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Student day 2012 4 1-2012
1) Cool temperature correlates with longevity in C. elegans, calorie restricted mice, Ames dwarfs, and long-lived humans. However, the underlying mechanisms were not understood. 2) Using C. elegans, the author found that genes in the pro-aging branches of the unfolded protein response (UPR) pathway, such as pek-1 and atf-6, negatively correlate with longevity, while inhibition of these genes increases lifespan. 3) Cool temperature may promote longevity by inhibiting the pro-aging branches of the UPR pathway, similarly to genetic inhibition of these genes. Understanding how cool temperature interacts with the UPR could provide insights into mechanisms of aging.
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Student day 2012 4 1-2012
- 1. Angela Sanchez Mentor: PamelaLarsen Pro-Aging Branches of the Unfolded Protein Response
- 2. • Short-lived • Inexpensive •Tractable model • Considerable discovery potential C.elegansas a model system forlongevity
- 3. C.elegansas a modelsystem forlongevity • Identify a long-lived intervention • Use worm to understand mechanism
- 4. Cool Temperature DoublesLifespan in C. elegans
- 5. Cool temperature correlateswith longevity Caloric Restriction (Lane 1996)
- 6. Cool temperature correlateswith longevity Ames Dwarf (Hunter 1999)
- 7. Cool temperature correlateswith longevity Long-lived human (Roth 2002)
- 8. Why does cooltemperature improve health and survival? Underlying mechanism not understood How can we use C. elegans to gain insight?
- 9. Classic Genetic Approachfor Mutants Screen for mutants that fail to be normal (WT) Mutants that fail to increase lifespan due to cool temperature daf-2 mutants!
- 10. Qa daf-2: Candidate forcool temperature mediated longevity Homologous to human insulin/IGF receptor Found to have increased lifespan daf-2 25˚C Wild type 25˚C daf-2 15˚C Wild type 15˚C Similar to benefit from cooler temperatures Attenuated benefit at 15˚C for daf-2mutants
- 11. Qa daf-2: Candidate forcool temperature mediated longevity daf-2 25˚C Wild type 25˚C daf-2 15˚C Wild type 15˚C SUGGESTS DAF-2 AND COOL TEMPERATURE ARE LINKED OVERLAPPING MECHANISM OF LONGEVITY
- 12. Understanding cool mediatedlongevity Microarray • Affymetrix C. elegans array • Unbiased genome wide snapshot • Comparing daf-2 mutants and wild type • Comparing response to temperature (15C vs 25C) • Look for +/- correlations with longevity UNFOLDED PROTEIN RESPONSE GENES NEGATIVELY CORRELATE WITH LONGEVITY
- 13. • Synthesis of •Membrane •Secreted •Glycosylation •Trimming •Post-translational Endoplasmic Reticulum Synthesizes Membrane and Secreted Proteins P
- 14. •Sorted to •Membrane •Organelle •Secreted Endoplasmic ReticulumSynthesizes Membrane and Secreted Proteins SECRETED LYSOSOME MEMBRANE
- 15. BiP Unfolded Protein Response IRE-1PEK-1 BiP Translation eIF2 Transcriptionof UPR Genes ATF-5 Misfolded Protein BiP Site 2 Protease XB P-1 CHOP
- 16. daf-2 has lowerlevels Temperature and Genotype Regulation of S2P MicroarrayExpressionLevels S2P WT daf-2 CHOP WT daf-2 hsp-3 WT daf-2
- 17. Cool temperature correlateswith longevity Long-lived human (Roth 2002)
- 18. Reduced Bip Levelsin Offspring of Long-lived Individuals Reduced Bip in other longevity promoting interventions?
- 19. Cool temperature correlateswith longevity Caloric Restriction (Lane 1996)
- 20. Reduced Bip levelsin Dietary Restricted Mice
- 21. Cool temperature correlateswith longevity Ames Dwarf (Hunter 1999)
- 22. Reduced Bip levelsin Growth Hormone Deficient Mice
- 23. Canonical Unfolded ProteinResponse IRE-1PEK-1 BiP ATF-5 Site 2 Protease CHOP PRO-AGING BRANCHES of UPR
- 24. Two BiP Isoformsin C. elegans: hsp-3 and hsp-4 IRE-1PEK-1 hsp-4hsp-3 Heatshock InducibleConstitutive/Non-Heatshock BiP •93% Identical 607/658 amino acids •hsp-4 is heatshock-inducible (>29°C) •hsp-3 constitutive
- 25. hsp-3 and hsp-4RNAi increase lifespan at 25˚C in wild type
- 26. Inhibition of UPRgenes increases lifespan at 25˚C in wild type%Survival Adult Age (days) Knockdown increases lifespan under warm conditions
- 27. Inhibition of Pro-AgingUPR and Cool Temperature: Same Mechanism of Longevity? Can we identify signaling pathways underlie cool longevity?
- 28. IRE-1PEK-1 Translation eIF2 Site 2 Protease CHOP ATF-5 BiP BiP BiP Inhibitionof Pro-Aging UPR and Cool Temperature: Same Mechanism of Longevity?
- 29. RNAi of UPRgenes does not alter lifespan at 15˚C Cool temperature promotes longevity by INHIBITION OF PRO-AGING UPR
- 30. Pro-Aging Branches ofthe Unfolded Protein Response IRE-1PEK-1 Site 2 Protease CHOPATF-5 BiPBEFORE AFTER ?
- 31. Pro-Aging Branches ofthe Unfolded Protein Response IRE-1PEK-1 Site 2 Protease CHOPATF-5 BiP ?
- 32. pek-1, ire-1, atf-6deletion mutants ire-1 (C41C4.4) ok799 Deletion atf-6 (F45E6.2) ok551 Deletion pek-1 (F46C3.1) ok275 Deletion
- 33. ire-1 deletion reduceslifespan at 25˚C ire-1 is reduces for wild type life span
- 34. hsp-3 and hsp-4RNAi rescues ire-1 lifespan hsp-3 and hsp-4 longevity DOWNSTREAM of ire-1
- 35. hsp-3and hsp-4 RNAirescues ire-1 to hsp-3/4 RNAi levels hsp-3 and hsp-4 RNAi longevity is INDEPENDENT of ire-1
- 36. pek-1 and atf-6deletion increases lifespan at 25˚C pek-1 and atf-6 are PRO-AGING
- 37. IRE-1PEK-1 hsp-4 hsp-3 PRO-AGING PRO-AGING hsp-4 Model ofPro-Aging Branches of the UPR
- 38. pek-1 deletion increaseslifespan and is hsp-3 dependent hsp-4 RNAi promotes longevity by pek-1 inhibition
- 39. pek-1 deletion increaseslifespan and is hsp-3 dependent hsp-4 RNAi promotes longevity by pek-1 inhibition
- 40. atf-6 deletion increaseslifespan and is hsp-3 independent Different response to hsp-3 RNAi in atf-6 and pek-1 deletion mutants
- 41. atf-6 deletion increaseslifespan and is hsp-3 independent hsp-4 RNAi promotes longevity by inhibition of atf-6
- 42. hsp-3 interacts withpek-1 to regulate longevity atf-6 and hsp-3 inhibition have an additive affect on lifespan pek-1 mutant longevity partially dependent on hsp-3
- 43. hsp-4 RNAi promoteslongevity by inhibition of atf-6
- 44. Can structural detailsof hsp-3 and hsp-4 predict UPR sensor Interaction?
- 45. Amino Acid DifferencesCluster In Two Distinct Regions Substrate Binding Domain hsp-4 more hydrophobic UPR Sensor Binding Domain
- 46.
- 47. Model of Pro-AgingBranches of UPR IRE-1PEK-1 hsp-4hsp-3 PRO-AGING PRO-AGING + + PRO-LONGEVITY DAF-2 Cool TemperatureWarm Temperature
- 48. Future Directions • Identifieda novel gene involved in physUPR • Identified novel genes with strong longevity signatures • Identified novel genes with strong aging signatures
- 49. Acknowledgements Committee MembersPast LarsenLab Members CollaboratorsFunding Dr. Shelly Buffenstein Dr. Ellen Kraig Dr. Merry Lindsey Dr. Jim Lechleiter Dr. Syuichi Takano Wendy Lee Dr. Hal Boylston Dr. Pamela Larsen NIA Training Grant Glenn Medical Foundation Ellison Medical Foundation Dr. Simon Tavaré Dr. Christina Curtis
Editor's Notes
- #5 452 823 1 1.67
- #10 452 823 1 1.67
- #28 More than 20% here. What is the molecular mechanism of cool longevity. MAKE OWN TEMPERATURE GRAPH TO MATCH FORMAT OF DAF-2 GRAPH. INCLUDE ALL DATA POINTS. Point out early death in daf-2 that wt 15C is better. 15C may bring health benefits mention this . Is this respose conserved?
- #44 You have an unbiased data set, but the experimental design allows you to parse results and understand more deeply. You can say why what is important in your conclusions. You do not have to wonder what if a different dose/severity of DR was used- we have two mild and three severe alleles. The multiple alleles all backcrossed to the same wild type allows you to accurately associate molecular changes with share phenotypes of known magnitude difference from WT and pleiotropic effects of the alleles with their unique profile of mutantness. Then put it together in general concepts to humans, how association of specific SNPs or polymorphisms or alleles with longevity or health or disease makes sense. E.g. which is Hispanic-like prone to diabetes, smaller stature, potential longevity.....How is a centenarian possible? analogy to daf-2 mutations opening an maximal life span option that is not possible for WT. Over 100 or over 110 years old are rare- absolutely what we that really know extent of pleiotropies of daf-2 mutants.