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DART talk. >> Again, welcome to the second
to have a guest speaker. Tonight, we're fortunate enough
of anatomy and neurology, Clivand is also the directorssor
of the Stem Cell and
By trade, he's a neuro stem cell Regenerative Medicine Center.
brain regeneration biologist, focusing on
His lab has extensive experience following disease trauma.
differentiation of human in the growth and
His lab uses novel molecular neural stem cells.
and detailed transplantation and cellular techniques,
In combination with the clinical programs to assess function.
and his colleagues are facility at Waisman Center, he
banks, which they hope might be devmoved into the clinic.ll
Please join me in welcoming Clive.
( applause )
introduction. So, I'm going to start kicking
off the discussion section. I'm happy to be here, and very
supportive of DART and what it's
And hi to all of you on campus setting out to achieve.
It's pretty bad out there. for braving the weather.
at home, nice and toasty and So if you're sitting out there
you're here, I hope you enjoy warm, watchit. this, or if
*So I thought to get into the
Svendsen, and I approve of this mood of tmessage.'m Clive
Most of you are going to be voting tonight.
And elections and politics are
that we do. ve*On campus, here, at theing
University of Wisconsin, we are developing, as mentioned in the
video that opened up the
hopefully one day prevent or session, new ways to treat and
And on campus, we have a cure different disease types.
in this area. wiI think across the country,ing
Wisconsin is probably one of the
medicine in stem cell biology. strongest in regenerative
formation of the UW Stem Cell and Regenerative Medicine
Center. These are the faculty that are
involved in the center. Many of their names you may be
familiar with.
We're all aiming to try and bring stem cells, in one way
learn about stem cell biology,
bring this to the clinic through and in another way, to try and
So that's my advertisement for this program.
Medicine Center, which DART is a the Stem Cell and Regenerative
We're happy to have DART on board to represent the students
at the UW.
thought I'd start with basic *So for the talk tonight, I
warmed up, because that's what I neuroscience, just to get you
I sort of fell into stem cells a am, a neurobiologist.
ago now, to ask a specific few years ago, around 15 years
disease called Parkinson's question about a neurological
I'll discuss that. Disease.
about stem cells, and how they Of course I'm going to talk
therapies. may be used in transplantation
some of the therapies that are And at the end, I'll discuss
cells. being thought about using stem
minefield for ethical issues. *TIt involves the use of fetal
tissue, in some cases, embryos
So as I go through the talk, I'm in others.
I consider are the main ethical goichallenges of this field.hat
I think that's quite
As young students, you should be appropriate.
and where we are with them. There's some very exciting new
data from the University of Wisconsin and Japan that
actually addresses some of these
I'd like to start with the 101,* ethwhat's in a brain.ly.
And this is my son. When I asked him what's in a
brain when he was nine, he said it's pretty simple.
From this cartoon comic, we have
everything. So it's not quite that simple.
When he got to be 12 or 13 years
this. oHe said he found some Web site
that had some exciting areas of the brain.
So I asked him what it was. He said that it's this.
That's not that exciting. Let's see what's inside.
He said, wait, dad, this is exciting.
Wow, that's great-- *Essentially, this shows you
what's inside your head.
watching this video, you're using parts of your brain.
The brain itself is one of the
universe. There are around 50 billion
individual cells in the brain.
second. And the structure of the brain
is such that it allows us to be who we are.
By now, it allows you to be a UW
And if your GPA is good, it'll student.
things after being a student. And if we get down to more the
nitty gritty, as we say in
brain, how many of you know how Emany cells are in the brain?
How many different types of cells?
Put your hand up if you do know.
hand up in here. There's nobody putting their
you. So that's a good sign, I'll tell
cells in the brain. TThe first type of cell is the*
neuron, which we all think about
of the brain. The neurons conduct impulses in
the brain.
shown here in blue. They have a lot of connection to
other cells around the rest of
So the neuron is the thing that we normally think about.
But if I want you to take home*
that neurons in the brain are important, but they have to have
support cells.
cells in the brain that comes Theralong with the neuron.port
One is oligodendrocyte, and the other is astrocyte.
Now the oligodendrocyte, does anybody know what an
oligodendrocyte does in the brain?
Okay, the oligodendrocyte is
This is a cartoon where you'll involved in remyelination.
The oligodendrocyte cell see what happens.
cells in the brain. pWithout the insulation, nerveve
cells would all crosstalk.
Multiple Sclerosis in which the insulation dies in the brain,
and you get the crosstalk, and
Sclerosis. you get the symptoms of Multiple
moment involve the loss of Other diseases I'll mention in a
*And we're now finding that neurons.
where they fit in, is they sit these cells, the astrocytes,
them. alonThey're rather like theurse
nursemaids for these very delicate neurons in the brain.
And the astrocytes are very important support cells for
maintaining neuron function.
in the brain, the neuron, the So therastrocyte and thef cells
oligodendrocyte.
is this man, Santiago Ramon y Cajal, who did some of the first*
drawings of the brain.
interesting chap. His papers are still cited
today, more than any other
later. neuroscientist, almost a century
His father, actually, wanted him He predicted many, many things.
Sorry, his father wanted him to be an artist.
to med school. to be a doctor and go
But his father won out, so he ended up going to med school.
And the first thing he did when he got to med school and got to
a lab, was to start drawing everything he saw.
He was a brilliant drawer.
interpreted from the brain what And the dwas going on.he did, he
So he made many predictions about the brain.
And the one that I'm going to focus on how is this depressing
thought.
certain number of neurons, which *That is, we're born with a
put alcohol in because we're in decrWisconsin, the drinkingd to
university of the universe, apparently.
And everything must die in the
Nothing can regenerate. This is pretty depressing if
you're a UW student and starting
We're losing brain cells every to get older.
That was what Cajal told us. day.
repair from stem cells as we get So what's the hope for brain
So neurological diseases, it's* into medicine?
There's also neurological worse than diseases.cells dying.
And many of you know these
Alzheimer's Disease causes a diseases.
And Reagan, unfortunately, got loss of memory.
Stroke, Margaret Thatcher, a this disease.
succumbed to stroke in England. She was the Prime Minister of
England.
Huntington's disease. All these affect different parts
of the nervous system.
on, the two that my lab work on, And the one I'm going to focus
disease. the first one is Parkinson's
who has Parkinson's disease. This chap is Michael J. Fox,
Parkinson's disease, which gives He set up a foundation based on
The other one is a horrible name, Amyotrophic Lateral
Sclerosis.
better known as Lou Gehrig's This is something, ALS, is
Lou Gehrig, of course, is the famous baseball player who
succumbed to ALS. *I'm not going to have time to
discuss our work in ALS.
Parkinson's as a model for stem I'm rcell therapy.o give
But a lot of the things I'm telling you about with
Parkinson's are applicable to other neurological diseases.
Maybe we can discuss that as we go through.
So what is Parkinson's disease?* This is the typical example of
an elderly gentleman with Parkinson's.
He has the stooped gait.
posture and movement is The basically restricted. the
And often, these patients have
It's caused by a loss of an arresting tremor.
*And if you can see, this is a brain section, a spinal cord, a
mid-brain section from a
And you see this black area Parkinshere.patient.
This is the dopamine neurons in the brain, pigmented black.
It's called the substantia
This is from a normal patient, nigra, this area of the brain.
This is from a Parkinson's without Parkinson's.
You can see these dopamine patient.
from the Parkinson's patient. n*And the result of losingtely
dopamine, so here's now a picture of the human brain.
And the dopamine neurons sit
brain. very deep in the base of the
aspect of the brain. This is looking at the frontal
the way up to here. This area of the brain is called
the striatum. This is where the cell bodies
are that I told you about, down here, in the brain stem.
So this is a cell body, and it
striatum. *These neurons release a
neurochemical called dopamine.
of like the oil in the engine of And dopamine, you have to think
Dopamine allows you to move. a car.
out dopamine, you have smooth When thesemovement.are pumping
So when I'm moving now, trying
making small amounts of dopamine, which is allowing me
to do this fluid movement.
something happens to those *What happens in Parkinson's,
That's a million dollar dopaquestion.ons.
We don't know why they die, but the dopamine neurons die in
Parkinson's disease.
get this syndrome of Parkinson's The dopamine goes down, and you
This is a chap we'll come back* disease.
to later who has Parkinson's
we've asked him to put his hand on his knee there and turn it as
fast as he can. I think you can see from this
video that he can hardly turn his hand, moving very slowly.
And he's got this resting tremor, where his hand is
bouncing up and down. *So this is the syndrome of
Parkinson's disease.
There's really no cure for We don't know what causes it.
There's drugs that you can take for Parkinson's disease that
reduce the symptoms, but the
drugs become less effective. disease keeps going and the
neurology have been trying to *implant stimulators, which we
don't know exactly how they work, but you can put this
little electrode in part of the brain and stimulate it, and get
good results in Parkinson's disease.
Again, that doesn't stop the
progression. continuation of the disease, the
after a certain amount of time. SoIt's not a therapy that isk
really treating the disease.
dopamine neurons? *So What if we could put newse
dopamine neurons back in the
So the question is, is brain brain?
So if you go back in history, and think what can we do, you
might think brain transplants,
least likely area to get into. unless you're Steve Martin, the
brains. We have the screw-cap technology
there.
brain transplantation. Actually, there is a history of*
paper in 1510. I picked this out from a British
transplant. It reads here, "A gentlemann
otherwise well, had the idea his
He went to the King," and in brain was rotten.
queens rather than presidents, England, we have kings and
physician] and myself to open his head, remove his diseased
brain and replace it with another.
We did many things to him, but
restore his brain." I hate to think what happened
to this poor chap.
whole brain transplantation, But this was a first attempt at
forward. which obviously, isn't the way
psychologically for the patient, That would be a big problem
think in a slightly different they had a whway.new brain, I
He might look pretty old, this
actually has a laptop computer. And he's actually got a special
mouse here, he's communicating, going home for dinner, I think.
So the key here is adult brain*
It's not like a kidney, where you can take it from the
deceased patient and put it into
We have to be more subtle in the another patient.
It is interesting, actually, brain, so putting new cells in.
cells that divide. that the adult brain does have
the brain. Neural stem cells are present in
thinking, there's a part of your So when you're all sitting here
There's two regions of the brain that's making new cells.
One is here, in the middle, called the subventricular
region. And one is here, in the
hippocampus region. The hippocampus is very
important for learning and
And there is a theory that new memory.
and are very important for neurons are born as you think
If you get depressed, it seems things like depression.
neurons in the brain. likeSo there are new neuronsess
produced in the brain, in both
However, if we wanted to take these out and grow make new
neurons from them, they don't grow very well in the culture
dish. And they don't make these
critical dopamine neurons that
disease. you need for Parkinson's
work very well for Parkinson's So adult tissues don't seem to*
dopamine neurons. So researchers then turn to the
fetal brain, rather than the
During development we start, as you know, as a little egg.
It's fertilized.
development, up here to 40 This is the timeline for human
The brain gets bigger and bigger weeks.
If you go in at around 21 weeks, during development.
And in Parkinson's, they you can isolate tissues.
weeks. actually go at around eight
human brain, which can be used You can isolate tissues from the
neurons. to generate these dopamine
rodents, in rats. ActSo here, I'm showing you ain*
picture of where you take the part of the brain that makes
dopamine neurons. If you remember from that
picture earlier.
that would make a dopamine This is the part of the brain
You take it out. neuron.
from this rat fetal tissue. You And then you mix it upces
in a tube.
not the whole brain, just these Thecells, back into the rat.lls,
So now you've given the rat new dopamine neurons.
And in those early studies, in*
now with Parkinson's disease. thYou see here are the neuronsat
of the dopamine positive, with
This is the other side of the area where the dopamine neurons
are.
that area called the striatum. This is where they project to,
is missing from the striatum. If And this animal has had amine
transplant. Can you see this area here has
dopamine in it? This is because that fetal
tissue cells went into the rat
making new dopamine neurons. brain, and lived, and started
they were developing normally. They were tricked into thinking
away from the fetal environment and put them into the adult rat
brain, and they started making
*So you say this is great for dopamine neurons.
What about Parkinson's patients? curing rats, making dopamine.
The first trial was done in England and Sweden, actually, in
1987. They did a brain transplant on
human patients. And even back then, there were
ethical issues about using
used in Sweden for this trial. aborted fetal tissue that was
experiment in patients. And this is an example of what*
happens in a human patient. Here's the drill hole in the
brain, where they took a drill
patient's brain. and made a small hole in the
injected the fetal tissue Tdirectly into the patient'sd
brain.
is a PET scan, showing that the Seefetal tissue has made newthis
dopamine neurons in this
So this patient has new dopamine Parkinson's patient.
And the neurons do release neurons in the brain.
In fact, there's been 350 patients in the world, including
in America, who received these
And many are doing very well in terms of reducing some of their
medication. But it's not a cure for
Parkinson's disease.
world of stem cells and *Treplacement for using fetale
tissue. The fetal tissue that is used is
basically unreliable. There are a lot of ethical
concerns of using aborted fetal tissues in many countries.
And you can't really control the
these cells. dopamine that's released from
ago, for alternatives to using So we're looking, and many years
get these transplants to work, fetal tissue, to see if we could
And stem cells are the things that, perhaps, would offer an
alternative for this. *Now, I'm going to move to the
third part of the talk, which is
this plays into this model of Parkinson's disease I'm talking
about, and Parkinson's disease
So just to jump now to a in general.
Wisconsin by Jamie Thomson. dAnd that is, if you take veryn
early embryos, when they're developing, at around five to
seven days, the embryo at this
It has an outer cell layer here, stage is the size of a pin head.
This golden area in the middle this purple area.
inner cell mass of the embryo. here, is an area called the
to remove that inner cell mass, and put the cells into a dish,
you can get those cells to grow as human embryonic stem cells.
They will divide, practically indefinitely in culture.
*These cells are what's called pluripotent.
They can make any lineage in the
They can make heart, brain, blood, bone.
They keep dividing. So they're kind of like the gold
dust, if you like, of stem cell biology.
They can divide in the dish.
conditions, they can make muscle And if you give them the right
nerve cells. cells, pancreatic cells, and
here in Wisconsin is actually And focusing on how to take on*
embryonic stem cells down these different routes.
And during these DART talks,
professors who are working on different areas of biology,
including heart and pancreas, and trying to see if these cells
can be used in a cell therapy way for these diseases.
Along with this, comes ethical*
You have to look carefully at this slide.
But there are ethical challenges
destroying them. with creating embryos and then
around the world. TAnd some countries just won'trn
condone destroying embryos in order to isolate these powerful
stem cells, which I'll tell you in a minute, can make dopamine
neurons for Parkinson's disease.
for alternatives. *IAnd one of the big questions
was, for these pluripotent kind of cells, do we need to use
embryos?
recently, yes. And the answer was, until
groups, one from Japan, Takahashi, et al, and Dr. Yu,o
here in Wisconsin, as part of Jamie Thomson's lab, discovered
that they could do a trick. It's quite amazing.
This goes back to every cell in your body, theoretically, has
the potential to make a new you.
the DNA, the whole DNA The cells in your body contain
beginning. complement that made you in the
divide, they just remember the So every cell, when your cells
were first born. amount of DNA you have when you
enough information to make a So, every cell in your body has
And many of you remember Dolly,* the sheep, proving that for the
first time in 1997. They took an adult cell from a
sheep, and they made it into a
That needed therapeutic cloning, whole animal.
cells are put into an egg, and reproductive cloning, where the
And they made a clone of that the egg waanimal.to divide.
But it proved that adults cells in your body have the potential
to make a whole new animal. What these two groups did, was*
they said, well, if that's true,
make a whole human being, theoretically, what if we took
cells, in this case, from a Parkinson's patient, or from any
person, and we got skin cells,
the skin and we grew them in a say, or fibroblasts from under
Now, skin cells, or fibroblasts, culture dish.
Normally, they can't make other can onlytypes of tissue.blasts.
What they did, was they put a number of genes, two of these,
Oct-4 and Sax-2.
this pluripotency. These are genes that control
cells, and make them very They conpowerful.ticity of
When they put these genes into
happened. That is, they appeared to turn
into what looked like embryonic stem cells.
*So now they turned these, induced pluripotent stem cells
So potentially now, you can take
these genes in and turn them a skin cell from a patient, put
*And the Japanese group, they into these pluripotent cells.
then be made into new epithelial showed that these cells could
the central nervous system. These cells could then be grown
and differentiated into neurons.
dopamine neuron using an adult *So now we can get to our
And what's particularly nice cell.
about transplants in a moment, about this is, and we'll talk
rejection. but there is an issue with
person to another, they'll If you transplant cells from one
You have to take strong immunosuppression.
Now if we could take cells from the same person and develop
embryonic, or induce pluripotent
source for transplantation back ceinto the same patient after a
doing all these tricks. So that's an important point.
*The other point is that actually, if you think about it,
you can now take cells from any patient with a disease that you
don't understand and grow those cells in a dish and make neurons
from that patient.
Parkinson's patient, make Sodopamine neurons, and askm a
questions about how the disease process works.
In the culture dish, you could
slow down the death of those could look at drugs, perhaps, to
So this is opening up a lot of neurons.
The advantage of going back to* different avenues.
these types of cells is the Papractical advantage.using
We can control, perhaps, the
the graft that you couldn't nucontrol with fetal tissue.n
And if you engineer those cells and do some tricks, you can
actually maybe tell them to release dopamine within the
transplant. So now your patient could take a
pill and switch on his dopamine or switch it off again.
Those are the sort of things
these types of cells. This is work from a colleague,h*
who's among one of the first
from human embryonic stem cell. It just shows you, I don't want
to go into the details. You go through different stages
from embryonic stem cells through to neural tissue, and
then he took the neural tissue
They're shown in red here. through to dopamine neurons.
developing. So he can take these embryonic
stem cell and hopefully soon, IPS cells, through to dopamine
neurons.
dopamine neurons. This has potential for replacing
fetal tissue. *I'll stop in a moment for
questions.
on two different things. But just to show you a warning
very powerful. Human embryonic stem cells are
great price. But with great power comes a
And the price is that these cells are so powerful, they may
keep dividing when you put them in the brain.
If there are any of the powerful
could form something called a embryonic steratoma. left, they
And in the brain, we're very
particularly in the spinal cord, worried about that, because
growing that can affect brain yfunction and cause pressure.
So we have to be very careful that these cells are not going
to form tumors. But this is a practical issue
that is being worked on.
working on resolving this issue Many groupsright now.e world are
But we can dopamine neurons.
why aren't we putting these So whacells into patients. and*
The story really ends a little
Because the results of the fetal sadly, I think.
completely successful. tissue transplants has not been
may have already realized, is we're replacing dopamine neurons
in the brain of the patient.
neurons in the transplant up But we're putting the dopamine
The dopamine neurons are all the here, in the striatum.
brain stem. Why don't we put the dopaminee
neurons back in the brain stem? Well, if you put them back in
the brain stem, they don't
back together. reconnect and put the circuit
back to this part of the brain. This is very important for cell
therapy in the brain, neuroma therapy.
You have to be able to
in order to get it to work. reeAnd what the patients aretion
showing, is it's nice if you put
because they're at where they thshould be for releasing thee,
dopamine. So they'll release the dopamine,
but the patients suffer some
full function or recovery, side effects and they don't have
circuit back. What we'd love to be able to do
is what I'm showing you now, you
on the screen, is put the cells cback here and then encouragen
them to make a new connection,
circuit. so they now they complete the
we're not there. People are working on that, but
animals or in Parkinson's We're nowhpatients.that yet in
Until we can get that circuit reconnected, I think it's going
to be difficult for dopamine
disease to work. neuron therapy in Parkinson's
ahead of us. So I think we have a lot of work
talk, I'm going to move more to a different approach, asking the
question, well, if we can't replace the dopamine neurons
correctly in the circuit, is
them and stop them from dying? there any way we can protect
talking about. This is neural protection I'm
questions, if anybody has a questions about Parkinson's.
( inaudible ) So around a small percent of
Parkinson's patients have an
This is common with many inherited form of the disease.
There's a mutation in a gene, which in a small percentage,
less than five percent of Parkinson's patients, that
mutation is known to cause
And there's around ten or 15 Parkinson's disease.
to cause Parkinson's. othBut if you add all thoseown
mutations up, there's still around 90% of Parkinson's
patients who we call sporadic,
mutation. wWe can't find a mutation.wn
It just happens.
Parkinson's, it's very difficult So when we look at the causes of
causing it. We can't seem to find a gene.
The most, what we're really coming down to in all
neurological diseases, I think, is that we all have different
sets of genes and risk factors. And that risk factor will
probably interact with your environment at some point in
your life.
smokes a lot of cigars. He's approaching 80 and he's
healthy.
inhales. He smokes cigars, he always
risk for lung cancer, and doesn't carry risk genes for
that. Other people can have a few
cigarettes and get lung cancer. They may have a lot of risk
factors for lung cancer.
like that. So I think Parkinson's is maybe
don't understand what those There arfactors are yet.we just
That's a good question. I wish we knew what caused
Parkinson's, because I wouldn't be spending all this time trying
to cure it. I'd be trying to work out if,
you know, the mechanism.
well, in 2008, will you know Twenty years ago, if you said
I would have said of course we what causes Parkinson's?
But we still don't know what will by then.
except in a few families, where causthere's a mutation.ase,
Yeah? ( inaudible )
That's a good point.
dopamine neurons back here, like These cells, when you put the
So they make dopamine and they I showed you.
But you're right, there's nothing to connect to them to
tell them when they should be sending their signals.
They're not in the right
They do get connections, but circuit.
connection. In fact, the patients often get
these movements called
So dopamine will be made by the new neurons, but it's not in the
circuit correctly.
controlling themselves. And some of the neurologists say
we now have Parkinson's disease and we also have Parkinson's
plus graft disease, which is a
we put these dopamine neurons in new disease completely because
But they can reduce the medication, that's the good
thing.
the movement, which is a bad But they lose control of some of
I think we need to understand most of the clinical trials with
with new dopamine have stopped
side effects of the transplant, putting the neurons in the wrong
place.
whatever fires them or drives Andthem isn't correct either.t,
They're outside the circuit.
they die, which is down in the If we can get them back where
Then we've got to make them put a new connection out to get the.
dopamine to the right place.
Well, what's interesting is there actually are, if you
get the cell body back, the old connections that came in
are still there.
If they find a new cell body They just sit there dormant.
can put a new one back, they back where it died before, if we
often in other models. will reconnect with it quite
the cell back there, they may get the appropriate connection
back. When the cell dies, whatever was
connected to it before sort of
But if you put a new cell in, it'll reconnect and reestablish
the whole circuit.
question. But I think, maybe, hopefully,
it was obvious from my answer.
Okay, I will move on into a slight detour.
I realized pretty early on that* this was going to be a
challenge, replacing neurons in the brain.
So this part of the story is
neurons, now you have to change your thinking here a little bit.
Remember I mentioned in the beginning there were different
types of cells in the brain.
Support cells are also very Neurons aimportant.portant.
What I'm going to tell you now
cells to provide drugs to the brain that are very difficult to
get in. What I'm going to talk about is*
this one. Glial derived neurotrophic
factor, GDNF. This is known to be a powerful
drug for Parkinson's disease. It's rather like fertilizer for
these dopamine neurons. If you put it on a dopamine
neuron, the dopamine neuron is
It sprouts fibers out. happy.
It protects neurons in the brain It survives better.
It has a long term effect in in many different models.
*So why aren't we using it in monkey models of Parkinson's.
If you give this drug, it doesn't penetrate the
blood-brain barrier.
It doesn't get into the brain. It gets stuck outside the brain.
So there are different ways. So we can't get it in.
so how about we just drill a We can't just give it as a pill,
in? hThat's sort of a neurosurgerymp
approach. And in fact, they did that.
They put a pump in a number of
GDNF directly into the brain. They put it into the ventricles
in the middle of the brain. And it turned out the GDNF still
can't get to where it needs to
part of the brain that has the get, which is down here, this
I showed you on the slide dopabefore. it.
So in Bristol, the neurosurgeon* colleague started now to put
this catheter directly into where the dopamine terminals are
to get the GDNF out.
protect the dopamine neurons and The GDNF will be taken back and
That was the theory. cause sprouting in the brain.
years ago. *So this was published a few
x-ray of one of the patients. WhatThis is in England.s an
There were five Parkinson's patients.
You can see from this x-ray, the
thin catheter that went into the neurosurgeon put tubes in with a
has lost its dopamine. brain, targeting the part that
to stimulate plasticity, if you So we're tryilike. get the GDNF
Plasticity means those dopamine* fibers, which are old and
pulling back, suddenly get the
they grow. fertilizer, like your grass, and
of fibers in the brain, Treinnervating areas that haveth
deinnervated and are sick.
This is a cross-section. *So this was the idea.
patients have to use. This pump goes under the skin.
You can see this chap here has
pump on this side. one pump on this side and one
to the tubing under the skin and This is a little bit of plumbing
This is the patient that we saw* before, with Parkinson's
disease.
patients in this initial trial He was actually one of the
This is him before. And if we look at him after the
GDNF therapy, we saw quite a
There were five in this group, dramatic effect in this patient.
effect once they got the GDNF. many of whom showed a similar
into the brain and allowing that The idea is the GDNF is going
occur. plasticity and regrowth to
development, when the system is GDNF ideveloping.during
This is reintroducing it, if you
All I want to show you on this* like, in an adult.
for Parkinson's, 70 is bad, 20 slide is that this is a score
You can see that when they is good.
around 70. started, these patients were up
they were down to around an And at taverage of 30.months,
They came down in their scores
video showed. f*As a biologist, I was morethe
interested in the biology of the
What we showed is that GDNF, in this section through the brain.
Now we're looking at a PET scan,
the brain. which shows dopamine activity in
before they received GDNF. This is before, one patient
patient. This is after, in the same
It's missing here. If you can look at this spot.
going up, because the fibers are Now the dopamine levels are
patient, in response to GDNF. So this was really encouraging
for us. This is another way of looking
at this data. The red spots mean an increase
in dopamine, on average of these
*One patient died of a heart attack three years after
receiving this therapy. We looked at his brain.
Inside the brain you can see
GDNF, around this area. The dopamine markers were all
increased.
get the GDNF. THe only had the pump in onet
side, this chap.
Again, this was published It didn't recently. increase.
We think GDNF is an interesting* molecule.
There was a larger trial done by
go into the details. Amgen, but I don't have time to
of the pump and delivery device, But there were some complexities
that delivers the GDNF to the and the sizbrain.he catheter
In the larger group of patients,
effects. they only saw very small
Amgen decided that they weren't going to pursue this any more
because the delivery techniques were awkward, and a number of
other reasons.
idea in your minds. *And that is, instead of using
these complex pumps and delivery
technique that's quite awkward through a pump, which is a
use cells to deliver the GDNF. and difficult, maybe we could
stem cells I've shown you, we Theput them in the brain bythe
injection.
integrate. And they'll migrate and
cells. Now, they'll provide support
out GDNF. *The cells we used for thesemp
studies were actually derived from the same post-mortem fetal
tissue that was used for the
We just used the cortex part of the brain that's not needed in
this fetal tissue to expand the cells.
We expand the cells.
which you'll hear about more Now we combine gene therapy,
cell therapy. late*How do we do it?th stem
We use the old enemy of man,
But we gut it. HIV, the lentivirus.
and we just leave in, and we We take out all the bad things
that lentivirus that we want. aIn this case, we gutted it ando
put in the GDNF gene, that we have said was very useful, into
the lentivirus. We then put the lentivirus onto
the stem cells and let it do its
And what it does, as you will note in your biology classes,
the lentivirus inserts its RNA
cells produce, normally, more into tcopies of itself.es the
But of course, we've engineered it so now the cells make GDNF
instead of lentivirus.
that makes GDNF. *So we now produce a cell line
And when you look at these cells I've colored them green.
pump out GDNF into the media in the dish, they actually
And now, what we want, a small in the culture dish.
This is in collaboration with a number of different people, both
in Switzerland and here in my lab.
*Now the cells that we want to pump out GDNF are very active.
These are neural stem cells.
video to show you what they look I just thought I'd show you this
So this sphere of cells here, this ball of cells, is called a
neurosphere.
individual stem cells. Within there, there are 100,000
neurospheres a substrate, something to stick to, we use
lamina. What happens is the sphere
sticks down and it starts to
And when you're watching this video, if you look around the
edge of the video, you'll see individual cells migrating out.
Up here, there's a good example of a few.
These are all individual brain
stem cells. What they're doing is they're
turning into neurons, astrocytes
talked about right at the and oligobeginning.s that I
They're doing it right in front of your eyes.
This is a time lapse movie over three days.
You can see the cells as they migrate out.
Yeah? ( inaudible )
The question is are they
because of the environment. differThe answer is yes. cells
If we have them in a different
as stem cells. environment, they keep growing
where we put them on a We put them in an environment
case, lamina, which is a substrate, which means, in this
And we take away the growth factors, which is this case, is
called EGF and FGF, that make them divide.
So now they start to
The environment is directing differentiate.
They stop dividing and their differentiation.
Those signals, we can control they staquite well.tiating.
That's why they're doing this.
neuronetwork on the dish. We're worried that if we leave
them too long, they'll start
These are human brains, basically, in the dish.
We might get arrested for creating brains in a dish.
We'll have to give them the New York Times to see if they can
read it, or the Daily Cardinal. So right now, we haven't created
any thinking monsters in the
But I will touch on that in a moment, because it's important
when we start talking about transplants.
*So this is what they do, they make cells that include neurons
and astrocytes, those two important cell types
that I mentioned before,
And along with a colleague who when they differentiate.
the stem cell bank that we have is thhere in Wisconsin, theor on
NIH-approved stem cell bank, we are working on making these
neurospheres, these cells I just
making big banks of it. *Now the GDNF isn't important
for that differentiation, back
Those cells want to turn into toneuro tissue.n.
We keep them dividing by putting
divide. But when they go into the brain,
or if we take away the things that make them divide, they
naturally want to make nerve
But we keep them dividing in the tissue, neurons and astrocytes.
stock of these cells for bank, so we now have a large
Any other questions at this point before I go to my last
series of slides? Yeah?
The microphone is coming around,
Is the GDNF going to make them into dopamine-releasing
neurons, or is it going to make all types of neurons.
Because, then wouldn't there be the question of is it going to
start making all this new brain tissue and screw up connections
that already exist? >> Great question.
You have to get away from dopamine neurons.
The cells, when you grow the
talking about, the fetal tissue, cells from the tissue I was
They don't make many neurons and you expand them.
They make many glial cells, or anymore.
Those astrocytes, when you put astrocytes.
in a minute, they make nearly them in all astrocytes. show you
They're no longer making
The astrocytes release the GDNF. neurons.
dopamine neurons that are still The left in the patient with the
Parkinson's disease.
neurons in anymore. We're trying to just use these
cells as delivery cells.
out the GDNF. So we're not trying to do neural
replacement.
this, you keep expanding them, Luckily, when you grow them like
while, especially after they stotransplantation. after a
Yeah? That's a great question.
You're following the talk very
Just in general, I'm saying, carefully.
factor. And when it's delivered, even if
it's delivered to certain places, and it's still going to
promote a lot of sprouting of the neurons that are in that
area, that might not even include dopamine neurons?
You want to be careful. And in fact, that's a good
point.
whole brain, you get aberrant If you jusprouting.NF to the
And for some of the early clinical trials, they put it in
the cerebral spinal fluid.
and another syndrome, which was Ia vibration and a tingling inin
the neck. So you have to target it.
We target it by putting some
slides in a minute. We put the cells right in the
area where the dopamine fibers
So the only fibers that really are.
Other fibers don't really rerespond too much to GDNF.es.
Or if they do, it's not in a bad
A really good point. way.
the drug in a very located area This isis very important.ease of
If you put it in the wrong area,
or bad plasticity, which can you might get aberrant sprouting
So we want to try to keep focused an area, and keep it
where the dopamine fibers are.
Okay, all quiet. All right?
transplants to address some of *So we'll move now to
For the transplantation part, again, I told you about fetal
tissue early on in the talk.
ago. Everybody remembers that long
thing with the neural stem cells Now we're going to do the same
Here they are, growing balls I talked about.
We basically incubate them in neurospheretripsom.sing GDNF.
And these experiments are actually being done today here
at the UW in our lab. We collect them.
We dissociate them to a single
neurospheres. And then we inject them into the
rat.
Parkinson's disease. These rats have induced
in one side of the brain in Sthese rats, like I showed yous
earlier on. The rats, actually, rotate in a
direction that goes away from where the dopamine neurons were
lost. This is a way of testing your
therapy.
it has the dopamine neurons that So if the rats rotate it means
If the rats go in a straight died.
dopamine neurons on both sides. So what we want to try to do is
put these cells in and get the
*This is from old work done in rathe last century.ng.
This is when you know you're
that. geIn 1997, we published somey
papers showing the cells can
This is a series of section through the rat brain.
And every black dot is a human
brain. cell that's gone into the rat
in the middle of the transplant. And you can see a focus of cells
migrated. If you look, you can see they've
migrated out into all areas of
They haven't gone into the the striatum.
other regions of the brain. So going back to your question,
we've managed to target the region of the brain that we need
the cells to release GDNF in, which is where the terminals are
for the dopamine neuron.
terminals might not react *somewhat to the GDNF, but it
doesn't seem to have a
These cells, just to go back detrimental effect.
don't make neurons. agThey're just making thesen,
astrocytes, or support cells. They're releasing the GDNF.
And if we look in more detail,*
Every black dot-- The beauty of doing human cells
to rats, and these are human
track them with specific neuro stem cells, is you can
In this case, we use a marker, markers.
So every black dot is a human the human cell.ar protein.
And where there's no black dots,
cells. like up here, there's no human
protein. The rat doesn't have this
We can see that the cells integrate into this part of the
brain.
graft. This animal had a rather large
place, just to show you, this is And if they get into the wrong*
crosses the midline. the corpus callosum, which
regions through white matter. The cSo you have to target other
efficiently, in order to get the
*This is just showing you now graft in the right place.
sections. This brown staining is showing
you where the GDNF is actually
So the GDNF is being made by the being expressed.
There's no GDNF on this side, because we didn't transplant the
animals on that side. We just did a transplant on this
side. So the cells have gone in,
they've migrated to fill out
pumping out this GDNF. this side, and now they're
ago. This work was done a few years
studies in the lab. *WIn this case, we've measured
these rotations I mentioned before.
These animals have had a
And the thing to look at is when transplant.
come down, which means the they have GDNF, their rotations
they're producing dopamine. fThese rats are no longerd
rotating in circles.
you'll see that the dopamine And if you look carefully,
GDNF released from the cells. It's not complete protection,e
but it's enough to have functional effects.
*I think with this therapy,
Parkinson's, slowing down the disease progression, or helping
the fibers that are left sprout more and be more plastic by
giving them this rejuvenation factor, if you like.
*This whole idea of putting
we call a chimera, which means cells in the brain, this is what
brain. it's a human animal kind of
And if you get enough human cells in an animal brain, the
question is, will it start
And it might sound funny to you thinkto a degree.uman.
But actually, this is a serious issue.
And we've been instructed by
particular has been told, that certain, and Stanford in
50% human brains, they have to if they get mice that have over
they might start thinking. stop the experiment, because
pretty small, having a rat or a Nomouse with a human brain.are
The difference in size is pretty
This is a human brain, not a enormous.
The difference in size is realenormous.ain.
And I think the rat brain,
so I don't think the rat will wenecessarily become human byls,
doing these chimeric
I think we're safe to use rats, particularly adult rats.
*However, I think the other
transplants, if you look at Alzheimer's, for instance, if
you replace enough neurons in the brain for cognitive
deficits, like Alzheimer's, you may change the personality of
the person. This is a whole other talk.
But if you have lost your memory, we put new stem cells
back in, now they're making neurons, the new neurons, you
might not get grandmother back, you might get somebody else.
That might be a good thing or a bad thing.
But it's an issue in terms of how replacing neurons, again, is
very complex. We're just trying to prevent
cells from dying. Finally, if you're putting stem*
cell transplants in the brain,
an issue. Because actually, if you go into
very early developing primates, we may end up with a primate
human chimera, which could
differently. actually start thinking
rules, actually restrict putting And in fact, NIH, the federal
primate embryos for that very human stem creason.ck into early
So I think there are limits on
But I think human cells to rat what we can do.
These debates are going on is probably quite safe.
*So that's the transplantation afor Parkinson's.
We can get the cells in.
So where are we, in terms of They make GDNF.
therapy? using this in a clinical
on in Parkinson's disease right Thenow, where those viruses Ing
talked about, they're using direct viral gene therapy.
They're putting a virus in the
something like GDNF. brain of patients that make
around, now there's about 30 These patients are walking
from direct gene therapy. *One of the issues using cellsn
to deliver these proteins to the
it, how do you switch it on and brain, is howoff?you regulate
What if GDNF is bad after two years?
The cells are going to be there.
out, whether you used a virus to They're going to be pumping it
whichever way you do it, how are put it in or the cells,
Right now, there is no way that if I put cells into your brain
that I could switch it off. GDNF would be made forever,
basically.
concerned in Parkinson's disease So I think we're kind of
problem. that this is going to be a
it. WSo a lot with the work in thete
lab, is looking at how we can switch on and off the GDNF
production. And there are molecular tricks
you can do. You can make an inducible motor,
it's called, for the gene.
drug and switch on and off GDNF That meanproduction.ve another
So the patients will take a tablet and then GDNF will come
out of the cells for about four hours.
That wears off and the GDNF shuts off again.
So this is really exciting, if
Because now we can put the cells we can get this to work.
take a second tablet in order to in, but the patients have to
*Drug companies love this activate that GDNF release.
for that tablet that switches becauthe gene on and off.oney
That way, you know, what you
the patient every day. want is a tablet that you give
commercially how to make money It's difficult to understand
once you put the cells in, the from these transplants, because
And there's no more to pay. geneYou just go home.ever.
So this is an interesting area. *So regulation is really
important.
are looking at monkey models But assuming we can get that, we
Primate Center at the University with Parkinson's here at the
We've been treating monkeys with of Wisconsin.
Parkinson's disease. MPTP, which gives them a form of
injections of GDNF secreting Then we've been doing stem cell
These are some examples from cells.
come out. sWe put cells in making GDNF.o
This is another rating scale, like I showed you before, but
this time for monkeys.
Parkinsonian symptoms because Andthey have this high score.ave
Then two of the monkeys come
transplant. The other one comes down a
little bit. It's just a pilot study.
But this comes down to a region that shows significant effects
after transplant in this model, but in a small group of animals.
We need to repeat this with a larger group.
*When we look at the monkey
this is GDNF staining. brain, which is much bigger,
different transplant locations. The graft were the same size as
the rat, but because the brain
it looks smaller. You can see we've targeted three
areas.
just targeted one area. *If you look at dopamine inve
those areas, wherever there are cells making GDNF, as shown
here, you get more dopamine markers, shown in this spot
here. So we've had plasticity and
regeneration of dopamine fibers growing toward those areas where
the GDNF is being made in the primate level.
So this is evidence that this technique could work in humans.
And again, we're just waiting for the regulation and release
data to come through before using it.
*So our final data slide will
said today. And I think one of the take home
messages is, for this Parkinson's talk, is that GDNF
released from stem cells can
Parkinson's. increase plasticity in
that are dying, regenerate. And we've got evidence that it
protects the ongoing cell death of the dopamine neurons in
models that we've used. Will it work on Parkinson's
disease?
the clinical trial. Nobody really knows that until
you go into patients. We don't have the best model of
Parkinson's in animals. We're assuming it'll do
something from all the data that
yet whether it will work. we If were generally just totand
overview what I've told you about these three cell types
that I started with in the
do, I think, for stem cell cartoon, what's really hard to
medicine for the brain, is the therapy and regenerative
with, which is replacing lost firneurons in the brain.rted
This is complicated.
cells, long-distance We need integration of the
It's really tough. connections.
myelinated cell, like an *If you want to take a
in between cell, it may be oligodendrocyte, which is an
With Multiple Sclerosis, people are talking about using these
cells to remyelinate. There are many locations of
these plaques in MS. It's complicated to think we could
target all of those in MS, but
that. thereThat gets easier.about
*The last thing, which I've
the astrocyte, from the neuro stem cell, modify it to produce
a growth factor like GDNF.
deliver this drug to the brain. I don't have time to talk about*
it, but we've got very
in ALS, which is a model of Lou Gehrig's disease, that this GDNF
can protect motor neurons very
And in fact, because ALS as a prognosis, is about four years
from the time of being diagnosed
really big need. with ALS to death, there's a
for ALS whatsoever, for Lou There'sGehrig's disease.ulators
There's a big need for this type
And I'm not worried about switching on and off the GDNF
for ALS patients. I think it would be fine to
leave it on.
the alternative, which is dying. The risks are minor compared to
approaches, depending on the So I think we have to modify our
*And so, that may be the first use of stem cells in the brain,
providing either astrocyte
There's some evidence in ALS support--
are sick. So just replacing the astrocyte
may help the motor neurons a lot
But if they're making growth in movement.
*Just to leave you a last factors, even better.
collected for prosperity. thoughThey looked at it.n was
They dissected it.
more neurons, obviously. They were expecting to see many
more astrocytes. And what they found was he had
brain, but the same number of He had more neurons.cells in his
So don't underestimate how
the brain. important support cells are in
astrocytes that we used to And it's turning out that these
actually modulate a lot of think, are next to this neuron,
transmission, synapses, how signals are sent through the
brain.
astrocytes might have a bigger So actually, just replacing the
imagined. This is a whole new area ofr
neuroscience that's just
now, is how neuro function is modulated by astrocytes, and how
this may be a way to get new
Maybe we can't turn everybody into Einstein, but I think it
would be interesting to see how we can use these to treat
diseases. *So to finish up my Waisman
Center slides, this is where I
It's a beautiful sunny day in work,Wisconsin today.oad.
This is slight artifact, I
I'd like to thank the many think, from the PowerPoint.
in my lab, who I've mentioned along the way, and collaborators
who are, again, from in this
Emborg, who did the primate case, from Uwork.ison, Marina
And in Lausanne, in Switzerland,
us with the viruses. Patrick Aebischer, who provided
important funding agencies. We get money from wherever we
can.
the Michael J. Fox Foundation, We've been funded by
National Institutes of Health, Department of Defense, the
to mention a few. tSo I'll stop there,
and take any final questions. And applause!
( applause
)