good afternoon well I'd like to start
today's talk off with a little riddle
and a little road or is what are these
three forms of life have in common
developing embryo a male Sonnenberg and
recovering stroke patient and the answer
is of course neurogenesis the developing
embryo has no Genesis to develop brain
spinal cord peripheral nervous system
the male songbird vote his vocal control
center generates recruits and promotes
neurogenesis as new songs are learned
his melody is a reflection of his of his
neurogenesis and a big lag Rafi of his
entire life and finally the recovering
stroke patient recovering stroke patient
as you know about one-third of stroke
patients do have some type of
spontaneous recovery and neurogenesis is
responsible for that so our goal here is
to try to promote all types of
neurogenesis and after Genesis for
stroke recovery
so the neuroscience laboratory here at
Henry Ford Hospital run by dr. Michael
chop has an embolic stroke rat model
this involves cannulating the external
carotid artery threading a catheter up
into the middle cerebral artery and
injecting a clot right into the middle
cerebral artery to induce to induce a
stroke which produces an infarct volume
and also measurable neurological
deficits what we can measure in the
laboratory now as you know there are
three main ways of treatment of stroke
the first is we cannulation through TPA
and through ia treatment
second one is neural protection which
has fallen out of favor and the other is
neural restoration neuro restoration is
treatment of intact prang Comeau cells
specifically neural progenitor cells
which exist in the southern trickler
zone and after stroke these cells tend
to migrate to the area of injury where
they evoke a repair response they evoke
neurogenesis angiogenesis synaptogenesis
oligodendrocytes and any other type of
Genesis so what we have here in from an
experimental stroke standpoint is we can
invoke neural restoration through two
primary methods that we've done in the
lab first is through drugs where they've
done lipitor inside NFL has shown a
neuro restorative process the second is
through and more importantly is
administration of Meza camel stromal
stem cells now these are isolated from
the bone marrow and there are progenitor
cells so what you can do is you can
isolate these cells grow them in the
culture dish go to that rat you just
stroked and inject them through the tail
vein and these cells were magically land
right into the area of the infarct
volume and produce a neuro restorative
process so what's going on these and
these mesenchymal cells will promote
neurogenesis angiogenesis
all orchestrating into neurite
remodeling and functional recovery so
this is an example this is one of the
examples of the test that we do we have
a mini way this is a middle-aged rat now
one of the tests is a sticky tab test
and what we do is you play sticky tabs
on the rats paws and a normal rat
doesn't like anything on its paws so
it's going to rip it right off in two to
five seconds however a rat that's had a
stroke is going to probably take at
least two minutes so as you can see here
when we've taken the rat we stroked them
we've waited a whole week whole week
before these cells are administered and
as low and behold you can see from this
graph there's significant improvement
that is observed over four months now
what is the mechanism of this recovery
well the mechanism is it's not so much
the mesenchymal stromal cell that's in
integrating itself into the brain what
it's doing it it generates and delivers
exosomes exosomes are nanoparticles
about 30 to 100 nanometers in size the
little lipid containers that contain all
sorts of goodies in there
synaptic proteins message RNA and more
importantly micro RNAs and then those
those exosomes are delivered to
astrocytes to the endothelial cells and
to the neurons to promote a repair
process now as you can see the
mesenchymal cell is the delivery
mechanism and not the FedEx truck that
you see and yet as you know good things
do come in small packages so what we
have here is one of those things that
inside that little package which I want
to mainly focus on four main Atok is
micro RNAs micro RNAs are small RNA
molecules about 20 nucleotides in length
which bind to message RNA and message
RNA as you know will go from message to
protein well bind to that message RNA
and inhibit translation of that protein
so what's interesting about micro RNAs
is you can have one micro RNA bind to
twenty different types of message RNA so
you have this extreme fine level of gene
regulation and that's also demonstrated
through life the more diversity you have
in micro RNAs the more complex your life
so the worm here the C elegans worm has
about 400 micro RNAs and the humans and
primates have about 3,000 micro rnas so
I found this very interesting because
when I looked at this I thought to
myself boy take away 2,000 of our micro
RNAs and we're just a worm so anyway so
anyway what we can do now instead of
injecting this mezzo karma cell into the
rat or potentially humans which we'd
like to see done is instead of taking
that cell injecting it which as you know
could promote some type of complications
coagulation problems why not just grow
these cells isolate the exosomes and
have the exosomes orchestrate
neurogenesis angiogenesis and neuro rate
remodeling so this is a different rat
but the same test an instead of
injecting mesenchymal stromal cells into
the
we went ahead and injected these
exosomes that we easily isolated in the
cell culture dish and lo and behold you
can see that they were injected one day
after the stroke and as you can see that
this recovery almost went back to
baseline moreover for those of you are
saying well rats a rat you know whatever
we this was done in a primate mato and
Boston University at by Tara Moore and
Doug rose and what they did is they have
a primate model where they can steer a
tactically induce a stroke just to
operate just the from the monkeys hand
induced a stroke right in his hand so he
has a deficit and then they administered
these exosomes and then you can see how
from these graphs that recovery occurred
twice as fast and recovery occurred
pretty much back to baseline so now we
can advance this technology now we have
these X's ohms and now what we can do is
we can transfect or we can take micro
rna's which we know which we know will
promote functional recovery and the one
micro RNA which we're going to focus on
is micro RNA 133 B 133 B has been found
to show improvement in spinal cord
injury models as well as Parkinson's
disease as a somehow affect a midbrain
dopaminergic secretion and what we found
is that when we take these mesenchymal
cells transfect micro RNA into these
cells into the mesenchymal cells they
will spit out these supercharged
exosomes
which have high levels of micro RNA 133
B and then we can go ahead and take
those exosomes and inject them into our
rat model now the the test on your right
no yeah and your left is a foot fault
test and what that is is that we put the
rat on a test tube rack and see normal
rat I'll be able to walk real easily
over the test tube rack where a rat
that's had a stroke is foot will
constantly fall through and what we can
do is we can quantify
by how many times his foot falls through
that rack and as you can see the
supercharged exosome with 133 be had
significant recovery over the control
X's ohms as well as the control
conversely on the other graph is a
modified neurological severity score and
this is a battery of tests where the rat
goes through a whole bunch of different
types of tests motor sensory
proprioception the rat is hung by the
tail and dropped and see if he lands
right he walks on a balance beam and
what you can see from that is the same
the same pattern you can see that there
is improvement over controls on all
these tests now how about we take that
rat brain and now we take that rap brain
and we take it out and suction it and we
stain from Milan because as you know
myelin is extremely sensitive to
ischemia so when you extremely sensitive
to ischemia
so which produces all these neurological
deficits that we can see and as you can
see in this in this slide that there is
pretty much twice the amount of myelin
density in the treated rats when
compared to control of the micro RNA 133
B and remember these are all X's ohms
which are given one day after stroke so
there's no time constraints so finally I
want to give the final experiment here
which was performed where we took the
chain reaction of exosomes and what we
did was we took X's ohms or we took
astrocytes and we grew them in oxygen
deprived conditions so this is kind of a
cell culture mimic of a stroke so we
grew these astrocytes and then what we
did is we took those message RNA
supercharged exosomes from the MSC s and
we incubated and with the astrocyte
and then after a while we were able to
isolate those exosomes
and lo and behold we found out
astrocyte kicked out even greater
quantities of these exosomes now one
thing you have to realize about
astrocytes astrocytes are considered the
mother of the neuron the astrocytes
wraps its foot process around the
neurons supports the neurons and cleans
up its messes after it clean cleans up
the neurons messes so what that does now
is you can show that the astrocyte when
it's gone under ischemic condition can
actually be helpful to the neuron and
promote this neuro restorative process
so as you can see when we take these
exosomes from the astrocyte
culture them with neurons you can see
that from this graph you can see that
the neurons have much greater growth
than compared to controls so in summary
we can take Bezeq comma strokes cells
supercharge them transfect them with the
micro RNA we that we know will help that
we'll know promote functional recovery
that will produce micro heroine and 133
supercharged exosomes
and that can treat the neuron or we can
go ahead and take that and incubate them
with astrocytes under these ischemic
conditions and she'll kick out even more
axes ohms with micro Naro 33b
to treat the affected neuron promoting
neurite increased neurite outgrowth so
in summary i just like to say that the
neuroscience laboratory is working hard
on this technology it's the forefront
we've also applying this technology to
brain trauma to dementia models to
hemorrhagic stroke models as well as
peripheral neuropathy due to diabetes
when we eventually hope to develop this
technology for clinical trials
hopefully here at Henry Ford Hospital
thank you very much
[Applause]