Smith-Lemli-Opitz...
Transcript of Smith-Lemli-Opitz...
![Page 1: Smith-Lemli-Opitz Syndromesellmangen564s14.weebly.com/uploads/2/6/0/1/26017356/slos_final_talk.pdfSmith-Lemli-Opitz Syndrome (SLOS) and the DHCR7 gene! Max Sellman! !! sellmangen564s14.weebly.com!!](https://reader030.fdocuments.in/reader030/viewer/2022040121/5ec7d73130997e484434c76f/html5/thumbnails/1.jpg)
Smith-Lemli-Opitz Syndrome (SLOS) and the DHCR7 gene!
Max Sellman!!!
sellmangen564s14.weebly.com!
!
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What is SLOS?!
Cholesterol metabolism disorder!!Developmental and debilitating!!Relatively common in populations of European origin!!
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Morphological symptoms of SLOS!
2/3 Toe Syndactyly ! Microcephaly!
Cleft Palate!
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Biochemical Symptoms of SLOS!
Failure to thrive!!Mental retardation!!Cholesterol deficiency!!Buildup of cholesterol precursors!7DHC and 8DHC !
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Treatment and management of SLOS!
No cure for SLOS!!Cholesterol supplementation!!Symptom control!
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DHCR7!
7-dehydrocholesterol reductase!
Last step in this cholesterol pathway!
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DHCR7 Gene Ontology!
Cellular Component!• Intracellular
membrane-bound organelle!
Molecular Function!• 7-dehydrocholesterol
activity!
Biological Process!• Lung Development!• Regulation of cholesterol biosynthetic
process!• Post-embryonic development!• Multicellular organism growth!• Cell differentiation!• Blood vessel development!• Regulation of cell proliferation!• Sterol biosynthetic process!
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DHCR7 is expressed in many cell types!
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DHCR7 is expressed in many cell types!
Brain
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DHCR7 is well conserved!
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DHCR7 is well conserved!
Human!
Zebrafish!
Chimp!
Mouse!
Cow!
Arabadopsis!
ERG4/ERG24- Ergosterol Biosynthesis!
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Human!
Zebrafish!
Chimp!
Mouse!
Cow!
Arabadopsis!
DHCR7 is well conserved!
ERG4/ERG24- Ergosterol Biosynthesis!Transmembrane Domain!
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SLOS is caused by mutation in DHCR7!
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SLOS is caused by mutation in DHCR7!
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DHCR7 does not work alone!
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DHCR7 does not work alone!
Sterol biosynthesis proteins!
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DHCR7 does not work alone!
Fatty-acid biosynthesis proteins!
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DHCR7 does not work alone!
SP1 is an important transcription factor!
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What are the factors that contribute to the SLOS phenotype?!
!Cholesterol deficiency!!Buildup of cholesterol precursors?!!Other effects?!
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What are the factors that contribute to the SLOS phenotype?!
!Cholesterol deficiency!!Buildup of cholesterol precursors?!!Other effects?!
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Hypothesis: The cholesterol deficit and buildup of cholesterol precursors in DHCR7- mutant mouse cells creates a stressed physiological environment in which changes to protein interactions and transcription occur!
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Specific Aims!
Aim 1: To identify proteins that are over- or under-expressed in Dhcr7 mutant mice.!
Aim 2: To determine conserved proteins that interact with DHCR7 across species. !
Aim 3: To establish the regulatory effects of a Dhcr7 mutation on gene expression throughout the cholesterol pathway.!
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Specific Aims!
Aim 1: To identify proteins that are over- or under-expressed in Dhcr7 mutant mice.!
Aim 2: To determine conserved proteins that interact with DHCR7 across species. !
Aim 3: To establish the regulatory effects of a Dhcr7 mutation on gene expression throughout the cholesterol pathway.!
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Dhcr7 mouse model!
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Identifying changes in expression in Dhcr7 mutant mice!
!Study brain, liver, and adrenal cells!!Use SILAC approach to quantify proteins in sample!!Compare healthy to SLOS tissues!!!
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Aim 1: Anticipated results!
Impact on expression of proteins with GO term “cholesterol biosynthesis”!!The unanticipated?!
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Specific Aims!
Aim 1: To identify proteins that are over- or under-expressed in Dhcr7 mutant mice.!
Aim 2: To determine conserved proteins that interact with DHCR7 across species. !
Aim 3: To establish the regulatory effects of a Dhcr7 mutation on gene expression throughout the cholesterol pathway.!
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Determining conserved DHCR7 interactions!
Query STRING for conserved interacting partners and complexes!
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Aim 2: Results!
Human DHCR7!
SP1!SREBF2!Associated Pathway !Proteins!
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Aim 2: Results!
Mouse Dhcr7!
Sp1!Srebf1/2!Associated Pathway !Proteins!
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Aim 2: Results!
Zebrafish dhcr7!
!srebf1!Associated Pathway !Proteins!
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Specific Aims!
Aim 1: To identify proteins that are over- or under-expressed in Dhcr7 mutant mice.!
Aim 2: To determine conserved proteins that interact with DHCR7 across species. !
Aim 3: To establish the regulatory effects of a Dhcr7 mutation on gene expression throughout the cholesterol pathway.!
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Establishing the effects of a Dhcr7 mutation on pathway expression!
Microarray experiments to generate expression datasets!!Gene Set Enrichment Analysis to find affected pathways!!
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Aim 3: Anticipated results and Future Directions!
Some variance in pathway expression from wild type!!!Most DHCR7 mutations in humans have no model!!
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References!1. http://www.smithlemliopitz.org/our-families/abigail-roberts/!2. Porter, F. D. (2008). Smith–Lemli–Opitz syndrome: pathogenesis, diagnosis and management. European Journal of
Human Genetics, 16(5), 535-541!3. http://en.wikipedia.org/wiki/File:Celldeath.jpg!4. http://www.nlm.nih.gov/medlineplus/ency/imagepages/17256.htm!5. http://indianpediatrics.net/may2004/may-512.htm!6. http://www.chori.org/Principal_Investigators/Shackleton_Cedric_HL/Pictures/shack_fig2.gif!7. http://www.metagenes.co/main/product/sloesterol!8. https://www.gymra.com/blog/how-healthy-are-eggs-what-are-the-health-benefits/!9. http://learn.genetics.utah.edu/content/disorders/singlegene/slos/images/GeneLocations.jpg!10. http://themedicalbiochemistrypage.org/slos.php!11. http://www.ohsu.edu/xd/health/services/doernbecher/research-education/research/research-labs/images/Stair_1.JPG!12. Witsch‐Baumgartner, M., Löffler, J., & Utermann, G. (2001). Mutations in the human DHCR7 gene. Human mutation,
17(3), 172-182.!13. http://www.cma.ca/multimedia/staticContent/HTML/N0/l2/cim/vol-24/issue-6/pdf/pg311.pdf!14. http://news.bbcimg.co.uk/media/images/66262000/jpg/_66262511_alex_and_daniel_slos.jpg!15. http://www.broadinstitute.org/files/shared/proteomics/images/silac1.png!16. http://www.broadinstitute.org/gsea/index.jsp!17. http://arrayit.com/rare_diseases.jpg!18. http://askmax.countrymax.com/!!Dhcr7 mouse: Yu, H., Wessels, A., Chen, J., Phelps, A. L., Oatis, J., Tint, G. S., & Patel, S. B. (2004). Late gestational lung hypoplasia in a mouse model of the Smith-Lemli-Opitz syndrome. BMC developmental biology, 4(1), 1.!!Cover Slide: http://www.bio.davidson.edu/projects/GCAT/tour/one_grid.jpg!!
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18!