Stem cells for the treatment of

g
disease
neurodegenerative
Jan A. Nolta, Ph.D.

Professor, Director of the Stem Cell Program and Institute for Regenerative Cures
University of California, Davis

Neurons generated from human

pluripotent stem cells in culture
Jeanie Liu, Nolta Lab

UC Davis Stem Cell Program Disease TeamsOver 145 Basic, Translational, and Clinical Faculty Investigators working together

Liver repair
Li
i and
d regeneration,
i
bioengineered
bi
i
d livers
li
Peripheral artery disease: revascularization to prevent amputation
Eye degeneration/blindness
Lung disease,
disease lung repair and regeneration
Skin: Non-healing ulcers, burn repair
Bone repair, osteoporosis, cartilage regeneration
Heart disease, infarction repair and stroke
Neurodegenerative (Parkinsons, Huntingtons, Alzheimers, ALS)
Neurodevelopmental disorders (autism spectrum, FXTAS, fragile x)
Kidney repair and regeneration
Bl dd repair,
Bladder
i bi
bioengineered
i
d bl
bladders
dd
Blood disorders, autoimmune disorders (Scleroderma, MS)
HIV treatment using gene-modified stem cells
Hearing, inner ear cilia repair
Tumor stem Cells, Cell-based immunotherapy for Cancer

Neuroscience research at UC Davis: programs and centers devoted to

advancing our understanding of the brain:

The Center for Neuroscience

The MIND Institute
The Center for Mind and Brain
The Alzheimers Disease Center
Center for Visual Sciences
The Imaging Research Center

 Adult stem cell therapies began in 1956

Bone
B
marrow ttransplantation
l t ti
Hundreds of thousands of
lives have been saved
through BMT
Our team has processed cells for
over 800 stem cell transplants
and
d has
h performed
f
d 18 stem
t
cell
ll
therapy clinical trials for tissue repair
and treating genetic disease

Types of stem cells

Adult Stem Cells

Pluripotent Cells

Blood forming (hematopoietic)

Embryonic

Mesenchymal (supporting cells)

Induced pluripotent stem cells

Why use adult stem cells?

Adult stem cells cannot create an entire tissue,
as can embryonic cells.
However, under the right conditions, adult
stem cells can act as the paramedics
paramedics of the
body, rushing to areas of hypoxia and rapidly
g revascularization, earlyy after tissue
causing
injury.

Mesenchymal Stem Cells (MSC):

Reparative adult stem cells that can act as excellent
d li
delivery
vehicles
hi l iin th
the b
body
d

BoneMarrowisharvested
From a normal, healthy donor
Fromanormal,healthydonor

Cellsexpandedinaclean
room facility
roomfacility

M
MesenchymalStemCells(MSC)
h
l St
C ll (MSC)

Expansion of MSC from bony spicules/trabeculae

Techniques detailed in:
Genetic Engineering of Mesenchymal Stem Cells. Kluwer
Nolta JA (Editor). Feb 2006

MSCs can be engineered

to secrete copious amounts
of protein factors for
delivery to other cells and
tissues in the body

MSC delivering a protein product (red) throughout the tissue

(Meyerrose et al, Nolta lab, Published in Stem Cells 2008).
A no MSC transplantation
p
B - MSC alone, making lower innate levels of the enzyme beta-glucuronidase
C MSC engineered to deliver the protein. The entire tissue is corrected.

Nolta lab:
Papers since 1989
using Gene- modified
Marrow Stromal
Cells (MSC) to
systemically produce
cytokines, enzymes,
and other factors
i vivo
in
i

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2.
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L. S. Sender, J. A. Nolta, J. A. Barranger, D. B. Kohn, Pediatr Res 25:145 (1989).

J. A. Nolta, L. S. Sender, J. A. Barranger, D. B. Kohn, Blood 75, 787 (1990).
J. Weinthal et al., Bone Marrow Transplant 8, 403 (1991).
J. A. Nolta, G. Crooks, R. Overell, D. Williams, D. B. Kohn, Exp Hem. 20, 1065 (1992).
M E
M.
E. Hanley
Hanley, J
J. A
A. Nolta
Nolta, R
R. Parkman
Parkman, D
D. B
B. Kohn
Kohn, Blood Cells 20,
20 539 (1994)
(1994).
J. A. Nolta, M. B. Hanley, D. B. Kohn, Blood 83, 3041 (1994).
J. A. Nolta, E. M. Smogorzewska, D. B. Kohn, Blood 86, 101 (1995).
J. A. Nolta, M. A. Dao, S. Wells, D. B. Kohn, Proc Natl Acad Sci U S A 93, 2414 (1996).
M. A. Dao, J. A. Nolta, Cytokines Cell Mol Ther 3, 81 (1997).
M. A. Dao, K. A. Pepper, J. A. Nolta, Stem Cells 15, 443 (1997).
M. A. Dao, C. H. Hannum, D. B. Kohn, J. A. Nolta, Blood 89, 446 (1997).
M. A. Dao, X. J. Yu, J. A. Nolta, Exp Hematol 25, 1357 (1997).
J.A. Nolta et al, in Current Protocols in Human Genetics, vol. 13, pp. unit 13.7. (1997)
M. A. Dao, K. Hashino, I. Kato, J. A. Nolta, Blood 92, 4612 (1998).
M. A. Dao, A. J. Shah, G. M. Crooks, J. A. Nolta, Blood 91, 1243 (1998).
D. C. Ertl, K Pepper, DB Kohn, and JA Nolta, Blood 92, 718 (1998).
M. A. Dao, J. A. Nolta, Int J Mol Med 1, 257 (1998).
M A
M.
A. Dao
Dao, J
J. A
A. Nolta
Nolta, Blood 92,
92 125a (1998)
(1998).
M. A. Dao, N. Taylor, J. A. Nolta, Proc Natl Acad Sci U S A 95, 13006 (1998).
M. Dao, J. Nolta, Leukemia 13, 1473 (1999).
J. Arakawa-Hoyt et al., Bone Marrow Transplant 24, 1167 (1999).
M.A. Dao, J. A. Nolta, Current Opinion in Immunology (1999).
E. Tsark, M. Dao, X. Wang, K. Weinberg, J. Nolta, J Immunol 166, 170 (2001).
J. A. Nolta et al.,, Leukemia 16,, 352 (2002).
(
)
M. A. Dao, J. Hwa, J. A. Nolta, Blood 99, 499 (2002).
X. Wang et al., Blood 101 (10) 4201 (2003).
G. D. Wu et al., Transplantation 75, 679 (2003).
T. E. Meyerrose, P. Herrbrich, D. A. Hess, J. A. Nolta, Biotechniques 35, 1262 (2003).
M. A. Dao, J. Arevalo, J. A. Nolta, Blood 101, 112 (2003).
Y. Sohara et al., Cancer Res 65, 1129 (2005).
T Meyerrose
T.
M
and
d J.
J N
Nolta,
lt G
Genetic
ti E
Engineering
i
i off MSC
MSC. Editor
Edit J.
J Nolta,
N lt Chap
Ch 2,
2 (2006).
(2006)
M. A. Dao, J. A. Nolta, Blood 109, 2373 (2007).
T. Meyerrose et al., Stem Cells 25, 220 (2007).
T. E. Meyerrose et al., Stem Cells 26, 1713 (2008).
G. Bauer et al., Mol Ther 16, 1308 (2008).
P. Zhou et al., Methods Mol Biol 430, 213 (2008).
J.Rich, I. Rosova, J. Nolta, L. Sandel, A. McAlinden. BBRC 372, 230 (2008).
I. Rosov, Link DC, Hess DA, Nolta JA. Stem Cells. 26:2173 (2008).
C. Sondergaard C, D Maxwell, C Weinheimer, A Kovacs, J Nolta. (2009).
P. Zhou, L. Wirthlin, J. McGee, G. Annett, J. Nolta, Semin Immunopathol (2009).
I. Rosov, D. Link, and J. Nolta. Tissue Engineering (2010).

1987-2010 Nolta lab: Development of models to study biosafety and efficacy of

engineered human Marrow Stromal Cells / Mesenchymal Stem Cells (MSC)

Decade-long
Decade
long biosafety studies:
Bauer et al., Nolta lab
Mol Ther 16, 1308 (2008)

Using
U
i M
Mesenchymal
h
l Stem
St
Cells
C ll as paramedics

di
engineered to treat Huntingtons disease

Clelland et al, 2008

HD is not a schema
schema like the prior slide. It is people, families, friends who suffer.
Please visit sites like that from our friend Chris, below, to help understand it.

www.huntingtonsdance.org

Chris Furbee

 Currently, no preventative or curative

treatments exist for HD
HD.
In addition to new drugs being developed
to treat different aspects of the disease
such as chorea, cellular therapy and gene
therapy provide the best options for
permanent cures.

In HD, a repeated sequence of DNA leads to a mutant RNA and

then a toxic protein that leads to the death of neurons.
This disease is tough to treat because we need to restore the neurons
but also to knock out the bad RNA and protein.
The field of RNA interference gives hope.

RNA
Interference

Studies Underway,
Working Toward Cellular Therapy
Trials
a s for
o Huntingtons
u t gto s disease
d sease
1. MSC to promote neurorestorative effects:
MSC and MSC/BDNF
2. MSC to continually secrete RNA interference
molecules to reduce levels of the toxic
huntingtin RNA and protein.

WHY plan to use Mesenchymal stem cells??

Strong safety profile

Neurorestorative effects
Relative ease of isolation and expansion

Neurorestorative effects of MSC

Reduce inflammation
Increase vascularization
Reduce death of damaged neurons
Restore synaptic connections between damaged
neurons
MSC can be safely delivered into the brain and
spinal cord, in small and large animal models
(Parr et al
al, 2007
2007, Pittenger 2008,
2008 and
Joyce et al, 2010)

 Following intravenous injection, only low levels

of human MSC cross the blood brain barrier in
chronic disease models (Meyerrose et al 2007).
To effectively combat most neurodegenerative
diseases we will need to deliver larger numbers
of MSC directly into the brain tissue
tissue.
We are validating the biosafety of catheterbased MSC delivery systems into the brains of
rodents and non-human primates.

Brain: the final frontier

MSC use the external surface of the vasculature as train tracks to migrate
throughout the brain tissue to deliver factors to damaged neurons

FDA Gives TCA Cellular Therapy Green Light to Proceed

with First ALS Adult Stem Cell Trial
TCA Cellular Therapy announced that the U.S. Food and Drug Administration
(FDA) has approved its adult mesenchymal stem cell protocol to conduct Phase I
clinical trials to treat Amyotrophic Lateral Sclerosis (ALS or Lou Gehrig
Gehrigss
disease).
MSC will be injected into the spinal cords of patients.

Important consideration:
RISK
vs.
vs
Potential
BENEFIT

Medium Spiny Neuron

Damaged/Lost in HD- they control movement, cognition and emotion

Kelly, Dunnet and Rosser, Biochem. Soc. Trans. (2009) 37, 323328

Damaged neurons can round up and retract axons: this prevents effective
signaling from cell to cell in the neural network

Mesenchymal stem cells can restore synaptic connections between neurons

by secreting factors (reviewed in Joyce et al, Nolta lab, 2010)

Adult Human Stem cells home

to sites of tissue injury

Injured
I
j
d
leg

healthy
leg

Labeled
stem cells:
48 hours
Capoccia et al, Nolta lab 2009

MSC migrate to damaged striatum after

implantation into the brain in an HD rat model

Sadan et al, 2009

MSC (dark spots, white arrows) migrated from injection site at the red arrow
to the area of striatal damage (white lesion)

Human MSC (pink) in the brain of immune deficient mouse

Intracranial MSC implant

8 human cells per 30 murine in this field

Sham implant
Wirthlin et al, Nolta lab, MS in prep 2011

 Our ongoing studies show that human

MSC iinjected
MSC,
j t d di
directly
tl iinto
t th
the b
brains
i off
immune deficient mice, survive for months
and migrate readily throughout the neural
tissue.
MSC are still present in robust numbers
and the brain tissue architecture is
unaltered.

Intracranial injection of human mesenchymal stem

cells
ll into
i t the
th mostt stringent
ti
t biosafety
bi
f t model
d l
available:

1. After 5 months, human mesenchymal stem

cells were still present in the brain tissue.
2. No tumors or other tissue abnormalities were
detected.
SAFETY WAS DEMONSTRATED
-Pre-IND
Pre IND paperwork
paper ork was
as filed
-written input was received from the FDA

Initial paperwork for the proposed HD clinical trial on its way to the FDA

Gerhard Bauer, Jan Nolta, Geralyn Annett

Working toward clinical trial for HD

f ll i dialog
following
di l with
ith FDA,
FDA IND submission
b i i and
d approvall

MSC from healthy donors implanted near the affected portion of the brain in
symptomatic HD patients.
patients

Evaluation of neuroprotective effects: slowing of disease progression as

measured by Total Functional Capacity score and delay in volumetric MRI
changes known to occur in HD
HD. Clinical improvement in severity of
movement disorders and cognitive impairment as measured by the Unified
HD Rating Scale (UHDRS) and a battery of cognitive tests.

Planned following the ongoing IND-enabling

IND enabling studies at UC Davis and
conversations with the FDA/ IND Application/ pending future FDA approval.

HD Disease Team: UC Davis: Wheelock, Nolta, Tempkin, Shalaie,Olson,

Annett Bauer,
Annett,
Bauer Walker
Walker, other GMP and clinical staff
staff, Jane Paulsen (MRI)

Inclusion and exclusion criteria have been delineated

Currently assembling advisory board and DSMB

St

Monitor patient for

Potential improvement in
UC Davis Movement
Disorders Clinic

Stem cell infusion

via catheter

 MSC will exert neuroprotective/ neurorestorative

effects in the brain. We can engineer them to
produce additional factors such as BDNF to
further delay striatal neuron death in HD, and we
are testing MSC biosafety in the brain.
However, the toxic mutant htt protein will still be
present in Huntingtons
Huntington s disease.
disease
To trulyy cure the disease the levels of this
protein must be continually reduced.

Small interfering RNA (siRNA) molecules bind to the RNA messages transcribed
from a specific gene and cause their destruction.
greatly
y reduced from that
If the siRNA is effective, the protein levels will be g
targeted message.

RNA
Interference

 Small interfering RNA (siRNA) strategies in mouse

models of HD have been shown by ten different
laboratories to specifically decrease mutant protein
expression and aggregation, while sparing the normal
protein and prolonging survival.

reviewed in:
Joyce N, Annett G, Wirthlin L, Olson S, Bauer G, Nolta J.
Mesenchymal stem cells for the treatment of
neurodegenerative disease.
Regenerative Medicine (2010) 5 (6), in press

How to deliver siRNA to the neurons

of HD patients?
Charged nucleic acids do not readily cross the
bloodbrain barrier.
Our strategy uses human mesenchymal stem
cells for sustained production and delivery of
anti-mutant
ti
t t HTT siRNA
iRNA (Olson,
(Ol
Wirthlin,
Wi hli Nolta,
N l UC
patent #2009-170, International patent filed 2009).
Funded by CIRM TR1-01257: (Nolta) translational grant

 Working with colleagues in the UC Davis Center for

Biophotonics we have shown that MSC infuse siRNA
Biophotonics,
directly into damaged cells through tunneling
nanotubules and exosomes.
This technology could have
far-reaching
g impact
p
into
any neurodegenerative
disorder where a protein
must be decreased
in the brain.

Olson et al,
al Nolta lab 2010

Human MSC migrate rapidly and interact with many cells, infusing them with factors
if needed. MSC Migration Video Courtesy of Scott Olson, Ph.D., Nolta lab

MSC to neuron transfer of alexa-dye labeled siRNA

standard microscopy (Pontow, Nolta lab 2010)

human MSC shown

transferring labeled
anti-huntingtin
ti h ti ti siRNA
iRNA
(red) into a target cell
(green)
From Scott Olson,
Nolta lab

Target cell (green) with anti-htt SIRNA (red) that had been
infused into it by a neighboring MSC. Scott Olson 2010

High-resolution visualization
Of MSC-mediated siRNA transfer
Scott Olson
Olson, Nolta lab 2010
Center for Biophotonics, UCD
CIRM TR1-01257

Transfer of siRNA from MSC into neural progenitors can cause significant
reduction in the levels of mutant htt protein in co-culture

1.2

muttHTT/actinraatio

1
08
0.8
0.6
0.4

ahttmscprotein/actin

0.2

agfpmscprotein/actin

0
0

4
6
Daysincoculture

10

O
Ongoing
i Testing:
T i
Efficacy - HD mouse models (R6/2, NSG/HD)
Biosafety

The siRNA transfer project

is funded by the
California Institute for
Regenerative Medicine
CIRM TR1-01257

Studies Underway at the level of GLP,

to Work Toward Cellular Therapy
Trials
a s for
o Huntingtons
u t gto s disease
d sease
1 MSC tto ttestt neurorestorative
1.
t
ti effects
ff t ((+//
BDNF).

2 MSC to
2.
t continually
ti
ll secrete
t siRNA
iRNA tto reduce
d
levels of the toxic htt RNA and protein.

UC Davis Institute for

Regenerative Cures

Opened March 2010

-109,000 SF renovated building
-Former state fair building
Over 200 scientists and
-Over
MDs working together

UC Davis/CIRM Institute for Regenerative Cures

Proposed
Auditorium

Toxicology
testing

Translational
Lab
GMP
Facility

Stem Cell
Imaging

Shell space

Stem Cell
Discovery
Lab
Blue area=$20 million
Of the $60 mil total cost
funded by CIRM

Clinical
Translational
Science
Center

Good Manufacturing Practice

( l
(clean
room)
) F
Facility
ilit
Hi
High
h flflexibility,
ibilit versatility
tilit
6 manufacturing labs
3 intermediate labs
Nolta, Bauer, Annett: two
decades of experience with
stem cell gene therapy trials
18 trials since 1994
Established contracts exist with Stanford, Industry, and other Institutions
MSC expansion is completed for phase 1 trials
iPSC expansion is underway (Stanford CIRM disease team grant for EB)

Stem Cell Clinical Trials Ongoing/

Pending at UC Davis

Non-union bone fractures Started in 2008

Cardiac infarction Started Sept. 2009

Peripheral vascular disease Started November 2010

Retinal Occlusion, causing blindness pending, IND submitted

Gene-Modified MSC for Huntingtons Disease pending, pre-IND

submitted and ALS pending

St
Stem
cell
ll Biological
Bi l i l bandages
b d
for
f non-healing
h li ulcers
l
pending
di

Liver disease pending

Cardiac patching with stem cell augmentation pending

UC Davis HD Team

Vicki Wheelock
Jan Nolta
Terry Tempkin
Geralyn Annett
Scott Olson
Louisa Wirthlin
Gerhard Bauer
Laurie Macintosh
Jeannine McGee
Karen Pepper
Heather Stewart
Alice Tarantal
Amal Kambal
Whitney Cary
Jeanie Liu
Gaela Mitchell
Catherine Nacey
Kari Pollock
Suzanne Pontow
Alice Tarantal
Matt Lindsey
Jeremy Tempkin

Advisors:
Leslie Thompson, UCI
Bob Deans, Athersys
Willie Mays, Athersys

Barcelona Team:
Jordi Munoz
Jordi Alberch
Josep Canals

Milan Team:
Elena Cattaneo

Funded by CIRM grant

TR1-01257

Thank you HD patient advocates, patients and their families

** *

*
** *

** *
*

= Huntingtons disease
team members

*
*

Funding THANK YOU!!!

California Institute for Regenerative
Medicine (Prop. 71)

National Institutes of Health

UC Davis CTSC p
pilot g
grant
Team KJ
Roberson Family
Private philanthropy

Thanks!!
Jan.nolta@ucdmc.ucdavis.edu