please tell me your idea about my slides.

1. Viral gene therapy of
“Multiple sclerosis”

3. Definition, Symptoms, Cuases:
 An autoimmune Inflammatory T cell dependent disease
that affect on myelin and distroy them in CNS.
 Weakness in limbs, loss of centeral vision, double vision,
tingling or pain in some parts of body, Electric-shock
sensations that occur with certain head movements,
Fatigue, Dizziness.
 ( all the symptoms are show up because of breakdown the
communication.)
 The cause of multiple sclerosis is unknown, some
reserchear’s blivied in a combination of factors, ranging
from genetics to childhood infections, may play a role.

4. Immune system:
T helper
• Th1 cytokines are pivotal in the induction of human
organ-specific autoimmune disease (ie Hashimoto's
thyroiditis, Graves' disease, rheumatoid arthritis,
insulin-dependent diabetes mellitus and MS) .
• Specialy Encephalitogenic T cell.

5. Important Risk Factors:
 Geoghraphic :
In both hemispheres, its prevalence increases with distance from the
equator.
 Ethnic:
Different populations and ethnic groups have a markedly different
prevalence of MS. The disease is especially common in Scotland,
Scandinavia, and throughout northern Europe.
 Genetics:
First, second and third degree relatives of people with MS are at
increased risk of developing the disease
MS. Some scientists theorize that MS develops because a person is born
with a genetic predisposition to react to some environmental agent.
 Viruses:
EBV, varicella zoster, Hepatitis vaccine

7. Perspectives in Gene Therapy
for MS:
Gene therapy for multiple sclerosis might include
different ‘human-grade’ vectors, which could be
used to deliver anti-inflammatory molecules as well as
neuroprotective agents into the CNS in a flexible and
useful way.

8. Therapies in Multiple Sclerosis:
anti-inflammatory therapies:
Deliver the ‘therapeutic’ molecules directly into the
CNS in order to inhibit blood-borne CNS-confined
Mononuclear cells acting as ultimate effector cells.
neuroprotective therapies:
Rescue surviving oligodendrocytes.
Induce oligodendrocyte progenitor migration and
differentiation into demyelinating areas.

9. Anti-inflammatory Gene
Therapies:
 Pro-inflammatory cytokines in MS:
Inf-Gama / (TNF-α/β)/ IL-2
downregulation of inflammatory mediators
Systemic delivery:
peripheral circulation encephalitogenic T cells.
CNS delivery:
directly into the CNS via the intraparenchymal or
intrathecal route.

10. Systemic Delivery:
Experimental Autoimmune
Encephalitis ( EAE) EAE
 IL-4 / IL-10 / TGF-β 1
 Autoreactive encephalitogenic
T cells
 Retriviral vector
 Improved symptoms.
 Note: opposite; TNF-α gene
Animal type:
mice and non-human primates
(rhesus monkeys).
 IL-6/ TNF- α/ IL-1b / IL-2 or IL10
 Autoreactive encephalitogenic
T cells
 vaccinia-virus-derived vectors
( Adenovirus)
 Improved symptoms.
 Note: opposite; IL-4
Animal type:
mice and non-human primates
(rhesus monkeys).

11. CNS Delivery:
1) EAE
2) Co-stimulatory molecule CTLA-4-human
immunoglobulin / IL-4
3) non-replicative adenoviral vector
 infect : ependymal layer and leptomeningeal cells
 produce a discrete amount of the transgene CSF for up
to 1 month after injection.
 improved EAE.

12. CNS Delivery continue:
 EAE
 IL-10/ IL-4
 adenoviral vectors
1) Delayed EAE onset
2) Decrease in clinical score
3) Decrease in perivascular inflammatory infiltrates and in the
number of macrophages infiltrating the CNS parenchyma and
the submeningeal spaces
4) Reduction in demyelinated areas and axonal loss.
 No toxic reactions/ without any interference with the proper
functioning of the immune system.
improved EAE.
 Animal type:
Mice

14. neuroprotective therapies:
1) transplantation of oligodendrocytes or o. precursors
 Transplantation experiments :
• Defined demyelinating areas of experimental animals.
• transplantation is not a useful tool in multifocal CNS
demyelinating diseases at present. ( MS)
2) Neurotrophic growth factors
• Migration/proliferation /differentiation
• Partially successful in EAE but not in MS patients.
3) promoting myelin restoration

15. Neurotrophic growth factors:
1) EAE
2) fibroblast growth factor (FGF)-II
3) HSV-1-derived vectors
 ameliorated EAE /without toxic reaction /inducing
oligodendrocyte precursor proliferation and migration
into demyelinating areas.
 FGF-II was only seen when administered for up to 4
weeks, After that induced reactive astroglyosis.

16. Using FGF-II gene :
CNS production of FGF-II (ependymal, choroidal and
leptomeningeal cells)
Decrease myelinotoxic cells (T cells and macrophages)
both in the CNS parenchyma and in the
leptomeningeal space.
Increas of the number of oligodendrocyte precursors
and myelin-forming oligodendrocytes in areas of
demyelination and axonal loss.

19. HSV-1 (d120) :
 Some of the limitations of the gene vector technology
so far employed in EAE/MS has been overcome by the
use of HSV-1-derived vectors :
• to be easily transferred within the CNS trough the CSF
Circulation.
• No major reactions of resident cells
• Intracellular machinery was used to produce the
heterologous gene (eg cytokine genes).

21. Conclusions:
Benefit:
• Cytokine/growth factor gene therapy based on
injection of HSV-1-derived vectors into the CSF space
might be a favourable approach for a chronic and
multifocal CNS disease such as MS.
1. high cytokine/growth factor levels in all CNS.
2. therapeutic effect persists after a single vector
administration.

22.  Our results indicate that intrathecal delivery of HSV-1-
derived vectors containing anti-inflammatory cytokine
genes may play a major role in the future therapeutic
armamentarium of inflammatory CNS-confined
demyelinating diseases

23. Problems:
 that rodent immunogene therapy cannot be transferred
easily to humans.
 Encephalitogenic autoreactive T cells can be isolated from
animal but in human the autoantigen(s) is still unknown.
 vaccinia virus adenoviral vectors are immunogenic and
they stimulate the immune sys and this feature makes it
hard to use them.
 HSV-1-derived vectors are short lasting and we should re-
inject them.

24. QUESTION?

25. Thanks for your attention

26. References:
1.Kieseier BC, Storch MK,
Archelos JJ, Martino G, Hartung
HP. Effector pathways in
immune mediated central
nervous system demyelination.
2. Calabresi PA, Fields NS,
Maloni HW, Hanham A,
Carlino J, Moore J et al. Phase
1 trial of transforming growth
factor beta 2 in chronic
progressive MS.
3. The Lenercept Multiple Sclerosis
Study Group and University of
British Columbia MS/MRI
Analysis Group. TNF
neutralization in MS: results of
a randomized, placebocontrolled
multicenter study.
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Adorini L. The ependymal route
to the CNS: an emerging gene
therapy approach for MS.
5. Selmaj KW, Raine CS. Tumor
necrosis factor mediates myelinand
oligodendrocyte damage in
vitro.
6. Shaw MK, Lorens JB, Dhawan
A, DalCanto R, Tse HY, Tran AB
et al. Local delivery of
interleukin 4 by retrovirustransduced
T lymphocytes
ameliorates experimental
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27. 7.Chen LZ, Hochwald GM,
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myelin basic protein-specific
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9. Croxford JL, O’Neill JK, Ali RR,
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MJ et al. Local gene therapy
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fusion protein in experimental
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simplex virus replication defective
multigene vectors for combination gene
therapy applications.
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Schaffer PA. Isolation and
characterization of deletion
mutants of herpes simplex virus
type 1 in the gene encoding
immediate-early regulatory
protein ICP4.

28. 13.Kuklin NA, Daheshia M,
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Modulation of mucosal and
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15. Furlan R, Poliani PL, Marconi
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16. Poliani PL, Brok H, Furlan R,
Ruffini F, Bergami A, Desina G
et al. Delivery of a nonreplicative
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1 vector engineered with the
IL-4 gene to the central nervous
system protects rhesus monkeys
from hyperacute autoimmune
encephalomyelitis.

29. 17. Ruffini F, Furlan R, Poliani PL,
Brambilla E, Marconi PC,
Bergami A et al. Fibroblast
growth factor-II gene therapy
reverts the clinical course and
the pathological signs of
chronic experimental
autoimmune encephalomyelitis
in C57BL/6 mice.
18. Mathisen PM, Yu M, Yin L,
Johnson JM, Kawczak JA,
Nishiyama A et al. Th2 T cells
expressing transgene PDGF-A
serve as vectors for gene
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