GENETIC BASIS OF DISEASES

PROF. H.A.NGGADA FMCPATH. DEPT OF HUMAN PATHOLOGY COLLEGE OF MEDICAL SCIENCES, UNIVERSITY OF MAIDUGURI

INTRODUCTION
Studies of genetics is taking us to the promise land. Engineer the genetic code, carrying with it the hope of
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alleviating or curing mutations. Much progress in medical genetics advances in molecular biology, involving recombinant DNA technology. Can excise human DNA by using restriction enzymes and to clone the excised fragments in appropriate cloning vectors.

When a known DNA fragment is used to detect whether

an unknown DNA fragment or m RNA carries complementary nucleotide sequences a DNA probe. Applications of DNA probe analysis is molecular hybridization. Hybridization can take place between complementary strands of either DNA and RNA; RNA hybrids, or between a DNA and RNA => DNA: RNA hybrid.

Some of these methods used are the southern blotting involves DNA: DNA hybridization. 4 DNA extracted from cell. Use of restriction enzymes.

Recognizes a specific sequence of base pairs and cut the DNA fragments. Place on gel electrophoresis. Denatured into single strands Transferred onto nitrocellulose membranes. Radiolabel led DNA probe applied. If any DNA fragment on the paper contains a sequence complementary to that of the probe, hybridization occurs.

Northern blotting Involves m- RNA : DNA hybridization

Dot blotting Does not involve restriction digestion RNA or DNA to be tested. Is applied onto nitrocellulose membrane and probed with a radio labeled DNA probe. Molecular probes are proving increasingly useful in the
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diagnosis of genetic as well as non genetic diseases. Last few years several cloned human genes have been induced to synthesize their products in unsuspecting bacteria large quantities of scared biologically active products. Like formation of recombinant growth hormone, factor VIII, thrombolytic agents and immunomodulators (Interleukin 2, Interferon).

Classic genetics
Traditional approach that has been successfully
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utilized to study a variety of inborn errors of metabolism such as Phenylketonuria and disorders of Hb synthesis. Common to these genetic disease is:
Knowledge of the abnormal gene product. Corresponding protein. When affected protein is known, then a variety of methods can be employed to isolate the normal gene. Clone it, and ultimately determine the molecular changes that affect the gene in the patient with the disorder.

Human diseases can be classified into 3 categories.

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Those

environmentally determined Genetically determine Both genetically and environmentally determine.

Terms used
Hereditary disorders derived from ones parent and
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are transmitted in the gametes through generations and therefore are familial. Congenital born with.

It should be noted that some congenital diseases are not genetic, e.g. congenital syphilis On the other hand, not all genetic diseases are congenital, e.g. Huntingtons disease, begin to manifest their condition only after 3rd or 4th decade of life.

Genotype - Phenotype
Genotype is defined as the genetic constitution of
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an individual and refers to which particular alternative version (allele) of a gene is present at a specific location (locus) on a chromosome. Phenotype is defined as the observed Structural, Biochemical and Physiologic characteristics of an individual, determined by the genotype, and refers to be the observed structural and functional effects of a mutant allele at a specific locus.

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PURINE/PYRIMIDINE: (A,G,C,U,T). DNA/RNA: (Sugar+ Phosphate Moiety +

Purine/Pyrimidine). REPLICATION: Reproduction of DNA. TRANSCRIPTION: Formation of RNA from DNA. TRANSLATION: Formation of protein on the mRNA molecule. Nucleus of somatic cell=DNA(Genome)=Arranged in Chromosomes(set)=46 (22 pair of Autosome/2 Sex Chromosome; XX/XY).

Genetic disorders fall into 5 major categories.

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Arising in chromosomal aberrations numerical Numbers - Structural 2. Those related to mutant genes of large effects 3. Diseases with multifactorial (polygenic) inheritance. 4. Single Gene Disorders with Non-classic Inheritance. 5.Mitochondrial Disorders
1.

The Normal Karyotype Cytogenetics (Study of genetics at the chromosomal level) 12

Human somatic cells are 46 chromosomes, these

comprises of 22 homologous pair of autosomes; and 2 sex chromosomes.

XX in the female XY in the male

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Study of chromosomes Karyotyping, which is the basic tool of the cytogenetist.

Chromosomal analysis.
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Use of blood cells / dividing cells. Culture cells in Phytohemagglutinin. Arrest of cell dividing after 72 hours (arrest of mitosis in

the metaphase stage) using colchicines Expose cells to hypotonic solution to swell up the cells. Fix cells on a slide Stain with Giemsa / Lisheman Arrange the cells base on their size and the position of the centromere. They are classified into seven groups. A G

Karyotyping
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A B C

D
E F G

13 45 6 12 X 13 15 16 18 19, 20 21, 22 Y

Writing of the Karyotype

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Karyotype are usually described using a short hand

system of symbols 46 XY. Total number of chromosomes first 47 Then sex chromosomes XY And finally description of any abnormality +21

47, XY, +21 Trisomy 21

Diagram showing a chromosome

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The Nature of the gene and the genetic code. (DNA)

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Cytosine

2 pyrimidine
Thymine Adenine

2 purine
Guanine The RNA is a single strand; Uridine replaces Thymine in the DNA

The Human Genome

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The human genome has approximately 38,000

genes, which are the individual units of heredity of all traits. Reproductive or germ line cells contain one copy(N) of this genetic compliment and are haploid, whereas somatic (non germ line) cells contain two complete copies (2N) and are diploid.

Genes
The genes are organised into long segments of
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deoxyribonucleic acid (DNA), which, during cell division, are compacted into intricate structures with protein to form chromosomes. Each somatic cell has 46 chromosomes (22 pair of autosomes, or non sex chromosomes), with 1 pair of sex chromosomes (XY in a male and XX in a female).

Genes
Germ cells (eggs, sperm) contains 22 autosomes
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and 1 sex chromosome for a total of 23 at fertilization, the full diploid chromosomes complement of 46 is again realized in the embryo.

DNA
The DNA molecule has 3 building block; a pentose
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sugar (deoxyribose), a phosphate group, and four types of bases, either purines (adenine and guanine) or pyrimidines (thymine and cytosine). These 4 bases form the alphabet of the genetic code. The basic subunit of DNA is the nucleotides composed of one deoxyribose, one phosphate group and one base

Structure of the DNA

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Phosphor

Phosphor

Sugar
Phosph Sugar Phosph

A----- T
T----- A

Sugar
Phosph Sugar Phosph

Sugar
Phosph Sugar

G----- C
C----- G A----- T

Sugar
Phosph Sugar

Phosph
Sugar Phosph Sugar

Phosph
Phosph

Mutations
24 Refers to a heritable alteration in the genome.

Some involves large segments of the genome and

therefore produce visible alterations in the structure of a chromosome. Many mutations, however, are submicroscopic. They may result in partial or complete deletion of a gene, or more often affect a single base. E.g. a single nucleotide base may be substituted by a different base a point mutation. Less commonly, one or two base pairs may be inserted or deleted from the DNA, alteration in the frame of the DNA strands.

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MUTATION: Permanent change in the DNA. Mutation can affect:

Germ cells=transmitted to progeny (inherited diseases). Somatic cells=not transmitted to progeny (important cause of cancer/congenital diseases). TYPES OF MUTATAION: Point mutation: Substitution. Missense. Non-sense. Frame shift: Insertion/Deletion. Trinucleotide repeat mutations: CATEGORIES OF GENETIC DISEASES: Mendelian (Single gene disorders). Polygenic /Multifactorial(Genetic/Environment). Chromosomal( Numeric/Structural). Single gene disorders with atypical inheritance: Triplet mutation. Mutations in mitochondrial DNA. Genomic imprinting.

Mutations within coding sequences. A point mutation (single base substitution) as seen in HbS. 26
DNA a chain 6th A.A _______________ C T C G A G ________________ ________________ C A C G T G ________________ s chain mRNA ______________ G A G A.A Glutamic acid

________________ G U G

Valine

This single A.A substitution alters the physiochemical properties of Hb, giving rise to sickle cell anaemia

A point mutation may change an Amino acid codon to a chain terminator or stop codon.
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38 39 40 Normal--------Thr-----------Gln------------Arg----------- A.A sequence A C C C A G A G G Codons

A C C U A G Mutant--------Thr--------STOP 38

A G G

Codons A.A sequences

This is seen in globin, where C is replaced by U

Prematured stoppage of the - globin gene transcription and the resulting short peptide is rapidly degraded. The affected individuals lack -globin chains severe form of anaemia, the o thalassemia.

1. CYTOGENETIC DISORDERS
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It takes the form of abnormal number of the

chromosomes or alteration in the structure of one or two or more of the chromosomes.

Numerical abnormalities Structural abnormalities

Normally 46 known as 2n or diploid number

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First meiotic reduction division in the formation of

gametes 23 or haploid Normal count 46XX / 46XY Exact multiple of the haploid number euploid If an error occurs in meiosis or mitosis that is not an exact multiple of 23, it is referred to as aneuploidy. Usual causes for aneuploidy are
Nondisjunction Anaphase lag

Chromosomes during Mitotic Divisions

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Chromosomes during Meiotic Divisions

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Nondisjunction. Have a look at the normal meiosis.

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Nondisjunction at gametogenesis gametes (n+1)

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Nondisjunction at gametogenesis gametes (n+1)

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In anaphase lag
one homologous chromosome in meiosis or one
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chromatid in mitosis lags behind and is left out of the cell nucleus, resulting to one normal cell and one cell with Monosomy. Monosomy or Trisomy involving the sex chromosomes, or even more bizarre aberrations are compatible with life and are usually associated with variable degrees of phenotypic abnormalities.

In anaphase lag
On the other hand, Monosomy involving an autosome
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generally represents a loss of too much genetic information to permit live birth or even embryogenesis; but a number of autosomal trisomies do permit survival. Mitotic errors in early development give rise to two or more populations of cells in the same individuals, a condition referred to as mosaicism. This can result from mitotic errors during the cleavage of the fertilized ovum or in somatic cells. Mosaicism affecting sex chromosomes is relatively common.

 In the division of the fertilized ovum an error

37 may lead to one of the daughter cells receiving 3 sex chromosomes, while the other receives only one 45,X / 47, XXX mosaic All descendant cells derived from each of these precursors will thus have either a 47, XXX count or a 45, OX count Turners syndrome. Autosomal mosaicism is less common. You can have but rarely. Loss of a nonviable cell in embryogenesis is tolerated mosaic. e.g. 46, XY /47, XY + 21 Down syndrome.

Structural Aberration
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Occurs from chromosomal breakage followed by loss

or rearrangement of material. Translocation a segment of one chromosome is transferred to another. One form of Balanced reciprocal translocation

Translocation
39 Phenotypically is normal, No material / information is loss. However, a balanced reciprocal translocation carrier is at risk of

producing abnormal gametes. They become unbalanced; subsequate fertilization with a normal gametes will produce an abnormal (unbalanced) zygote spontaneous abortion or malformed child. Phenotypically is normal, No material / information is loss.

Another form of translocation is the Robertsonian translocation.

Translocation
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Much informations are not lost; the small one

carries little genetic information that this loss is compatible with a normal phenotype. It can be seen in Downs syndrome You can have other forms of translocation, such as Deletion loss of a portion of chromosome. May be terminal or interstitial46, XY, 16 p-

Trisomy 21 (Downs Syndrome)

Nigeria 1/ 800 live birth US 1 / 700 live birth Has an association with increase in maternal age

Nondisjunction

95%
47,XX +21 47,XY +21

Translocation 4%

Mosaic 1%
46,XX/47,XX,+21

46,XX 14 + t (14q21q) 46,XY 22 ,+t (21q22q)

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Features
44 Mental retardation 80% IQ between 20 80

(mostly b/w 45-55) Hypotonia 80 100%

Flat facial profile, oblique palpebral fissures Epicanthic folds Cardiac defects 40% - septal defect

Abnormal GI 15% oesophageal and duodenal

atresia Susceptibility to infection, acute leukemia

Features
45 Abundant neck skin, Broad and / or short trunk

Dysplastic and low set ears Hyperflexibility, lack of Moro reflex Single horizontal palmer crease Dysplastic middle phalanx

Sexual development retarded Prolonged jaundice, Polycythemia Later an increased tendency to thyroid dysfunction.

Trisomy 18 (Edwards Syndrome)

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Incidence 1 / 4000 live birth

Associated with increased maternal age

Nondisjunction

47, XX, +18 90% Features Mental retardation , FTT, Prominent occiput Micrognathia, low set ears, Hypertonia

Mosaic 46, XX/ 47, XX+18 10%

Features continue
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Flexion of finger (2nd and 5th digits overlapping 3rd

and 4th ) Rocker bottom feets Cardiac dextroposition and Intestinal defects Short sternum and small pelvis Death often caused by cardiac failure of pneumonia Perinatal and postnatal growth retardation

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Trisomy 13 (Pataus Syndrome)

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Incidence 1/ 6000 live birth

Associated with maternal age

Nondisjunction 80%- 47, XX, +13 Translocation 10% 46, XX, - 14, +t (14q13q)

Mosaic

--10% 46, XX /47,XX, +13

Feature
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Microcephaly Mental retardation, scalp defect Microphthamia, cleft lip and palate

Polydactyly
Abnormal ears Extensive visceral defects

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CYTOGENETIC DISORDER INVOLVING SEX CHROMOSOMES

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Effects of X chromosome

Genetic disease associated with karyotypic changes

involving the sex chromosomes are far more common than those related to autosomal aberrations. Furthermore, imbalances (excess or loss) of sex chromosomes are much better tolerated than are similar imbalances of autosomes. They are often difficult to diagnose at birth, and many are first recognized at the time of puberty. The higher the number of X chromosome in both male and female, the greater the likelihood of mental retardation.

 Has slight association with maternal increase of 53

Klinefelters syndrome

age 1 / 850 male birth 47, XXY ; 46XY/47 XXY FEATURES Testicular atrophy and azospermia Eunuchoid body habitus Increased in sole-to-os pubis length Gynecomastia

Turners syndrome
1/3000 female birth, 45X, 46,XXp54

Features Primary amenorrhea Infertility, webbing of neck, Cubitus Valgus Peripheral lymphedema, Broad Chest and wide spaced nipples, low post hair line Coarctation of aorta Normal IQ

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Mendelian Disorders
Result of expressed mutation in single genes of large
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effect transmitted by Autosomal dominant, recessive, or X linked. You have the homologous loci the identical pair of loci alleles If a mutant gene occurs at a locus in one chromosome but not at the homologous locus of other person is heterozygous If the mutant gene does not affect the heterozygous state, it is called recessive gene AA mutant gene, phenotypically -normal

Mendelian disorder contn.

If the mutant gene affect the heterozygous state, it is a

dominant gene. AA mutant gene, phenotypically affected A person having the same mutant gene at both homologous loci, is homolozygous for that gene. AA Homozygous Autosomal recessive gene manifest their clinical effect only in the homozygote

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Dominant Inheritance
When a gene produces its effect whether it is present 58

either upon one or upon both chromosomes of the pair concerned. Homozygous Heterozygous Parents A A A A Gametes
fertilization

AA Homozygous affected

AA Heterozygous affected

offspring

Dominant Inheritance
Every affected person has an affected parent 59 The gene must have come from one or both

Affected persons married to normal, on the average,

affected and normal offspring in equal proportion. Heterozygous Normal Parents A A A A Gametes
Fertilization

AA

AA AA AA normal heterozygous affected

offspring

Autosomal Dominant
The normal children of affected person, when they in 60

turn marry a normal person, will have only normal children. Examples

Achondroplasia Dentinogenesis imperfecta Huntington chorea Marfan syndrome Neurofibromatosis Polycystic kidney (all types) Retinoblastoma Tuberous sclerosis

Autosomal recessive
Must produce its effects only when present upon both 61

chromosomes. Only abnormal gene alone produces no effect. AS A A Parent gametes

AA AA Normal

fertilization AS AS offspring's Affected, but phenotypically normal, heterozygote

Affected person must receive the gene for the condition from both parents.
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AS A S A S

AS

Parents Gametes

fertilization
AA

AS

Normal

AS SS offsprings heterozygous homozygous not affected affected looks normal

Only when a heterozygote happens to marry another heterozygote can the homozygote appears.
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Examples Adrenogenital syndrome Albinism 1 antitrypsin deficiency Cystic fibrosis Galactosemia Haemoglobinopathies Hurler syndrome

III.X-Linked Disorders:
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All sex linked disorders are X-linked. Most X-linked disorders are X-linked recessive and

are characterized by:

Transmitted by heterozygote females carriers only to their sons(hemizygous for X chromosome). Random inactivation of X-xsome occurs in females. An affected male does not transmit disorder to sons but all daughter are carriers. Sons of heterozygous women have 1 in 2 chance of receiving the mutant gene.

1.BLOOD:

Chronic Granulomatous Diseases. Hemophilia A and B. XLD G-6-PD. Agammaglobinaemia. X-linked SCID. Wiscott-Aldrich Syndrome. Diabetes insipidus. Lesch nyhan syndrome. Fragile X syndrome. XLD Duchenne muscular dystrophy. XLD
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2.IMMUNE:

3.METABOLIC:

4.NERVOUS:

5.MUSCULOSKELETAL:

IV. X-LINKED DOMINANT

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Few X-linked dominant diseases and they are

characterized by:

Inheritance pattern is characterized by transmission of the disease to 50% of sons and daughters of an affected heterozygous female. An affected male can not transmit the disease to his sons but all daughters are affected

 SEX- LIMITED TRAIT

These appear in only one sex. Eg baldness which is

limited to the male

MULTIFACTORIAL INHERITANCE(POLYGENIC):
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Disorders with multifactorial inheritance are the

consequences of the interplay between two or more mutant genes combined with environmental factors. Multifactorial inheritance controls many physiologic attributes:

Height,Weight,Hair Color and Blood Pressure.

Multifactorial inheritance underlies many congenital

malformation and common disorders such as;

Diabetes,Hypertension,Gout,Coronary Heart Disease,Scizophrenia,Bipolar disorders.

CONT
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Disorders with multifactorial inheritance exhibit these

characteristics: Although the risk of expression is conditioned by the number of mutant genes inherited, Environmental influences significantly modify the risk of expression; hence the Concordance rate even in- identical twins- is only 20-40%. Risk of concordance of the disorder in first degree relative is 2%-7%. If two children in a family have the disorder the risk of recurrence in subsequent offspring rises to 9%.

Examples: Diabetes mellitus (Insulin dependent) Diabetes mellitus (Insulin independent) Hypertension Leprosy Pyloric sclerosis Spina bifida Schizophrenia etc

4. SOMATIC CELL GENETIC DISORDERS

Here mutations arise in fertilized ovum, which will be

transmitted to all daughter cells. Cancer provides examples of this. Examples: Chronic myeloid leukemia Wilms tumour Breast cancer Retinoblastoma Colon cancer etc

5. MITOCHONDRIAL DISORDERS
Human mitochondria each have about 10 copies of double

stranded DNA which are self replication and they encode for several enzymes involved in oxidative phosphorylation. As mitochondria are found in the cytoplasm, they are transmitted in the ovum from a mother to all her children. A few clinically important disorders have been discovered to result from their mutations. Examples are: Lebers hereditary optic neuropathy Mitochondrial myopathies