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1 Introduction
Schwann cells 2 What are Schwann cells?
3 Why use Schwann cells for spinal cord repair?
Transplantation for 4 Spinal cord injury models
Spinal Cord Repair 5 Schwann cells promote axonal regeneration
6 Schwann cells promote remyelination
7 Schwann cells promote functional recovery
Akkradate Siriphorn 8 Schwann cells vs. other cell types
4938035 SIMP/D 9 Combination Strategies
March 25th, 2008
10 Conclusion

CNS axon growth

During development, CNS neurons extend wildly.
Growth cone

Introduction
Growth
cone

Target
Once the critical period for the formation of specific
1 2 3 4 5 6 7 8 9 10 connection ends, CNS plasticity in higher vertebrate greatly
diminished.
3 4

CNS injury = Permanent deficit Spinal cord injury (SCI)

Christopher Reeve: Horse accident (1995)

The patient with SCI generally experience little functional recovery

5 and suffer from permanent functional deficit

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CNS Vs. PNS axonal damage CNS Vs. PNS response to axotomy

CSPG = Chondroitin sulfate proteoglycans

PNS CNS Axotomy RAG = Regeneration-associated gene

Central nervous system Peripheral nervous system

Myelin
Myelin--derived inhibitor CSPG Macrophage clearance
Bunger’s band
Reactive astrocyte Schwann cell
RAG induction
Glial scar proliferation
In contrast to the CNS, Peripheral nervous system (PNS) axons
maintain their plasticity beyond the developmental phase and Failed axonal Successful axonal
remain capable of axonal regeneration after injury. regeneration regeneration
8

CNS graft PNS bridge

Anderson (1954), Love (1956)
David and Aguayo, 1981

Spinal cord graft Peripheral nerve

Injured
Injured Spinal cord
Spinal cord
PNS bridge
CNS bridge
CNS axons failed to regenerate into the graft The CNS environment suppresses axonal regeneration, but in
permissive PNS environment, many CNS axons can regenerate.
9

PNS bridge without Schwann cells Schwann cells as a hope

Smith and Stevenson, 1988
In animal models,
Grafting SCs promote
Peripheral nerve
• Axonal regeneration
without Zhang (2007), Dinh (2007), Rasouli (2006),
Schwann cells Pearse (2004), Timmer (2003)
• Remyelination
Papastefanaki (2007), Girard (2005), Pearse (2004)

Injured • Functional recovery

Pearse (2007), Li (2007), Papastefanaki (2007), Pearse
Spinal cord (2004), Timmer (2004)

PNS bridge SNI

CNS axons failed to regenerate into the graft
SCs is an important
Schwann cells play an important role in candidate for future spinal
promoting CNS axonal regeneration cord repair
11 12

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What are Schwann cells?

What are Schwann cells?

1 2 3 4 5 6 7 8 9 10
13 14

Schwann Vs. Oligodendrocyte Origin of Schwann cells

15 16

Migration and Myelination of SCs Two types of Schwann cells

Myelinating SCs

Ensheathing SCs

17 18

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Schwann cell promote axon growth

Axon Intracellular
CNS & PNS
Why Schwann cells become signaling
Axonal growth
an important candidate for Integrin or regeneration
TrK
spinal cord repair? ECM Produced
CAMs
by SCs
Neurotrophic
factors

Schwann cell
1 2 3 4 5 6 7 8 9 10
19 Adapted from Dezawa et al. (2002) 20

Schwann cells can expand in vitro Autologous transplantation

Peripheral Nervous System

in vitro

Autologous transplantation
• minimize immunological rejection
• obvious ethical issues
21 22

Spinal cord injury models

Transection Contusion Demyelination

Spinal cord injury models

• Surgical knife • Micro-balloon • X-radiation and

• Microscissors • Aneurysm clip injection of
• Computer controlled ethidium bromide
contusion devices

1 2 3 4 5 6 7 8 9 10
23 24

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Computer controlled impactor

NYU impactor
SCs transplantation
promote
axonal regeneration

1 2 3 4 5 6 7 8 9 10
25 26

SCs transplantation Transection injuries

promote Transection
axonal regeneration
1. Axonal regeneration in transection injury

2. Axonal regeneration in contusion injury Schwann cells

Semi--permeable polymeric channels

Semi
3. Molecular mechanism of SCs directed Chau et al. (2004), Hsu et al. (2005)

regeneration Polyacrylonitrile/polyvinylchloride
Polyacrylonitrile/polyvinylchloride (PAN/PVC)
Xu et al. (1995), Xu et al. 1999)

Poly beta hydroxybutyrate (PHB)

1 2 3 4 5 6 7 8 9 10 Novikova et al. (2008)
27 28

SCs in transection injuries Axon regeneration through SCs

seeded channel
Type of regenerated
Author Method Type of lesion Donor
axon Xu et al. (1995)
Schwann cells + exogenous Transection Adult rat - propriospinal
PAN/PVC + SCs
Xu et al. (1995) BDNF and NT-3 (T9-T11) - serotonergic
- brainstem axons
Human Schwann cells + Transection Human PNS - corticospinal
Guest et al.
guidance channel + (T9-T10) - propriospinal
(1997)
methylprednisolone - sensory
Fouad et al. Schwann cells + OEG + Transection Adult rat - serotonergic
(2005) chondroitinase (T8)
Schwann cells bridge + OEG Transection Adult rat - corticospinal
Ramon-Xueto et
(T9) - serotonergic
al. (1998)
- propriospinal
Hurtado et al. D15A-SCs + guidance Transection Adult rat - Axon regeneration
(2006) channel (T9-T10) (not classified)

SCs + guidance channel Transection Adult rat - propriospinal

Xu et al. (1997) (T8) - serotonergic PAN/PVC + SCs 22 day
- noradrenergic
29 30

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Axon regeneration through SCs Human SCs transplantation promote

seeded channel CNS axon regeneration
Xu et al. (1995) cont. Adult rat SCs Guest et al. (1997) Human SCs
Neurofilament

PHA-L
31 32

D-15A SCs seeded with guidance SCs + biodegradable channel

scaffold promote axonal regeneration
Novikova et al. (2007)
Hurtado et al. (2006) GFP + D15A Schwann cells
NF

5-HT 5-HT
6 week GFP-SCs

CGRP CGRP

D-15A = bifunctional NT-3/ BDNF-like molecule 33 34

SCs transplantation Contusion injuries

promote
axonal regeneration
1. Axonal regeneration in transection injury

2. Axonal regeneration in contusion injury

3. Molecular mechanism of SCs directed
regeneration

1 2 3 4 5 6 7 8 9 10
35 Onose et al. (2007) 36

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Blood supply to the spinal cord Pathological feature of contused

spinal cord injuries

37 38

SCs in contusion injuries SCs transplantation into subarachnoid

space promote axonal regeneration
Author Method Type of lesion Donor Type of regenerated axon
Martin et al. Schwann cells Contusion Adult rat DRG - propriospinal Firouzi et al. (2006)
(1996) (lower thoracic) - sensory
Takami et al. Schwann cells alone or Contusion Adult rat - corticospinal
(2002) Schwann cells + OEG (T9) - propriospinal
transplantation - serotonergic
- ascending sensory
Pearse et al. Schwann cells + Contusion Adult rat - serotonergic Silver staining
(2004) elevation of cAMP (T8)
Schaal et al. Schwann cells Contusion Adult rat - reticulospinal Control SCs transplantation
(2007) (C5)
Golden et al. D15A-transduced SCs Contusion Adult rat - Supraspinal
(2007) (T8) - propriospinal
Firouzi et al. SCs transplantation into Compression Neonatal rat - axonal regeneration
(2007) subarachnoid space (T10) (not classified)

39 40

D15A-transduced SCs promote axonal

growth
SCs transplantation
promote
Golden et al. (2007)
axonal regeneration
1. Axonal regeneration in transection injury

2. Axonal regeneration in contusion injury

3. Molecular mechanism of SCs directed
regeneration
D-15A = Bifunctional NT-3/
BDNF-like molecule
1 2 3 4 5 6 7 8 9 10
41 42

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SCs and axonal regeneration SCs and axonal regeneration

• Laminin • Laminin
ECM • Collagen ECM • Collagen
Chernousov (2000), Dubovy (2004), Wen (2006) Chernousov (2000), Dubovy (2004), Wen (2006)

• β1-integrin • β1-integrin
Cell • NCAM Cell • NCAM
adhesion adhesion
molecule • N-cadherin molecule • N-cadherin
Schwann cells Addock (2004), Previtali (2003) Schwann cells Addock (2004), Previtali (2003)

• NGF • NGF
Trophic • BDNF Trophic • BDNF
factor • NT-3 factor • NT-3
• CNTF • CNTF
• GDNF • GDNF
Yamamoto (1993), Mirsky (1999), Verderio (2006) Yamamoto (1993), Mirsky (1999), Verderio (2006)
43 44

Myelin inhibiting molecules Nogo

Hanila (2008), Filbin (2003), Gao (2003)

Neuron
MAG
Nogo
OMgp Nogo
Nogo

NgR Nogo

Nogo ER
Nogo
Nogo

Nogo Nogo
Signal transducer
(P75 or other protein)
Neuron Nogo
ER
Nogo

Rho
Nogo

Nogo
ROCK
Axon Growth
Inhibition

MLC
MLCK Oligodendrocyte Oertle (2003)
45

Neurotrophin inhibit inhibition SCs and axonal regeneration

Hanila (2008), Filbin (2003), Gao (2003)
• Laminin
ECM • Collagen
MAG Chernousov (2000), Dubovy (2004), Wen (2006)
Nogo
OMgp
Neurotrophin
NgR
• β1-integrin
Trk Cell • NCAM
adhesion
molecule • N-cadherin
Schwann cells Addock (2004), Previtali (2003)
Signal transducer
(P75 or other protein)
Neuron • NGF
PDE4 Rho Trophic • BDNF
cAMP ERK factor • NT-3
Axon Growth ROCK
Inhibition • CNTF
PKA • GDNF
CREB MLCK Yamamoto (1993), Mirsky (1999), Verderio (2006)
MLC
Transcription 47 48

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Cell adhesion molecules Cell adhesion molecules

Bixby et al. (1988) SCs monolayer + neuron

Control Anti-Integrin

anti-Integrin
anti-Integrin + anti-L1
+ anti-L1 + anti-Cadherin 49 Kokolva et al. (2000) 50

SCs and axonal regeneration Extracellular matrix

• Laminin
• Collagen Intracellular
ECM Chernousov (2000), Dubovy (2004), Wen (2006) signaling
CNS and PNS
Integrin axon
• β1-integrin
Cell • NCAM
adhesion ECM Produced
molecule • N-cadherin by SCs
Schwann cells Addock (2004), Previtali (2003)

• NGF
Trophic • BDNF
factor • NT-3
Schwann cell
• CNTF
• GDNF
Yamamoto (1993), Mirsky (1999), Verderio (2006)
51 Adapted from Dezawa et al. (2002) 52

1 Introduction
2 What are Schwann cells?
3 Why use Schwann cells for spinal cord repair?
SCs transplantation
4 Spinal cord injury models promote
5 Schwann cells promote axonal regeneration remyelination
6 Schwann cells promote remyelination
7 Schwann cells promote functional recovery
8 Schwann cells Vs. other cell types
9 Combination Strategies 1 2 3 4 5 6 7 8 9 10
10 Conclusion 54

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SCs transplantation Remyelination in transection model

promote Transection
remyelination
1. Remyelination in transection model

2. Remyelination in contusion model

3. Remyelination in demyelination model

1 2 3 4 5 6 7 8 9 10
55 56

SCs promote remyelination SCs transplantation

Bamber et al. (2001) promote
remyelination
1. Remyelination in transection model

2. Remyelination in contusion model

3. Remyelination in demyelination model

1 2 3 4 5 6 7 8 9 10
57 58

Remyelination in contusion model SCs promote remyelination

Contusion Golden et al. (2007)
Control SCs

59 60

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SCs transplantation Demyelination models

promote
remyelination Normal Demyelination
1. Remyelination in transection model

2. Remyelination in contusion model

3. Remyelination in demyelination model

1 2 3 4 5 6 7 8 9 10
61 62

SCs promote remyelination SCs promote remyelination

Lankford et al. (2002) cont.

Nucleic acid-
chelating
agent

Lankford et al. (2002) 63 64

SCs promote remyelination SCs promote remyelination

Honmou et al. (1996) X-EB Honmou et al. (1996) cont.
Normal Demyelination
Intact

X-EB
SCs β-gal

SCs

65 66

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Human SCs transplantation repair

conduction properties
Kohama et al. (2001)
SCs transplantation
promote
functional recovery

1 2 3 4 5 6 7 8 9 10
67 68

SCs transplantation Open-field locomotor test

promote
functional recovery
1. Open-field locomotor test
2. Rotarod test
3. Von Frey hair test
4. Possible mechanism of functional recovery
BBB: Basso-Beattie-Bresnahan Locomotor score
1 2 3 4 5 6 7 8 9 10 BMS: Basso mouse score
69 70

SCs graft into subarachnoid space cAMP + SCs

Firouzi et al. (2006) Pearse et al. (2004)

SCs + cAMP * *
* *
SCs transplant * *

Control Injury control

Medium

71 72

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PSA-NCAM SCs graft SCs transplantation

Papastefanaki et al. (2007) promote
functional recovery
* 1. Open-field locomotor test
*
2. Rotarod test
PSA-NCAM-SCs
Vector-SCs 3. Von Frey hair test
4. Possible mechanism of functional recovery
control
1 2 3 4 5 6 7 8 9 10
73 74

Rotarod test BDNF-,NT-3 transduced primate SCs

Girard et al. (2005)

75 76

SCs transplantation Von Frey hair test

promote
functional recovery
1. Open-field locomotor test
2. Rotarod test
3. Von Frey hair test
4. Possible mechanism of functional recovery

1 2 3 4 5 6 7 8 9 10
77 78

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SCs + OEG + ChaseABC SCs transplantation

Found et al. (2005)
promote
functional recovery
1. Open-field locomotor test
2. Rotarod test
3. Von Frey hair test
4. Possible mechanism of functional recovery

1 2 3 4 5 6 7 8 9 10
79 80

Mechanism of functional recovery Mechanism of functional recovery

Bradbury and McMahon (2006) 81 Bradbury and McMahon (2006) 82

Mechanism of functional recovery

Comparison between
Schwann cells and
other cell types

1 2 3 4 5 6 7 8 9 10
Bradbury and McMahon (2006) 83 84

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Cell types that can graft in SCI Olfactory ensheathing glial (OEG)
SCs OEGs ESC NSC Macrophage Fibroblast

To fill cyst
cavity     
To replace
neuron  
To
remyelinate    
To create
regenerating
environment
     
SCs = Schwann cell, OEGs = olfactory ensheathing glial,
ESC = Embryonic stem cell, NSC = neural stem cells
Zhang (2007), Papastefanaki (2007), Li (2007), Pearse (2007), Schaal (2007), Golden (2007), Guo (2007), Andrews (2007),
Kimura (2005), Pallini (2005)Found (2005), Feng (2005), Akiyama (2004), Pearse (2004), Takemi (2002), Menei (1998),
Guest (1997), Martin (1996) 85 Thuret, Moon and Gage (2006) 86

OEG promote spinal cord repair OEG Vs. Schwann cells

CST regeneration Takami et al.
Keyvan-Fouladi et al. (2003) (2002)

Fast-blue
7mm distal
87 88

OEG Vs. Schwann cells OEG Vs. Schwann cells

Takami et al. Control
(2002) cont. Takami et al.
(2002) Cont.
SC * * * *
SC/OEG
SCs

OEG medium
OEG

89 90

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Purified OEG do not form myelin LacZ-OEG do not associate with

in vitro myelinated axon in vivo
Schwann/DRGN OEG/DRGN Boyd et al. (2004)

Plant et al. (2002) 91 92

Endogenous Schwann cells invade SCs Vs. OEG

and remyelinate demyelinated axon
Olfactory
Property Schwann cells
Black et al. (2006) ensheathing cells
Produce neurotrophic factor
1 year after X-EB and cell adhesion molecules
Support axonal growth

Proliferate and migrate after

axon injury
Integration with astrocyte

In vitro myelination

In vivo myelination ????

Functional recovery
P0 = Schwann cell myelin
MOG = Oligodendrocyte myelin Wewwetzer (2002), Williams (2004), Lakatos (2000), Lakatos (2003), Boyd 2004), Woodhall (2003), Cui (2003)
93 Sims (1994), Lu (20002) 94

Scenario of spinal cord injury

Combination strategies

1 2 3 4 5 6 7 8 9 10
95 Klussmann and Martin-Villalba (2005) 96

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Elevation of cAMP + SCs SCs + ChaseABC

Pearse et al. (2004)
Control SCs SCs + cAMP

Rolipram + db-cAMP 97 Chau et al. (2004) 98

SCs + OEG + ChaseABC SCs BDNF, NT-3 promote axonal outgrowth

Bamber et al. (2001)

Found et al. (2005) 99 100

NT3-transduced SCs + neural stem cell

Guo et al. (2007) CMEP CSEP

Normal control
Conclusion
SCI control
NSCs

SCs + NSCs

NT
NT--3-SCs+ NSCs
1 2 3 4 5 6 7 8 9 10
60 days 101 102

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Conclusion

• SCs = the most promising cell types for repair injured

spinal cord

• Ability to obtain highly purified and large population Thank You

= opportunity for clinical application
For Your Attention!
• In animal models, SCs promote
• axonal regeneration
• remyelination
• functional recovery

• Combinational strategies are required

103

105 Hall and Springer (2004) 106

Pathophysiology of SCI Primary cause of dysfunction is the

axonal interruption

Normal

Dysfunction: from interruption of

local neuron
Major cause of dysfunction

Dysfunction: from axonal

interruption in upper level

Normal
107

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Prior exposure to neurotrophins block SCs – Axon reciprocal relationship

inhibition of MAG on neurite growth
Proliferation
Differentiation
Axon
Direct contact Diffuse molecule

Cell signaling Diffuse molecule

Direct contact Schwann cell

Nakai et al. (2006), Dezawa et al. (2000)
Cai et al. (1999) 109 110

SCs transplantation and elevation of

cAMP promote axon regeneration

Pearse et al. (2004)

SCs alone SCs+ elevation of cAMP

p75/5-HT p75/5-HT
111

19