Cellular Apoptosis And Disease
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Cellular apoptosis and disease
Zhao Mingyao BMC.ZZU
2006 ， 06
Cell signal transduction change
cell signal
Apoptosis ↑or↓or loss→disease
Concept of apoptosis • ——autonomic and ordered cell death controlled by gene or called as programmed cell death (PCD)
model: the nematode, drosophila and mouse
Flow Cytometer
nematode drosophila
mouse
research model
12000
publications
10000 8000 6000
rapid up-shift period
4000 2000
silent period
0 1970 1975 1980 1985 1990 1995 2000 2005
Thirty years on apoptosis
Classical Apoptosis model
• Nematode : total 1090 cells in young ,959 cells in adult, 131 cells lost ? • Time : 1 h r • Gene : 14 gene known
Section1 Instruction of apoptosis
1.differences between necrosis and apoptosis
Strong stimulation
Passive ATP(-) 溶酶释出，
? 炎症反应
Weak ~
Active new protein ATP↓ Apoptotic bodies
Comparison between apoptosis & necrosis
DNA electrophoresis necrosis
+
apoptosis
–
famous nucleosomal ladder
2.the biological meanings of apoptosis
Holes are forming ?
Mitochondrio-nuclear AIF translocation in the interdigital cells of the mouse embryo
Inthehumans, By time I was born, as more me had thanadult survived. It was as many 1011ofcells die died in each each day andnoare wonder I cannot remember; during time went to through brain body by of other cells. replaced The mass cells we lose eachthat year is Iclose our entire after brain for nine months, finally contriving the one model that could weight! be human, equipped for language.
Apoptosis importance ①Maintain normal development and growth:remove excess cell ② Maintain homeostasis : remove damaged, mutant senile cell ③Active defense function: block replication of cells infected by virus
3. apoptosis process (period)
course of cellular apoptosis 1.Transduction of apoptosis signal starting signal Ca2+ 、 cAMP 、 ceramide
2.activation of apoptosis gene express enzymes and other substance
3.execution of cellular apoptosis DNase and Caspases
4.removal of apoptotic cell engulfed by near cell ( Mφor other cell )
4.the two major biomedical changes in the courses of apoptosis
(1)Endogenous Dnase activation and its effect • A series cellular ST activation • Execution of chromosome DNA cleavage
• Fragment of DNA ： classic ladder pattern ？
chromosome Endonucleases first cleave 5min
300 kb pieces 50kb 90min 180~200bp
or its whole folds fragment
(2) Caspases activation and its effect • A group of protease ( 13 members ) cysteine-containing aspartate-spicific protease
caspases
note:Cysteine 半胱 aa Aspartate 天冬 aa
Caspases function ①Deactivating inhibitor of apoptosis: as Bcl-2 ②Hydrolyzing pro structure, cellular decomposing, apoptotic body ③ Hydrolyzing the related active proteins : make them to gain or lose function
Section 2 Regulation of cell apoptosis
1.Signal pathway and effectors of apoptosis
DNA damage
p53
[Ca2+]i dyshomeostasis
ROS
bax
Cyto C leakage Bad Bcl-XL-Apaf-1
TNFa, Fas
Adaptor (FADD,TRADD)
Bid Apaf- cyto c- dATP
Caspase 8
Caspase 9 Caspase 3 Cell apoptosis
Signal pathway of apoptosis
2. regulators of apoptosis • (1)the central role of mitochondria in apoptosis DNA damage
[Ca2+]i dyshomeostasis
ROS
Bax open p53
PTP VDAC
bax
Bcl-XL closing Cyto C leakage Bad
Bid
Apaf- cyto c- dATP Caspase 9
mitochondria damage lead to apoptosis
PTP opening
(permeability transition pore)
Inducing factor →Δ Ψm↓ PTP permeability↑→apoptosis starting factor （ Cyt. C ， Apaf ， AIF ） released Abbreviation note: •PTP----permeability transition pore •Apaf----apoptosis protease activated factor •Cyt.C----cytochrome C •AIF---- apoptosis inducement factor
Deleterious network hypothesis of cellular apoptosis –
PTP opening
–
+
–
Caspase inhibitor
Apaf+Cyt.C
–
Mt Δ Ψm↓
Bcl-2
AIF Pro-Caspase 9
Caspase-9 +
Inactivity DNnase
+ Ca2+
Pro-Caspase3 DNase activated DNA rupture
Caspase-3
+ Protein hydrolyzed
apoptosis
(2)bcl-2 family pro • Bcl-2, Bcl-XL, Mcl-1 promote survival (anti-apoptotic pro) • Bax, Bad, Bak, Bid induce death (pro-apoptotic pro)
cell
bcl-2 • • • •
Binding Apaf-1 Prevent release of cyto C Regulator of Ca2+ homeostasis antioxidant
• Inhibiting ~ ： bcl-2 • Promoting ~ ： fas ， P53 • Dual direction regulating ~ ： cmyc ， bclx
Bcl-2 —— 229aa （ mouse 236aa ） distributing in endomembrane surface Bcl-2----B cell lymphoma/leukemia-2
(3)IAP pro • (IAP)/BIRP gene family: XIAP, c-IAP-1, c-IAP-2, ILP-2, NAIP, Livin and Apollon • block apoptosis directly interacting with initiator and effector caspases preventing their proteolytic processing and enzymatic activity BIRC# for "baculoviral IAP repeat-containing".
BIRC# for "baculoviral IAP repeat-containing".
The "Inhibitor of Apoptosis Proteins" family Characterization of IAP family members • BIRC1 (Neuronal apoptosis inhibitory protein; NAIP) • BIRC2 (API1; HIAP2; cIAP1; MIHB) • BIRC3 (API2; HIAP1; cIAP2; MIHC) • BIRC4 (XIAP; API3; MIHA; ILP) • BIRC5 (Survivin; API4; TLAP) • BIRC6 (Apollon; BRUCE) • BIRC7 (MLIAP; KIAP; Livin) • BIRC8: (ILP-2; TIAP)
•
******Inhibitory factor ： cytokine— IL-2 ， NGF hormone— ACTH 、 testosterone , estrogen other— Zn2+(zinc), phenobarbital , Cysteine protease inhibitor, EB virus, neutroamino acid
3.Inducers of apoptosis • (1) apoptosis induced by death signal TNFa, Fas
Adaptor (FADD,TRADD)
Caspase 8
apoptosis signal pathway cell Induction signal
many pathways SAPK/JNK
ceramide RR ROS
NF-kB,JNK/AP-1 pathway P53 gene
+ mtDNA
bax or down regulating bcl-2
(2) apoptosis induced by DNA damage DNA damage
p53
bax
Cyto C leakage Bad Bcl-XL-Apaf-1
Bid Apaf- cyto c- dATP
oxidative stress induce apoptosis? • • • •
P53 gene activation activating Ca2+ /Mg2+ -dependent Dnase Ca2+ influx↑ activating NF-kB and Ap-1 （ Fas)
(3)Other apoptosis triggers
1)Inducing factor ： hormone, GF↑↑, GC physic and chemic factor immune microbe
2)P53 molecular policeman?
Blocking IGF+R + Bax
Bcl-2↓
G1 Cyclin B
M
p53
Cyclin D
S Cyclin A+CDK
Cyclin A+CDK1
G2
3) [Ca2+ ] dyshomeostasis • activating Ca2+ /Mg 2+ -dependent Dnase • activating nuclear transcription factor related to apoptosis • Ca 2+ promoting to exposing enzymolytic sites between nuclear ribosomes
Section 3 abnormal cell apoptosis in diseases
1.tumor insufficiency of apoptosis • Bcl-2 express↑ • P53 mutation or loss
2. immune diseases (1) Autoimmune diseases • Hashilmoto’s thyroiditis (HT ) Fas/FasL mediated apoptosis • SLE
(2)AIDS HIV
CD4+ cell damaged selectively
of CD4+
mechanism lymphocyte apoptosis Cell level
CD4+
syncytia
Effect and target effect
CD4+
HIV Apoptotic body Signal substance
AICD
HIV induce CD
+ 4
apoptosis Receptor,molecule level
gp120- R Fas-R↑ TNF
OFR tat Host cell
CD4+cell
Mφ
3. Heart failure myocyte hypertrophy and widening of interstitial spaces due to depostion of collagen, inflammatory cells and amyloid.
atherosclerosis, AS Insufficiency and excess of apoptosis exist together
oxLDL ↑ activated plt ↑ Ag II hypertension
Endothelia ~↑ Smooth ~ insufficiency (proliferation↑>apoptosis↑ ）
Endothelia barrier↓
4.Alzheimer disease, AD Neuron retrogression —— loss of neuron in hippocampus and basal nuclei • Loss of Cholinergic neurons reachs 30%~50% ， involved cortex
AD mechanism disease cause such as OFR
neuron Ca2+ inward flowing↑ activating related genes promoting β- amyloid content ↑ Tan pro overphosphorylation
Section 5 regulating apoptosis in treating diseases
meaning of apoptosis in prevention and treatment of diseases • (1) Correctively administration of the related factors to ~ • Inhibiting ~ ： AD---- NGF acute T cell leukemia ： low dose • radiation 、 TNFα promoting ~ : heat therapy of local tumor ； high temperature 43℃ 、 30min prostate glands cancer ： removal of • testicle(testes), testosterone↓
(2) Interfere apoptosis ST • FAS/FASL ST system ： adriamycin stimulation • SPP （ metabolism products of ceramide ST system ）： transducting proliferation signals ， antiapoptosis→AIDS ， AD
(3) Regulating related gene to apoptosis • transfection of wild type P53 gene
(4) Control related enzyme to apoptosis • DNase ： Ca 2+ /Mg 2+ -dependent Ca 2+ ↑ activation ； Zn 2+ ↑inhibiting • Caspases: Ca2+ directive or indirective activation
(5)Prevent Mt Δ Ψm↓ • Cyclosporin A,immune inhibitor blocking Δ Ψm↓ ， its depriver without immune inhibiting effect, but stabilize Δ Ψm, prevent↓
• • • •
A simple experiment A smart observation A further thinking Show a very different formation of cellular death and it`s abstruse meaning of life review founding the process of cellular apoptosis
THE 2002 NOBEL PRIZE WINNER
Sydney Brenner
H. Robert Horvit
John E. Sulston
1/3 of the prize
1/3 of the prize
United Kingdom
United Kingdom
USA
The Molecular Sciences Institute Berkeley, CA, USA
Massachusetts Institute of Technology (MIT) Cambridge, MA, USA
b. 1927 (in Union of South Africa)
nematode
b. 1947
Apoptosis gene
The Wellcome Trust Sanger Institute Cambridge, United Kingdom b. 1942
cell lineages of the nematode
Sydney Brenner receiving his Nobel Prize from His Majesty the King at the Stockholm Concert Hall. Photo: Hans Mehlin, Nobel e-Museum
THE NOBEL DIPlOMA
support & cooperation in my teaching
more success on the way of study & work in future Zhao Mingyao(Jasper) 2006-06-08
Zhao Mingyao BMC.ZZU
2006 ， 06
Cell signal transduction change
cell signal
Apoptosis ↑or↓or loss→disease
Concept of apoptosis • ——autonomic and ordered cell death controlled by gene or called as programmed cell death (PCD)
model: the nematode, drosophila and mouse
Flow Cytometer
nematode drosophila
mouse
research model
12000
publications
10000 8000 6000
rapid up-shift period
4000 2000
silent period
0 1970 1975 1980 1985 1990 1995 2000 2005
Thirty years on apoptosis
Classical Apoptosis model
• Nematode : total 1090 cells in young ,959 cells in adult, 131 cells lost ? • Time : 1 h r • Gene : 14 gene known
Section1 Instruction of apoptosis
1.differences between necrosis and apoptosis
Strong stimulation
Passive ATP(-) 溶酶释出，
? 炎症反应
Weak ~
Active new protein ATP↓ Apoptotic bodies
Comparison between apoptosis & necrosis
DNA electrophoresis necrosis
+
apoptosis
–
famous nucleosomal ladder
2.the biological meanings of apoptosis
Holes are forming ?
Mitochondrio-nuclear AIF translocation in the interdigital cells of the mouse embryo
Inthehumans, By time I was born, as more me had thanadult survived. It was as many 1011ofcells die died in each each day andnoare wonder I cannot remember; during time went to through brain body by of other cells. replaced The mass cells we lose eachthat year is Iclose our entire after brain for nine months, finally contriving the one model that could weight! be human, equipped for language.
Apoptosis importance ①Maintain normal development and growth:remove excess cell ② Maintain homeostasis : remove damaged, mutant senile cell ③Active defense function: block replication of cells infected by virus
3. apoptosis process (period)
course of cellular apoptosis 1.Transduction of apoptosis signal starting signal Ca2+ 、 cAMP 、 ceramide
2.activation of apoptosis gene express enzymes and other substance
3.execution of cellular apoptosis DNase and Caspases
4.removal of apoptotic cell engulfed by near cell ( Mφor other cell )
4.the two major biomedical changes in the courses of apoptosis
(1)Endogenous Dnase activation and its effect • A series cellular ST activation • Execution of chromosome DNA cleavage
• Fragment of DNA ： classic ladder pattern ？
chromosome Endonucleases first cleave 5min
300 kb pieces 50kb 90min 180~200bp
or its whole folds fragment
(2) Caspases activation and its effect • A group of protease ( 13 members ) cysteine-containing aspartate-spicific protease
caspases
note:Cysteine 半胱 aa Aspartate 天冬 aa
Caspases function ①Deactivating inhibitor of apoptosis: as Bcl-2 ②Hydrolyzing pro structure, cellular decomposing, apoptotic body ③ Hydrolyzing the related active proteins : make them to gain or lose function
Section 2 Regulation of cell apoptosis
1.Signal pathway and effectors of apoptosis
DNA damage
p53
[Ca2+]i dyshomeostasis
ROS
bax
Cyto C leakage Bad Bcl-XL-Apaf-1
TNFa, Fas
Adaptor (FADD,TRADD)
Bid Apaf- cyto c- dATP
Caspase 8
Caspase 9 Caspase 3 Cell apoptosis
Signal pathway of apoptosis
2. regulators of apoptosis • (1)the central role of mitochondria in apoptosis DNA damage
[Ca2+]i dyshomeostasis
ROS
Bax open p53
PTP VDAC
bax
Bcl-XL closing Cyto C leakage Bad
Bid
Apaf- cyto c- dATP Caspase 9
mitochondria damage lead to apoptosis
PTP opening
(permeability transition pore)
Inducing factor →Δ Ψm↓ PTP permeability↑→apoptosis starting factor （ Cyt. C ， Apaf ， AIF ） released Abbreviation note: •PTP----permeability transition pore •Apaf----apoptosis protease activated factor •Cyt.C----cytochrome C •AIF---- apoptosis inducement factor
Deleterious network hypothesis of cellular apoptosis –
PTP opening
–
+
–
Caspase inhibitor
Apaf+Cyt.C
–
Mt Δ Ψm↓
Bcl-2
AIF Pro-Caspase 9
Caspase-9 +
Inactivity DNnase
+ Ca2+
Pro-Caspase3 DNase activated DNA rupture
Caspase-3
+ Protein hydrolyzed
apoptosis
(2)bcl-2 family pro • Bcl-2, Bcl-XL, Mcl-1 promote survival (anti-apoptotic pro) • Bax, Bad, Bak, Bid induce death (pro-apoptotic pro)
cell
bcl-2 • • • •
Binding Apaf-1 Prevent release of cyto C Regulator of Ca2+ homeostasis antioxidant
• Inhibiting ~ ： bcl-2 • Promoting ~ ： fas ， P53 • Dual direction regulating ~ ： cmyc ， bclx
Bcl-2 —— 229aa （ mouse 236aa ） distributing in endomembrane surface Bcl-2----B cell lymphoma/leukemia-2
(3)IAP pro • (IAP)/BIRP gene family: XIAP, c-IAP-1, c-IAP-2, ILP-2, NAIP, Livin and Apollon • block apoptosis directly interacting with initiator and effector caspases preventing their proteolytic processing and enzymatic activity BIRC# for "baculoviral IAP repeat-containing".
BIRC# for "baculoviral IAP repeat-containing".
The "Inhibitor of Apoptosis Proteins" family Characterization of IAP family members • BIRC1 (Neuronal apoptosis inhibitory protein; NAIP) • BIRC2 (API1; HIAP2; cIAP1; MIHB) • BIRC3 (API2; HIAP1; cIAP2; MIHC) • BIRC4 (XIAP; API3; MIHA; ILP) • BIRC5 (Survivin; API4; TLAP) • BIRC6 (Apollon; BRUCE) • BIRC7 (MLIAP; KIAP; Livin) • BIRC8: (ILP-2; TIAP)
•
******Inhibitory factor ： cytokine— IL-2 ， NGF hormone— ACTH 、 testosterone , estrogen other— Zn2+(zinc), phenobarbital , Cysteine protease inhibitor, EB virus, neutroamino acid
3.Inducers of apoptosis • (1) apoptosis induced by death signal TNFa, Fas
Adaptor (FADD,TRADD)
Caspase 8
apoptosis signal pathway cell Induction signal
many pathways SAPK/JNK
ceramide RR ROS
NF-kB,JNK/AP-1 pathway P53 gene
+ mtDNA
bax or down regulating bcl-2
(2) apoptosis induced by DNA damage DNA damage
p53
bax
Cyto C leakage Bad Bcl-XL-Apaf-1
Bid Apaf- cyto c- dATP
oxidative stress induce apoptosis? • • • •
P53 gene activation activating Ca2+ /Mg2+ -dependent Dnase Ca2+ influx↑ activating NF-kB and Ap-1 （ Fas)
(3)Other apoptosis triggers
1)Inducing factor ： hormone, GF↑↑, GC physic and chemic factor immune microbe
2)P53 molecular policeman?
Blocking IGF+R + Bax
Bcl-2↓
G1 Cyclin B
M
p53
Cyclin D
S Cyclin A+CDK
Cyclin A+CDK1
G2
3) [Ca2+ ] dyshomeostasis • activating Ca2+ /Mg 2+ -dependent Dnase • activating nuclear transcription factor related to apoptosis • Ca 2+ promoting to exposing enzymolytic sites between nuclear ribosomes
Section 3 abnormal cell apoptosis in diseases
1.tumor insufficiency of apoptosis • Bcl-2 express↑ • P53 mutation or loss
2. immune diseases (1) Autoimmune diseases • Hashilmoto’s thyroiditis (HT ) Fas/FasL mediated apoptosis • SLE
(2)AIDS HIV
CD4+ cell damaged selectively
of CD4+
mechanism lymphocyte apoptosis Cell level
CD4+
syncytia
Effect and target effect
CD4+
HIV Apoptotic body Signal substance
AICD
HIV induce CD
+ 4
apoptosis Receptor,molecule level
gp120- R Fas-R↑ TNF
OFR tat Host cell
CD4+cell
Mφ
3. Heart failure myocyte hypertrophy and widening of interstitial spaces due to depostion of collagen, inflammatory cells and amyloid.
atherosclerosis, AS Insufficiency and excess of apoptosis exist together
oxLDL ↑ activated plt ↑ Ag II hypertension
Endothelia ~↑ Smooth ~ insufficiency (proliferation↑>apoptosis↑ ）
Endothelia barrier↓
4.Alzheimer disease, AD Neuron retrogression —— loss of neuron in hippocampus and basal nuclei • Loss of Cholinergic neurons reachs 30%~50% ， involved cortex
AD mechanism disease cause such as OFR
neuron Ca2+ inward flowing↑ activating related genes promoting β- amyloid content ↑ Tan pro overphosphorylation
Section 5 regulating apoptosis in treating diseases
meaning of apoptosis in prevention and treatment of diseases • (1) Correctively administration of the related factors to ~ • Inhibiting ~ ： AD---- NGF acute T cell leukemia ： low dose • radiation 、 TNFα promoting ~ : heat therapy of local tumor ； high temperature 43℃ 、 30min prostate glands cancer ： removal of • testicle(testes), testosterone↓
(2) Interfere apoptosis ST • FAS/FASL ST system ： adriamycin stimulation • SPP （ metabolism products of ceramide ST system ）： transducting proliferation signals ， antiapoptosis→AIDS ， AD
(3) Regulating related gene to apoptosis • transfection of wild type P53 gene
(4) Control related enzyme to apoptosis • DNase ： Ca 2+ /Mg 2+ -dependent Ca 2+ ↑ activation ； Zn 2+ ↑inhibiting • Caspases: Ca2+ directive or indirective activation
(5)Prevent Mt Δ Ψm↓ • Cyclosporin A,immune inhibitor blocking Δ Ψm↓ ， its depriver without immune inhibiting effect, but stabilize Δ Ψm, prevent↓
• • • •
A simple experiment A smart observation A further thinking Show a very different formation of cellular death and it`s abstruse meaning of life review founding the process of cellular apoptosis
THE 2002 NOBEL PRIZE WINNER
Sydney Brenner
H. Robert Horvit
John E. Sulston
1/3 of the prize
1/3 of the prize
United Kingdom
United Kingdom
USA
The Molecular Sciences Institute Berkeley, CA, USA
Massachusetts Institute of Technology (MIT) Cambridge, MA, USA
b. 1927 (in Union of South Africa)
nematode
b. 1947
Apoptosis gene
The Wellcome Trust Sanger Institute Cambridge, United Kingdom b. 1942
cell lineages of the nematode
Sydney Brenner receiving his Nobel Prize from His Majesty the King at the Stockholm Concert Hall. Photo: Hans Mehlin, Nobel e-Museum
THE NOBEL DIPlOMA
support & cooperation in my teaching
more success on the way of study & work in future Zhao Mingyao(Jasper) 2006-06-08
