Presentación de PowerPoint - SEQC · Anabolism Catabolism. Ubiquitin proteasome system Main...
Transcript of Presentación de PowerPoint - SEQC · Anabolism Catabolism. Ubiquitin proteasome system Main...
Autofagia: el sistema de reciclaje que
necesitan todas nuestras células
CIB-CSIC, Madrid, Spain
CSICCONSEJO SUPERIOR DE INVESTIGACIONES CIENTÍFICAS
Caloric restriction
Caffeine
Green Tea
Rapamycin
Resveratrol
Metformin
Spermidine
Lithium Exercise
Hypoxia
Torin-1
Trehalose
What have all these treatments have in common?
Anti-aging
Caloric restriction
Caffeine
Green Tea
Rapamycin
Resveratrol
Metformin
Spermidine
Lithium Exercise
Hypoxia
Torin-1
Trehalose
What have all these treatments have in common?
Autophagy
• ¿Qué es? Tipos / Métodos estudio
• Funciones: implicaciones en fisiología
• Enfermedad y Envejecimiento
Autofagia
Ubiquitin proteasome system
Main catabolic pathways in cells
Autophagy-lysosomal system
Lysosome
Acid
hydrolases
Autofagia
Auto
Fagia= comer
Proceso de digestión intracelular y
reciclaje de los componentes celulares
Discovery of autophagy
Christian de Duve 1974
Lysosomes
Autophagosomes
Autophagy 1963
Atg genes 1992
Yoshinori Ohsumi 2016
Catabolic mechanism
Regulated by
hormones
Functional Groups of Atg Proteins(Y. Ohsumi, M Thumm, DJ Klionsky)
1. Protein Kinase Complex
Atg1, Atg13 , Atg17 , Atg11
2. PI3 Kinase Complex
Atg6 (Beclin 1), Atg14 , (Vps15 , Vps34)
3. Recycling pathway, others
Atg2 , Atg9 , Atg18
4. Atg12 Conjugation System
Atg12 , Atg7 , Atg10 , Atg5 , Atg16
5. Atg8 Lipidation System (PE)
Atg8 (LC3), Atg4, Atg7 , Atg3
Initiation
Nucleation
Expansion
Sealing
Atg genes
Diseases
Neurodegeneration
Muscle disorders
Aging
Infectious diseases
…
Autophagy: the molecular era
New tools for quantitative Autophagy
Image analysis SISTEMA EXPERTO DE ANÁLISIS CUANTITATIVO DE IMÁGENES PARA EVALUACIÓN AUTOMATIZADA DE ENSAYOS CELULARES DE AUTOFAGIA Y APOPTOSIS
WimScratch
for Wound Healing Assay
WimTube
for Tube Formation Assay
WimTaxis
Automated Cell Tracking
WimSprout
for Sprouting Assay
WimCounting
for cell and nuclei counting nuclei
WimAutophagy
Autophagy in cells and tissues
WimApoptosis
Apoptosis in cells and tissues
Tamaño (px)
Frecuencia
Tamaño medio autofagosoma: 8 px
Tamaño mín autofagosoma: 3 px
Tamaño máx. autofagosoma: 11 px
Tamaño cluster: >12 px
AP
Cluster
WimAutophagy
Autophagy in cells and tissues
Cells transfected with GFP-LC3Cells transfected with the Tandem GFP-RFP-LC3Immnunodetection of LC3 with antibodies
Epiflourescence/confocal microscopy
WimAutophagy
Autophagy in cells and tissues
HeLa si RNA scramble-GFPsi RNA scramble Atg7-GFP
Ct N/L
20
60
Num
eb
r o
f a
uto
phago
som
e/ ce
ll
Ct N/L
100
*Scramble
Atg7 KD
*
GFP
Degradation of protein aggregates
Physiological functions of autophagy
Basal autophagyInduced autophagy
Elimination of damaged organelles
Elimination superfluous organelles
Degradation of protein aggregates
Physiological functions of autophagy
Cell viability during starvation
Basal autophagyInduced autophagy
Metabolic adaptation
Elimination of damaged organelles
Elimination superfluous organelles
Degradation of protein aggregates
Physiological functions of autophagy
Cell viability during starvation
Degradation of intracelullar pathogensAntigen presentation
Basal autophagyInduced autophagy
Metabolic adaptation
Elimination of damaged organelles
Elimination superfluous organelles
Degradation of protein aggregates
Physiological functions of autophagy
Cell viability during starvation
Degradation of intracelullar pathogensAntigen presentation
Basal autophagyInduced autophagy
Metabolic adaptation
Protection against DNAdamage
Elimination of damaged organelles
Cell death
Elimination superfluous organelles
Degradation of protein aggregates
Diseases associated to altered autophagy
Cell viability during starvation
Degradation of intracelullar pathogensAntigen presentation
Basal autophagyInduced autophagy
Metabolic adaptation
Protection against DNAdamage
Elimination of damaged organelles
Cell death
Elimination superfluous organelles
Cancer
Neurodegeneration
Infectious diseases
Chronic inflammation
Heart diseases
Cancer
Cancer
Aging
Diabetes, obesity
Respuesta adaptativa al estrés
Mecanismo citoprotector P Boya et al., Mol Cell Biol 2005
González-Polo et al., J Cell Sci 2005
HCQ
Monensin
Bafilomicin A1
Lamp 1,2 -/-
siRNA Lamp1,2
3-MA
siRNA Atgs
Caloric restriction
Caffeine
Green Tea
Rapamycin
Resveratrol
Metformin
Spermidine
Lithium Exercise
Hypoxia
Torin-1
Trehalose
What have all these treatments have in common?
Anti-aging
Lausanne octobre 2008
Caloric restriction (rhesus monkeys):
Colman et al. 2009 Science 325, 201
Rapalogues and TOR inhibitors (mice, drosophila):
Harrison et al. 2009 Nature 460, 392
Lee et al. 2010 Science 327, 1223
Autophagy and anti-ageing
Aging = decreased autophagic pathways
- Fusion problems (reduced macroautophagy and CMA)
- Decreased mRNA expression of Atg genes
- Accumulation of pigments--- Lipofuscin
- Inadequate turnover of damaged mitochondria---- ROS
- Energy compromise
Autophagy
Damaged
mitochondria
Aggregated
proteinsER-stress
ROSproduction Lipolisis Increase
ATP production
Decreased Ageing
Autophagy
Damaged
mitochondria
Aggregated
proteinsER-stress
ROSproduction Lipolisis Increase
ATP production
Decreased Ageing
Take home message!
Alumni
Lorena Esteban-Martinez
Lucia Garcia Ledo
Esther Seco Martin
Fengjuan Wang
Marta Mauro
Rita Marques
Kike Iglesias
Marián Mellén
Patricia Vázquez
Sergio Hernández
Marta Fierro
Current members
Elena Sierra-Fliardi
Ana Serrano Puebla
Beatriz Villarejo Zori
Raquel Gomez Sintes
Ignacio Ramirez Pardo
Natalia Rodriguez-Muela