Genetics of Vascular Diseases Jake Lusis 3-730 MRL [email protected].
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Transcript of Genetics of Vascular Diseases Jake Lusis 3-730 MRL [email protected].
![Page 2: Genetics of Vascular Diseases Jake Lusis 3-730 MRL Jlusis@mednet.ucla.edu.](https://reader036.fdocuments.in/reader036/viewer/2022062423/56649db15503460f94a9f7cc/html5/thumbnails/2.jpg)
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210
410 200 210 180 430
420190460490
Carl Miller, ~1938, Oslo County Hospital, Norway
Inheritance of:
Cholesterol level (mg/dl):
Tendon xanthomas:
Early coronary artery disease (under 55 years of age):
330 210
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Tendon Xanthomas
Cosegregation of triad of xanthomas, high cholesterol and early heart disease
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210190 400
210 430 490420 180
170420
Inheritance of:
Cholesterol level (mg/dl):
Tendon xanthomas:
Early coronary artery disease (under 55 years of age):
210 230
810 420
This individual died from severe CAD at age 7
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Lecture delivered in Sweden Dec 9, 1985 when they received the Nobel Prize in Physiology. Published in
Science, 232, 34-47, 1986
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Experiments with 125I-LDL and skin fibroblasts
skin fibroblasts from normal and FH individuals
add labeled 125I-LDL ( )
incubate to allow binding
wash away unbound LDL and follow label
- 125I-LDL on surface can be displaced from cells by anionic polymers
- 125I-LDL that has been internalized cannot be displaced from cells
- 125I-LDL that has been degraded to amino acids is soluble in tricholoroacetic acid
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FH homozygote
Normal
Experiments with fibroblasts from normal and FH individuals
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Sequential steps in LDL receptor pathway: Discovery of receptor mediated endocytosis
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Degradation of LDL by lymphocytes and following injection
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Cloning of the Human LDL Receptor
cultured skin fibroblastsisolate plasma membrane
purify such that a single protein remains upon electrophoresis
extract proteins with detergent and subject to chromatography
125I-LDL bound
digest the protein and sequence the resulting
peptides
-ASN-PHE-THR-SER-based on the amino acid
sequence, synthesize corresponding oligonucleotide
probes
….CACTCGACAGCGAAC...
probe cDNA and genomic libraries.
Confirm identity of clones by sequencing
and expression studies
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Domain structure of the LDL receptor
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Five classes of LDL receptor mutations
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Development of drugs to treat hypercholesterolemia based on concepts learned from FH studies
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Combined heart-liver transplant in an individual with homozygous FH
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Lessons from familial hypercholesterolemia
• Studies of an uncommon disease (FH) can have important implications for a common disease (coronary artery disease)
• Genetics studies of clinical traits can lead to an understanding of basic biologic mechanisms (receptor mediated endocytosis, cholesterol homeostasis) as well as new therapies (statins).
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Francis Collins (1995)
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Schematic of the approach generally utilized for positional cloningSchematic of the approach generally utilized for positional cloning
FamiliesFamilies PhysicalPhysical
MappingMapping
andand
CloningCloning
FinerFiner
GeneticGenetic
MappingMappingLinkageLinkage
AnalysisAnalysis++
TranscriptTranscript
IdentificationIdentification
CandidateCandidatecDNAscDNAs
MutationMutation
SearchSearch
..A A C T C..A A C T C
..A A G T C..A A G T C
BACsBACs
NormalNormal
MutationMutation
GeneticGeneticMarkersMarkers
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Marfan Syndrome -autosomal dominant disorder characterized by weakened connective tissue -most life-treatening is aortic dissection (an aneurysm of the aorta) -other symptoms include mitral valve prolapse, emphysema-like lung disease, and ocular and skin disorders -patients tend to be tall and lanky, with very long limbs, fingers and toes
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Fibrillin 1 Gene Mutated in Marfan Syndrome Patients(A)Single-strand conformational polymorphism screen(B) DNA sequence
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Letter
Nature Genetics 33, 407 - 411 (2003) Published online: 24 February 2003;
Dysregulation of TGF- activation contributes to pathogenesis in Marfan syndrome
Enid R. Neptune1, 2, Pamela A. Frischmeyer2, Dan E. Arking2, Loretha Myers2, Tracie E. Bunton3, Barbara Gayraud4, Francesco Ramirez4, Lynn Y. Sakai5 & Harry C. Dietz2, 6
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Lung histopathology and morphometry of mice deficient in fibrillin-1.
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Figure 3
Active TGF-B expression and signaling in lung tissue of mice deficient in fibrillin-1.
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Neutralizing antibody to TGF- rescues lung maturation in mice deficient in fibrillin-1.
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Mice deficient in fibrillin-1 have preserved lung cell proliferation but greater apoptosis.
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Model
Fibrillin deficiency
TGF not sequestered
TGFB activated
Apoptosis in lung, bone overgrowth, etc.
A lesson: Structural matrix elements serve crucial regulatory roles in cytokine activation and signalling
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Identification of Mendelian and complex trait genes 1980-2002
0
200
400
600
800
1000
1200
1400
1600
1800
1980 1985 1990 1995 2000
Year
Number of Mendelian genes
0
10
20
30
40
50
60
70
80
90
100
Number of complex trait genes
Mendelian traits
All complex traits
Human complex traits
Glazer, et al. (2002) Science
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Nature Genetics 34, 383 - 394 (2003)
VEGF is a modifier of amyotrophic lateral sclerosis in mice and humans and protects motoneurons against ischemic deathDiether Lambrechts1, Erik Storkebaum1, Masafumi Morimoto1, Jurgen Del-Favero2, Frederik Desmet1, Stefan L Marklund3, Sabine Wyns1, Vincent Thijs1, Jörgen Andersson3, Ingrid van Marion4, Ammar Al-Chalabi5, Stephanie Bornes6, Rhiannon Musson4, Valerie Hansen5, Lars Beckman3, Rolf Adolfsson7, Hardev Singh Pall4, Hervé Prats6, Severine Vermeire1, Paul Rutgeerts1, Shigehiro Katayama8, Takuya Awata8, Nigel Leigh5, Loïc Lang-Lazdunski9, Mieke Dewerchin1, Christopher Shaw5, Lieve Moons1, Robert Vlietinck1, 10, Karen E Morrison4, Wim Robberecht1, Christine Van Broeckhoven2, Désiré Collen1, Peter M Andersen3 & Peter Carmeliet1
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VEGF Polymorphisms
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Lesson
Basic vascular biology studies (regulation of VEGF expression in response to hypoxia) can lead to an understanding of a complex disease