cancer immunotherapy from bench to clinic_mohamed labib salem ,ppt

Cancer Immunotherapy from bench to clinicMohamed Labib Salem, PhDMohamed Labib Salem, PhDProf. of Immunology, Faculty of ScienceDirector, Center of Excellence in Cancer ResearchTanta University, Egypt

[email protected]

The 1st Conference of Society of Pathological Biochemistry &Hematology, Faculty of Science, Menofia University, EgyptFeb 16, 2016

Tug of warTug of warProf. Mohamed Labib Salem

The smart interaction

Immune cells

Cancer Cells

Cancer cellsHost Tumor

microenvironmentProf. Mohamed Labib Salem

Prof. Mohamed Labib Salem

The main focus of my research interest

• Understanding the regulatory mechanisms behind escape of cancer and virus from immunity:– myeloid-derived suppressor cells (MDSC)– CD4 regulatory T cells– Cancer Stem cell

• Develop new anti-cancer immunotherapeutic strategies:– The use of cytokines and TLR agonists to enhance T

cell function– The use of TLR agonists to enhance dendritic cell

based vaccination


Talk outline

• Cancer Immunotherapy• Failure of Cancer therapy• Suppressive cells• Preclinical data• Clinical data


Markers of Non-Self

Non-self leukocyte

Antibody

Epitope Class I MHC protein

Epitope

Antibody

Antigen

Antigen

Bacteria

Non-self nerve cell

SARS virus


The Beginning of Cancerous Growth

Underlying tissue


Conventional anti-cancer therapy

Surgery

Immunotherapy

Radio-therapy

Chemo-therapy


Anti-tumor immune responses


Immune surveillance/editing of tumors


Cancer Therapy In clinic

• Surgery: Effective but doesn’t cure tumor

metastasis or prevent tumor recurrence

• Radiotherapy/Chemotherapy: Effective but doesn’t prevent tumor

recurrence besides its serous side effects


Cancer Therapy In clinic and clinical trials

• Immunotherapy: Can cure local and distant tumors

and prevent tumor recurrence through developing tumor-specific memory T cell responses


Effectors that can kill Cancer Antibody

Helper T cell

Natural killer cell

Macrophage

Cytotoxic T cell

Cancer cells


??

?

?

?

Typical Events Essential for Anti-tumor Immunity


Types of Cancer Immunotherapy

• Antibodies

Cytokines (IL-12, IFN-Cytokines (IL-12, IFN-a)a)

Adoptive T Cell Adoptive T Cell Therapy in Therapy in combination with combination with chemotherapychemotherapy

VaccinesVaccinesProf. Mohamed Labib Salem

Adoptive Cell Therapy“Adoptive= from donor to recipient”

1. Adaptive cell-based therapy–CD8+ (cytotoxic) T cell therapy– CD4+ (helper) T cell therapy– LAK cell therapy

2. Innate cell-based therapy–Dendritic cell therapy– NK cell therapy

3. Progenitor cell therapy– Bone marrow therapy– Stem cell therapy


s.c./i.d./i.v./i.t.injection

Dendritic Cell based anti-tumor vaccination


Nature Reviews Immunology 2; 227-238 (2002)



Antibody-based targeted immunotherapy

Antibody

Breast cancer cell

Growth factor

Herceptin blocks receptor

Growth slows

Radioisotope

Antigen

Lymphoma cell Lymphoma cell

destroyed

Herceptin


mAb-based therapies against liver cancer (40 clinical trials on www. ClinicalTrials.gov)• Bevacizumab/Avastin®: vascular endothelial growth factor

A (VEGF-A)• Cetuximab/Erbitux®:Epidermal growth factor receptor)• Cixutumumab (insulin-like growth factor 1 receptor [IGF-

1R])• MEDI-575: Platelet-derived growth factor receptor)• CT-011: Programmed Death-1 (PD-1)• CP 675/tremelimumab (CTL antigen-4)• RO5323441: Placenta growth factor (PGF)• HGS1012/mapatumumab: TRAIL-R1).• GC33 (glypican-3[GPC3]): is the only mAb targeting a HCC-

specific tumor antigen. Prof. Mohamed Labib Salem

T cell

T-Cell Receptor Gene Transfer

T cell

A tumor-reactive

T cellExogenous

TCR

Endogenous TCR

Rubinstein MP, Salem ML, et al., J Immunology 170:1209-17, 2003Rubinstein MP, Salem ML et al., Cancer Gene Ther. 2009 Feb;16(2):171-83

Gene therapy


Immuno-therapy with TCR-engineered T cells


Gene Transfer of Immunostimulatory molecules


Procedure of Adoptive T cell transfer therapy

Grow T cells With IL-2

Test T cell function

Chemo or IrradiationBefore T cell transfer

Infusion of T cells + vaccine

Clonal expansion of T cells with IL-2 & CD3

Tumor excision


29

Cancer Immunotherapy


Why does cancer Why does cancer therapy fail?therapy fail?


Causes of Failure of Cancer ImmunotherapyIntrinsic/extrinsic mechanisms


Causes of Failure of Cancer Immunotherapy1- Intrinsic mechanisms

• Tumor cells originate from self (i.e. not foreign)• Tumor-specific T cells are absent. • Tumor-reactive immune cells do not

localize to the tumor.• T cells fail to proliferate and persist in

response to tumors.


Clinically Un-detectable

LocalizedDisease

AdvancedDisease

1:10,000 T cells 1:50 T cells 1:2 T cells

Dise

ase

Burd

en

Causes of Failure of Cancer Immunotherapy High tumor cells/T cell ratio


0

2

4

6

8

1 3 5 7 10 14 21 30 60 120

CD8+

T c

ell r

espo

nse

Days Post Vaccination

Rapid expansion Rapidcontraction

Lowmemory

10

12

Tumor antigen alone

Tumor antigen + Danger signal

Causes of Failure of Cancer Immunotherapy Rapid contraction of T cells upon vaccination


• Tumor favor expansion of Regulatory cells that inhibit tumor-specific T-cell activities

• Downregulation of antigen expression in tumor cells.

• Secretion of immunosuppressive factors. • Death-receptors such as CD95 and TRAIL

receptor are mutated or lost entirely.

Causes of Failure of Cancer Immunotherapy Extrinsic mechanisms


Tumor-induced Immune Dysfunction

MDSC

IL-10

Arginase

ROS, RNS

IDO: depletes L-tryptophan, results in T cell unresponsiveness

Inhibitory/death R:s

Inhibitory/death receptors:FasL

TGF-β PGE2

ROS, RNS: reactive oxygen species results in T cell un-responsiveness

TGF-β and or IL-10 fromTreg, MDSC, and tumor

cells suppress T cells

Treg

IDO

MDSC/DC

Arginase: depletes L-arginine, results in T cell unresponsiveness


The current Dogma

Every type of myeloid and lymphoid derived cells can be found in a stimulatory or an

inhibitory (regulatory) status

All depends on the surrounding microenvironment


Regulatoryimmune cells

Immunity is controlled by the balance between stimulatory/regulatory cells

TregMDSCNKT cellsB cellspDCs

Stimulatory immune cellsDendritic cellsCD4+ T cellsCD8+ T cellsNK cells


Suppression of anti-tumor immunity by myeloid-derived suppressor cells (MDSC)


Mechanisms of the immune-suppressive Mechanisms of the immune-suppressive effects of MDSCeffects of MDSC

Prof

. Moh

amed

Lab

ib S

alem

Don’t work hard, work intelligent, and choose your direction

Our strategiesOur strategies1. Targeting cancer stem cells2. Blocking regulatory cell expansion

and functions3. Enhancing T cells survival and

function using IL-124. Enhancing dendritic cell expansion

and functions using TLRs.5. Combination of 2 + 3


ENHANCING T CELL SURVIVAL AND FUNCTION BY IL-12

Cancer Immunotherapy


Pmel

Spleenand LN

Ag IL-12 D3 Wash and reculture + IL-2

D7

Early Pmelsham Late Pmelsham

In-vitro culture system and gating strategy

Phenotypic and functional

analyses

Early Pmel12 Late Pmel12


Days after tumor implantation

Tum

or s

ize

(mm

2 )

Anti-tumor activity of CD8+ T cells programmed by different survival cytokines against solid B16 melanoma


Anti-tumor activity of CD8+ T cells programmed by different survival cytokines against metastasized B16 melanoma


1. Brief conditioning of anti-tumor CD8+ T cells in vitro with IL-12 enhance their survival phenotype and function.

2. IL-12-programed CD8+ T cells regress tumor in absence of vaccination.

3. Preconditioning the recipient hosts with cyclophopsphamide is a must.

4. IL-12 programs CD8+ T cells to acquire stem cell phenotype.

5. CD8+ T cells with stem cell phenotype resist cytotoxic effects of chmotherapy

Conclusion


Understand and optimize chemo-immunotherapy

Building on the success of IL-12 based immunotherapy


Expansion of DCs during restoration phase from CTX treatment

CD

11c

CD11b

5.6 ± 1.1 3.5 ± 0.9 6.4 ± 0.8 18.7 ± 4.125.2 ± 1.1

Lymphopenic phase Restoration phase

0200400

600800

1000

12001400

0 2 6 10 14Days post CTX treatment

# C

D11

c+ CD

11b+ c

ells

(106 /L

)

Day 0 Day 2 Day 6 Day 9 Day 12


Is dose-dependent. Induction of proliferation of dendritic

cell progenitors in bone marrow. Induction of production of myeloid cell

mobilizing factors (GM-CSF, G-CSF, M-CSF, Flt3L, and chemokines). Flt3 and CCR2 signaling pathways are

critical.

Mechanisms of the CTX Induced expansion of dendritic cells

Salem et al., 2010 J ImmunologySalem et al., 2010 Cell Immunology Prof. Mohamed Labib Salem

Phases post CTX treatment relative to adoptive T cell therapy (Salem CII 2010)

• Creation of a space niche due to the induced lymphopenia

• Homeostatic expansion of T cells• Elimination of regulatory cells• Elimination of cytokine

competition• Microbial translocation• Activation of dendritic cells

• Cellular recovery from lymphopenia• Less of lymphopenia• Less homeostatic proliferation of T cells• Expansion of immature dendritic cells

3 6 9 12 15 18

Days after CTX treatment

Num

ber o

f den

driti

c ce

lls

Lymphopenic phase Recovery phase

0-1

ACT Antigen priming+ TLR agonists

Antigen boosting+ TLR agonists

20

DCs

DonorT cells


Activation of post CTX expanded DCs is essential for augmenting CD8+ T cell responses

+-+---Poly I:C++++--gp100++--+-CTX++++--B16

+-+---++++--++--+-++++--

# D

Cs

in P

BL

(106 /L

)

# D

Cs

in D

LNs

(106 )

0

0.03

0.06

0.09

0.12

0.15

0.18

0

250

500

750

1000

1250

1500

0

0.05

0.1

0.15

0.2

0.25

0

500

1000

1500

2000

2500

# pm

el-1

in D

LNs

(106 )

0.02% 3.1% 0.6%

39.4%

0.04%0.05%

0.1% 0.2%4.5% 2.1%0.1%

12.5%#

pmel

-1 in

PB

L (1

06 /L)

6.0%14.9%

7.6%

18.2%

14.6%

6.5%

1.9% 1.7%3.6%

4.6%

9.1%

2.2%


Vaccination at the peak of post CTX DC expansion improves survival of tumor-bearing host

0 10 20 30 40 50 60 70 80 900

25

50

75

100

Days post tumor challenge

Perc

ent s

urvi

val

PBS

PmelSham/PBS

PmelIL12/PBS

PmelSham/CTX

PmelIL12/CTX


Vaccination at the peak of post CTX DC expansion induces effective anti-tumor immunity

050

100150200250300350400450

12 14 17 20 22 24 26 28 32

Days post tumor inoculation

Tum

or a

rea

(mm

2 )

PBS (No T cells)CTX (No T cells)

CTX/Vac + IL-2

CTX/Vac+ poly(I:C)

PBS/Vac + poly(I:C)


T cell responses after CTX/vaccine/poly(I:C) prevent tumor from growth


Cancer immunotherapyCombination, Combination, Combination …..

Nature 446, 964-966 (26 April 2007)Prof. Mohamed Labib Salem

Funded agentsFunded agents• National Cancer Institute (NCI), National

Institute of Health (NIH), USA• Hollings Cancer Center, Medical

University of South Carolina, USA• Chain Reaction for Brest Cancer, USA• Tanta University Research Fund• Science and Technology Development

Fund (STDF)


ACKNOLWEDGEMENTSTanta University, EgyptAmir A Alkhami, PhD Randa Al-Naggar, PhD Wael Attia, PhDSabry EL-Naggar, PhD Sherif Zidane, PhD Said Hamaad, MDMohamed Elshanshoory, MD Mohamed Attia, MD Mona Zidan, MScShaima Sobhy, MSs, Asmaa Shaaban, MSc Sohaila Galal, MSc

Suez Canal University, EgyptAhmed Khafagy, PhD

Medical University of South CarolinaDavid J. Cole, MD William Gillanders, MD Mike Nishimura, PhDAndre Kadima, MD Yian Chen, PhD Osama Naga, DDElizabeth Little, BSc Rick Peppler, MS Narender Nath, PhDGuillermo Rivell, MD Sabry EL-Naggar, PhD Amir A Alkhami, MS

University of Miami, Miller School of MedicineMarcela Montero, PhD Alberto Montero, MD

University of California in San Diego, USAMark Rubstein, PhD


THANK YOU

Prof. Mohamed Labib Salem, PhDProf. Mohamed Labib Salem, PhDTanta University, EgyptTanta University, Egypt