Sam Rhine - Genetic Update Conferences - www.samrhine.com Genetics of Common Human Traits and Diseases….. Cloning and Stem Cells…..New Medical Applications I. CHROMOSOMES - CYTOGENETICS: A. Metaphase Spread - Karyotype - Autosomes - Sex Chromosomes - XX and XY 1. Chromosome Anatomy - Centromere - p-arm - q-arm - telomeres - chromatids 2. Chromosome Identification - Chromosome Banding - unique sets of stripes B. Syndrome: a recognizable pattern of anomalies, a group of signs or symptoms, that occur together and characterize a particular abnormality C. Chromosome Syndromes: 1 in 160 live births / 50% of a miscarriages MCA babies (Multiple Congenital Anomalies) 1. Trisomy 16 - most common chromosome syndrome - always miscarries 2. Trisomy 21 - Down Syndrome - 1/800 - Dr John Langdon Haydon Down - 1866 3. Trisomy 18 - Trisomy 13 - Monosomy X = Turner Syndrome - Klinefelter Syndrome II MONOGENIC INHERITANCE - SINGLE GENE PAIR - MENDEL'S RULES of INHERITANCE A. Johann Gregor Mendel - Monk at St. Thomas Monastery in Brno, Czech Republic B. Monogenic Reference Sites: 1. OMIM - http://www.ncbi.nlm.nih.gov/sites/entrez?db=omim 2. Genes and Disease - http://www.ncbi.nlm.nih.gov/books/NBK22183/?depth=2 C. AUTOSOMAL DOMINANT INHERITANCE - AD 1. Brachydactly - Huntington Disease - Neurofibromatosis - Marfan Syn - Alzheimer D. AUTOSOMAL RECESSIVE INHERITANCE - AR 1. Cystic Fibrosis - Sickle Cell Anemia - Tay-Sachs Disease E. X LINKED RECESSIVE INHERITANCE - XLR 1. Hemophilia Type A & Type B - Duschenne Musc Dys - Fragile X - Happiness Genome and Genes Review: I. INTRODUCTION: Genome / Genes / Gene Control A. Genome: the sum total of all the genetic information for any biologic organism 1. ~1.5% - 'Coding' DNA 2. ~98.5% - 'Non Coding' DNA..…'Junk' DNA B. Gene: 1. 'Coding' DNA Genes - carry the genetic code to make a protein 2. 'Non Coding' DNA Genes - carry a code to make RNA only C. GENE CONTROL = GENE REGULATION - ON / OFF MECHASNISMS 1. Transcription Factors - turn transcription ON and OFF at the promoter 2. microRNA = miRNA - negative regulators - turn genes OFF at 3' UTR 3. Epigenetics - 'Epi' means upon / on top of / above and beyond the DNA Epigenetic control mechanism acts on chromatin - Open or Closed Chromatin D. ENCODE - ENCyclopedia Of Dna Elements - new information as of September 5, 2012 1. International Consortium - 32 Research Institutions - 442 Consortium members 2. New Genome Findings: a. the 'Junk' contain 'Docking Sites' - 2,890,000 - new gene control areas b. sncRNAs - small non coding RNAs and lncRNAs - long non coding RNAs: INTRODUCTION: GENOMIC MEDICINE - NEW ERA of HUMAN GENETICS A. use knowledge about the entire GENOME - study POPULATIONS of people 1. Genomic Medicine - there is no 'Normal'…..no normal human genome a. 'Variants' - millions of differences among individuals in a population b. 'Mutation' - DNA variant that is pathologic / causes disease or cancer 2. Why don't we study the genetics of…..Common Conditions: Adult Type 2 Diabetes / Macular Degeneration / Hypertension High Cholesterol / Heart Disease & Stroke / Cancer & Tumors BiPolar Disorder (Manic/Depressive) / ADHD - Attention Deficit / Alzheimer Alcohol Dependency / Autism Spectrum - ASD / Epilepsy / Asthma / etc. …..COMMON DISEASES…..but COMPLEX GENETIC TRAITS C. HYPOTHESIS that might Explain Complex Diseases: 1. 'COMMON DISEASES / COMMON VARIANTS HYPOTHESIS' 2. Persons with the same Common Disease…..have unique set of DNA Variants in Common 3. HERITABILITY: H - the proportion of Phenotype variation due to Genetics 4. HUMAN QUANTATIVE TRAITS: Height, blood pressure, blood glucose levels 5. Flipping Pennies Model: polygenes are additive - not dominant or recessive 6. Medical Conditions - Threshold effect: 7. TWO perfectly NORMAL parents can have an AFFECTED child!! D. What would it take to find the Polygenes for Common Diseases? H G P / SNPs & CNPs E. GWAS - GenomeWide Association Studies - New England Journal Med July 8, 2010 1. Evaluate the entire genome of thousands of people and see if any SNPs might be Associated with a particular Common Disease 2. GWAS Example: T2D - T1D - Crohn's - Obesity - Height - Blood Lipids - Autism 3. CNPs / CNVs - Copy Number Polymorphisms = Copy Number Variantas Normal - 2 copies / CNP = 1 copy or 3 copies, etc. 4. de novo CNVs account for 5-8% of autism simplex cases - paternal age effect 5. Obesity & Underweight / Macrocephaly & Microcephaly / Autism & Schizophrenia Examples of CNVs and Reciprocal Variants - Phenotypic Opposites 6. YOUR PERSONAL GENOME EVALUATION - personal chip - fetal chip Cloning and Stem Cells…..New Medical Applications I. Human Embryology - formation of the fetus = Applied Embryology A. Embryonic Stem Cells…..give rise to four major branches B. HSCs - Hematopoetic SCs / MSCs - Mesenchymal SCs ESCs - Endodermal SCs / NSCs - Neural III. Reproductive cloning: Oocyte Retrieval / Enucleation / SCNT - transfer donor somatic nucleus Artificial Activation of the egg / ET - Embryo Transfer into the uterus of a surrogate mother IV. Therapeutic cloning: Human Oocyte Retrieval / Enucleation / SCNT - transfer donor somatic nucleus Artificial Activation of the egg / grow in lab for 6 days - develop a cell line "Ethical" Pluripotent Stem Cells …..REPROGRAMMING 1. iPS for: Huntington Dx, Parkinson Dx, Type 1 Diabetes, 2. huESCs from donated blastocysts for Dry MD and Stargardt's 3. RP - Retinitis Pigmentosa - Most Common Inherited Vision Loss 4. Huntington Disease - AD - CAG Triplet Repeat Mutation 5. Also iPSCs for: Alpha-1-Antitrypsin Deficiency / Hutchison Guilford Progeria Friedreich's Ataxia / Fragile X Syndrome / Rett Syndrome - Autism 6. Man-Made Organs: Teeth / Optic Cup Retina / Cerebral Cortex Primitive Hypothalamus / Primitive Cerebellum / Heart Vein / Trachea Cartilage / Cardiac Fibroblasts make new Cardiomyocytes / Fetal Heart Valve Osteogenesis Imperfecta / Finger Regeneration / burger from muscle stem cells Made rat pancreas in a mouse embryo…..Human Pancreas?? 7. Yamanaka - iPSCs Stock Cell Project in Japan will make iPSCs available for most of the Japanese population