syndromes. practice: lessons from pediatric cancer susceptibility syndromes. Journal of Clinical Oncology. Journal of Clinical Oncology. 2006; 24: 3799-3808.2006; 24: 3799-3808.

Mirchandani, K., D’Andrea, A. The fanconi anemia/BRCA pathway: Mirchandani, K., D’Andrea, A. The fanconi anemia/BRCA pathway: a coordinator of cross-link repair. a coordinator of cross-link repair. Experimental Cell Research. Experimental Cell Research. 2006; 312: 2647-2653.2006; 312: 2647-2653.

Pettigrew, A. Personal Interview. January 2007.Pettigrew, A. Personal Interview. January 2007. Tischkowitz, M., Ameziane, N., Waisfisz, Q., De Winter, J., Harris, Tischkowitz, M., Ameziane, N., Waisfisz, Q., De Winter, J., Harris,

R., Taniguchi, T., R., Taniguchi, T., D’Andrea, A., Hodgson, S., Matthew, C., D’Andrea, A., Hodgson, S., Matthew, C., Joenje, H. Bi-allelic silencing of the fanconi anaemia gene FANCF Joenje, H. Bi-allelic silencing of the fanconi anaemia gene FANCF in acute myeloid leukaemia. in acute myeloid leukaemia. Br J Haematol.Br J Haematol. 2003; 123: 469-471. 2003; 123: 469-471.

Tischkowitz, M., Hodgson, S. Fanconi anaemia. Tischkowitz, M., Hodgson, S. Fanconi anaemia. J. Med. J. Med. Genet. Genet. 2003; 40:1-10. 2003; 40:1-10.