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Genetics and Mechanisms of

Prepared by: Morad Merwan Abed
Supervised by: Dr. Basim Ayesh

Hemophilia: derived from haima meaning blood, and
philia meaning affection

The word hemophilia introduced by Hopff at

University of Zurich in 1828
Blood does not clot normally
Bleed for a longer time NOT more profusely or
There are 3 known types (A, B, C)

The Royal Disease

Queen Victoria (18371901) was a carrier

Her son Leopold suffered
from frequent hemorrhages
before dying from a brain
hemorrhage at 31
Queen Victorias daughters
passed the disease to the
Spanish, German and
Russian Royal Families


Sex-linked recessive
genetic disorder
affecting X chromosome
(type A & B)
Only females can be
carriers; men are
affected or unaffected

Clinical Manifestations

Hemarthroses (bleeding into muscles and joints ),

deep muscle hematomas, hematuria and easy
Mild forms may be asymptomatic
Moderate forms usually bleed after surgery or trauma
Severe forms show spontaneous bleeding

Brain hemorrhaging is a frequent cause of death


Diagnosis prompted by unusual bleeding/bruising or

positive family history
2 approaches to genetic diagnosis:

Linkage analysis (to track defective X-chromosome in family)

Direct mutation detection

Must be confirmed by specific factor analysis or by tests

that reveal the presence of a dysfunctional protein


Primary Hemostasis

Begins after injury to blood vessel endothelium

Platelets immediately plug the site
Called the platelet plug

Secondary Hemostasis

Occurs simultaneously to primary

Proteins called coagulation/clotting factors
respond to complex cascade forming fibrin strands
the coagulation cascade
Strengthen platelet plug

Coagulation Cascade

Divided into 3 pathways:

Contact activation (intrinsic) pathway

Tissue factor (extrinsic) pathway
Common pathway

Contact Activation (intrinsic)

CS: collagen surface
HMWK: high
molecular weight

PL: phospholipids


Upon injury blood vessel is damaged

Plasma is exposed to negatively charged surfaces
such as collagen which activates the pathway
Hemophilia A, B and C affect factors VIII, IX and XI

Tissue Factor (extrinsic)


Following damage to blood vessel, tissue factor is

Tissue factor is a lipoprotein present in many tissues
including high concentrations in brain, lung and
placental tissues.


The Common Pathway


Intrinsic and extrinsic pathway come together at

factor X to start the common pathway
Results in a clot being formed by fibrin monomers

Hemophilia A
Factor VIII Deficiency

Most common type

Frequency of 1 per 10,000 males
In 1/3 of cases there is no family history
Patients with mild to moderate hemophilia may not
exhibit abnormal bleeding until later in life
>80% of children with severe hemophilia experience
clinically significant bleeding during the first year of
Once children with severe hemophilia begin to walk
they typically experience recurrent hemarthroses

Genetics of Hemophilia A

Inherited as an X-linked recessive disorder

Mutations including a variety of deletions, insertions,
missense, nonsense and splice site mutations have
been reported
Commonly an inversion of intron 1 and 22
Sporadic cases result from de novo mutations

Factor VIII Gene and Protein

F8 Intron 22 Inversion

This split F8 gene cannot encode

functional FVIII protein; severe hemophilia A always results.

F8 Intron 22 Inversion Analysis

Results from homologous intrachromosomal

Inversion mutation occurs de novo once per
10,000 male meioses
Every ejaculate contains at least one sperm
with a F8 intron 22 inversion mutation
Responsible for 45% of severe hemophilia A

F8 intron 1 inversion

F8 Intron 1 Inversion Analysis

Similar to intron 22 inversion

900 bp region 5 to F8 gene crosses
over with homologous region in intron 1
Results in F8 gene lacking a promoter
and first exon
Responsible for approx 2% of severe
hemophilia A

Intrachromosomal inversions
cause 50% of cases of severe
hemophilia A

The majority of non-inversion cases of

hemophilia A, and nearly all cases of
hemophilia B result from point
mutations, affecting a single nucleotide.

Examples of Point Mutation

-Cys Arg Lys Lys Thr Gln- Normal

-Tyr Arg Lys Lys Thr Gln- Missense
-GTC TGA AAA AAA ACG CAG-Val Arg Lys Lys Arg Met- Frameshift

Ways to Eliminate FVIII Activity

(severe disease)

Intron 1 or 22 inversion
Delete part of gene
Insert extra nucleotides
Nonsense mutation
Splice site defect
Missense mutation at strategic amino

Ways to Reduce FVIII Production

(moderate/mild disease)

Missense mutation, less important

amino acid
Splice site defect
Most families have a private mutation
Mutation not identified in ~2% of

Hemophilia B
Factor IX Deficiency
Prevalence: 1 per 50 000 males
70% of cases-true deficiency of coagulation factor IX
30% of cases-presence of a dysfunctional protein
severe (IX activity <1% of normal)
moderate (IX activity 1-5%)
mild (IX activity 5-25%)

Hemophilia B: Genetics

The gene for Factor IX is located on the longarm of the X-chromosome

Mutations: deletions, insertions, inversions and
point mutations (represent 90% of all
All regions of Factor IX gene have been

Factor IX
single chain vitamin K-dependent glycoprotein
415 amino acids

Factor IX Gene and Protein

There are no mutations equivalent to the

inversions seen in hemophilia A.
The same general profile of non-inversion
mutations is seen as in hemophilia A, with
one important exception, hemophilia B
The majority of patients have point
mutations, most resulting in missense

Hemophilia B Leiden

Most hemophilia is lifelong disorder of same

Small proportion of hemophilia B patients
have FIX levels which increase at puberty
Hemophilia B Leiden 3% of hemophilia
This is the only situation where hemophilia
gets better

Hemophilia C
Factor XI deficiency

In the USA, it is thought to affect 1 in 100,000 of the

adult population
Distinguished from Hemophilia A and B by the
absence of bleeding into joints and muscles
Occurrence in both sexes .

Hemophilia C: Genetics

The gene encoding FXI is located on the distal arm of

chromosome 4 (4q35)
The deficiency is inherited as an autosomal recessive

Mutations of the FXI gene:

Majority are missense mutations
Nonsense mutations
Splice site mutations

Factor XI: Pathway


Factor Replacement therapy

isolated from human blood plasma


Plasma-derived factor VIII and IX are made from human

plasma pooled together then separated into different

a genetically engineered cell line made by DNA

technology, called recombinant factors

The human factor VIII (or IX) gene is isolated through

genetic engineering which is inserted into non-human
It is cultured then purified for human use

Treatment Frequency

Depending on disease severity, factor

replacements can be given daily, weekly,
monthly or almost never


infusion of factor concentrates immediately after the

beginning of a bleed to stop the bleeding quickly


hemophiliacs receive factor concentrates one or more

times a week to prevent bleeding

Treatments (cont)

Desmopressin synthetic drug which is a copy of a

natural hormone
Treats mild/moderate hemophilia A
Acts by recruiting Factor VIII to the site of blood
vessel damage
Used by injection or as nasal spray
Cyklokapron and Amicar
Treat both hemophilia A and B
Dont actually help form a clot, only help hold a
clot in place therefore, cannot be used instead of
desmopressin or replacement therapy
Available as tablets


38 year old female

Uncle died from the dz
Brother with hemophilia lieden
Risk for having a child with the dz??


Hemophilia occurs when the blood does not clot normally; an

affected individual bleeds for a longer time, not more profusely
or quickly
Clinical manifestations could include hemarthroses, deep muscle
hematomas, hematuria, easy bruising and spontaneous
bleeding, while brain hemorrhage is a frequent cause of death.
Hemophilia A and B are caused by rare recessive traits, with the
defective gene (the Factor VIII and IX, respectively) located on
the X-chromosome. As a result males with a defective X
chromosome exhibit hemophilia, while females (with 1 mutant
copy) are carriers of the mutation (but do not exhibit overt
Mutations could be due to deletions, insertions, missense,
nonsense, splice site, and point mutations.
Hemophilia C is a Factor XI deficiency inherited as an autosomal
recessive trait. The gene Factor XI is located on chromosome 4
and therefore can occur in both sexes

Summary (cont)

It is also distinguished from Hemophilia A and B by the absence

of bleeding into joints and muscles.
The normal clotting pathways are a series of reactions activated
by factors that then catalyze the next reaction in the cascade,
ultimately resulting in cross-linked fibrin.
Therefore a deficiency in these factors (VIII, IX) lead to
improper clotting
Treatment: Factor VIII or IX Replacement Therapy

It is isolated from human blood plasma or genetically engineered cell line

made by DNA technology, called recombinant factors
Depending on disease severity, factor replacements can be on-demand or

Desmopressin treats hemophilia A

Cyklokapron and Amicar treats hemophilia A and B

Thank you