The Pathogenesis of Diseases from Genetic and Genomic Point of View Oliver Rácz and František Ništiar Institute of Pathological Physiology Medical School, Šafárik University 2009 – 2016 20. 5. 2018 gene16a

26th june 2000 is neither the beginning nor the end of the way 5 years before term (1990 - 2005) The race is over, victory for Craig Venter. The genome is mapped* - now what ? Not a discovery! A very important technological result and competition is always useful. all is based on Mendel‘s and Watson‘s & Crick‘s discoveries in XIXth XXth century *3*109 letters 20. 5. 2018 gene16a

Mendel, Watson, Crick & the medical genetics of XIXth – XXth century Mendel‘s laws are valid also today Watson & Crick provided the material basis of these laws (central dogma of molecular biology) Mendel‘s laws in medicine can be applied to monogenic diseases – long list, relatively rare What is the genetics of diabetes, hypertension, coronary heart disease, Alzheimer disease ? 20. 5. 2018 gene16a

White flower from red parents ??? 20. 5. 2018 gene16a

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Quidditch ball for Harry Potter ? 20. 5. 2018 gene16a

We discovered the secret of life, let’s have a beer! 20. 5. 2018 gene16a

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Central dogma of molecular biology (cca. 1965) Replication Transcription Translation Transformation DNA 2 DNA RNA PROTEIN A B 20. 5. 2018 gene16a

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Central dogma of molecular biology? – not so simple DNA 2 DNA RNA PROTEIN A B 20. 5. 2018 gene16a

Central dogma of molecular biology? Epigenetics ? Role of RNA? Regulation of transcription Transcription factors, etc. Methylation, acetylation… Regulation of RNA editing Alternative splicing Regulation of RNA transport Regulation of translation siRNA, tRNA modifications Postsynthetic modifications 20. 5. 2018 gene16a

Central dogma of molecular biology 20. 5. 2018 gene16a

Central dogma of molecular biology (now) – role of RNAs Replication Transcription Translation Transformation DNA 2 DNA RNA PROTEIN A B 20. 5. 2018 gene16a

We and our relatives Organism Genome size chromosomes/ genes Homo sapiens 3 000 000 000 23/30 000 Mus musculus 2 600 000 000 20/30 000 D. Melanogaster 137 000 000 4/13 000 C. Elegans 97 000 000 6/19 000 S. Cerevisiae 12 100 000 18/6 000 E. Coli 4 600 000 {1}3 200 HIV virus 9 700 9 A. Thaliana 100 000 000 ?/25 000 20. 5. 2018 gene16a

It is a little more complicated DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B DNA 2 DNA RNA PROTEIN A B 20. 5. 2018 gene16a

What is not “genetic” The number of human genes is as low as 30 000 the small worm C. elegans has 20 000 genes the mouse has as many genes as we, also with very similar function The mystery is in complexity and networking: 230000 >>>> 220000 (possible on/off states) And a number of surprises around transcription and translation (miRNA, tRNA modifications) It is mapped but do we understand it? GENETICS = HEREDITY GENOMICS = EVERYTHING 20. 5. 2018 gene16a

GENES AND THE ENVIRONMENT GENOME ENVIRONMENT A SEVERE MONOGENIC DISEASES NEGATIVE AND POSITIVE ENVIRONMENTAL FACTORS physical chemical biological nutrition life style B POLYMORPHISMS C GENETIC RISK 20. 5. 2018 gene16a

GENES AND THE ENVIRONMENT GENOME ENVIRONMENT A THE DNA IS A DEAD MOLECULE COMMUNICATES WITH THE ENVIRONMENT THROUGH EPIGENETIC MECHANISMS DNA methylation Histone modification siRNA SEVERE MONOGENIC DISEASES NEGATIVE AND POSITIVE ENVIRONMENTAL FACTORS physical chemical biological nutrition life style B LESS IMPORTANT MUTATIONS C GENETIC RISK 20. 5. 2018 gene16a

Genes and diseases in practical medicine XIXth Century: symptom Þ diagnosis sugar in urine = diabetes XXth Century: symptom ß etiopatogenesis Þ diagnosis autoimmune destruction of b cells = dm Type 1 XXIst Century: symptom ß genes and environment ß susceptibility + overeating = subtypes of dm Type 2 20. 5. 2018 gene16a

Mutations changes of genetic information THREE PRINCIPAL POSSIBILITIES changes in genome not compatible with life development and diversity disease or increased risk fo disease* THE BASIC DIFFERENCE FOR HEREDITY: somatic and germ cell mutations genome, chromosomal and gene mutations no genes for diseases! – sickle cell, Alzheimer, diabetes... 20. 5. 2018 gene16a

Everything is clear? CTT ACT GCC GGT TCG..........TGA EACH AMINOACID IS CODED BY THREE CONSECUTIVE NUCLEOTIDES (TRIPLETS, CODONS) CTT ACT GCC GGT TCG..........TGA Leu Thr Ala Gly Ser............STOP OK, silent, missense, nonsense, frameshift AND THE NONCODING SEQUENCES ? 20. 5. 2018 gene16a

Everything is clear? BUT CTT ACT GCC GGT TCG..........TGA Leu Thr Ala Gly Ser............STOP BUT IN THE GENOME THERE IS MORE NONCODING (REALLY NONCODING?!) SEQUENCES AS CODING INTRONS IN THE GENES REGULATION (SWITHCES) REPETITIVE SEQENCES (IN MAN UP TO 50 % ) LINE 6000 bp, SINE 100-300 bp, DIRT (PSEUDOGENES, FOREIGN GENES) TELOMERES (STABILISATION OF CHROMOSOMES) 20. 5. 2018 gene16a

Gene mutations and SNPs* Point mutations in exons Frameshift mutation in exons (1,2,4,5...) Small deletion of triplets (3,6...) Bigger deletions – transition to chromosomal aberrations Mutations or polymorphisms in regulatory parts, introns, genes for r-tRNA = SNP, 10*more than the classic mutations Variability of repeated sequences - markers Dynamic mutations – triple repeat mutations *SINGLE NUCLEOTID POLYMORPHISM 20. 5. 2018 gene16a

CLASSIC MONOGENIC DISASES Autosomal recessive (AR) Sickle cell disase, other Hbpathies, thalassemias Cystic fibrosis Inborn errors of metabolism Autosomal dominant (AD) Polydactylia Familial hypercholesterolemia Polycystic kidney disease X chromosome linked diseases Heamophilia A, B Daltonism 20. 5. 2018 gene16a

Autosomal recessive (AR) Two healthy (carrier heterozygotes) hidden in the population Probability of ill (homozygote) children 1:4 (25%) High number of relatively rare diseases, often fatal Autosomal dominant One ill heterozygote parent Probablity of transfer to next generation 50% Survival is better – “low penetration and expression” 2 * why X chromosome linked diseases Healthy heterozygote mother Probablity of ill sons 50 % (Charles, William, George) 20. 5. 2018 gene16a