Immune Modulation, Monitoring and Metabolism
Week 1
HC 2 30-10
Intro and overview on monitoring and modulation
Immunotherapy
Type of treatment in which the immune system is modulated (enhanced or suppressed)
with the ultimate goal of curing/alleviating a disease
Can be used against cancer, but also in auto-immunity
Microenvironment: analyse all cell types and cytokines to identify key biomarkers
Cell-cell interactions: track how immune cells interact with cancer cells, focusing on
checkpoint proteins
Immunophenotype: profile immune cells types and states using minimal sample sizes
Signalling & function: understand how treatments alter signalling pathways and
immune responses
Why studying single cells?
It is important to study single cells due to the heterogeneity of biological systems. In
these systems minority cell populations play important roles in disease progression,
regeneration and immune defence. These cells can be cancer stem cells, tissue-
specific stem cells and diverse immune cells. Bulk analysis can miss these minority
cells, so single-cell technologies are essential to: identify tumour driving cells
• Identify tumour driving cells
• Understand tissue repair mechanisms
, • Decode immune responses
• The immune system is a classic example for the need of single cell technologies,
because its dynamic cellular heterogeneity is essential for fighting off the variety
of attacking pathogens.
Understanding tissue heterogeneity
By only using bulk analysis, you would miss a lot of information, you need single-cell or
spatial profiling to truly understand tumour biology
Immunotherapy modalities
The pillars of cancer care
1. Surgery – earliest form of treatment
2. Radiotherapy
3. Cytotoxic chemotherapy
4. Targeted therapy
5. Immunotherapy
Cancer care has progressed from broad, invasive methods
to highly targeted and immune-based approaches
Current approaches to cancer therapy and the specific hallmarks of cancer
Don’t learn this, just to give insight into
therapies
Passive vs active immunotherapy
Based on the ability of the immunotherapy to activate or reactivate the host immune
response against malignant cells.
• Passive:
o Delivers immune components directly
, o Works independently of the patient’s immune activation
▪ Tumour-targeting monoclonal antibodies (mAbs)
▪ Adoptively transferred T cells
• Active:
o Stimulates the patient’s own immune system to attack cancer
o Requires immune engagement for effectiveness
o educates the immune system to fight against the target (tumour or
pathogen)
▪ Anti-cancer vaccines
▪ Checkpoint inhibitors
Can also classify the immunotherapies as:
• Antigen-specific
o Precisely targets known cancer markers
▪ Vaccine
▪ CAR T cell
• Broad (non-antigen specific)
o Stimulate the immune system without targeting a specific antigen
▪ Immunostimulatory cytokines
▪ Checkpoint blockers
General classification
• Cell based therapies
• Molecular therapies
• Biological agents
• Microenvironment modifiers
Don’t think you have to know all of this, just
to show that immunotherapy is not a single
strategy, but have different approaches to
engage the immune system in different
ways to fight cancer
Tumour targeting antibodies
• Refers to:
o Signal disruption: mAbs that specifically alter or block the signalling
functions of receptors expressed on the surface of malignant cells. E.g.,
Anti-EGFR → stops tumour growth by blocking signals
, o Neutralizing signals: mAbs that bind to, and hence neutralize, trophic
signals produced by malignant cells or by stromal components of
neoplastic lesions. E.g., anti-VEGF
o Targeting tumour-associated antigens: mAbs that selectively recognize
cancer cells base in the expression if “tumour associated antigen” (TAA),
i.e., an antigen specifically (or at least predominantly) expressed by
transformed cells but not (or at least less so) by their non malignant
counterparts. E.g. Antig-CD20 in CLL
Anti-EGFR, mechanism of action
Inhibits pathways that are for cell proliferation, by inhibiting this the tumour cells will go
into apoptosis. EGFR normally binds ligands triggering downstream pathways like RAS,
RAF, MEK or ERK. These pathways promote cell proliferation, survival and angiogenesis.
By having mAbs that bind EGFR the ligand interaction will be inhibited which will block
these downstream pathways.
Anti-VEGF, mechanism of action
Block angiogenesis. VEGF family proteins bind to receptors on endothelial cell
triggering angiogenesis. By blocking VEGF you block angiogenesis.
Functional variants of mAbs
• Signal blockers: Naked mAbs that inhibit signaling pathways required for the
survival or progression of malignant cells, but not of their non-malignant
counterparts:
o Cetuximab (EGFR-specific mAb)
• Death receptor activators: Naked mAbs that activate potentially lethal
receptors expressed on the surface of malignant cells, but not of their non-
transformed counterparts:
o Tigatuzumab (TRAILR2-specific mAb)
• Immune conjugates: i.e., TAA-specific mAbs coupled to toxins or radionuclides:
o Gemtuzumab ozogamicin, an anti-CD33 calicheamicin conjugate
o The toxins or radionuclides enhance tumour killing
• Effector activators: Naked TAA-specific mAbs that opsonize/tag cancer cells
and activate antibody- dependent cell-mediated cytotoxicity (ADCC), antibody-
dependent cellular phagocytosis (ADCP), and complement:
o Rituximab (CD20-specific mAb)
o Rituximab is against B cell malignancies
• Bispecific T-cell engagers (BiTEs): i.e., chimeric proteins consisting of two
single-chain variable fragments from distinct mAbs, one targeting a TAA and one
specific for a T-cell surface antigen:
o Blinatumomab (CD19- and CD3 BiTE)
Week 1
HC 2 30-10
Intro and overview on monitoring and modulation
Immunotherapy
Type of treatment in which the immune system is modulated (enhanced or suppressed)
with the ultimate goal of curing/alleviating a disease
Can be used against cancer, but also in auto-immunity
Microenvironment: analyse all cell types and cytokines to identify key biomarkers
Cell-cell interactions: track how immune cells interact with cancer cells, focusing on
checkpoint proteins
Immunophenotype: profile immune cells types and states using minimal sample sizes
Signalling & function: understand how treatments alter signalling pathways and
immune responses
Why studying single cells?
It is important to study single cells due to the heterogeneity of biological systems. In
these systems minority cell populations play important roles in disease progression,
regeneration and immune defence. These cells can be cancer stem cells, tissue-
specific stem cells and diverse immune cells. Bulk analysis can miss these minority
cells, so single-cell technologies are essential to: identify tumour driving cells
• Identify tumour driving cells
• Understand tissue repair mechanisms
, • Decode immune responses
• The immune system is a classic example for the need of single cell technologies,
because its dynamic cellular heterogeneity is essential for fighting off the variety
of attacking pathogens.
Understanding tissue heterogeneity
By only using bulk analysis, you would miss a lot of information, you need single-cell or
spatial profiling to truly understand tumour biology
Immunotherapy modalities
The pillars of cancer care
1. Surgery – earliest form of treatment
2. Radiotherapy
3. Cytotoxic chemotherapy
4. Targeted therapy
5. Immunotherapy
Cancer care has progressed from broad, invasive methods
to highly targeted and immune-based approaches
Current approaches to cancer therapy and the specific hallmarks of cancer
Don’t learn this, just to give insight into
therapies
Passive vs active immunotherapy
Based on the ability of the immunotherapy to activate or reactivate the host immune
response against malignant cells.
• Passive:
o Delivers immune components directly
, o Works independently of the patient’s immune activation
▪ Tumour-targeting monoclonal antibodies (mAbs)
▪ Adoptively transferred T cells
• Active:
o Stimulates the patient’s own immune system to attack cancer
o Requires immune engagement for effectiveness
o educates the immune system to fight against the target (tumour or
pathogen)
▪ Anti-cancer vaccines
▪ Checkpoint inhibitors
Can also classify the immunotherapies as:
• Antigen-specific
o Precisely targets known cancer markers
▪ Vaccine
▪ CAR T cell
• Broad (non-antigen specific)
o Stimulate the immune system without targeting a specific antigen
▪ Immunostimulatory cytokines
▪ Checkpoint blockers
General classification
• Cell based therapies
• Molecular therapies
• Biological agents
• Microenvironment modifiers
Don’t think you have to know all of this, just
to show that immunotherapy is not a single
strategy, but have different approaches to
engage the immune system in different
ways to fight cancer
Tumour targeting antibodies
• Refers to:
o Signal disruption: mAbs that specifically alter or block the signalling
functions of receptors expressed on the surface of malignant cells. E.g.,
Anti-EGFR → stops tumour growth by blocking signals
, o Neutralizing signals: mAbs that bind to, and hence neutralize, trophic
signals produced by malignant cells or by stromal components of
neoplastic lesions. E.g., anti-VEGF
o Targeting tumour-associated antigens: mAbs that selectively recognize
cancer cells base in the expression if “tumour associated antigen” (TAA),
i.e., an antigen specifically (or at least predominantly) expressed by
transformed cells but not (or at least less so) by their non malignant
counterparts. E.g. Antig-CD20 in CLL
Anti-EGFR, mechanism of action
Inhibits pathways that are for cell proliferation, by inhibiting this the tumour cells will go
into apoptosis. EGFR normally binds ligands triggering downstream pathways like RAS,
RAF, MEK or ERK. These pathways promote cell proliferation, survival and angiogenesis.
By having mAbs that bind EGFR the ligand interaction will be inhibited which will block
these downstream pathways.
Anti-VEGF, mechanism of action
Block angiogenesis. VEGF family proteins bind to receptors on endothelial cell
triggering angiogenesis. By blocking VEGF you block angiogenesis.
Functional variants of mAbs
• Signal blockers: Naked mAbs that inhibit signaling pathways required for the
survival or progression of malignant cells, but not of their non-malignant
counterparts:
o Cetuximab (EGFR-specific mAb)
• Death receptor activators: Naked mAbs that activate potentially lethal
receptors expressed on the surface of malignant cells, but not of their non-
transformed counterparts:
o Tigatuzumab (TRAILR2-specific mAb)
• Immune conjugates: i.e., TAA-specific mAbs coupled to toxins or radionuclides:
o Gemtuzumab ozogamicin, an anti-CD33 calicheamicin conjugate
o The toxins or radionuclides enhance tumour killing
• Effector activators: Naked TAA-specific mAbs that opsonize/tag cancer cells
and activate antibody- dependent cell-mediated cytotoxicity (ADCC), antibody-
dependent cellular phagocytosis (ADCP), and complement:
o Rituximab (CD20-specific mAb)
o Rituximab is against B cell malignancies
• Bispecific T-cell engagers (BiTEs): i.e., chimeric proteins consisting of two
single-chain variable fragments from distinct mAbs, one targeting a TAA and one
specific for a T-cell surface antigen:
o Blinatumomab (CD19- and CD3 BiTE)
