BMSC3101 Inherited

Disorders

Dr. Al Pickles

a.r.pickles@leeds.ac.uk
Sex of this individual?
Characteristic banding pattern on the chromosome is due
to staining differences; light indicates areas of gene-rich
areas as euchromatin stains lightly and contains actively
expressed genes
Top = p
Lower = q
Central point of
each chromosome
= centromere
{
Autosomal dominant
Affects both males & females equally
Transmitted from one generation to the next
All forms of transmission are observed i.e.
male-male, male-female, female-male,
female-female

Autosomal recessive
Disorder affects males & females equally
Usually only affects individuals in one
generation in a single sibship (brothers &
sisters), not occurring in previous and
subsequent generations
Consanguinity in the parents provides further
support for autosomal recessive inheritance
Paternal Age
0
1
2
3
4
5
24 29 34 39 44 47
Age
R
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R
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Achondroplasia
Achondroplasia
USA
Aperts
Marfan
Case of sporadically occurring
disorders new autosomal dominant
mutation
X-Linked recessive

Affects males
No male-male transmission
Females may manifest the condition

X-Linked dominant

Affects males & females, females more
frequently
Females less severely affected than males
Females transmit to son & daughter, males
only transmit to females = all affected

Mutations
Definition: A heritable alteration or change in the
genetic material
Genomes between any two individuals are
99.9% identical any two people differ by 1
nucleotide in every 1000 = 3 million diff
Can occur following exposure to mutagens, but
majority occur spontaneously following errors in
DNA replication & repair
Usually only cause problems when in coding
areas

Somatic cell mutation can not be
transmitted to offspring
Gonadal tissue or gamete mutation
transmitted to future generations
Estimated that each person carries up to 6
lethal recessive mutant alleles if
homozygous then would be serious
Harmful alleles constitute the genetic load
of the population
Types of mutations
Traditionally two types:
- gross structural chromosomal changes
- submicroscopic changes in one or more
nucleotides
Until recently mutations considered to be
transmitted unaltered termed fixed or stable
mutations
More recent are dynamic or unstable mutations
undergo further alteration as they are
transmitted in families & explains unusual
patterns of inheritance.
Fixed/stable point mutations: Classified
according to DNA molecular changes

Majority of these differences are called:
Single Nucleotide Polymorphisms (SNPs
pronounced snips) (also called
substitutions)
Involves replacement of single nucleotide
by another

Deletions Involves loss of one or more
nucleotides.
If occurring in coding sequences it disrupts the
reading frame

Insertions Involves the addition of one or more
nucleotides into a gene, same disruption can
occur as above

Dynamic/unstable mutations
New class, important cause of inherited disease in
humans
Consist of triplet repeat sequences which in
affected people occur more frequently
Increase in these, termed triplet amplification
identified as the mutational basis for single gene
disorders such as Huntingtons
Mechanism not clear, triplet repeats below a
certain number are consistently transmitted
during mitosis and meiosis
However, above a certain number, they are
transmitted unstably usually with increase or
decrease in repeat number = unstable mutations
Triplet repeat sequences usually take place over
a number of generations within a family=
unusual inheritance patterns (anticipation),
increasing severity
Mutation effects on proteins:
STRUCTURAL
Mutations sub-divided into two main groups
depending on the effect on the polypeptide
sequence of the encoded protein:

- Synonymous

- Non-synonymous

Synonymous/silent mutations
Mutation that doesnt alter the polypeptide
product of a gene
SNP, due to degeneracy of the code will
often result in another triplet which codes
for the same amino acid
Leads to no alteration in the resulting
protein
Non-synonymous
Term for mutation leading to alteration in
the encoded polypeptide
Occur less frequently than synonymous
Change in protein will lead to abnormal
function = disease and occur in three main
ways:
- Missense
- Nonsense
- Frameshift
Missense
SNPs can lead to coding for a different amino
acid and subsequent altered protein
Can lead to gross reduction or complete loss of
biological activity = non-conservative substitution
Protein can retain normal biological activity (e.g.
enzyme action) but has altered characteristics
such as stability or pH
SNPs that result in replacement of amino acid
but without effect = conservative substitutions
Nonsense
Substitution that leads to generation of a
stop codon
Results in premature termination of
translation
Unlikely to retain biological function of the
protein, particularly if resulting in loss of
functional domain
Frameshift
If mutation is insertion or deletion which
are not multiple of three = disrupts the
reading frame
Amino acid sequence will mean the
proteins function will be abnormal
These type may also result in a stop
codon immediately after the mutation
Mutation effects on proteins:
FUNCTIONAL
Mutations exert phenotypic effect through
either loss or gain of function
Loss of function
Result in reduced activity (hypomorph) or
decrease stability (null allele/amorph) or
even loss of gene product
Inherited enzyme function reduction usually
X-linked recessive as normal allele would
be enough to compensate..
Heterozygous state such as this where
phenotypic effect is seen is termed
haploinsufficiency ( e.g. familial
hypercholesterolaemia)
Number of autosomal dominant disorders
have basis of this
Homozygous mutations in this state result
in more severe phenotypic manifestations
of the reduction in protein function
(surprise, surprise).
Gain of function
Result in increased levels of gene expression or
development of new functions of gene product
- E.g. expanded triplet repeat mutations of
Huntington gene cause aggregations in the CNS
leading to the features of the disorder
Expression
is a statement about the features of the
condition, many conditions have variable
manifestations.

Penetrance
is a measure of how many individuals who
carry the mutant gene will manifest
features of the condition (heterozygotes).
Example: Neurofibromatosis
Starting point: neurofibromin

Background
First described in the 18
th
Century
Most common genetic disorders in
humans (some thought was cause for
original elephant man was actually
Proteus syndrome
Two main types NF1 & NF2; NF1 more
common by far with incidence of 1:3000
births

Clinical features
Small pigmented skin lesions (caf-au-
lait spots) & small soft fleshy growths
Caf-au-lait spots first appear in
childhood, increasing in size & number
until puberty
Diagnosis requires 6 spots>5 mm
Late childhood = neurofibromata;
benign tumours usually on the skin,
increasing with age
Neurofibromatosis

Clinical manifestation
Also may include:
- axillary freckling, macrocephaly, mild
developmental delay & Lisch nodules (small
raised pigmented hamartomata or iris)



Mostly NF1 individuals enjoy a normal healthy life
Small number develop more major complications:
epilepsy, CNS tumour or scoliosis
Genetics
Autosomal dominant inheritance
Mutation in the neurofibromin (NF1)
gene.
Located at 17q11.2





Spans 350 kb containing 56 exons.

Gene product
Neurofibromin protein: very
large protein (350 kD);
structural homology to
GTPase activating protein
(GAP) important role in
signal transduction;
downregulates RAS activity
(GTP-binding protein)

The GAP activity of
neurofibromin resides in a
segment which represents
only 10% of the protein and
remains the only clearly
defined biochemical
function of the protein
Normal role: NF1 gene functions as a tumour
suppressor
GAP-related domain (GRP) interacts with RAS
proto-oncogene product
NF1 plays an important role in cell growth and
differentiation
Mutations
>100 identified including deletions, insertions and
substitutions
Most lead to severe truncation or complete
absence of protein loss of function!
Large deletions = severely affected with more
intellectual impairment + increased
neurofibromata
Almost complete penetrance by 5 yrs old
Expression variable amongst family
members (identical twins not so =
modifying genes at other loci?)
50% of all NF1 cases are a result of new
mutations (m.r. = 1:10,000 gametes)


Treatment
At present no cure for NF1
Drug therapy aimed at upregulating
neurofibromin GAP activity or
downregulating RAS may be beneficial
Difficulty in applying a systemic drug to
diverse range of tissues and CNS
Requires better understanding first
cloning studies
Module information
Your seminars should be similar but
contain more detail YOU DO NOT
NEED TO KNOW ABOUT NF1 FOR
THE EXAM, IT IS PURELY
INSTRUCTIONAL

All you need to know will be explained
in the session on Friday i.e. groups,
topics, seminars/reports etc etc.