Clinical Cytogenetics: The
Chromosomal
Basis of Human Disease
Le Minh Thong PhD.
Chromosome abnormalities
• 1 of every 150 live births
• Leading cause of intellectual disability and pregnancy loss
• Seen in 50% of first-trimester and 20% of second-trimester
spontaneous abortions
• Specific genes can affect cytogenetic phenotypes
In this chapter:
• Abnormalities of chromosome
• Number
• Structure
• Sex chromosome abnormalities
• Cancer
• Other disease
• New technology in cytogenetics
CYTOGENETIC TECHNOLOGIES
46,XY,t(7;10)(p22;q23)
• Put the root tip in acid
• Soak in the dye
• Press on the slide
• Watch under microscope
[Link]
Partial trisomy of the distal part of 10q: A report of two Egyptian cases
Karyogram and karyotyping
• To identify the number, shape and size of chromosomes of
cells
Medical dictionary
Karyotyping: other cases
Chromosome banding techniques
Q Banding
Solid Staining
R Banding
G Banding
Other selective staining techniques
C (centromere) Banding Silver Staining
High Cytogenic Resolution Banding
Prophase cells
Band level: ~ 500 -> 800 bands
Resolution: 2 – 3 Mb < 5 Mb
Karyotype nomenclature
• By Size
• By band pattern
• By centromere pattern
• Metacentric
• Acrocentric
• Sub-metacentric
Karyotype nomenclature
Karyotype nomenclature
• Male: 46,XY
• Female: 46,XX
• Region:
• 14q32: second band in the third region of the long arm of
chromosome 14
Fluorescence in Situ Hybridization (FISH)
• Can use metaphase,
prophase and interphase
(faster, no need to culture
cell)
• Higher resolution: ~ 1 Mb
• Detect delete and extra
chr/region
• Multiplex ~ spectral
karyotyping
Small arrow: Chr 17
Large arrow: 17p
Spectral karyotyping
Comparative Genomic Hybridization
• Array CGH:
• High resolution 50 – 100 kb or only one gene
• No need culture cell
• Less required DNA
Chromosome abnormalities
• Chromosome number
• Polyploidy
• Triploidy
• Tetraploidy
• Aneuploidy
• Autosome: Trisomy 13, 18, 21
• Sex chromosome aneuploidy:
X-, XXY, XXX
• Chromosome structure:
• Translocation: balance,
unbalance
• Deletion
• …..
Polyploidy
• Triploidy: 69,XXX (Y also)
• Rate: 1/10000 in birth but
15% in conception: loss at 2
first trimesters
• Shortly died
• Almost by dispermy Triploidy
• Also by using a polar body
• Or by meiotic failure
• Tetraploidy: 92,XXXX (Y also)
• Very rare in birth or
conception
• Short life
• Caused by meiotic failure
Mosaic triploidy
Tetraploidy
15 months old baby with tetraploidy
Autosomal aneuploidy
• Aneuploidy = not euploidy (23 chr x 1 or 2 or 3 or 4 …)
• Usually only one chromosome affected
• Missing – monosomy
• Extra (duplicated) – trisomy
• Common cause: Nondisjunction in meiosis I or II
• Autosomal aneuploidy normally die except:
• Trisomy 13, 18, 21
• But not monosomy
Nondisjunction in meiosis
Down syndrome – Trisomy 21
John Langdon Down – 1866
Down syndrome
• Prevalence: 1/700
• Characteristic appearance of the face
• Short neck, limp (and flat)
• Hypotonia (decrease muscle tone)
• 3% closure or absence of anus
• 40% heart defect
• IQ 40 – 60
• 20 time higher leukemia
• Infertile in male
• ~ 95% due to mother extra chr
• 2- 4% mosaicism – tissue specific
• Important gene: DYRK1A - a kinase gene that causes learning and memory defects
when it is over-expressed
“Lastly, a recent clinical trial has shown that
epigallocatechine gallate (EGCG), a DYRK1A inhibitor, given
to young patients with DS improved visual recognition
memory, working memory performance and adaptive
behaviour.”
Edwards syndrome – Trisomy 18
• Trisomy 18 (47,XY,+18)
• Prevalence 1/6000 ~ 5% of cases from conceptus (95% died, stillborn)
• Characteristic facial features
• Heart defect
• ~ 50 % die at first year
• Only ~ 8% get to 12 year olds
Patau syndrome – Trisomy 13
• Trisomy 18 (47,XY,+18)
• Prevalence: 1/10000
• Malformation at nervous system
• Heart defect
• 95% die at 1st year
• Due to mother site
Trisomies, Nondisjunction, and Maternal Age
• Increase in nondisjunction
among older mothers
• Nearly all of a female’s oocytes
are formed during her
embryonic development (45
year old female have 45 year old
eggs)
• Other factors (but not clear in
human):
• Hormone level
• Cigarette
• Alcohol
• Radiation
Sex Chromosome Aneuploidy:
Monosomy of the X Chromosome (Turner
Syndrome)
• 45,X, women
• Short stature, sexual infantilism (less 20 cm)
• Triangle-shaped face
• Webbing of the neck
• Chest is broad and shield-like Henry Turner 1938
described the disease
• Congenital heart, kidney defect
• Normal intelligence
• Infertile (Some have baby)
• Hormone therapy (growth hormone, estrogen)
• 40% have mosacism (45,X/46,XX >> 45,X/46,XY)
• 60 - 80% of monosomy due to the lacking X chr in
sperm
• 1 - 2% of conception but seen 1/2000 - 1/3000
(99% died in conception)
• Main gene: SHOX (expressed on X and Y short tip)
Klinefelter Syndrome - 47,XXY - male
• 1/500 to 1/1000 male births
Described the
• taller than average syndrome in 1942
• long arms and legs
• small testes (10 ml in vol)
• sterile
• 1/3 developed breast => cancer Harry Klinefelter
• Normal intelligence
• 15 % mosaicism
• Also 48,XXXY; 49,XXXXY (more X
more severe)
• Testosterone theraphy
Trisomy X - 47,XXX - female
• 1/1000
• benign consequences
• sometimes have sterility
• mild cognitive disability
• Due to nondisjunction from mother
• More X more severe
Wiki
47,XYY Syndrome -male
• Taller than average
• Reduce IQ
• Incidence in the male prison
population (1/30 >> 1/1000)
• Behavioral disorders: hyperactivity,
attention deficit, learning
disabilities.
CHROMOSOME ABNORMALITIES AND PREGNANCY
LOSS
• Spontaneous pregnancy loss is common in humans (1/3, after
implantation)
• 10 – 20% abnormalities in chromosome => 95 % lost before term
• 50 % trisomy (trisomy 16 is most common, but never come)
• 20 % monosomy
• 15 % triploids
• Remain: tetraploids & structure abnormalities
• How about gametes?
• Egg (from unused IVF): 20 – 25% missing or extra, 1% structure abnormalities
• Sperm (fused with hamster egg condensed): 3 – 4%, 5 % structure abnormalities
• Incidence increased follow age
ABNORMALITIES OF CHROMOSOME STRUCTURE
• Balance
• Unbalance
• Cause
• Unequal crossover
• Chromosome breakage during
meiosis
• Ionizing
• Radiation
• Viral infection
• Other chemicals
Translocation: Reciprocal Translocations
• Mutually exchange between non-
homologous chromosomes
• The carrier normally normal
• The offspring:
• Normal
• Duplication – partial trisomy
• Deletion
Translocation: Robertsonian Translocations
• Short arm loss and long arm fuse to
single chromosome
• Occur in acrocentric chromosome
(13, 14, 15, 21, 22)
• Usually no harm: short arm less
materials
• The carrier: 45 chromosomes in
cell, unaffected
• The offspring: extra (Down) or
missing long arm of acrocentric
chromosome
• Most common 45,XY or
XX,der(14;21)(q10;q10)
Deletions
• Terminal deletion
• Interstitial deletion
• Loss materials ~ several genes
(microscopy abservable)
• Example: Cri-du-chat syndrome
• Distal short arm of chromosome 5
• 46,XY,del(5p)
• 1 in 50,000 live births
• Intellectual disability (IQ 35)
• Abnormal crying
• Normal apperrance
Microdeletion Syndromes
• Detected by high resolution FISH or aCGH (< 5 Mb)
• Example: Prader–Willi syndrome 70% cases caused by microdeletions of
15q
• Also microduplication
• Caused by multiple repeated sequences at boundary of the dup/del
Subtelomeric Rearrangements
• High density of genes in telomere
• Telomeric rearrangement normally causes disease
• Most common 1p36 deletion (monosomy) (1/5000 births) =>
intellectual disability, developmental delay, seizures, hearing
impairment, heart defect
Uniparental Disomy
• 30 % cases of Prader-Willi syndrome
• Abnormal imprinting (both copies imprinted and not expressed)
Duplications
• Seen in offspring of reciprocal translocation carriers
• Unequal crossover during meiosis
• X-linked color vision loci
• Produce less serious consequences
Ring Chromosomes
• Deletion in both chromosome tips and ligate
to the ring
• Ring structure is not favored in mitosis =>
loss in some cell monosomy mosaicism
• Any chromosome can be ring, most
common: 14 and 22 chr
46,X,r(X)
Inversions
• Due to 2 breaks of chromosomes
and reinsertion by repair system
• Pericentric inversion (with
centromere)
• Paracentric inversions (without
centromere)
• Example: inversion that interrupts
the factor VIII gene produces
severe hemophilia A
• Other consequences: Induce
deletion or duplication in meiosis
Isochromosomes
• Wrong axis devide of chromosome
propagation or Robertsonian translocation
• Autosome – lethal
• Seen in X isochromosome
• (46,X,i[Xq]: Turner
CANCER CYTOGENETICS
• Take place in somatic cell
• chronic myelogenous leukemia (CML): reciprocal translocation between
chromosomes 9 and 22 increase tyrosine kinase
• And more than 100 rearrangement for 40 type of human cancers
CHROMOSOME INSTABILITY SYNDROMES
• Chromosome very easy to be breaked
• May relate to repair system defect
• induce cancer
• Many protein encoded by nuclear genes and go to mitochondria, how
about mitochondria situation in case of polyploid peoples?
• Why many chromosome abnormalities lead to heart defect
• How to detect mosacism?
