APOPTOSIS

Dr. Shahzad Ali Jiskani

Assistant Professor
Department of Pathology
Chandka Medical College, SMBBMU
 Introduction
 Apoptosis, or programmed cell death, is highly
regulated process that allows a cell to self-degrade for
the body to eliminate unwanted or dysfunctional cells.

 During apoptosis, the genome of the cell will fracture,

the cell will shrink, and part of cell will disintegrate
into smaller apoptotic bodies.

 In human adults, 50-70 billion cells die each day

 A pathway of cell death induced by a tightly regulated
suicidal program, in which the cells destined to die,
activate enzymes that degrade cell’s own nuclear DNA
and nuclear and cytoplasmic proteins.

 The word ‘’apoptosis’’ means ‘’dropping off’’ or

‘’falling off’’
 Characteristics

Controlled by specific genes

Fragmentation of DNA

Fragmentation of nucleus

Blebs form and apoptotic bodies are released

Apoptotic bodies are phagocytized

 Apoptosis is also a protective mechanism, directing
lysis of virus-infected cells, foreign cells or incipient
neoplasm.

 Excess cell death can contribute to acquired immune

deficiency syndrome (AIDS) and neurodegenerative
diseases like Alzheimer’s and Parkinson diseases, and
ischemic injury

 Too little death could lead to cancer, persistent viral

infection or autoimmune disorders
 Role of Apoptosis in Immune System

 Immature lymphocytes that bind to autoantigens are

eliminated by apoptosis

 Apoptosis in thymocytes is associated with the

elimination of self-reactive clones of developing T cells
following their interaction with antigens in thymus
 Inducers of Apoptosis and Apoptotic
Signaling

Activators of apoptosis:
• Tumor necrosis factor-α (TNF-α)
• Fas ligand (FasL)
• Transforming growth factor-β (TGF-β)
• Bax
• Glucocorticoids
• Aberrant oncogene expression (e.g., c-myc)
 Physiological Apoptosis

Embryogenesis and fetal development

Hormone dependent involution

Immature lymphocytes

Epithelial cells in GI tract

Elimination of self-reactive lymphocytes

Death of cells that have served their function

 Pathologic Apoptosis

DNA damage due to radiation, chemotherapy etc.

Accumulation of misfolded proteins leads ER stress which

ends with apoptosis

Cell death in viral infections that induce apoptosis e.g., HIV,

adenovirus, hepatitis etc.

Organ atrophy after duct obstruction

 Genes that Regulate Apoptosis

 The ‘point of no return’ in apoptosis is reached when

caspases become enzymatically active in cleaving
target proteins
 Regulators Executioners
Triggers
DNA damage p53 Apaf-1

Cytokine Death domain Caspases

starvation factors

Hypoxia Bcl-2 family

Detachment Myc/oncogenes

Temperature Cytokine-
responsive kinases
Death receptor
Cytochrome c
 Mechanism of Apoptosis

Death receptor (extrinsic) pathway

Mitochondria (intrinsic) pathway

Execution phase

Removal of dead cells

 Intrinsic – Mitochondrial Pathway
 Increased mitochondrial permeability with release of pro-
apoptotic molecules into cytoplasm (cytochrome c)

 Synthesis of anti-apoptotic molecules (Bcl-2) promoted by Growth

factors

 When cells are deprived of growth factors or subjected to stress

anti-apoptotic molecules (Bcl-2) are lost

 Mitochondrial membrane becomes permeable and protein that

activate caspase leak out
 Extrinsic – Death Receptor Initiated
Pathway
 Death receptors are members of tumor necrosis factor
receptor family and a related protein called Fas (CD95)

 These molecules contain a death domain

 Fas and TNFα Receptor – Mediated
Apoptotic Signaling
 Fas receptor (CD95) belongs to family of receptors including
TNF and nerve growth factor (NGF) receptors that utilize
related signaling pathways to regulate cell proliferation,
differentiation or death.

 Fas serves as a receptor on cell surface for a ligand (FasL) and

the cross-linkng of FasL to Fas receptor triggers apoptosis on
target cells

 The Fas apoptotic pathway is implicated in eliminating

unwanted activated lymphocytes or virus-infected cells
 Executioners of Apoptosis: Caspases

 Caspases (Cysteinyl aspartate – specific proteinases) are family

of proteases contain cysteine at their active sites.

 These proteases play critical roles in executing programmed

cell death
 Inhibition of caspases is also one of the major
strategies adopted by viruses to elude their destruction
through suicide of infected cell.

 Leakage of cytochrome c from mitochondria might

serve as key signal to activate procaspases and initiate
biochemical events of death.
 Execution Phase

 The intrinsic and extrinsic pathways converge to a

caspase activation cascade

 Synthesized as inactive precursors; activated by

proteolytic cleavage

 Family of at least 12 proteases

 How Caspases Disassemble a Cell?

 Cleave structural proteins leading to nuclear

breakdown

 Converts cytoplasmic DNase to active form

 DNase cause characteristic internucleosomal cleavage

of DNA
 Removal of Dead Cells

 Dying cells secrete factors that recruit phagocytes.

 This facilitates prompt clearance before they undergo

secondary necrosis.

 Dead cells disappear without a trace and do not

produce inflammation
 Anti-Apoptosis Pro-Apoptosis
• Bcl-2 • Bax
• Bcl-XL • Bad
• Bcl-W • Bid
• Mcl-1 • Bok
• A1 • Bik
• Bak
Anti-Apoptotic Genes

Bcl-
Bcl-2
XL
E1B
 Morphological Changes

Cell shrinkage

Nuclear fragmentation

Chromatin condensation

Chromosomal DNA fragmentation

Formation of cytoplasmic blebs and apoptotic bodies

Phagocytosis
 Apoptotic Bodies
 Round oval mass of intensely
eosinophilic cytoplasm

 Fragments of dense nuclear

chromatin