13.11.2007 gene1.ppt 1 The Pathogenesis of Diseases from The Pathogenesis of Diseases from Genetic and Genomic Genetic and Genomic Point Point of View of View Oliver Oliver R ácz ácz and František and František Ništiar Ništiar Institute of Pathological Physiology Institute of Pathological Physiology Medical School Medical School , Šafárik , Šafárik University University 13.11.2007 gene1.ppt 2 26th 26th june june 2000 2000 is is neither the beginning neither the beginning nor nor the the end end of of the way the way 5 5 years years before before term term (1990 (1990 - 2005) 2005) The race is The race is over, over, victory for Craig Venter victory for Craig Venter. The genome is mapped The genome is mapped* - now what now what ? Not Not a a discovery discovery! A A very very important important technological technological result result and and competition competition is is always always useful useful. all all is is based based on on Mendel Mendel ‘s and ‘s and Watson Watson ‘s & ‘s & Crick Crick ‘s ‘s discoveries discoveries in in XIXth XIXth XXth century XXth century *3*10 9 letters
24
Embed
The Pathogenesis of Diseases from Genetic and Genomic ...€¦ · The Pathogenesis of Diseases from Genetic and Genomic Point of View ... 13.11.2007 gene1.ppt 3 Mendel, Watson, ...
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
13.11.2007 gene1.ppt 1
The Pathogenesis of Diseases from The Pathogenesis of Diseases from Genetic and GenomicGenetic and GenomicPoint Point of Viewof View
OliverOliver RRácz ácz and František and František NištiarNištiar
Institute of Pathological PhysiologyInstitute of Pathological Physiology
Medical SchoolMedical School, Šafárik , Šafárik UniversityUniversity
13.11.2007 gene1.ppt 2
26th 26th junejune 2000 2000 isis neither the beginning neither the beginning nor nor the the end end of of
the waythe way�� 5 5 yearsyearsbeforebefore term term (1990 (1990 -- 2005) 2005)
�� The race isThe race isover, over, victory for Craig Ventervictory for Craig Venter ..
�� The genome is mappedThe genome is mapped** -- now whatnow what ??
�� Not Not a a discoverydiscovery!!
�� A A veryvery importantimportant technologicaltechnologicalresult result and and competitioncompetitionisis alwaysalwaysusefuluseful..
�� allall isis basedbasedon on MendelMendel‘s and ‘s and WatsonWatson‘s & ‘s & CrickCrick‘s ‘s discoveries discoveries in in XIXth XIXth XXth centuryXXth century
*3*109 letters
13.11.2007 gene1.ppt 3
Mendel, Watson, Crick & the medical Mendel, Watson, Crick & the medical geneticsgeneticsofof XIXXIX thth –– XXXX thth centurycentury
�� MendelMendel‘s ‘s lawslaws in in medicinemedicinecancan bebe appliedapplied to to monogenicmonogenicdiseasesdiseases–– longlong list, list, relativelyrelatively rarerare
�� TheThenumbernumberofof humanhumangenesgenesisis asaslowlow as as 30 00030 000�� thethesmallsmallwormworm C. C. eleganseleganshas 20 000 has 20 000 genesgenes
�� thethemousemousehas has asasmanymanygenesgenesasaswewe, , also with very similar also with very similar functionfunction
�� TheThemysterymysteryisis in in complexitycomplexityand and networkingnetworking: : 223000030000>>>> 2>>>> 220000 20000 (possible on/off states)(possible on/off states)
�� It is mapped but do we understand it? It is mapped but do we understand it?
�� We know the keys of the piano but are we able to We know the keys of the piano but are we able to play even simple composition on it?play even simple composition on it?
�� GENETICS = HEREDITYGENETICS = HEREDITY
�� GENOMICS = EVERYTHINGGENOMICS = EVERYTHING
13.11.2007 gene1.ppt 18
GGEENNESES AAND THE ENVIRONMENTND THE ENVIRONMENT
GENOME ENVIRONMENT
SEVERE MONOGENICDISEASES
LESS IMPORTANT MUTATIONS
GENETIC RISK
NEGATIVE ANDPOSITIVE
ENVIRONMENTALFACTORS
physicalchemicalbiologicalnutritionlife style
13.11.2007 gene1.ppt 19
MutationsMutationschanges of genetic informationchanges of genetic information
�� THREE PRINCIPAL POSSIBILITIESTHREE PRINCIPAL POSSIBILITIES1.1. changes in genome not compatible with lifechanges in genome not compatible with life
2.2. development and diversitydevelopment and diversity
3.3. disease or disease or increased risk increased risk fofo disease*disease*
�� THE BASIC DIFFERENCE FOR HEREDITY:THE BASIC DIFFERENCE FOR HEREDITY:–– somatic and germ cell mutationssomatic and germ cell mutations
�� genome, chromosomal and genome, chromosomal and genegenemutationsmutations
no genes for diseases! – sickle cell, Alzheimer, diabetes...
13.11.2007 gene1.ppt 20
Gene mutations and Gene mutations and SNPsSNPs**
�� Point mutations in Point mutations in exons exons –– silent, silent, missensemissense(AA change) and nonsense (stop)(AA change) and nonsense (stop)
�� FrameshiftFrameshiftmutation in mutation in exonsexons(1,2,4,5...)(1,2,4,5...)�� Small deletion of triplets (3,6...)Small deletion of triplets (3,6...)�� Bigger deletions Bigger deletions –– transition to chromosomal transition to chromosomal
aberrationsaberrations�� Mutations in regulatory parts, Mutations in regulatory parts, intronsintrons, , genes for genes for rr--
�� X chromosome linked diseasesX chromosome linked diseases–– hemophilia A, B; hemophilia A, B; daltonismdaltonism
�� and von and von WillebrandWillebranddisease, factor V disease, factor V LeidenLeiden, , hereditary hereditary hemochromatosishemochromatosis......
13.11.2007 gene1.ppt 22
Sickle cell diseaseSickle cell disease�� Clinical description Clinical description ≈ 1910, Hb abnormality, 19401910, Hb abnormality, 1940�� PaulingPauling/ Ingram / Ingram -- 1 AA change in 1 AA change in ββ chainchain�� Point mutation Point mutation –– Glu Glu → Val on 6th place (GAG/GTG)
� Decreased solubility of Hb in low pO2� Rigid, deformed red cells in venous blood� Thrombosis, decreased life span of Er, hemolysis,
icterus, anemia - HYPOXIA
Epidemiology: 8 % of black people in USA are heterozygotes; 1:400 homozygotes
5 – 20 % heterozygotes in some parts of Africa
13.11.2007 gene1.ppt 23
And the other And the other haemoglobinopathieshaemoglobinopathies??
�� Theoretical number Theoretical number –– astronomicalastronomical�� Known Known ≈≈ 500, 500, very rarevery rare�� Hb C = same point as Hb S but lysine, mild Hb C = same point as Hb S but lysine, mild
haemolysishaemolysis. . HbSC heterozygotesHbSC heterozygotes�� Different types: labile, low and high oxygen affinity, Different types: labile, low and high oxygen affinity,
methemoglobinmethemoglobinformation, etc.formation, etc.�� Why is sickle cell disease relative common?Why is sickle cell disease relative common?�� Plasmodia Plasmodia malariaemalariaedo not like Hb S!do not like Hb S!�� AA dies on malaria, SS on sickle cell diseaseAA dies on malaria, SS on sickle cell disease�� AS have relative advantage for survivalAS have relative advantage for survival
13.11.2007 gene1.ppt 24
Occurrence ofOccurrence ofHbHb S in the worldS in the world
13.11.2007 gene1.ppt 25
13.11.2007 gene1.ppt 26
The molecular structure of humanThe molecular structure of humanHbHb
13.11.2007 gene1.ppt 27
Normal and Normal and sickledsickledred cellsred cells
13.11.2007 gene1.ppt 28
The pathogenesis of sickle cell diseaseThe pathogenesis of sickle cell disease
�� Deletion of smaller or bigger parts of Deletion of smaller or bigger parts of αα or or ββ gene gene region region
�� (or mutation in regulatory parts, nonsense (or mutation in regulatory parts, nonsense mutation and mutation and intronintron splicing mutations)splicing mutations)
�� ββ deletions deletions –– back to back to embryonalembryonalHb F if possibleHb F if possible�� αα deletions deletions –– 2*2 = 4 genes!2*2 = 4 genes!�� αααα αααα norm, healthynorm, healthy�� αααααα 1 deletion, clinically not manifest1 deletion, clinically not manifest�� αααα 2 deletions, clinically mild/not manifest2 deletions, clinically mild/not manifest�� αα thalassemiathalassemiaHb Bart = Hb Bart = γγ44
�� no no αα hydrops fetalishydrops fetalis
13.11.2007 gene1.ppt 33
Alpha thalassemiasAlpha thalassemias
ε Gγ Aγ Ψβ δ β
ζ Ψζ Ψα α2 α1
2-krát
13.11.2007 gene1.ppt 34
Beta thalassemiaBeta thalassemia
ε Gγ Aγ Ψβ δ β
ζ Ψζ Ψα α2 α1
13.11.2007 gene1.ppt 35
Cystic fibrosisCystic fibrosis
�� Woe Woe to to that child which when kissed that child which when kissed on on the the forehead tastes forehead tastes salty. salty. He is bewitched and soon He is bewitched and soon must diemust die
�� AndersAnders, 1938 , 1938 –– cystic fibrosiscystic fibrosisofof pancreaspancreas�� FarberFarber, 1945 , 1945 -- mukoviscidosismukoviscidosis�� SR 3 SR 3 centrescentres–– BA, BB, KE (cca 60)BA, BB, KE (cca 60)�� 1/21/266 heterozygotheterozygoteses, 1/2, 1/2736, (1/676 736, (1/676 marriagesmarriages))�� Increased NaCl is sweatIncreased NaCl is sweat, , thick secrets of glands in thick secrets of glands in
bronchibronchi, , pancreaspancreas, GIT , GIT -- organ organ failurefailure, , deathdeath�� Disorder of Disorder of reverse chloride reverse chloride and and water water transporttransport
13.11.2007 gene1.ppt 36
Cystic fibrosisCystic fibrosiscompetitioncompetitionWillamson etWillamson etalal., London ., London 1987 1987 -- missmiss
LapLap--Chee Tsui Chee Tsui a spol, Toronto, 1989 a spol, Toronto, 1989 -- hithit
�� VVery different situation compared ery different situation compared to Hb Sto Hb S–– TheThefaultyfaulty protein protein waswasnot not knownknown–– The localization of the The localization of the gene gene was unkownwas unkown
�� Genetic linkage with an Genetic linkage with an enzyme enzyme polymorphism polymorphism --chromosome 7 (chromosome 7 (classicalclassicalgeneticsgenetics))
�� Further markersFurther markers, , narrowing down of the narrowing down of the regionregion�� 4 4 clonesclones, 1 , 1 complementarycomplementarywith cDNAwith cDNA** of of a a
protein protein from sweat glandfrom sweat gland�� LocalizationLocalizationand sequenation of the and sequenation of the genegene
*cDNA = mirror of mRNA for a synthesized protein
13.11.2007 gene1.ppt 37
Cystic fibrosisCystic fibrosis
�� Different from haemoglobinopathiesDifferent from haemoglobinopathies::
–– No No known known protein protein involvedinvolved
–– Unknown site Unknown site for for mutationmutation
�� Genetic linkage with an Genetic linkage with an enzyme enzyme polymorphism polymorphism located located on on chch. 7. 7
�� Further markers in theFurther markers in theregionregion
�� 4 4 clonesclones, 1 , 1 is complementary is complementary to a to a sequence from sequence from sweat glandsweat gland
�� the the gene gene is foundis foundandandsequencedsequenced
�� 1989, chromosome 7 1989, chromosome 7 -- a a big big gene gene with with 24 24 exonsexons
�� Codes Codes a a big transmembrane big transmembrane protein protein -- Cl channelCl channel
�� More More than than 101000 00 mutations found in the mutations found in the genegene
BUTBUT
�� 60 % 60 % patients have patients have a triplet a triplet deletion deletion -- omission omission a a Phe Phe on 508th on 508th place of place of proteinprotein
�� additionaladditional15 % 8 15 % 8 other mutations other mutations ((also in intronsalso in introns))
13.11.2007 gene1.ppt 39
The structure of chloride transporter coded by The structure of chloride transporter coded by CFTR geneCFTR gene
�� Queen VictoriaQueen Victoriaandandher her descendantsdescendants
�� Family of the last Russian Czar NicolausFamily of the last Russian Czar Nicolaus((Alexandra Alexandra -- 4 4 daughters and one affected sondaughters and one affected son))
�� Absolute deficiency of factor Absolute deficiency of factor VIIIVIII
�� 1/10000 1/10000 boysboys, , one third one third are are new mutations in their new mutations in their ancestors ancestors ((during meiosisduring meiosis))
�� High number of mutationsHigh number of mutations, , the the most most common common form is an form is an inversion inversion with with 0 0 activity of factoractivity of factor
13.11.2007 gene1.ppt 42
HHemophiliaemophiliaAA
13.11.2007 gene1.ppt 43
Structure of factor VIII and IX genes and proteins Structure of factor VIII and IX genes and proteins (with (with vWfvWf))
F VIII F IX
13.11.2007 gene1.ppt 44
Other coagulopathiesOther coagulopathies
�� Haemophilia Haemophilia B B -- similar similar to Ato A
�� Haemophilia Haemophilia C C -- AR heredityAR heredity
�� All other factors All other factors -- very rarevery rare
�� Von Willebrand disease Von Willebrand disease -- AD; AD; mild or mild or asymptomaticasymptomatic, , heterogeneousheterogeneous
�� vW factor is vW factor is a a big big protein protein with multiple function with multiple function --stabilizes factor stabilizes factor VIIIVIII
�� Bleeding when associated with other Bleeding when associated with other circumstances circumstances ((acetylsalicylic acidacetylsalicylic acid))
13.11.2007 gene1.ppt 45
An An „„ upside downupside down“ “ coagulopathycoagulopathy
Hereditary thrombophiliaHereditary thrombophilia�� Point Point mutation in factormutation in factorV (V (LeidenLeiden))�� TheTheproteinproteinis is resistantresistanttoto thrombolytic thrombolytic
inactivationinactivation. . �� Part Part of common european heritage of common european heritage (2(2--7 %)7 %)�� Elevated Elevated risk risk of venous thrombosisof venous thrombosis::�� VV = 1; VV = 1; vV vV = 7; v= 7; vvv = 80; = 80; �� Manifestation in association with other Manifestation in association with other