PEDIGREES CONTINUED 25/2/2013

PEDIGREES CONTINUED
Pedigrees

have been used historically to track inheritance patterns. This has helped to localise genes to chromosomes, providing the means for treatment, either pharmacological, surgical or gene therapy. It provides information for genetic counselling allowing risks within families to be calculated and indicating which family members should be tested. Also used in forensic medicine for determining the source of DNA material eg paternity testing.

PEDIGREES CONTINUED FROM LAST WEEK

Always

have a key Autosomal dominant Autosomal recessive Sex- linked X or Y Mitochondrial DNA (maternal only) Today: X-linked dominant X-linked recessive Non Medelian X-linked Inheritance pattern for a familial translocation

SEGREGATION OF ALLELES IN X-LINKED INHERITANCE

This presentation includes: Segregation of alleles in X-linked inheritance DMD gene tracking in a family X chromosome tracking in a pedigree

From the NHS National Genetics Education and Development Centre www.geneticseducation.nhs.uk

X-linked inheritance where the mother is a carrier

Father

(Unaected)

Mother (Carrier)

Parents

Gametes
At conception

Daughter

Daughter (Carrier)

Son

Son (Aected)

http://health.allrefer.com/health/ duchenne-muscular-dystrophy-xlinked-recessive-genetic-defects.html

Duchenne Muscular Dystrophy (DMD) is a progressive genetic muscle wasting disorder. It is an X-linked condition caused by a mutation in the DMD gene. This gene encodes the dystrophin protein and lies on the short arm of chromosome X (p21.2).

Duchenne Muscular Dystrophy

X-Linked condition caused by a mutation in the DMD gene on Xp21.2 A milder form of the condition is known as Becker muscular dystrophy Deletion of large sections of the gene in 61% cases, duplications 13%, 26% point mutations

Women have two X chromosomes, and therefore if they have only one X chromosome with a mutation in the DMD gene, the other copy of the gene may be able to compensate. Up to about 20% of girls who carry a DMD mutation are thought to be affected to some extent. This is usually due to skewed X chromosome inactivation. The effects in girls are generally milder than those seen in boys, but a few girls have shown similar disease severity to boys.

Clinical features of Duchenne Muscular Dystrophy (DMD)

Children with Duchenne MD may need a wheelchair by the age of 10. They can also develop scoliosis, in which their spine begins to curve sideways. This can lead to one shoulder or hip being higher than the other. By the mid-teens, some people with Duchenne MD will develop dilated cardiomyopathy. This is where the condition affects the heart muscles, causing the chambers of the heart to enlarge and the heart walls to become thinner. By late teens or early twenties Duchenne MD can begin to cause respiratory (breathing) problems. The condition can affect the intercostal muscles (muscle tissue between the ribs) and the diaphragm (the main muscle between the chest and the abdomen that is used during breathing). Once the heart and respiratory muscles are damaged, Duchenne MD becomes life-threatening. In most cases, someone with Duchenne MD will die from heart or respiratory failure before they are 30 years old.

Once a boy is identified as being affected by DMD (Duchenne Muscular Dystrophy) or BMD (Becker muscular dystrophy milder form), their mother can be assessed to see if she also carries the mutation, and if she proves to be a carrier, then other potential adult female carriers in her family can be tested, as well as any possibly affected male offspring (i.e. cascade screening). About a quarter to a third of mutations are de novo. These cases occur in families with no known history of the disease WHY test? timely initiation of interventions (e.g. physiotherapy, corticosteroids) identification of behavioural and cognitive issues avoidance of exposure to harmful interventions (e.g. general anaesthetics) identification of cohorts for studies of novel therapies provision of timely information to parents for subsequent reproductive decision making and to allow time to make emotional and practical preparations associated with the diagnosis.

X chromosomes only are shown

X-linked recessive

This pedigree pattern can be explained by deducing the inheritance of the maternal X chromosomes

X chromosomes only are shown

X-linked recessive

or

An equal chance of being a carrier or not

Although the deletion for DMD can be large, it is not visible microscopically on a chromosome. Therefore molecular techniques are used to detect the the presence of a deletion or a mutation within the Dystrophin gene.

Introduction: multiplex PCR

http://www.dmd.nl/DMD_mPCR.html Multiplex PCR is a technique in which several PCR-products are amplified in one PCR-reaction. 1. Denaturation of DNA at high temperatures 2. Hybridisation to commercially prepared probes in the region of the DMD gene. 3. Amplification of the product through multiple cycles. 4. Visualisation on an electrophoresis gel that separates the DNA according to size Deletions detected by failure to amplify the hybridised region. Known mutations detected.

Tracking the inheritance of the gene causing Duchenne muscular dystrophy through the family

X-chromosomes shown

Therefore, Anne is not a carrier for Duchenne muscular dystrophy

Legend: Size: size of PCR product (in base pairs). Exon: exon of DMD gene present in PCR fragment.

Tracking the inheritance of the gene causing Duchenne muscular dystrophy through the family

X-chromosomes shown

Therefore, Anne is not a carrier for Duchenne muscular dystrophy

Lyonisation X inactivation
It is not necessary for both X chromosomes to have all their genes active. Therefore one of each X chromosome pair becomes inactive shortly after fertilisation. The process is random. Methylation occurs when the paternal and maternal chromosome are switched off at random in the early embryo. This happens to all homologous chromosomes. The Xist (X inactive specific transcript) gene produces an RNA that coats the chromosome and silences the genes. The chromatin is tightly wound around the histone molecules, also preventing genes from being active. All this is a process to control gene function and the protein produced, ie gene dosage. The only genes not switched off on the inactive X chromosome are those that are the same as on the Y chromosome and termed pseudoautosomal.

X- linked Dominant Inheritance

http://en.wikipedia.org/wiki/X-linked_dominant_inheritance

Congenital Generalised Hypertrichosis (CGH) X-Linked Dominant

Originally thought to be autosomal recessive, now known to be Xlinked dominant. Gene localised to Xq24-q27

X-linked dominant disorders are characterised by: expression in both sexes, but with a greater incidence in females due to the greater number of X chromosomes the female may be homozygous or heterozygous for the affected gene this can only be elucidated from the family pedigree - while the male can only be heterozygous the pedigree mirroring that of autosomal dominance. The only difference is that a positive father will give the condition to all of his daughters, but not his sons, whereas a positive female will transmit the trait to half of her sons and half of her daughters affected males having a uniform severity of disorder, while females are affected to different degrees Presently, there only a few known human X-linked dominant traits. With the exception of the Xg blood group, all are rare. Examples are: Xg blood group Alports syndrome vitamin D resistant rickets CGH congenital generalised hypertrichosis Rett's syndrome Fragile X syndrome . Can be considered non mendelian inheritance

NONMENDELIAN INHERITANCE Some clinical presentations do not fit the classical patterns of mendelian inheritance and represent examples of nontraditional or non-mendelian inheritance. These include triplet repeats, genomic imprinting, mosaicism, and mitochondrial inheritance.

The fragile X syndrome is said to be the most common heritable cause of mental retardation after Down's syndrome. It is caused by a dominant X-linked gene with a penetrance of only 50% in females. The gene which is most commonly responsible is FMR-1 (familial mental retardation 1). The disease occurs when the expression of FMR-1 is disrupted by: a large number, more than 230, of trinucleotide repeats a deletion (rare) It seems that loss of fmr-1 results in the fragile X syndrome.

Culture with folic acid deficient media. Block stain. CGC repeats detected using molecular techniques

The vast majority of fragile X patients have been shown to carry an expanded CGG repeat array at the fragile X A site (FRAXA) Normal individuals have alleles with between 6 and 52 CGG repeats, whereas fully affected fragile X patients have an allele with greater than 200 repeats. Nonpenetrant fragile X family members carry so-called premutation alleles in the range of 50 to 200 repeats

For those diseases associated with relatively small expansions, for example, less than 150 repeats, allelic status in all individuals may be determined rapidly by polymerase chain reaction (PCR) amplification of the region containing the repeats and sizing by polyacrylamide gel electrophoresis. For fragile X syndrome, a similar approach can also be used to identify normal alleles and small expansions

Fragile X Pedigree Premutation carrier Note male and female premutation carriers, plus affected individuals with full mutation

There is an increase in trinucleotide repeats with every generation.

Fragile-X Pre-pubertal features: normal growth but large head - greater than 50th percentile delayed attainment of developmental milestones tantrums, hyperactivity and autism Post-pubertal low IQ (20-70) long face, prominent forehead, large ears, large jaw macroorchidism Other features ophthalmologic - strabismus orthopaedic - pes planus & joint hyperextension dermatologic - soft, smooth skin cardiac - mitral valve prolapse Fragile X should be considered for all children with developmental delay of unknown cause.

The Y-linked inherited conditions are rare because there are only a small number of Y-linked genes outside the pseudoautosomal region or without a functional X homologue. A pedigree would show only male-to-male transmission, as no other transmission is possible, and one would never see affected females.1, 2 The bestknown example of a Y-linked condition is male infertility related to deletions of genes on the Y-chromosome (deleted in azoospermia). This condition usually results from new mutations, as complete infertility precludes any transmission, but rare males with some preserved fertility can transmit the mutation.

The meaning of "de novo" genetic variation

Sometimes, a disease-associated genetic variation is newly generated in an individual. In other words, an inherited disease can suddenly appear within a member of a previously unaffected family. The descendants of the affected family member are then at high risk of inheriting the newly generated familial mutation. In addition, the siblings of the affected family member may also be at increased risk of the disease, since the familial mutation may, in fact, have been generated in one of the patients parents ie, one of the parents could be a mosaic for the genetic variation. It is therefore important to consider genetic testing of the index patients siblings even if the genetic variation found in the index patient cannot be detected in either parent.