Harnessing

THE
cell IMPORTANCE
death mechanisms OF CELL
toDEATH
improve IN
therapy
CANCER
Gianmaria Liccardi
 OUTLINE:

1. Why is Programmed cell death important.

2. What is Programmed cell death and how it
works.
3. Programmed cell death and cancer
 OUTLINE:

1. Why is Programmed cell death important.

2. What is Programmed cell death

3. and how it works.

4. Programmed cell death and cancer

 Cell Death Survival

Inflammation Inflammation

Tissue Homeostasis
Tissue Repair

LOSS OF THIS BALANCE INDUCES

PATHOPHYSIOLOGICAL DISEASE LIKE CANCER
 Cell death and development

• Between 50 and 70 billion cells die each day due to apoptosis in

the average human adult. For an average child between the ages
of 8 and 14, approximately 20 billion to 30 billion cells die a day.

• Without apoptosis, human gut can grow up to 12 miles in length

• Whole epithelial lining in our body changes every 23 days

• Tissue homeostasis is achieved by the correct balance between

cell division (mitosis) and cell death.
 Survival Cell Death

Inflammation Inflammation

Cancer Hallmark Chronic Inflammatory Diseases

-Inflammatory Bowel Diseases

-Psoriasis

-Rheumatoid Arthritis
 Cancer is a multifactorial disease
• Cancer cells proliferate uncontrollably
• Cancer cells loose their ability to die
• Cancer therapy aims to:
– inhibition of cancer cell proliferation and or
cancer cells death
Cancer is a mutltifactorial disease
• Cancer cells proliferate uncontrollably
– Extensive proliferation induces hyperactive
inflammation causing tissue malfunction and
adaptation
Cancer is a mutltifactorial disease
• Cancer cells loose their ability to die
– Loss of tumour suppressor genes: such as p53
– Hyper- activation of membrane receptors: such
as EGFR
– Hyperactivation of prosurvival signaling
pathways such PI3K/AKT
– Loss of cell death machinery components
– Over expression of anti apoptotic genes
Cancer is a mutltifactorial disease
• Cancer therapy aims to:
– inhibition of cancer cell proliferation and/or
cancer cells death
 Anti-cancer strategies
Chemotherapies
Precision
Mitosis drugs
Mitosis
poisons
Immuno-related
intervention Oncogene

Surface receptors Radiotherapy

Treatment Efficiency
green = apoptosis
red = necroptosis
Activation of
Cell death Programme Cell Death
 ACTIVATION CELL
DRIVER OF SURVIVAL DEATH
PATHWAYS

TARGETING MOLECULAR THERAPY

LOSS OF CELL DEATH
COMPONENTS
Sharma et al. Nature Reviews Cancer 7, 169–181 (March 2007) | doi:10.1038/nrc2088
 OUTLINE:

1. Why is Programmed cell death important.

2. What is Programmed cell death

3. and how it works.

4. Programmed cell death and cancer

 How many different ways to die?

1. Apoptosis
2. Necroptosis
3. Pyroptosis
4. Ferroptosis
5. Autophagy
6. Parthanatos
 When one cell death platform is not
functional/disabled
We can activate another one to re-sensitise
cells to therapeutic treatment
 Programmed Cell death
Lytic forms of cell death

Apoptosis Necroptosis Pyroptosis Ferroptosis

Caspase1
Caspases MLKL Caspase4-5 (11 in mouse)
Lipid peroxides
GasderminD
 Programmed Cell death
1. Necrosis: Accidental cell death from
acute injury
• NECROPTOSIS: programmed
necrosis :cellular burst via
membrane pore formation
2. Apoptosis: Programmed cell death –
active cell process characterized by:
• DNA fragmentation
• Chromatin condensation
• Nuclear and cell fragmentation
• Apoptotic cells and cell fragments are
recognized and phagocytized by
macrophages and neighboring cells, and
rapidly removed from tissues
• Caspase dependent

• Necroptotic cells swell ad lyse; the

contents are released into the extracellular
space and cause inflammation
• Caspase independent
APOPTOSIS
INTRINSIC APOPTOSIS:
- mitochondrial membrane permeabilization

EXTRINSIC APOPTOSIS:
- DEATH LIGAND MEDIATED (FasL, Trail) and TNF
 VIDEO

Please refer to part 1 of lecture 2

recording to watch the apoptosis movie. It
is located at about 20:37 into the
recording.
INTRINSIC APOPTOSIS
INTRINSIC APOPTOSIS- key players

• In mammalian cells caspase-9 pro

apoptotic caspase is activated by
binding to Apaf-1 in a protein complex
called Apoptosome

• Cytochrome c is also required, which is

release from mitochondria
INTRINSIC APOPTOSIS- key players
• Bcl-2 is an inhibitor of apoptosis firstly
identified as an oncogene

• Mammalian cells encode about 20

proteins related to Bcl-2 in three
functional groups

• Some inhibit apoptosis while others

induce caspase activation
WHY IS THIS IMPORTANT IN CANCER?
 Cell Death priming status
Death stimuli (BH3 profiling technique)
Pro-apoptotic members
BH3 proteins Tony Letai
(Harvard, Boston)
BH3 Primed
mimetics

Unprimed
Bcl-2
Anti-apoptotic Unprimed
Mitochondrial membrane members Primed
permeability

Bcl2

Apoptogenic
factors
ANTI -APOPTOTIC PROTEINS LIKE BCL2, BCLXL AND
MCL1 ARE HIGHLY EXPRESSED IN CANCER AND
THE SURVIVAL OF MANY CANCER CELLS OF
DIFFERENT ORIGIN DEPENDS ON THEIR ABILITY TO
SEQUESTERS HIGH LEVELS OF THEIR PRO-DEATH
COUNTERPART
WHAT HAPPENS IF INTRINSIC APOPTOSIS
IS DISABLED?
Programmed Cell death

Necroptosis Apoptosis

Immunologically active
 Inflammatory diseases

Apoptosis
-Panayotova-Dimitrova D., et al, 2013, Cell
Reports
-Weinlich R., et al, 2013, Cell Reports
-Rickard J A., et al, 2014, eLIFE
-Gerlach B., et al, 2011, Nature
Necroptosis
-Dannapel M., et al, 2014, Nature
-Bonnet C M., 2011, Immunity
-Duprez L., 2011, Immunity
-Welz, P S., 2011, Nature

RIPK1
RIPoptosome/complexII

Chronic Inflammatory Diseases

-Inflammatory Bowel Diseases

-Psoriasis
-Rheumatoid Arthritis
-Cancer
 SM

TAK1i
z-VAD

IKKi

CHX
Annibaldi & Meier, Trends in Mol Med, 2018
The Ripoptosome as a ‘stress-sentinel’

Genotoxic stress
Microtubule poison
Inflammatory
signals
cIAP2
cIAP1 XIAP brake
brake

- Chronic intestinal inflammation

- Skin inflammation
- Susceptibility to viral infection
- Mitosis Unrelated to inflammatory signals
- Cancer
Casp8 and RIPK1 levels predict patient
survival
 OUTLINE:

1. Why is Programmed cell death important.

2. What is Programmed cell death and how it
works.
3. Programmed cell death and cancer
RESISTANCE TO- extrinsic Apoptosis
• Casp8 mutation or loss
• Iaps overexpression
• Trail receptor deficiency
Programmed Cell death

Necroptosis Apoptosis

Immunologically active
RIPOPTOSOME
Please refer to the recording, part
2 of lecture 2 to view the
ripoptosome movie. It is located
about 17:42 into the recording.
Casp8 and RIPK1 levels predict patient
survival
Immunogenic cell death in cancer treatment

Cytotoxic T cells

Dendritic cells

-Tumour antigens

-Immunogenic factors

Durable anti-tumour response

CONCLUSION:

1. Balance between cell death and survival

ensures tissue homeostasis
2. Hyper-inflammation induces many
pathophysiological diseases such as
Cancer
3. Apoptosis: caspase dependent cell death
– intrinsic and extrinsic
4. Necroptosis: Caspase independent
modality of cell death
CONCLUSION 2:
1. Cancer cells addiction to: oncogenes
leads to inhibition of cell death:
• P53 mutation/ loss
• Pi3K mutation
• EGFR mutation
• BCL2/MCL1 overexpression
• Casp8/loss or mutation
2. Reactivation of cell death modalities as
well as reactivation of immunogenic cell
death can lead to durable anti-tumour
response