1. Apoptosis and Diseases

2. 1.Concept 2.Major pathways 3.Key molecules 4.Apoptosis-related diseases Insufficient apoptosis in diseases Excessive apoptosis in diseases Coexistence of insufficient and excessive apoptosis in diseases 5.Principles of treatment Contents

3. History of Cell Death Research

5. Using C. elegan as a model

6. A highly regulated active cell death characterized by cell shrinkage and nuclear condensation. pre-programed, cascade events ATP, gene expression Morphology opposite to necrosis Apoptosis or Programmed Cell Death What is the difference of programmed cell death and apoptosis?

7. Apoptosis N ecrosis NaturePhysiological or pathological; specific Pathological, accidental StimulusMildStrong BiochemistryActive, energy-dependent, new protein synthesis Passive, energy-independent, no protein synthesis DNA Morphology Specific degradation, ladder (180-200 bp) Intact, shrinkage, condensation Random degradation Lysis, swelling InflammationNoYes Apoptotic bodyYesNo Gene regulationYesNo Apoptosis and Necrosis

8. Initiation Regulation Execution Phagocytosis Physiological, e.g., growth factors, estrogen; virus; chemicals. Inhibitory Factors Stimulatory Factors Physiological, FasL; Pathological, glutamate, free radicals; therapeutical, herb. Conserved Apoptotic Process

9. Conserved apoptotic paradigms in C. elegans, Drosophia, and mammals

10. Death receptor-mediated apoptotic pathway Mitochondria-mediated apoptotic pathway Nuclear-mediated apoptotic pathway Apoptotic Pathways

12. death-inducing signaling complex Fas-associating protein with death domain Apoptotic Pathways

13. Mitochondrial Membrane Permeabilization (MMP) in Apoptotic Process

14. ER and Apoptosis

15. Cross-talking among Organelles and Molecules in Apoptosis

17. 1. Activation of endonuclease 2. Activation of c aspases DNA ladder Collapse of cell and nucleus Executors

18. Role of Endonuclease 180-200 bp H1 Zn Zn 2+ CaMg Ca 2+ Mg 2+ Endonuclease

19. Role of Caspases (Focal adhesion kinase) (p21-activated kinase 2) (Ste20-related kinase)

20. Phosphatidylserine (PS) receptor (PSR) acts as a ‘tickle’ receptor for uptake of apoptotic cells

21. 2. Bcl-2 family ： anti-apoptotic: Bcl-2 (B cell lymphoma/leukemia), Bcl-X L pro-apoptotic: Bax, Bad Key Molecules 1.Caspases: Caspase-3,Caspase-8,Caspase-9, etc.Caspases: Caspase-3,Caspase-8,Caspase-9, etc. 3. Others: Apaf-1(apoptosis activating factor-1), cytochrome C, IAPs, p53, etc.

22. Table 1. Caspase-deficient miceKnockout Phenotype Caspase-1 Viable; impaired processing of IL-1; resistant to endotoxic shock. Caspase-2 Viable; excess numbers of female germ cells; oocytes resistant to chemotherapeutic drugs; B lymphoblasts resistant to granzyme B; accelerated death of facial neurons during development and of sympathetic neurons deprived of NGF. Caspase-3 Lethality at 3–5 weeks of age; defective neuronal apoptosis; T cells resistant to antigen-induced death; abnormal apoptotic morphology in dying cells. Caspase-8 Lethality around E12.5; hyperemia and abnormal heart muscle development; MEFs resistant to TNF, Fas and DR3 but sensitive to UV irradiation, etoposide, staurosporine, serum deprivation. Caspase-9 Perinatal lethal; impaired neuronal apoptosis; ES cells, MEFs and thymocytes generally resistant to intrinsic death stimuli such as DNA damage, though resistance depends on cell type. Caspase-11 Viable; impaired processing of caspase-1, IL-1; resistant to endotoxic shock. Caspase-12 Viable; embryonic fibroblasts are resistant to ER stress.

24. Apoptosis-related Diseases

26. Growth Apoptosis Balance of Growth and Apoptosis

27. Autoimmune disease, Tumor, virus infection, etc Proliferation Apoptosis Insufficient Apoptosis in Diseases

28. (1) Tumor Pathogenesis for tumor: stimulated cell proliferation inhibited cell apoptosis Cell survival > cell death in diseased tissue Etiologically, cell apoptosis is actually one of the natural anti-carcinogenic mechanisms

29. (2) Autoimmune diseases The lesion is caused by attack of auto-antibody or sensitized T cell to self-antigen. Normally, T cells against auto-antigen are eliminated by apoptosis during the development. When the negative selection is deregulated (thymus diseases), T cells survive and abnormally proliferate, then attack self tissue, lead to autoimmune diseases.

30. Mechanism of autoimmune diseases — Disrupted apoptosis of self-reactive cell Point mutation of FasL Insertion mutation of Fas Structural abnormity of FasL Decreased expression of Fas protein Escape the negative selection of self-reactive T cells Autoimmune diseases

31. Rheumatoid arthritis It is caused by decreased apoptosis and increased proliferation of arthral cell ; Increased IL-1 and TGF-β1 and decreased Fas expression, which inhibit apoptosis; Increased Bcl-2 、 Bcl-X L, which increased the threshold of apoptosis; Resistance of T-cells to apoptosis.

32. AIDS, neurodegenerative diseases, aberrant myocardial ischemic- reperfusion Excessive Apoptosis in Diseases

33. （ 1 ） Acquired Immune Deficiency Syndrome —AIDS HIV infection increased Fas gene expression gp 120 glycoprotein expression + receptor in CD 4 lymphocyte infusion of infected CD 4 cell leads to syncytin formation produce tat protein (enhance Fas expression) secret TNF CD + 4 T- lymphocyte apoptosis AIDS

34. (2) Cardiovascular diseases Cell death induced by ischemia-reperfusion Apoptosis Necrosis Early stage Later stage Peripheral region of infarct Center of infarct Mild ischemia Severe ischemia Chronic Acute

35. Possible mechanism (myocardial cell apoptosis induced by ischemia-reperfusion): (1) oxidative stress; (2) calcium overload; (3) p53 gene activation; (4) death receptor Fas, TNF over expressed. Cardiovascular diseases (cont.)

36. Heart failure: Myocardial cell diminishes in pressure- overload-induced heart failure Possible mechanisms: Oxidative stress; cytokines; ischemia; hypoxia; pressure or volume overload, neural-endocrine system deregulation; Lead to myocardial cell apoptosis

37. Alzheimer disease ， Parkinson disease ， Huntington disease ， multiple sclerosis Factors involved in neuronal apoptosis:  amyloid peptide, calcium overload, oxidative stress and neuronal growth factor insufficiency, etc. Lead to neuronal cell apoptosis （ 3 ） Neurodegenerative diseases

38. Coexistence of Excessive and Insufficient Apoptosis in Diseases

39. oxidative LDL platelet activation Ag Ⅱ hypertension excess apoptosis insufficiency in in endothelium smooth muscle atherosclerosis Coexistence of Excessive and Insufficient Apoptosis

40. Diseases Mechanism Apoptosis Heart failure Ischemia, inflammation, etc. AIDSHIV infection of T 4 cell AD, PDIschemia, inflammation, etc CancerP53, Bcl-2 Autoimune diseases Autoreactive T cells or B cells Atherosclerosis Endothelial cell, muscle cell Apoptosis and Diseases

41. Gene Affected disease Tumor necrosis factor Familial periodic fever syndrome receptor 1 (TNF-R1) Fas (CD95; Apo-1) Autoimmune lymphoproliferative syndrome type I(ALPS I), malignant lymphoma, bladder cancer Fas ligand Systemic lupus erythematodes (only one case identified) Perforin Familial hemophagocytic lymphohistiocytosis (FHL) Caspase 10 Autoimmune lymphoproliferative syndrome type II (ALPS II) bcl-10 Non-Hodgkin’s lymphoma p53 Various malignant neoplasms Bax Colon cancer; hematopoetic malignancies bcl-2 Non-Hodgkin’s lymphoma c-IAP2 Low-grade MALT lymphoma NAIP1 Spinal muscular atrophy Apoptosis Genes mutated in Diseases

42. Principle of Treatment

45. Thank you