Cell Death Flashcards

1
Q

What is the function of cytochrome P450 ?

A
  • Many compounds not excretable, need to be conjugated into something excretable
  • Oxidising them with P450 then conjugating them with something else to make them more soluble and hence excretable
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2
Q

Distinguish apoptosis from necrosis.

A

APOPTOSIS

  • May be pathological or physiological
  • Single cells
  • Biochem events: Energy-dependant fragmentation of DNA
  • Cell membrane integrity maintained
  • Morphology: cell shrinkage and fragmentation
  • No inflammatory response
  • Dead cells phagocytosed by neighbouring cells

NECROSIS

  • Always pathological
  • Groups of cells
  • Biochem events: Abnormal ion homeostasis
  • Cell membrane integrity lost
  • Morphology: Cell swelling and lysis
  • Inflammatory response present
  • Dead cells phagocytosed by inflammatory cells
  • Causes include ischaemia, trauma, metabolic
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3
Q

Identify the main kinds of necrosis, where each occurs, and the appearance of each.

A

Coagulative necrosis: In most tissues. Firm pale area with ghost outlines

Colliquative necrosis: In brain. Dead area liquefied.

Caseous necrosis: In tuberculosis. Pale yellow semi-solid material.

Gangrenous necrosis: necrosis with putrefaction (following vascular occlusion or certain infections). Black

Fibrinoid necrosis: microscopic feature in arterioles in malignant hypertension

Fat necrosis: following trauma (mass) or pancreatitis (multiples white spots)

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4
Q

Is programmed cell death the same as apoptosis?

A

No. Apoptosis is morphological whilst PCD is about intent.

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5
Q

Give examples of PCD.

A
  • Embryology – lumen of tubes
  • Response to growth signals – menstrual cycle
  • Inflammation – resolution, death of neutrophils
  • Immune defence – T and Natural Killer cell responses
  • Tumour prevention – prevent mutation
  • Autoimmune disease – self destruct
  • HIV AIDS – HIV and activated T cell death
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6
Q

Describe the main molecules involved in DNA fragmentation in apoptosis.

A

Caspases fragment DNA
Caspases fragment Poly (ADP ribose) polymerase (PARP) which is a repair enzyme
Caspases also fragment Inhibitor of Caspase activated DNAase=ICAD (results in the activation of CAD which “breaks up DNA during apoptosis”)

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7
Q

How does PARP relate to cancer treatment ?

A

In cancer, anticancer drugs/radiation try to damage DNA. If damage DNA, can chose to die by apoptosis but doesn’t die often because of repair enzymes

PARP inhibitors ineffective on their own but given with agents to damage DNA, show real potential, prevent repair (replicate what the cell is doing when it decides to do apoptosis)

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8
Q

How does recognition of cells undergoing apoptosis by macrophages and non-professional phagocytes take place ?

A

After fragmentation, membrane flips inside which means on the outside (outer leaflet), expose Phosphatidylserine which can be recognised by macrophages (recognise membrane bound fragments with abnormalities)

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9
Q

What are the two pathways to apoptosis ? Describe the main features of each.

A

EXTRINSIC

  • Trigger comes from outside. Typically through “death” receptors, which transduce messages inside cell
  • T cells may trigger this

INTRINSIC
-Oxidative stress (may be metabolic, poisoning, energy failure, DNA damage and p53)

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10
Q

What are possible triggers of the extrinsic pathway of apoptosis ?

A

Cytokines or ligand binding (eg ligand on cytotoxic T cell)

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11
Q

Explain the role of cytokines in triggering the extrinsic pathway of apoptosis.

A

TNF family (produced by macrophages in tumours) is ligand found on cytotoxic T cells.
Binds to CDC95 (=Fas) receptor
Receptor then trimerises
Death domain gets exposed which results in cascade of procaspases which amplify one another (activated by fragmentation), eventually leading to caspapases (which then cleave nuclear proteins, cytoplasm etc.)

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12
Q

Explain the role of T cells in triggering the extrinsic pathway of apoptosis.

A

Cytotoxic T cell contain a couple of things:

  1. Granzyme (kind of like a caspase), which activates caspase. Directly introduce granzyme into target cell, it activates caspases. Fragments the cells.
  2. Perforin, which perforates cell membrane

Combo of those two results in fast track to destruction through cytoplasmic activation

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13
Q

Identify examples of pathological mechanisms in which T cell mediated extrinsic apoptosis is involved.

A

Viral infection

Transplantation rejection

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14
Q

Describe the main features of the intrinsic pathway of apoptosis.

A
  • Triggered by cellular stress, specifically mitochondrial stress caused by factors such as DNA damage and heat shock.
  • Upon receiving the stress signal, the proapoptotic proteins in the cytoplasm, BAX and BID, bind to the outer membrane of the mitochondria to signal the release of the internal content. BAK, another proapoptotic protein that resides within the mitochondria, is also needed to fully promote the release of cytochrome c and the intramembrane content from the mitochondria
  • Following the release, cytochrome c forms a complex in the cytoplasm with ATP and Apaf-1, an enzyme.
  • Following its formation, the complex will activate caspase-9, an initiator protein. In return, the activated caspase-9 works together with the complex of cytochrome c, ATP and Apaf-1 to form an apoptosome, which in turn activates caspase-3, the effector protein that initiates degradation.
  • Besides the release of cytochrome c from the intramembrane space, the intramembrane content released also contains apoptosis inducing factor (AIF) to facilitate DNA fragmentation
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15
Q

What are the components of an apoptosome ?

A

A heptameric apoptotic protease activating factor 1 (Apaf-1)-cytochrome-c complex.

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16
Q

Identify the function of the apoptosome.

A

The apoptosome recruits and activates caspases that cleave intracellular substrates and ultimately lead to cell death by apoptosis.

17
Q

What is the physiological response to radiation damage to DNA (possibly due to oxygen free radicals) ? Which part of this response may be affected in cancer ?

A
p53 activated (sensory of damage in DNA) and forms tetromer.
Makes 3 classes of genes: 
1. stop cell cycle, 2. increase expression and function of DNA repair enzymes and 3. increase expression of genes leading to apoptosis

P53 is affected in cancer

18
Q

Identify examples of pro-apoptotic factors and anti-apoptotic factors.

A

Pro-apoptotic factors: BAX

Anti-apoptotic factors. Bcl2

19
Q

Explain importance of balance between pro an anti apoptotic factors in influencing apoptosis.

A

Members of the Bcl2 family dimerise. These dimers allow for control of sensitivity of cells to death (e.g. DNA damage to B cell will often lead to aptopotsis whereas in neurons will not usaully lead to apotpsosis).

BCL2 dimers: stops cytochrome C leaking out of mitochondria (prevents apoptosis)

BAX dimers: strong death signal (lets cytochrome C leak out, hence forming apoptosome and resulting in apoptosis)
E.g. some genes activated by p53 will be pro-death (through extra activation of Bax)

20
Q

What is the normal function of Bcl2 ? Hence, what is a potential effect of abnormal Bcl2 expression ?

A

Bcl2 keeps cell alive so if overexpressed, a cell that should die does not die and becomes cancerous

21
Q

How is abnormal expression of Bcl2 generally due to ?

A

This may occur in translocation of promoter of one gene on coding bit of bcl2, resulting in overexpression of bl2, and leading to dimerisation.

22
Q

Does Bcl 2 result in increased proliferation ?

A

No, just stops cells dying

23
Q

Which pathologies may abnormal Bcl2 expression be found in ? Is there any treatment available for this ?

A

Typically in follicular lyphoma, B cell malignancy

Treatment: Anti BCL2 therapy (binds to Bcl2 molecules)

24
Q

What are the different ways in which cell death is regulated ?

A
  • Caspases
  • Bcl2 and Bax expression
  • p53
  • Inhibitor of apoptosis=IAP (mop up caspases): need to be able to bring caspases in apoptosome to generate high enough concentration of caspases to overcome inhibitors
  • Anti-IAPs: block IAPs
25
Q

Where did the name caspase come from ?

A

C because cysteine in active site
asp because cleaves after aspartate
ase because it is a protease

26
Q

Identify some of the molecules cleaved by caspases, and the result of each cleavage.

A

Cleave

  • ICAD: destroy genetic information
  • PARP: prevents DNA repair
  • lamin: breaks down nuclear architecture
  • keratin: breaks down cytoplasmic architecture
27
Q

Describe the process of activation of caspases.

A

“Dimerization and often oligomerisation of pro-caspases, followed by cleavage into a small subunit and large subunit.
The large and small subunit associate with each other to form an active heterodimer caspase”

28
Q

What are cancers due to, with regards to intrinsic and extrinsic pathways of apoptosis ?

A

Cancers due to failure or activation of both intrinsic and extrinsic pathways, because intrinsic and extrinsic pathways communicate (If start chain of events from outside, inside will also contribute).

29
Q

Explain the significance of survival factors in apoptosis, providing an example.

A
  • Survival factors (e.g. growth factors) are extrinsic
  • If gets signals from inside telling it to die (eg BCL2) but also survival factor from outside telling it to grow (growth factor), will decide not to die
  • E.g. Hepatitis B/C first cause of liver cancer in the world: kills liver cells but also leads to chronic inflammation which brings in growth factors which, compete against factors requesting for apoptosis to happen, and thereby lead to proliferation (resulting in cancer)
30
Q

How does nutrition relate to apoptosis ?

A

With higher caloric intake, increased number of growth factors (so less likely to get rid of cells when you need to)

31
Q

Give examples of apoptosis gone wrong.

A
  • Autoimmune disease (apoptosis when do not require it)
  • Cancer (no apoptosis when require it)
  • Neurodegeneration (apoptosis when do not require it e.g. dementia)
32
Q

What components of the apoptosis pathway may be targeted in the following pathologies ?

  • Lymphoma
  • Cancer
  • Alzheimer’s
A

Lymphoma: Bcl2 (decrease it)
Cancer: IAP (decrease it)
Alzheimer’s: Caspase 3 (decrease it)

33
Q

What is Pyroptosis ?

A

A modification of necrosis and apotposis (features of both), exploited by one particular organism through innate (non-specific) receptors.

34
Q

How is pyroptosis triggered ?

A

Microbial trigger eg Salmonella

35
Q

What are examples of features of apoptosis and necrosis in pyroptosis ?

A

– Caspase 1 activation, not caspase 3
– Nuclear fragmentation but not cytoplasmic blebbing
– Pro-inflammatory

36
Q

What types of receptors are involved in pyroptosis ?

A

Innate (non-specific) Pattern recognition receptors

– NOD like and Toll like receptors

37
Q

What is Anoikis ? Is it most similar to necrosis or apoptosis ?

A

Death after losing contact with basement membrane/extra cellular matrix.
It has apoptosis morphology.

38
Q

What occurs when anoikis fails ?

A

Metastasis