Genetic in Clinical Setting Part II

NEUROMUSCULAR
DISORDERS

Tri Indah Winarni, MD, PhD

Faculty of Medicine, Diponegoro University
Study Objectives
• Memahami mekanisme jenis mutasi “trinucleotide
repeat expansion/TREs”How the length matter?
• Gene switch off/epigenetics/imprinting (loss of function)
• Gain of function (protein mutant)
• Instability and transmitting the diseases from carrier to affected
• Memahami implikasi mekanisme TREs pada beberapa
penyakit neuromuscular
• Fragile X syndrome (hypermethylation cause switch
off the gene)
• Friedreich ataxia
• Spinocerebellar ataxia (SCA)
 EPIGENETIC, DEFINITION AND FUNCTION
• Variations in gene expression that are not caused by changes in
DNA sequence (imprinting center)
• Epigenetic mechanisms regulate all biological processes from
conception to death organized by epigenome
There Are Two Type of Epigenetics
1. DNA methylation by the addition of a methyl (CH3) group
to Cytosine, will modifying the function of the genes (gene
expression). When a CpG island is methylated the gene is
turned off
2. Histone post-translational modifications: adding methyl group
making DNA coil more tightly (not accessible) or adding acetyl
group/demethylation making DNA more loose (accessible)
 A. Histones can undergo phosphorylation (Ph), methylation (Me), and acetylation (Ac), among other
chemical modifications. B. DNA molecules are methylated by the addition of a methyl group to carbon
position 5 on cytosine bases which maintains repressed gene activity. C. mRNA is translated into a protein
product, but this process can be repressed by binding of microRNAs (miRNA), a noncoding RNA (ncRNA)
Gene Expression and Epigenetic Role
• Gene is a factory that produce protein as requested by human
body
• Gene (blue print) contains DNA sequence who has specific function
• Not all gene are active in the same time
• Epigenetic (imprinting) regulate to switch on and switch off the
gene internally (physiologist) and externally/environmentally
(pathologist)
• Environment can interact with the gene and stimulate OR
suppressed gene expression/translation, this
(imprinting/epigenetics) can be inherited from parent
• Fear of dark or height
• Feeling anxious on specific events/situations
Inherited Epigenetic in coping to stressors
Impact of mutation on protein function
• Loss of Function
• Complete loss of the protein: null, loss-of-function
• Fragile X syndrome  gene silencing (NO protein product)
• Friedeich Ataxia, longer the repeats, the lower the level of frataxin
and the more severe the phenotype  gene silencing (no protein
product)
• Gain of Function
• Increase in the protein’s function, called activating mutations
• Almost always dominant
• Spinocerebellar Ataxia expanded polyglutamine (PolyQ)  mutant
protein
• Myotonic Dystrophy  CTG repeat cause new RNA properties
Fragile X syndrome (FXS)
• The most common cause of X-linked inherited intellectual disabilities
• Trinucleotide Repeat Expansion TRE’s disease, CGG repeats in UTR
FMR1 gene:
• Common allele (4-54)
• Premutation allele (55-200), called carrier  instability started
• Full mutation allele CGG (>200) will lead switch off the gene and results lack
of Fragile X Mental Retardation Protein
• FXS characteristics
• Behavioral, autism-like symptoms
• Physical, large and prominence ears, elongated facial feature, hyperextensible
joint/connective tissue laxity, and macroorchidism (≥ 25 ml)
• Prevalence: males 1 in 2500, females 1 in 4000
Why the repeat length matter?
• Determine instability of the gene (>54
the instability is started)
• The bigger the repeat the bigger the
expansion
• 55-59 repeats  5.4% expansion to full
mutation
• 100-109 repeats  97% expansion to full
mutation
• Cause switch off the gene when the
number above threshold (200 repeats)
Molecular Mechanism of FXS

Epigenetics mechanism
Fragile X syndrome (FXS)
 Friedreich Ataxia (FRDA)
• The most common autosomal recessive inherited young onset of ataxia
(neurodegenerative disease)
• GAA repeats expansion in intron 1 FRDA gene (chromosome 9q13)
• 6-36 GAA repeats (normal individuals)
• 70 to 1700 repeats (FRDA patient), most commonly 600 to 900 GAA
• The severity of the disease correlates with the number of repeats
• The basis of disease is impairment in mitochondrial function due to loss of
frataxin (a mitochondrial protein for cellular iron homeostasis)
• A young-onset progressive ataxia with sensory loss, scoliosis, and
cardiomyopathy occurring before age 25
• Co-morbid conditions: hearingloss, motor weakness, and diabetes
• Prevalence: 1:50.000 in Caucasian populations
Molecular mechanism of Friedreich Ataxia

Epigenetics mechanism
 Figure 1. Pathological mechanisms involved in FRDA and associated therapeutic strategy. FRDA is
caused by a GAA

Hum Mol Genet, Volume 19, Issue R1, 15 April 2010, Pages R103–R110,
https://doi.org/10.1093/hmg/ddq165
The content of this slide may be subject to copyright: please see the slide notes for details.
Spinocerebellar Ataxia (SCA1)
• Autosomal dominant inherited neurodegenerative diseases due to
the expansion of
• CAG repeats in exon 8 ATXN1 gene (chromosome 6p22.3)
• age of onset and disease severity are correlated with the length of the repeat
expansion
• Disease mechanism: the formation of an abnormal elongation of the
polyglutamine (polyQ) residues in the mutant protein  impaired
protein-protein interactions  formation of insoluble amyloid
inclusions in neurons
• Characteristics: gait ataxia (impaired balance) and uncoordinated
movements, Purkinje cell death and cerebellar atrophy
• Prevalence: 1 in 100,000 population (varied in some popualtions)
 Formation of an abnormal elongation of the
polyglutamine (polyQ) residues in the mutant
protein  neurons inclusions