CAR T cell therapy is a type of cancer immunotherapy treatment that uses immune cells called T cells that are genetically altered in a lab to enable them in locating in destroying cancer cells more effectively.

1. Programming CAR T- Cells to kill Cancer

2. CONTENT
S
Introduction
1
History
2
Basic overview of
CAR T- cell
therapy
3
Basic cellular and
Humoral
immunity
4
Purpose for CAR T
Cell
development
5
Chimeric Antigen
Receptor Design
6
Engineering of
CAR T- Cell
Receptor
7
CAR gene
Transduction in
T-Cell
8
Upgrade
Generation
Of CARS
9
General
processes
Of CAR T cell
10
Benefits of
CAR T-cell
Therapy
11
Disadvantages
Of CAR T-Cell
Therapy
12
References
13
Prospective and
Future Challenges
14
Conclusion
15

3. INTRODUCTION
 CHIMERIC ANTIGEN RECEPTOR (CAR) ARE GENETICALLY ENCODED ARTIFICIALLY
FUSION MOLECULE THAT CAN REPROGRAMMED THE SPECIFICITY OF PERIPHERAL
BLOOD POLYCLONAL T-CELLS AGAINST A SELECTED CELL SURFACE TARGET.
 CARS ARE GENETICALLY ENGINEERED RECEPTORS THAT COMBINE THE SPECIFIC
BINDING DOMAINS FROM A TUMOUR TARGETING ANTIBODY WITH T-CELL SIGNALLING
DOMAIN TO ALLOW SPECIFICALLY TARGETED ANTIBODY REDIRECT T-CELL ACTIVATION.

4. HISTORY
 In the year 2010 the first successful cancer treatment with CAR-T
was for an advanced follicular lymphoma patient and was reported
by the lab Steven Rosenberg M.D, PhD chief of surgery branch in
NCI centre for Cancer research.
 On August 30, 2017 tisagenlecleucel (Kymriah) was the first CAR-
T cell immunotherapy approved by the FDA.
 It was approved for children and young adults aged 25 and under
who relapsed or were not responding to therapy for acute
lymphoblastic leukaemia (ALL).
Steven
Rosenberg
tisagenlecleucel
(Kymriah

5. BASIC OVERVIEW OF CAR T CELL THERAPY
 CAR-T cell therapy is form of immunotherapy that uses specially alter T-cell (a part of
immune system) to fight cancer.
 A sample of a patient’s T-cell are collected from the blood then modified to produce special
structure called Chimeric Antigen Receptor (CARs) on their surface.

6. BASIC CELLULAR AND HUMORAL IMMUNITY

7. PURPOSE FOR CAR-T
CELL DEVELOPMENT
 Escape tumour surveillances
mechanisms:
 Low immunogenicity.
 Antigen modulation.
 immuno- suppression by tumour cells
T regulatory cells.
 Induction of lymphocytes apoptosis.
 Defects in mechanism of MHC-I
production
can render cancer cells invisible to CD-
8 cells.

8. CHIMERIC
ANTIGEN
RECEPTOR
DESIGN
 The overall structure of a CAR consists
of four domains joined in series, namely
:
i. An antigen recognition domain
(targeting moiety)
ii. A hinge/spacer
iii. Transmembrane element
iv. A signalling end domain

9. 1. THE TARGETING MOIETY-:
 The CAR ectodomain determines target specificity and most commonly
contain elements derived from a monoclonal antibody.
 Co-express both antibody variable heavy (VH) and light chains (VL) in two
separate polypeptide chains, thereby creating a single chain variable
fragment (scFv).
2. THE HINGE/SPACER AND TRANSMEMBRANE DOMAIN-
:
 Play a predominantly structural role in the CAR.
 Some reports have suggested that different hinge regions might critically
control surface expression levels, construct stability and antigen binding
affinity , which directly influence the efficiency of CAR redirected effector
functions .

10. 3. THE CAR SIGNALING DOMAIN
 First generation CAR T cells contain a
single T-cell activating domain , most
commonly derive from the zeta chain of
the TCR/ CD3 complex .and CD3z
alone provides a sufficiently potent
“signal 1” from its three immunoreceptor
tyrosine based activation motifs (ITAMs)
to substitute for the global signal
provide by the entire CD3 complex.
 the second and third generation CARs
have been developed one or two co-
stimulatory domains , respectively are
placed in series with CD3z . Typically
CD28 is included as a co-stimulatory
domains as this provides an early
second signal and promotes highly level
IL-2 secretion , Resistance to apoptosis
.

11. ENGINEERING OF CAR-T CELL RECEPTOR -:
CAR = specificity of antibody target recognition
+
effector mechanisms of T cell

12. CAR GENE TRANSDUCTION IN T-CELLS-:
1. Construct a CAR
gene
2.Inserted CAR gene into a
vector
3.Transduce and expand patient T-cell
ex-vivo

13. UPGRADE GENERATION Of CARS

14. GENERAL PROCESSES OF TCR/CAR T-CELL

15. BENEFITS OF CAR T-CELL THERAPY
 HLA –independent antigen
recognition
 Active in both CD4+ and
CD8+ T-cells
 Target antigen include
proteins, carbohydrates
and glycolipids
 Rapid generation of tumor
specific T cells
 Minimal risk of
autoimmunity or GVHD
(Graft-versus-host disease)

16. DISADVANTAGES
0F CAR-T-CELL-:
 Cytokine release syndrome
CRS (symptoms like high fever,
nausea, capillary leaky
syndrome )
 Anti IL-6 overcome the CRS
toxicity
1. Neurotoxicity (i.e. confusion ,
seizures or severe headaches)
2. Cost USD $475,000 Novartis
which is equal to
Rs.30,400,000

17. REFERENCES
• Programming CAR-T cells to kill cancer
Louai labanieh, Robbie G.Majzner and Crystal L.Mackall
• Bird, R. E. et al. Single-chain antigen-binding proteins. Science
242, 423-426 (1988)
• Sadelain, M.Chimeric antigen receptors: a paradigm shift in
immunotherapy. Ann. Rev. Cancer Biol., 447-466, (2017)
• Liu, E. et al. Use of CAR-transduced natural killer cells in
CD19- positive lymphoid tumours. N. Engl. J. Med. 382, (2020)

18. PROSPECTIVES
 There are several challenges include CAR T-cell development,
manufacturing and clinical trials like protocols pre-conditioning patient, CAR
T-cell formulation, quality and persistence.
 Novel therapy started to incorporate small molecules, which proved to
augment T-cell function and simultaneously inhibit the malignant population,
in the next few years, we are likely to witness increase efficacy of CAR T-
cells for solid tumors- a major current focus in this field
FUTURE CHALLENGES
 Immunosuppressive microenvironment
 Improving efficiency and persistence

19. CONCLUSION
 Cd19 CAR T-cells have demonstrated success in refractory B-
cell ALL and NHL.
 CRS and ICANS are common side effects that can occur.
 Some side effects persist after CAR T-cells for a bit longer, but
eventually recover.
 Active research ongoing in CAR T-cells for other cancer cells.

20. THANK YOU