CELL DEATH/APOPTOSIS
Dr. Deeba S. Jairajpuri, Ph.D.
Cell death
• All cells eventually die by necrosis or apoptosis
• Necrosis is a passive pathological process induced by cell
injury, and can occur for a group of cells. There is rupture
(breakdown) of the cell membrane and spilling of the
cytoplasm and may induce inflammation (movement of
immune cells at the site of necrosis).
• Apoptosis is an active, non-pathological (normal
physiological, as essential as division) process affect
individual cells without affect neighboring cells or induce
inflammation. Cell membrane show blebbed (bulging of the
plasma membrane) appearance
• Disturbance of the apoptotic pathway may result in cancer,
autoimmune disease or neurodegenerative disorder
Apoptosis and necrosis
Necrosis
• Passive pathological process induced by acute injury or disease
• Group of cells in a site of injury undergo necrosis e.g. as a result of
physical injury
• Cells increase in volume and lyse (burst) releasing their intracellular
content inducing a potentially damaging inflammation to the neighbor
cells
• It is completed within several days
• Gangrene : a type of necrosis caused by insufficient blood
supply. It may occur after an injury or infection, or in people suffering
from any chronic health problem affecting blood circulation
(Diabetes and long-term smoking increase the
risk of suffering from gangrene).
Cell
death
Apoptosis
• A cell, receives signals for survival or it
vanishes.
• Programmed cell death (apoptosis): In the cell
deprived of survival factors, a suicidal program
is activated leading to cell death.
• Apoptotic cells shrinks in size but do not lyse, their plasma
membrane remain intact, part of it bud off (bleb), became
asymmetrical, loss attachment with neighbor tissue.
• Its chromatin condenses,
• the cell fragmented into vesicles called apoptotic bodies which
contain: cytosol, condensed chromatin and organelles
• During apoptosis, a Phosphatidylserine,
(phospholipid) in the inner leaflet of lipid bilayer,
actively held facing cytoplasm by the enzyme
flippase, will invert by scramblase (floppase) and
became exposed on cell surface using ATP.
• When the phosphatidylserines flip to the
extracellular surface, they act as a signal for
phagocytes cells (e.g. macrophages) to engulf and
degrade the cells (reducing the risk of
inflammation)
• Phagocytes cells also release certain chemicals like
cytokines (IL-10) and transforming growth factor
(TGF-b) that inhibit inflammation.
Necrosis Apoptosis
• Effect is detrimental (harmful) • Effect is beneficial
• Causes always pathologic • Causes can be physiologic or
• Cellular swelling pathologic
• Membranes are broken • Cellular condensation
• Requires ATP • Membranes remain intact
• Cell lyses, resulting in an • Requires ATP
inflammatory tissue • Cell is phagocytosed, no
reaction tissue reaction
• whole areas of the tissue • individual cells appear
are affected affected
Biologic significance apoptosis
Function:
- Elimination of damaged cells. Cells subjected to
unrepairable DNA damage, infected by virus,
starving, exposed to radiations or toxins.
- p53 halts the cell cycle and stimulate apoptosis
via Bax protein.
- Removal of apoptotic cells is important because
this saves nutrients for other healthy cells to
consume and prevent spreading of viruses from
infected cells to normal healthy cells.
p53 enhance Bax expression
DNA Damage
Development
- Apoptosis is used during development of the embryo.
- There are several errors due to excessive proliferation and
differentiation.
- In developing nervous system approx. half of the generated nerve cells
are immediately undergo apoptosis
- Selective apoptosis of tissues between developing digits(fingers and
toes), must occur to separated fingers and toes. This is called as organ
sculpting. Incomplete apoptosis can result in abnormal structures
- Development of mature immune system also needs apoptosis e.g. to
remove auto-reactive T-cells (those immune cells that will attack cells
of the body), negative selection (rejection)of such cells happen in the
thymus and apoptosis is induced in them.
Homeostasis Requires Apoptosis
• In normal adults, the number of cells kept
constant by balance between cell division and
cell death.
• Billion of cells die every hour in bone marrow &
epithelia.
• Non-functioning or damaged cells are removed
and replaced by new ones to keep the balance
• If equilibrium is disturbed, abnormal growth
(tumors) or loss of cells will occur.
Initiation of apoptosis
By formation of Apoptosome:
- Unrepairable cell damage initiates death program
- Bax is a pro-apoptotic protein which is activated by p53
and it gets inserted in the mitochondria membrane to act
as a channel for exit of cytochrome c
- Once in the cytoplasm, cytochrome c triggers formation
of a large complex protein, the apoptosome which is a
combination of Apaf-1, Cytochrome c and caspase 9
that destroys cell proteins and DNA by using caspase
proteolytic cascade, causing apoptosis (cell death).
Apoptosome
Death receptors
• Initiate apoptosis in a cell under the influence of
external signals
• These receptors have cytoplasmic sequence known as
death domain (DD), which initiate apoptosis
• The Death Domain of an active death receptor to
activate caspase 8 or 10 which through series of
reactions can induce apoptosis.
• Example, if cells are infected with virus, Fas death
receptor stimulate apoptosis of the infected cells
• The apoptosis initiated by death receptors by binding
of ligands (like immune cells) is called the extrinsic
pathway whereas the one initiated by internal cellular
signals is called intrinsic pathway.
Fas death receptor
Caspase family of proteases
(cysteine-aspartic protease)
• Are a family of proteases, they are majorly involved
in apoptotic cell death
• The members of this class of proteases have
cysteine-aspartic acid present at their active site
• Synthesized as pro-enzymes (inactive) and are
activated whenever needed by post-translational
modification
Classification of caspases
• About 11 members found in humans, classified
based on function, some not involved in apoptosis
• Caspase 1 involved in cytokines maturation
• Caspases 4 & 5 involved in inflammation
• Caspase 14 important in skin development
• Remaining caspases (2, 3, 6, 7, 8, 9, 10)involved
in apoptosis, grouped into initiators and effectors
apoptotic caspases
Apoptosis caspases
Initiator caspases:
• Caspase 2, 8, 9 & 10. CARD domain (caspase recruitment
domain) in caspase 2 & 9 and DED domain in (8 & 10) which
enable them to interact with their substrates which are the
effector caspases
• They cleave and activate inactive effector caspases
Effector caspases:
• 3, 6 & 7, are also called as executioner caspases, bind to and
cleave important proteins within the cell causing apoptotic
demise of the cell
Caspase cascade:
• Is a sequential proteolytic activation of caspases in orderly
fashion during the apoptosis initiation. The cascade can be
activated by various stimuli including the apoptosome, death
receptors etc.
• Caspase inhibitors regulate the process.
Deregulation of apoptosis causing Cancer
• Insufficient apoptosis is seen in cancer or autoimmunity
• Tumour cells(cancer cells) can acquire resistance to apoptosis by :
Ø Expression of anti-apoptotic proteins Bcl-2 over-expression in
follicular B-cell lymphoma; over-expression of IAPs in different types
of cancers including neuroblastoma
Ø Inactivation of pro-apoptotic genes BAX mutation; APAF-1 in
melanomas(skin cancers)
Ø Alteration of p53 pathway by mutation in p53 gene making it non-
functional.
