Cell Death

BIOCHEM-700

Finals
Two types of cell death
Necrosis Apoptosis
1. Pathological, Results from damage by 1. A continuous, controlled process
an external agent, e.g. infection or 2. Physiological or Pathological, In an adult,
injury apoptosis maintains cell numbers, balancing the
production of new cells by mitosis and the death
2. Swelling, lysis
of older cells
3. Causes rupturing of cells and leakage 3. Condensation, cross-linking
of their contents into surrounding
tissues 4. Cells are destroyed in a way that does not cause
leakage of their contents into surrounding
4. As a result, the non-specific immune tissues
response occurs and leads to
5. So, the non-specific immune response does not
inflammation
occur and there is no inflammation
5. Passive
6. Active
6. Dissipates 7. Phagocytosed
7. Externally induced 8. Internally or externally induced
 Overview of Apoptosis
• Evolutionarily conserved
• Occurs in all multicellular animals studies (plants too!)
• Stages and genes conserved from nematodes (worms)
and flies to mice and humans
An example of
apoptosis
Life cycle of a frog

The tail of this developing

adult disappears as a result
of apoptosis
Apoptosis occurs in humans too
• Embryological development
• In the fifth week, our hands and feet develop as
paddle-like structures
• Ridges develop in these paddle-like structures, which
become our fingers and toes
• Apoptosis causes the death of cells between these Selection (eliminates non-functional cells):
ridges, allowing our individual fingers and toes to
become separate
• In childhood
• Between the ages of 8 and 14, the average child
loses between 20 and 30 billion cells each day as a
result of apoptosis
• In adulthood
• The average adult loses between 50 and 70 billion
cells each day as a result of apoptosis
Immunity (eliminates dangerous cells):

Self antigen
recognizing cell

Organ size (eliminates excess cells):

The role of caspases
• Caspases are a family of proteases (i.e. protein-digesting enzymes)
• They are synthesised in a cell’s cytoplasm as inactive procaspases
• Once activated, caspase molecules have several effects, for example, they:
• activate more procaspase to caspase
• hydrolyse structural proteins in the cytoplasm, e.g. actin
• hydrolyse structural proteins in the nucleus, e.g. lamins
• activate DNA-hydrolysing enzymes
• As structural proteins are hydrolysed:
• the nucleus breaks up
• the cell shrinks and forms extensions, called blebs
• Small membrane-bound fragments (apoptotic bodies) bud off from these
blebs and are engulfed by phagocytic white blood cells
 Apoptosis observed
This time-lapse sequence covers a period of about 5 minutes
The cytoplasm is shown in red and the nucleus in blue
The cell and nucleus both shrink as their proteins are hydrolysed
The cell forms blebs that break off into small fragments (apoptotic bodies)
Caspase cascade
• As each new active caspase molecule is formed it, in
turn, activates more procaspase molecules.
• A cascade (or amplifying reaction series) develops
• 1 caspase molecule
• 
• 2 caspase molecules
• 
• 4 caspase molecules
• 
• 8 caspase molecules
• 
• 16 caspase molecules
What activates caspase initially?
Intrinsic pathway (via Mitochondria) This stress could be caused by
• An intracellular signal in response to stress factors such as:
• radiation
• Mitochondria release substances into the cytosol of the cell • heat
• One of these is cytochrome c, one component of the electron • nutrient deprivation
transport chain of aerobic respiration • viral infection
• Stress to a cell can cause the mitochondrial membranes to become • an increase in the calcium ion
more permeable so that cytochrome c leaks into the cytosol concentration of the cytoplasm
• binding of glucocorticoids to
• In the cytosol, cytochrome c causes hydrolysis of inactive
procaspase to active caspase receptors on the nuclear envelope

Bax protein
cause the
release of
Cytochrome c.
Intrinsic pathway is
mainly controlled by
Bcl2 and IAPs.
 The action of cytochrome Release of cytochrome c from
mitochondria

c
Cytochrome c binds to second
Cytochrome c binds with ATP and with protease activating factor-1
mitochondria-derived activator of caspase
(Apaf-1) to form an apoptosome
(SMAC)

SMAC binds to proteins that inhibit

Apoptosome binds with, and activates,
apoptosis (IAPs) and so allows
procaspase
action of caspases

Active caspase starts caspase Active caspase starts caspase

cascade cascade
 2 3 4
1
Complementary

Extrinsic
protein on surface
of killer T cell
binds to ‘death

pathway
The
receptor’ on
surface of target
extrinsic
cellpathway - Uses
Adaptor proteins
extracellular death ligands
recruit and
( Fas ligand, tumor necrosis factor TNF) to activateactivate
‘death receptors’ which pass the apoptotic signal procaspase
to
molecules Active caspase
initiator caspases (e. g. capsase 8 ) and to the molecules start
executioner caspases e. g. caspase 3; caspase 7). the caspase
cascade
 The extracellular signal molecules

• Mitogens, which stimulate cell division, primarily by

relieving intracellular negative controls that otherwise
block progress through the cell cycle.
• Growth factors, which stimulate cell growth (an
increase in cell mass) by promoting the synthesis of
proteins and other macromolecules and by inhibiting
their degradation.
• Survival factors, which promote cell survival by
suppressing apoptosis
 Inhibition of Caspase Activation by Trophic Factors

• In the presence of a trophic factor,

caspases are maintained in their inactive
pro-forms.
• In neurons, NGF causes Bad to be
phosphorylated and inactivated via the
PI-3 kinase/PKB signaling pathway.
• Under these conditions Bcl-2 can inhibit
the activity of Bax.
• This prevents Cyt c release from
mitochondria and blocks the activation
of procaspase 9 by Apaf-1.
 Caspase Activation in the Absence of Trophic Factors
• The mechanism by which caspases are
activated in the absence of a trophic factor
(e.g., NGF) is illustrated in Fig.
• In the absence of the trophic factor, the Active
pro-apoptotic protein Bad inhibits the anti-
apoptotic protein Bcl-2.
• As a result, Bcl-2 cannot inhibit the activity of
the Bax pro-apoptotic protein.
• This results in release of cytochrome c (Cyt c)
from mitochondria.
• Cyt c binds to the Apaf-1 adaptor protein, and
this triggers procaspase 9 to undergo
activation.
• Caspase 9 then cleaves and activates
procaspase 3.
• Caspase 3 cleaves substrates leading to
alterations that result in cell death.
• Caspase activation and cell death can also be
triggered by binding of death signals, such as
tumor necrosis factor (TNF) to cells (not
shown).
 Apoptosis and Health
APOPTOSIS: Role in Disease
The balance between cell proliferation and cell death is linked to
human health. Here are two examples.

TOO MUCH: Tissue atrophy

In neurodegenerative diseases, such as Alzheimer’s
disease, Thin skin apoptosis causes the progressive loss
of neurones.

TOO LITTLE: Hyperplasia

Atherosclerosis and Cancerous cells have mutations
that stop them reacting to chemical signals that
would normally start apoptosis. As a result, they
over-proliferate.
 APOPTOSIS: Role in Disease
Neurodegeneration
Neurons are post-mitotic (cannot replace themselves;
neuronal stem cell replacement is inefficient)

Neuronal death caused by loss of proper connections,

loss of proper growth factors (e.g. NGF), and/or
damage (especially oxidative damage)

Neuronal dysfunction or damage results in loss of synapses

or loss of cell bodies
(synaptosis, can be reversible; apopsosis, irreversible)

PARKINSON'S DISEASE
ALZHEIMER'S DISEASE
HUNTINGTON'S DISEASE etc.
 APOPTOSIS: Role in Disease
Cancer
Apoptosis eliminates damaged cells
(damage => mutations => cancer

Tumor suppressor p53 controls senescence

and apoptosis responses to damage

Most cancer cells are defective in apoptotic response

(damaged, mutant cells survive)

High levels of anti-apoptotic proteins

or
Low levels of pro-apoptotic proteins
===> CANCER
 APOPTOSIS: Role in Disease
AGING
OPTIMAL FUNCTION (HEALTH)
Aging --> both too much and too little
apoptosis
(evidence for both) APOPTOSIS

Too much (accumulated oxidative AGING

damage?)
---> tissue degeneration

Too little (defective sensors, signals?

APOPTOSIS
---> dysfunctional cells accumulate Neurodegeneration, cancer, …..
hyperplasia (precancerous lesions)