Hello everyone
We are going to Guam right now
by the way, why we have to departure at dawn?
because of flight time
Look at those wings
Hey Friends.
If I go up to the sky, I can find a bird
I prepared this headphone for the long flight
Are you exciting now?
Of course
See you later
I rode in an airplane to go to Guam
The flight will take off now
I feel strange
RIWON! We are in the cloud
The cloud is so beautiful
How do you feel?
Feel great!
Look at the beautiful sea
(Landing)
Daddy! Is that OK that the airplane is rained on now?
Now we arrived at Guam
Let's go to hotel
How was your first flight?
What is on the tree?
Fig?
Arrived
We are arrived at Dusitthani Hotel
By the way, The storm is coming
For more infomation >> RIWON's first flight to GUAM. Kids react. Travel video. Family Fun. RIWORLD - Duration: 4:36.-------------------------------------------
EastEnders fans confused by Ben Hardy's accent - Duration: 4:01.
EastEnders fans confused by Ben Hardy's accent
The former EastEnders star came on to chat to Holly Willoughby and Phillip Schofield about his role on The Woman in White.
But its his time as part of the EastEnders cast that made him a recognisable actor in the UK.
Since leaving the London-based soap in 2015, he jetted over to Hollywood and scored the role as Archangel in X-Men: Apocalypse.
Ben, 27, was born in Bournemouth but it seems viewers have never got to hear his real accent.
On This Morning, Ben sounded posh according to viewers, a far cry from cockney character Peter Beale.
One person tweeted: I dont remember Peter being this posh. Another said: Oh Im Ben Hardy.
Im very posh.
F*** off. A third wrote: Ben Hardys posh accent has taken me aback. While someone else added: Ben Hardy sounds very posh for someone who grew up in Dorset..
"Its a great job and theres a lot of security being at EastEnders" Ben Hardy Ben mentioned his stint in Soap Land, saying: Its a great job and theres a lot of security being at EastEnders.
It was a scary thing to [leave], but it was a leap Id have to take or Id have always been wondering what I couldve done. He did, however, have a bit of a gripe with the soap and the amount of times he had to take his top off.
Ben said: I was young and did what I was told back then.
There was a scene where I took off my hoodie, then took off my t-shirt, to then give her my t-shirt and put my hoodie back on.
I couldve just given her my hoodie..
His adversity to strip off saw him make a behind-the-scenes decision on The Woman in White.
Ben explained: I was presented with the option of nice Victorian pyjamas or to be topless and I chose the former.
I think it felt that for this piece it would have been a bit gratuitous, maybe. The Woman in White tells the story of Walter (Hardy) who meets a mysterious escaped asylum patient on Hampstead Heath.
He becomes determined to find out who she is and why she was locked up, as he thinks she was wrongly imprisoned.
The show airs Sunday nights at 9pm on BBC One, while This Morning airs 10.30am on ITV.
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Academy of Art University - W...
For more infomation >> Academy of Art University - W...-------------------------------------------
Audi A1 1.4 TFSI Pro Line S - Duration: 1:09.
For more infomation >> Audi A1 1.4 TFSI Pro Line S - Duration: 1:09. -------------------------------------------
Audi A1 1.4 TFSI Pro Line S - Duration: 1:09.
For more infomation >> Audi A1 1.4 TFSI Pro Line S - Duration: 1:09. -------------------------------------------
Home Video 2 2 - Duration: 27:21.
For more infomation >> Home Video 2 2 - Duration: 27:21. -------------------------------------------
真相大白了！公正党候选人爆出真相来了！原来根本都不是他的错，都是他们在搞鬼？ - Duration: 11:21.
For more infomation >> 真相大白了！公正党候选人爆出真相来了！原来根本都不是他的错，都是他们在搞鬼？ - Duration: 11:21. -------------------------------------------
La famille secrète d'Avicii : sa compagne dévoile son bonheur brisé - Duration: 3:24.
For more infomation >> La famille secrète d'Avicii : sa compagne dévoile son bonheur brisé - Duration: 3:24. -------------------------------------------
RIWON's first flight to GUAM. Kids react. Travel video. Family Fun. RIWORLD - Duration: 4:36.
Hello everyone
We are going to Guam right now
by the way, why we have to departure at dawn?
because of flight time
Look at those wings
Hey Friends.
If I go up to the sky, I can find a bird
I prepared this headphone for the long flight
Are you exciting now?
Of course
See you later
I rode in an airplane to go to Guam
The flight will take off now
I feel strange
RIWON! We are in the cloud
The cloud is so beautiful
How do you feel?
Feel great!
Look at the beautiful sea
(Landing)
Daddy! Is that OK that the airplane is rained on now?
Now we arrived at Guam
Let's go to hotel
How was your first flight?
What is on the tree?
Fig?
Arrived
We are arrived at Dusitthani Hotel
By the way, The storm is coming
-------------------------------------------
PSG: Unai Emery annonce son départ - Duration: 1:30.
For more infomation >> PSG: Unai Emery annonce son départ - Duration: 1:30. -------------------------------------------
Play doh numbers. Video for kids. Purple - Duration: 10:35.
Play doh numbers. Video for kids. Purple
-------------------------------------------
skribbl.io: Mal´n wa ma - Duration: 1:47:35.
For more infomation >> skribbl.io: Mal´n wa ma - Duration: 1:47:35. -------------------------------------------
STORYTELLING HIGH STRINGS ►NEW WAY◄ SYNTH FX STREET {FREE} Hip Hop Beat ☯️Sacred Beatz☯️ - Duration: 4:17.
For more infomation >> STORYTELLING HIGH STRINGS ►NEW WAY◄ SYNTH FX STREET {FREE} Hip Hop Beat ☯️Sacred Beatz☯️ - Duration: 4:17. -------------------------------------------
Guam #11 The last day of Guam trip, United airline [#26] - Duration: 5:55.
For more infomation >> Guam #11 The last day of Guam trip, United airline [#26] - Duration: 5:55. -------------------------------------------
To Life The Movie For more infomation >> To Life The Movie-------------------------------------------
唱跳实力兼备！D七少年团《Red Alert》练习室版及翻唱音源公开 - Duration: 2:32.
For more infomation >> 唱跳实力兼备！D七少年团《Red Alert》练习室版及翻唱音源公开 - Duration: 2:32. -------------------------------------------
La famille secrète d'Avicii : sa compagne dévoile son bonheur brisé - Duration: 3:24. For more infomation >> La famille secrète d'Avicii : sa compagne dévoile son bonheur brisé - Duration: 3:24. -------------------------------------------
13 HIDDEN UPDATE FEATURES THAT 'Clash Royale' DIDN'T TELL US! - Duration: 10:57.
13 HIDDEN UPDATE FEATURES THAT 'Clash Royale' DIDN'T TELL US!
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This Horribly Wounded Stray Was Hiding At A Stranger's House Then Rescuers Tried To Save Him - Duration: 5:32.
if there's a country with no shortage of stray dogs its India in fact the nation
is estimated to have more than anywhere else in the world in 2015 there were in
the region of 30 million of these street dogs
despite the spate and neuter efforts of various charities there are several
reasons for the existence of such large volumes of homeless dogs for one the
amount of garbage left on the streets ensures that food is plentiful for
strays another factor is the decline of vultures in India fewer vultures mean
that's more animal carcasses are available to feed Street dogs there's
also the fact that it's hard for animal neutering groups to keep up with the
amount of drunks in need of sterilization the result is that dogs
line the streets and this has unfortunate consequences for instance
the number of rabid dog attacks on humans is incredibly high in India a
study in 2017 predicted that 7 million Indians are treated for rabies annually
consequently a lot of people consider dogs as pests or vermin and as a result
it can be hard for strays to find out when they're sick or injured yet despite
this a lot of street dogs just seek sustenance affection and companionship
from humans one of the organizations working towards changing the situation
for Indian strays is animal aide and limited or AAU the Rajasthan based
charity is never short of canine patients in its hospital animal its
mission is to both bring relief to suffering animals and to awaken
compassion among people the group states on its Facebook page showing everyone a
path for action is where we begin and one example of its efforts relates to a
homeless drug that the organization called Jupiter Jupiter was found by some
kind-hearted citizens when he was hiding under the stairs in a stranger's house
no doubt they couldn't help but notice the gory injury that the poor dog had
suffered they were caring enough to call a AU and look after the dog until
rescuers arrived On February 20th 2018 AAA you uploaded a
video showing their first encounter with Jupiter it begins with two people
presumably the ones who had discovered Jupiter
a you to the house in question when the rescue team see the poorer dog his
wounds are both obvious and whirring Jupiter curls upon the floor with a
large bloody wound visible on the back of his neck he's either too exhausted or
too racked with pain to move when a rescuer approaches him even with the
agony that he must be enduring though Jupiter shows no signs of aggression the
wounded dog gingerly accepts the biscuits that a rescuer offers him and
Jupiter allows his new friend and pet him closer inspection reveals what looks
like a makeshift collar tied around his neck but whether it's a mark of
ownership or abuse is anyone's guess on February 21st 2018 AAU posted an account
of the incident on Facebook his trusting response to his rescuers made us wonder
if on some level he was just waiting for help it read his sort of fool eyes told
us everything we needed to know about his pain in the Facebook post the
rescuer is very conscious that Jupiter needs help fast the dog is at risk of
dying without emergency treatment next the post footage reveals that Jupiter
doesn't even object when his rescuer picks him up and carries him out the
building the AAU staff then carefully puts you but in the back of their truck
when the team drove Jupiter back to the Animal Hospital
however they realized just how serious his wounds were the dog was closer to
death's door than they had thought Jupiter's injury was bigger than initial
impressions and in fact stretched all the way up to the base of his head the
veterinarians concluded that the wound was most likely a bite from another dog
furthermore they were counted on Facebook how the cut was infested with
maggots the AAU staff stated now it's time for this boy to start healing
doctors began by taking steps to stop the infection yay you described the
process on Facebook as to breeding the wound flushing removing maggots they
also applied a drip to help with Jupiter's recovery and bandaged up his
need nagesh it wasn't the last time but the vets would need to perform such a
procedure either indeed they would repeat it many times afterwards and the
rescue video reveals that Jupiter was well looked after in between his
treatments in the post footages viewers can see a carers at two
as he enjoys a good meal the road to recovery was a long one for Paul Jupiter
though and it took six weeks for him to heal but under the tender loving care of
a a you ended stuff the dog's wounds eventually closed in the next time but
the video shows him Jupiter looks like a completely different dog he seems happy
and healthy and appears to be enjoying all the pets and kisses that he's
receiving from the AAU staff Jupiter's gentle nature is obvious in the way that
he rests his head on his carers arms when a a you first met Jupiter of course
he was close to death this sweet boy might have stayed hiding until he died
the staff had stated but Jupiter seemed to know he was on the path of healing
from the first touch by the rescue team hey you added on Facebook he's
definitely a new boy in an angel through and through I hope you liked this video
then please hit the subscribe button in play con bell icon to never miss the
video from our Channel
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কিলোগ্রাম - Duration: 14:45.
For more infomation >> কিলোগ্রাম - Duration: 14:45. -------------------------------------------
What's The Holiest Place On Earth - Duration: 3:31.
What�s The Holiest Place On Earth
by Conscious Reminder
Different people have predicted the apocalypse on different dates.
It was on one such D-day when all the planets had aligned perfect, when all these predictions
were going to come true, and when all those with spiritual powers were saying that life
would never be the same again.
Many were trying to get to place that are powerful spiritually so that they could stay
safe.
There were also crowd spending all their money and then some because they thought the economy
was going to come crashing down anyway.
Others were bidding a sad farewell to those close to them.
I was travelling through Europe, completing a spiritual pilgrimage with some other people.
One small girl who was just 8 years old was scared because she had been told that it was
Doomsday.
She asked our guide if all of us could go to a place where we would be safe from all
kinds of danger.
The guide merely took her hands and told her that she herself was a place of safety and
power.
As long as she dwelt on her own spirituality, she would be protected no matter where she
goes.
Those words will always stay with me.
I�ve traveled quite a lot and I�ve been fortunate enough to see the most magnificent
holy places that the Earth has to offer.
I�ve seen the Pyramids in Egypt, Machu Picchu, the vortices of Sedona, Mt. Shasta, Mt. Fuji,
Jersusalem, and other spots we have deemed holy.
I have loved all the pilgrimages I�ve been on and each place has inspired a sense of
awe and wonder within me.
But when I was reading �A Course in Miracles�, a particular line made me stop and think.
It read: �The holiest spot on earth is where an ancient hatred has become a present love.�
The book says that because we are spiritually aware, our geographical location does not
matter.
The condition of our spirit is what makes us stronger and protects us.
Wherever we can feel God is a Holy site.
We can reach awakening by strengthening our relationships and replacing all negative emotions
with love.
In all our pride, we believe that if just make some changes, everything will work out.
But there is no point quitting your job without sorting out the problems that are driving
you away.
They will just follow you to your new job.
These external changes won�t work unless we change the way we think.
After we have healed ourselves, everything else will fall into place.
The world will appear to us as we view it.
If we believe that there is safety and kindness in this world, that is the world we will find.
All reality begins in our imaginations so it is up to us to choose what we project.
It is definitely a one of a kind experience every time you visit a new sacred spot but
rather than trying to find enlightenment in far off lands, we should be looking for it
within ourselves.
-------------------------------------------
Affaire Jeremstar : Pascal Cardonna promet de nouvelles révélations sur Snapchat - Duration: 3:26.
For more infomation >> Affaire Jeremstar : Pascal Cardonna promet de nouvelles révélations sur Snapchat - Duration: 3:26. -------------------------------------------
EPA Looks Other Way In Environmental Racism Cases - Duration: 7:40.
A judge recently ruled that the EPA had improperly delayed investigating toxic pollutants from
power plants that were affecting minority communities all over this country.
It's what's commonly referred to as environmental racism.
When I saw this article on this, Farron, the thing that startled me the most is these environmental
racism kinds of cases, they surround so many parts of our society.
In other words, in our town, in Pensacola, Florida for example, they built a sewage plant
right in the middle of an area where it's the highest minority populations.
They do this all over the country, whether it's a Superfund site.
They will build right next to high minority population, whether it's a power plant, that's
spewing all types of chemical in the environment, whether it's an oil refinery.
This is not just our town.
Probably if somebody's watching this and they drove around, they would find the same thing
exists in their city.
When you read this, what was your reaction?
Absolutely.
I think this issue of environmental racism is something people don't understand.
They don't understand that it is the minorities that are most affected by these corporate
pollutants.
It is the poor communities of any color as well, but this particular case was brought
because the EPA has a duty, according to the Supreme Court, to investigate any kind of
claim of environmental racism or any kind of pollutant.
They only have a matter of days to do it.
180 days.
180 days.
20 years in some of these cases that were just heard.
20 years it had been going on before the EPA even, well they never looked at it.
They'd been sitting there for 20 years, so the judge in this case says, "No, you absolutely
have to do this."
This is, they said it was environmental racism under Title VI of the Civil Rights Act, a
violation of the Civil Rights Act because they were essentially targeting, the corporations
targeting these communities and then the EPA turning a blind eye towards it because they
don't want to have to look into it.
Okay.
This stretched, Clinton, Bush and Obama.
Yeah those were the administrations.
Republican, Democrat, you know I get so tired.
God Almighty, I get so tired of getting these emails from these emails from these Democrats
who believe or Republicans who believe that their party is the solution.
It's both sides.
Yeah.
Obama was just as bad as Bush.
Bush was just as bad as Reagan.
Reagan was just as bad as Clinton on this.
25 years this has been going on.
Now look, there's nothing equivocal about this statute.
The statute addresses this.
It says, look, when we look at how, where we're putting these problematic type issues,
oil refineries, sewage plants, Superfund sites, power plants, what we're finding, and this
is what, this is why this started.
They were finding cancer clusters.
They were finding asthma and respiratory clusters all around these kinds of places.
They're finding neurological clusters.
They were finding, they were finding leukemia, non-Hodgkin's lymphoma around these kinds
of areas, so they said this is more than just environmental racism.
This is absolute medical health racism.
The thing that struck me about this story, to me that struck me the hardest was the EPA
comes back and says, "Well, you know we didn't do anything because they were moot issues.
We took care of them."
Well they didn't take care of them sometimes for a decade.
Under the law, you have something called a writ of mandamus that we've talked on this
show about.
Writ of mandamus or writ of provision is when you file a lawsuit and say this agency isn't
doing what they're supposed to do, and as a result, we got to do something about it.
It is again, the EPA is ordered to do these.
During the Bush administration, the EPA was, much like it is now, not doing their job at
all.
Yeah.
At least under Obama they gave us a little kind of appearance of doing their job.
They did it enough to not get in trouble basically, but during the Bush years, they weren't doing
anything.
They weren't investigating anything.
They weren't enforcing Clean Air or Clean Water Act rules, so the Supreme Court, under
Republican rule, ruled that hey, you have to do your job.
You have a constitutional authority, ability and duty to go out there and enforce the rules
of your agency and to fulfill them to the best of your ability.
Here, we've seen that, that decision came out while all of this was going on and the
EPA still did nothing.
I mean some of these are from 1994, is how far back these environmental racism cases
go.
It's for, these wood-burning factories that just spew pollutants into the air.
You've got coal ash facilities that are highly toxic and nobody is even talking about the
effects of toxic coal ash.
Right.
They target these communities.
They really do.
You're not seeing this in the gated communities.
Look at Flint, Michigan.
I was just going to go to Flint, Michigan.
Why did it happen in Flint, Michigan?
Why did they get the poisoned water?
Because they had no political clout, because they couldn't go meet with the mayor.
They couldn't meet with the City Council.
They couldn't meet with the House of Representatives or the Senate.
These folks can.
Exxon can go do that.
Monsanto can go knock on the governor's door and say I want to talk to you about this.
What is the access of these ... What's the possibility of that kind of access?
Here's one disappointment to this case.
To me the case really was about pattern in practice, okay.
Because you could take what happened in any one of these cases, whether it was Flint,
whether it was Texas, California, didn't make any difference.
This plays out all over the country, and it always ends up that it's about access.
It's about influence.
It's about affluence.
For us to look at it and say, oh gee.
There was absolutely no reason why Obama, of all people, in that last administration
couldn't have taken this, because there's still a ton of these cases still out there
and the EPA is still taking no action on it.
It just ... You know a crazy thing?
Now, full disclosure.
I'm not Republican.
I'm not Democrat.
I'm an Independent, have been for 35 years, okay?
There was a time though, think of who put in place the EPA?
Richard Nixon, of all people.
I mean, Tricky Dick gave us the EPA, all right.
Ronald Reagan took it away, but the point is, you can't look at parties and say, "Oh
this party's better than the other party," because it simply doesn't work out like that.
The math doesn't work.
That's really interesting to point out too, because there was a time when Republicans
were the, the conservation party.
I mean, Teddy Roosevelt was huge.
Right.
On preserving and protecting the environment.
Nixon came along, gave us this.
Gave us the Endangered Species Act.
But today, it's what can we give the corporations?
Where can they put their toxic sites?
They do it a lot of times, too, they offer them big tax breaks.
Yes.
To go put it in the poor ...
To put more pollution there.
Yeah, yeah.
Keep it away from the wealthy folk, and you put it over here, and we'll give you money.
Yeah.
To build in these communities.
Yeah, pretty interesting twist on politics there.
-------------------------------------------
To moon - Original - Duration: 3:20.
This is for when you are not with me.
For when you fly above the sea.
Then I'll remember what you said.
Look at the sky so we will be..
Under the same moon.
The same moon.
I've loved you since that I was two, we still have lots of things to do.
More kitchen shows, and games, and jokes, so look at the sky then we will be...
Under the same moon, the same moon..
The distance won't make me forget, how much I miss our roller skates.
the distance won't make me forget...
Calle Princesa y Carders...
This is for when you are not with me.
For when you fly above the sea.
Then I'll remember what you said.
Look at the sky so we will be..
Under the same moon.
The same moon.
the same moon,
the same Moon..
-------------------------------------------
Cette huile est miraculeuse : elle fait repousser les cheveux et les sourcils - France 365 - Duration: 8:40.
For more infomation >> Cette huile est miraculeuse : elle fait repousser les cheveux et les sourcils - France 365 - Duration: 8:40. -------------------------------------------
RIWON's first flight to GUAM. Kids react. Travel video. Family Fun. RIWORLD - Duration: 4:36.Hello everyone
We are going to Guam right now
by the way, why we have to departure at dawn?
because of flight time
Look at those wings
Hey Friends.
If I go up to the sky, I can find a bird
I prepared this headphone for the long flight
Are you exciting now?
Of course
See you later
I rode in an airplane to go to Guam
The flight will take off now
I feel strange
RIWON! We are in the cloud
The cloud is so beautiful
How do you feel?
Feel great!
Look at the beautiful sea
(Landing)
Daddy! Is that OK that the airplane is rained on now?
Now we arrived at Guam
Let's go to hotel
How was your first flight?
What is on the tree?
Fig?
Arrived
We are arrived at Dusitthani Hotel
By the way, The storm is coming
-------------------------------------------
Play doh numbers. Video for kids. Purple - Duration: 10:35.Play doh numbers. Video for kids. Purple
-------------------------------------------
WEBINAR: Inner Ear Gene Therapy – Recent Advances and Clinical Perspectives - Duration: 1:00:50.
Good evening, everyone. Thank you so much for joining us tonight for Inner Ear Gene
Therapy Recent Advances and Clinical Perspectives with Dr. Lukas Landegger. Thank you so much,
doctor, for joining us tonight. We appreciate it.
Before we get started, I want to say thank you to Darcy Kriens of Alternative Communication
Services for providing CART tonight. Everyone should have captions available right now,
if you click the closed caption button at the bottom of the screen, it should turn green
and you should see captions.
Tonight, you can hit the raise hand button if you have a comment or question, but I would
really prefer that you use the Q & A if you have a comment or question for me, and that's
the same place where you can pose a question to Dr. Landegger at the end of his presentation,
and I will post those questions verbally to him so that becomes part of our CART transcript.
Dr. Landegger earned his medical degree from the Medical University of Innsbruck in Austria.
He served as a military doctor in the Austrian Army. He joined the Molecular Neuro otology
Biotechnology Laboratory at Mass Eye and Ear in 2013.
Two days ago, Dr. Landegger completed the Boston Marathon, so I'm anxious to hear more
about that and I would not even be walking if I did that two days ago, but I'm really
glad that you're here alive and well and you finished, and I can't wait to hear about that,
too. So go ahead, and I'll let you get started.
>> LUKAS LANDEGGER: First of all, good evening. Thank you so much to everybody involved in
the organization of this talk. Thanks to the Hearing Loss Association of America for giving
me this platform. While I was preparing this talk, I kind of really had to figure out who
the audience was, and so primarily it will be patients from my understanding, however,
there will be some professionals in the audience as well, so what I try to do with these slides
is that I have a lot of information in the slides themselves. However, I really try to
walk the patients through it and highlight some of the most important information in
red so that the professionals can kind of follow up on this information and kind of
go through the primary sources, so go to all these papers that are mentioned on the slides.
I still want to give guidance to the patients, so as patients don't be overwhelmed by the
slides. I really will try to walk you through it step by step.
In this talk I'll try to present, obviously, inner ear gene therapy studies that have been
done here in our lab and other labs around the world. We'll try to just present some
of these most recent advances and give you an idea where we stand and when these potentially
could become available for clinical use. So to give you an overview of the talk we'll
first start with the definition of hearing loss and what the current standard of treatment
is. Then we will talk about inner ear gene therapy or gene therapy in general. Then we'll
briefly discuss different approaches and how genes could be targeted, primarily viral vectors
but also CRISPR Cas9. Some of them might have heard about the gene scissors that have been
mentioned in a lot of newspapers recently. We'll discuss certainly mouse models that
have been rescued, so mouse models of human disease that can be rescued with gene therapy,
and we'll briefly talk about stabilization of cells versus restoration of cells, which
is particularly important for age related hearing loss. The last part is what are the
hurdles on the way to the clinic as we said. Hearing loss, as we all know not as we all
know, but we know it's a big problem and it's the most common sensory deficit in humans.
Just last month the World Health Organization released a fact sheet on deafness and hearing
loss, and these numbers are really incredible if you haven't seen them before the estimate
is over 5% of the world's population has disabling hearing loss, and this number is expected
to basically double up until 2050 with about 900 million people being affected then. In
addition to that, about 1.1 billion young people are at risk of hearing loss.
And the reason for hearing loss are multifold, and can include genetic causes, complications
at birth, infectious diseases and ear infections, certain drugs especially chemo-therapeutic
drugs such as cisplatin or aminoglycosides which are certain types of antibiotics, noise,
aging, et cetera. An overview of how many patients are affected per life decade, you
see newborns at about 0.2%, school children goes to about 0.4% and then 16% in over 18
year olds, 34% in 65 to 69 year olds, and eventually we go up to 72% in octogenarians,
which is really remarkable. To understand hearing loss, we have to first
understand how hearing works in general. We have sound that comes in through the external
auditory canal, hits the eardrum, and the sound is then transmitted through the ossicles,
the smallest bones in the human body, to the fluid filled inner ear, specifically the stapes
or stirrup, that's the smallest one in the human body and connects to this oval window,
and then a fluid wave is created, but it's mechanical transduction and kind of tilts
or deflects the hairs of certain cell types that are called hair cells.
Once these hair cells or these hairs deflect, the stimulus is changed from a mechanical
stimulus to an electrical stimulus, and that electrical stimulus is then forwarded by the
auditory nerve to the brain where we then actually hear.
In this schematic, you see the cochlea where we hear, which is the snail shaped structure
here, and then we also have the vestibular part of the human inner ear that is responsible
like is the balance part of the human inner ear and you see the semicircular canals here
important for rotational movements, and we can see the utricle and saccule, altogether
five organs. They're responsible for acceleratory movements, either horizontally or vertically.
Then we can, based on where the hearing loss occurs, we can divide these types of hearing
loss into either conductive or sensorineural hearing loss.
Conductive loss means everything to this oval window here basically. So all the bones and
canal, et cetera, and we have multiple treatment options for that. However, the sensorineural
part of the system, the cochlea or inner ear and auditory nerve, right now it's basically
just the cochlear implant, although it's a really remarkable device and we'll talk about
the kind of a lot of pros, but the few things that are not perfect yet regarding this device
later in the talk. Another thing that could be used is an auditory
brainstem implant in case the nerve is missing, but we won't really talk about that in this
presentation. So the current treatment options are hearing aids. You're all familiar with
those and what they do. There's a lot of models, but we won't talk about this in this presentation.
Basically they simply amplify sound. Then we can change the sound transmission
itself and here so called middle ear prostheses or middle ear active implants are important
because for example, if we miss the ossicles, we can replace them by titanium prostheses,
so it's relatively straightforward, or these middle ear active implants.
What they do is they have a little microphone here behind the ear, and this microphone detects
all the frequencies that come in and then filters these frequencies and you have a little
magnet in the middle ear that attaches to the ossicles. And through the different frequencies
that are detected, the number of vibrations or kind of the vibration intensity itself
is determined, and that can then, again, facilitate hearing.
Then cochlear implants, you might have heard about them or even have one of them, and they're
really remarkable devices. What you have is, again, this outside microphone that detects
the different frequencies, and instead of manipulating the ossicles, what this device
does is that you have a coil that goes into the snail shaped structure, so into the cochlea,
and this device has certain outlets where you can kind of shoot electricity out that
directly stimulates the auditory nerve, and so you kind of circumvent these hair cells
that usually make the transition from the mechanical stimulus to the electrical stimulus.
It's really great that this actually works. What is gene therapy? So the idea is that
you kind of introduce normal genes into cells, and with those you can then replace missing
or defective genes so that the cells work again as they should. There's about 100 genes
that cause non-syndromic hearing loss that could potentially be targeted with gene therapy.
When we compare our field - the field of ENT, you know, ears nose and throat, to ophthalmology
it's relative sad because right now there's about 30 gene therapy trials for 10 diseases
of the retina (so the structure in the back of the eye); however, there's only one for
severe to profound hearing loss going on. The ophthalmologists already have one drug
FDA approved since December of 2017. So we really have to catch up in this way.
This is an image that shows how a virus transduces a cell, so how a virus gets taken up by a
cell. This is an adenovirus. Most of the talk is about an adeno associated virus, which
is a relatively similar virus, and the main difference is with adenovirus you can cause
a transient expression of the gene, with adeno associated virus usually the gene you want
to replace stays there pretty much until the cell dies, so pretty much forever. At least
that's the assumption right now - at the moment we don't really have the data in clinical
trials that show how long. There have been studies that have shown this expression for
several years over the decades. And this adenovirus attaches to the cell and
is taken up by the cell, and the virus itself cannot replicate or double itself basically
so what it needs is the machinery of the cell to actually kind of – yeah, replicate itself.
So what it does is that it delivers the gene that it has been loaded with into the inner
part of the cell, and then the effects take place that we could then potentially use for
the gene therapy. So what we briefly have to talk about genetics
as well, because those have implications for therapy, and so basically every human gets
one gene, so it has two copies of a certain gene. And one is received by the mother, and
the other one is received by the father. And if just one of those two has to be affected,
then we are talking about a dominant disease. So one of the two has to be affected so that
the patient has the disease, and then it's a dominant disease. However, if both have
to be affected, then it's a recessive disease. So dominant would mean my father gives me
a faulty gene, my mother gives me a healthy one = disease. Recessive disease means both
my mother and my father have to give me the faulty gene, so I have the disease. That is
important because that means how we can or that playing a role regarding how we can treat
these diseases. So the simple gene addition would be relevant
for a recessive disease, because in recessive cases, as we've just discussed, we basically
have no functioning gene, and so the delivery of a copy of a normal gene could then just
kind of make the cell function again. That's what has been done in most animal studies
so far. However, in a dominant case we can also use
so called gene we cannot use the gene addition therapy or strategy, but here we'd have to
use a gene disruption therapy because here we have this dominant gene that will independently
if there's a normal copy will still produce the faulty kind of protein, the faulty mechanism,
and so we kind of have to target that somehow. So by targeting that, the remaining gene can
then take over. And the last part is the gene editing part
where with a recessive or dominant missing, that's a completely irrelevant mutation. If
you have a minor part affected, you can then use a so called CRISPR Cas9, so the gene scissors,
again, to replace that specific part of the gene. The functional gene addition is also
something that we'll skip for this talk. So how would the gene therapy work specifically
for the inner ear? Well, we've talked about like how the hearing works, and so if we cut
a piece of the cochlea out here and look into it, then we see three fluid filled chambers.
The names are irrelevant, but in the scala media, we have the inner hair cells and three
rows of outer hair cells. One row of inners and three rows of outers. The inner hair cells
make this transition from the mechanical stimulus to the electrical stimulus, while these outer
hair cells have more of an amplifying function. And in one human ear we have about 15,000
hair cells, and it's the same cell type in the vestibular system, so the five organs
we discussed before with the utricle and et cetera. All of these have these stereocilia
on top and the hairs give them their characteristic look and name and we'll see a few pictures
afterwards. What's interesting in birds, for example,
these cells regenerate without any problems, while in mammals including humans they don't.
There's just one exception. In 2014 researchers found out in neonatal mice, newborn mice,
some of these hair cells do regenerate, and here supporting cells come into play. We'll
talk about them later. Those are the cells right next to the hair cells here.
And one potential surgical approach for gene therapy is the round window here. That's also
the place where usually the cochlear implant is introduced, at least nowadays is introduced.
So that would be a relatively straightforward process for the injection.
We've talked about the success story of the cochlear implant that has really enabled so
many patients to actually learn language and everything without any problems, however,
there's still some issues, namely the natural sound perception (so sound perception in general
is not that great), frequency sensitivity (enjoying music, for example, is relatively
hard), and speech discrimination in noisy environments. In a bar, for example, with
a lot of background noise, it's really hard with these devices to actually filter out
that one voice. The goal of gene therapy is to kind of restore natural hearing.
And this is the mouse inner ear, the mouse cochlea. It's relatively similar to the human
ear. We have the turns up here where we hear with the hair cells, and we have the oval
window here and the round window that could be used for gene therapy and is being used
for gene therapy by a lot of groups. It's right here. I'm sorry, I'm just trying to
remove this bar here from my screen. Okay. So the first study that I'm trying to
or I'll present is the VGLUT3 study, and it's one of the first rescue studies in mouse models.
This VGLUT3 is a receptor in inner hair cells and that's what we discussed in the last few
slides. It's right there at the connection between inner hair cells and the auditory
nerve. Mice lacking this transporter are actually
deaf, and it's only relevant in a few human patients but what makes this model interesting
or what made these models interesting is these adeno associated viruses that I mentioned
before actually specifically target these inner hair cells, so that's what needed here.
What's a relevant fact is these mice in general usually develop hearing around two weeks after
birth, whereas, humans are born with hearing. So that is interesting in terms of timing
afterwards because potentially that would mean that if a mouse is injected immediately
after birth, in humans we have to go into the womb or like the baby would have to be
injected in the womb. And if we see that these mouse pups who are
lacking the transporter are injected with VGLUT3, then they actually did not go deaf.
So this is a slide that or a figure that will be used in several of these studies, and what
you can see here on this Y axis is the loudness level. Down here it's like a whisper basically,
and up here it's like if a jet engine would start right next to you, so very loud.
These curves that you can observe are functions of basically an objective hearing test, so
once you see the curve, you can tell that the mouse heard that. You can see in wild
type mice, regular mice, healthy mice, we have these curves somewhere around here. In
knockout mice, the mice affected by this disease where the transporter is not there, we don't
have any response at all, so those are basically deaf. In these rescued animals where the VGLUT3
was injected with a virus, we can actually get responses again. It means that these mice
heard because the VGLUT3, the specific transporter, was introduced into the inner hair cells,
and in a different way that is depicted here, you can see 95 decibels is really, really
loud - no response at all while the wild type animals and the injected animals are actually
pretty good. So that was a really remarkable paper that came out in 2012.
What other functional rescue studies have there been? Quite a few published in the last
few years. However, what is common or what is like the same in all of them is that they
all talk about the inner hair cells and outer hair cells are really hard to target and you
can see that in these images where you can appreciate that all of these viruses kind
of just target the inner hair cells while the three rows of outer share cells that should
be out here somewhere remain dark. What is that dark part? So everything that's green
lights up with so the lighting up means that it's GFP, green fluorescent protein. If a
virus transduces a cell, gets into a cell, you can tell what it expresses, and basically
it expresses this green fluorescent protein and then the cell lights up. That is something
that researchers can use to determine where the vector goes.
For the rest of the talk, you can remember green cells are good, because that means that
the virus got into the cell, and we could potentially deliver something into the cell
including the healthy gene. So in this 2011 study, you can see that they tried five different
serotypes, and for all these adeno associated viruses, these inner hair cells were the most
effectively transduced cochlea cell types. So it's good that it worked in this specific
disease model before, but the difficulty is to get these outer ear hair cells I mentioned
before. Here a virologist comes into play, Luk Vandenberghe
and the institute affiliated with it comes into play. His lab looked at a computer model,
synthetic AAV, so they looked at the predicted ancestor of adeno associated virus types 1,
2, 8, and 9 and those are commonly used viruses. What they were looking for was that ancestor
because everybody has had a cold, and this adenovirus is actually the common cold virus.
The adenovirus and adeno associated viruses are relatively similar, so our immune system
actually recognizes the similarities and neutralizes some of these adeno associated viruses immediately
before they can target the cells they should target.
His lab hypothesized that this novel ancestor, predicted ancestor could actually circumvent
this pre existing immunity. They did a lot of high throughput screening, in vitro, in
vitro means in the petrie dish, and injected them into mice. So you can see AAV2, AAV8,
so these conventional adeno associated viruses in these columns, and Anc80 is the new virus
and you can see expression in the liver, muscle and retina, and Anc80 seems to outperform
all these other conventional adeno associated viruses.
But nobody had tried that in the ear, so we did that in the ear and we tried it on something
called cochlear explants. These are like microdissections of these mouse inner ears, and we can grow
them and culture and add certain viruses or whatever we want to them and see what they
do. Then in red, you can see hair cells, so a
specific hair cell marker. In blue you see a marker for neural structures, and in green
we have, again, this GFP, so the green fluorescent protein, which means that the virus went there.
Every column here represents one of the conventional viruses, and here the last two columns on
the right are Anc80, so that's this new virus. What you can see is that Anc80, so this new
adeno associated virus, really outperforms all of the other adeno associated viruses,
and we were very happy when we saw that for the first time. Specifically, it was not only
at the level of inner hair cells but also outer hair cells and supporting cells. I'll
show that to you in a few slides later. You can see that these micro-dissections are
really tiny, so it's really my thumb next to it. You see these white, little dots, those
are the explants. So mouse inner ears are not really very big. We then also tried this
together with or this work was primarily done in Jeff Holt's lab at Harvard Medical School.
They injected mice with the virus, the most promising vectors we identified in vitro in
the Petri dish. Anc80 still outperforms all the conventional viruses. They did a bunch
of studies regarding how the uptake of the virus would change the cells, and it was normal
in terms of how the cells reacted and how these animals then heard and then had these
objective hearing tests in a way. Again, for you to appreciate how tiny everything is,
we have a mouse pup here, and that's where you the injections then take place, so you
have to expose the round window back there. It takes about 10 minutes per pup once you
establish the approach. Some more images of this GFP expression. You
see along the whole length of the cochlea, we get a lot of viral expression in inner
hair cells and also the outer hair cells that could not be reached with the conventional
adeno associated viruses are finally transduced with Anc80 with a maximum follow up up to
a month and the expression was stable which was interesting because mice live between
2 to 3 years, something like that. It depends on the strain. So a month is a pretty substantial
amount of time. With the in vivo injections we showed that
from the very apex or the very top of the cochlea up until the very base of the cochlea,
so up all the way to the bottom of the cochlea, so throughout the cochlea we had a lot of
expression of this virus, which is very promising, and it was so strong that we sometimes even
saw it on the other side of the ear. Then we were wondering, how does it actually get
there? For that we used brain slices of the mice, so we cut the brain like this. You can
see the A here, so the section is called an axial section with this human head. For the
mouse in front here, we have the snout while here we have the cerebellum, which is the
posterior part of the brain back here. You can see that this is where we get the predominant
GFP expressions of green fluorescent protein expression, and you can see that these cells
here take it up, and so what we think is what happened is that the virus travels to the
other side through a structure that's called the cochlear aqueduct, which is a connection
between the fluid of the inner ear and the cerebrospinal fluid - that's the fluid where
the brain and spinal column kind of swims in it. That is actually known in rodents that
this is relatively patent, so we have to figure out how far that is translatable to larger
animals, because that would be another hurdle until it goes into clinic because you want
to avoid that it actually goes into the brain. We also looked at the vestibular system and
what you see in mouse tissue that we get a lot of green cells, so it's very positive
- we can actually reach them and what's very interesting is it was done by collaborators
in London, we also got it into human tissue. In some surgeries you actually have go through
the inner ear, and what they do in this case or to resect certain tumors you have to go
through the inner ear and you can get access to the precious human tissue. You see that
we have this excellent expression also in human tissue, so it seems to be a promising
candidate for clinical studies. Excuse me. This is another study that used Anc80 in adult
animals. In 7 week old animals they were injected through the posterior semicircular canal.
That could be an approach used in humans, specifically the lateral semicircular canal
that seems to be accessible. What you can see here is that, again, in these
adult animals that couldn't really be transduced at all until now with this new virus, you
get a lot of outer hair cells at the apex, not so much at the base (not so much at the
bottom of the cochlea). Still relatively promising. Other collaborators at the Harvard Medical
School used this virus in a model of Usher syndrome. That was Holt's group and Géléoc's
group. What Usher syndrome is, it's the leading cause of deaf blindness and is inherited recessively.
Both genes have to be affected then. What you can see in this figure is that it's a
scanning electron micrograph, so it's the highest resolution of the the highest resolution
of basically the common microscopes that you can see.
What you can see on the left side here is a wild type animal, so a healthy animal. What
you see in the middle is a diseased animal, so that's an animal with Usher syndrome. What
you see on the right side here is an animal that has been injected with this Anc80 that
had had protein that is missing had this gene that is missing in these animals, and you
can see that the hairs to the stereocilia that give the hair cells their name, are really
similar to these wild type healthy cells compared to the Usher cells.
So that was really remarkable these cells could be rescued to such an extent. They performed
many additional experiments and specifically performed these hearing tests, objective hearing
tests again and what you can see here is that the control animal, so the Usher animal with
the disease, did not show any thresholds at all with the hearing, so the louder it gets,
deaf animals no responses, while here on the right side - the animals that were injected
did actually have relatively nice hearing. For some of them the hearing was as good as
for wild type animals, as for healthy animals. Several other studies have come out since
then and like before them or around the same time, and you they targeted different Usher
models, so there's a lot of different subtypes. They all showed most of them showed rescue,
however, none of them was as substantial as with this novel virus presumably because the
standard adeno associated viruses did not target, or cannot target, the outer hair cells.
Are there any other approaches that allow viruses to target more hair cells? Yes.
Another lab here at Harvard is working on something called exosomes. They're small vesicles,
and for patients I would describe them as "bubbles filled with information" and
secreted from cells. They enable communication between cells. These other cells then take
exosomes up to process it, and these viruses have hijacked this approach to actually get
into the cells more easily. What this lab is doing now, and a very clever
strategy, is they package these conventional adeno associated viruses into these exosomes
and can target more cells with this approach. Then this CRISPR Cas9, these gene scissors
that we briefly had mentioned before, in a very interesting study that was published
at the end of the last year, they looked at they had a mouse model that lacked Tmc1. That's
something I'll explain in a minute. I want to show you this here in this frog
hair cell you see these stereocilia, so the "hairs" of the hair cells. What you can
see here is something called tip links. These are the little bridges between the "hairs"
of the hair cells and this Tmc1 and this transmembrane channel-like gene family 1 is part of this
gene complex here that is important to cause the kind of or the movement of these hair
bundles. In 2002 a study group created a mouse with a TMC mutation, and it showed that it
led to slow degeneration of the hair cells. They named this mouse Beethoven mouse, which
is not a great name because the composer had a completely different type of deafness. This
mutation is also relevant to humans and has been described in a Chinese family.
What these researchers did is they injected so called Beethoven mouse pups and compared
them to controls and this protein RNA complex, so CRISPR Cas9, targets only the affected
copy of the gene without influencing the other gene. If we can get the affected copy of the
gene, the diseased copy of the gene kind of out of the way, then in theory the normal
gene should take over and the cell should be functioning again. That's what these researchers
hypothesized. What you can see here on the right side, so
again, we have inner hair cells here, and then three rows of outer hair cells out here
all along the length of the cochlea. On the right side you see all the cells are still
viable, while here on the left side the uninjected animals, so these Beethoven mice, do have
degeneration at the lower part of the cochlea or the apex (or the top) is kind of still
there. Then these injected animals actually have preserved cells all throughout the cochlea.
So it was a substantial rescue after the injection. Then the researchers obviously again assessed
the hearing, and what you can see here, again, loudness on the Y axis - so loudest level
up here, very quiet level down here. What you can see is that these injected animals
actually heard better than the uninjected animals - deaf in some frequencies at least.
However, the rescue was not as good as for the uninjected animals, healthy animals. However,
this study is really proof of concept that this gene disruption be might be a potential
strategy of treatments of some form of this dominant hearing loss.
So far we only talked about stabilization. What about restoration actually? That is a
different an interesting difference between genetic hearing loss and age related hearing
loss because for age related hearing loss, we probably need a mix of gene therapy, molecular
therapy and stem cell therapy or just focus on yeah, there's a lot of overlap between
the three fields. As I said before in some of these newborn
mice, it is actually possible to make a transition from the supporting cells to hair cells, some
of these mice still regenerate hair cells. And usually that's through the switch of supporting
cells into hair cells, and that's been studied extensively and several different signaling
pathways have been identified by researchers all over the world.
In a relatively recent paper -- and that is lost after like the maturation of the mice
- so in adult mice, you cannot transition supporting cells to hair cells anymore. However,
in a relatively recent paper that also came out last year, a group actually tried to combine
several of these factors, and then was able to make the switch from supporting cells to
hair cells, so you can see here in blue the hair cells, the inner hair cells and three
rows of outer hair cells and here after noise damage they would be lost and could be regenerated
after the mix of different factors. So this group shows that for the first time in adult
animals, which is very relevant for age related hearing loss. The only clinical study at the
moment that targets or that is focused on gene therapy in the inner ear that I mentioned
before is actually targeting ATOH1. That's a study that the lead investigator,
the principal investigator is Hinrich Staecker in Kansas, but they also have sites at Johns
Hopkins in Baltimore and at Columbia in New York.
So to summarize, gene therapy is a potential solution to restore kind of natural hearing
and hopefully millions of people affected by this hearing loss, especially hereditary
hearing loss. Anc80 seems to be potent viral vector for cochlear gene therapy and several
mouse models that could be rescued and the best results for the major deafness genes
at the moment seem to be achieved with Anc80, because with Anc80 you can also reach the
outer hair cells and not just the inner hair cells.
Gene editing with CRISPR Cas9 is feasible as this last study showed that was published
at the end of last year, but now the really important part for the patient and also for
the physicians that are confronted with these questions all the time: "What are the hurdles
on the way to the clinic?" As we saw in the mouse model with the expression
in the cerebellum (so in the brain) - it's really necessary that there are studies in
larger animal models, specifically for dosing, because the inner ear is so much bigger in
larger animals and humans compared to mice, and also the safety issues that I mentioned
before. It's kind of the last step prior to starting multiple independent human experiments.
And what was interesting in the study that also came out last year was that a group showed
that they could actually inject a sufficient volume into the inner ears of rhesus monkeys
without worsening this objective hearing. They have these ABRs where they can then detect
the wave forms and see if the animal heard it or not.
I recently was attending a talk by an investigator working in ophthalmology, and he said that
the vector correlation, vector result correlation, is under 30% between mice and humans, while
it's over 75% between monkeys and humans. You can really see the result.
Like, I'm also not a big fan of large animal experiments, but if you look at these results,
then it really seems to be necessary to have a larger animal model to be sure that what
goes into human studies that is, yeah, associated with so much like risk, et cetera, can actually
work and in vivo as well and humans as well. Another question is can you specifically target
certain cell types? A way to do that is to give the vectors a kind of a different key.
This key is called promoter scientifically, because for some diseases you just want to
target inner hair cells for some diseases and you want to target outer hair cells for
some diseases and you want to target neuronal structures. If you have the keys for all the
different cell types, it's nice to be able to kind of customize a treatment for every
patient. For these adeno associated virus vectors,
the size of the gene you can actually put into them, there are some approaches to try
to circumvent that problem. Excuse me. Try to circumvent the problem with very promising
results that haven't been published yet, but I heard somebody talk about it the other day
at a conference. Yeah, there's several options that hopefully can avoid this issue. And then
the time window is another very important thing that I was talking about.
The degeneration of cells progresses in several animal models or in many animal and human
models, human diseases. So the question is, do you really have to treat patients in the
womb, or is it sufficient to kind of do it after birth? Would the results be better in
the womb, and if it has to be in the womb it's associated with a lot of risks and it's
really, really tricky to actually get that into the inner ear.
Then also the treatment of age related hearing loss. Are these results from the one mouse
study actually translatable, and what are the results from this multi centered trial
I mentioned before beyond the gene therapy trial in humans at the moment. And what's
really exciting is most gene therapy labs have now ordered this virus, and we really
hope to accelerate the translational research, and then there's several more applications
where definitely more research is necessary. And with that, I'd like to thank all the collaborators
that have worked with us on gene therapy projects and all our funding agencies and as Nancy
said, I did run the Boston Marathon two days ago, and it was really horrible weather. If
you want to support our research, then you still have time until April 30th if you click
on that link you can read some more about me and why I decided to kind of try this marathon.
It was my first marathon. Yeah, I finished it. I was very happy with
the time as well for these conditions, and thanks a lot for your attention. I'm happy
to answer any questions. >> NANCY MACKLIN: Congratulations on that.
There are a few questions that have come in that are very interesting. Your presentation
was very interesting. It provides so much hope, so promising, this research. Lauren
said, if you get a cochlear implant, are you not a potential candidate for gene therapy?
>> LUKAS LANDEGGER: That is an excellent question, and that was also a big discussion about,
yeah, 20 years ago or so. I wasn't part of that discussion, obviously, but back then
people started to implant both ears, so in Europe so primarily the first candidates only
received one sided cochlear implants, and then about 20 years ago or so in Europe primarily
they started to implant both sides of the both ears of the patient.
When that first started, then some researchers said this is madness, and you have to preserve
one ear for gene therapy. Then the surgeons asked, well, when will it be ready? Then they
said, yeah, the maximum is five years or so. That was 20 years ago. It's really hard to
make like any predictions when it will be ready, so I am not sure how many questions
I answered there, because that's usually a standard question that I get.
>> NANCY MACKLIN: Right. >> LUKAS LANDEGGER: Typically if there is
a cochlear implant in place in that ear, you'd rather not inject it at the moment at least.
However, in the future if there still are these supporting cells left, then potentially
if these approaches worked to really make the switch from supporting cell to hair cell,
and then potentially they might be candidates, but it's a tricky question to answer.
You look at the so in the slides I also posted the link to this current gene therapy trial,
and the criteria for the patient selection are very strict. In the first studies you
really have to determine what works in a very small patient pool, and then if it works in
those patients, then you can kind of expand it and try to include more patients.
>> NANCY MACKLIN: I think that's probably several people in the audience that would
like to just become in the clinical trial right now. Go from mouse to humans.
>> LUKAS LANDEGGER: Also after the big paper was published with the virus, I received a
lot of e mails and all the collaborators received a lot of e mails. People really appreciated
that people are willing to really participate in these trials, and hopefully we will be
there soon. Right now, unfortunately, there's only this one trial with very limited criteria.
You can definitely check out the website and see if you are a candidate for this.
>> NANCY MACKLIN: Okay. So we've identified about 100 genes associated with hearing loss.
Do these genes include both inner and outer hair cell information?
>> LUKAS LANDEGGER: It depends. It really depends on the disease. So, as I said, this
VGLUT3 is a specific inner ear hair cell problem, and that's why they fixed it with the AAVs
and they target it. However, a lot of diseases affect both inner and outer hair cells in
that these conventional vectors seem not to work so well. That's why this new vector seems
to be better specifically for these diseases for the animal models.
>> NANCY MACKLIN: Tony asked, he said, I have hereditary hearing loss on my father's side.
I'm considering getting genetic testing to identify the genes responsible for my hearing
loss. Do you think getting the tests could be beneficial at this time?
>> LUKAS LANDEGGER: I mean, it might be beneficial in terms of the prognosis for him. I mean,
that's personally, because based on the gene that's affected, it might tell you how will
I hear in like ten years? How will I hear in 15 years and so on? You can kind of see
it from the father's side already. Yeah, it's tricky to like say anything about it without
seeing the patient, and I haven't my clinical training is very limited at this point, so
I still have to finish my residency and so on. So I'm kind of hesitant to answer this
question and would rather recommend seeing somebody who really has experience with hearing
and genetics in the human background and geneticists, specifically, like genetic counseling.
>> NANCY MACKLIN: Fair enough. John says, it is my understanding that for people who
have lost hearing over a period of decades that changes have taken place in the brain.
For example, reduction of volume of gray matter in the cortex. If you are successful in restoring
hair cells, will the changes in the brain gradually be reversed?
>> LUKAS LANDEGGER: That is an excellent question as well. The question regarding tinnitus,
for example, the ringing in the ears and what the current hypothesis of tinnitus basically
is, is that if there's not enough information coming in from the ears, then the kind of
central gain is just like amped up, and then the brain kind of creates this noise itself.
In tinnitus even if you cut the auditory nerve that connects the inner ear and brain, then
it still doesn't go away. So it's probably not something that's caused just by the inner
ear itself. Again, from a limited clinical experience, but in patients that receive hearing
aids, they usually do better with the tinnitus as well. So some of it might come back, but
I think if you really have had have not done anything about a hearing loss for decades,
then it might be hard to actually do something with this information that the brain all of
a sudden receives. >> NANCY MACKLIN: All right. Ken said, are
patients with Meniere's disease candidates for gene therapy? I understand the hair cells
die off in Meniere's patients. Do they regenerate? >> LUKAS LANDEGGER: That's also a good question.
We look at the vestibular system, so in some of the genetic diseases the vestibular function
is affected. So in these mouse studies, for example, all of these mice were dizzy as well.
So there's some very interesting tests that you can do with mice to figure out whether
they're dizzy. You can kind of film them from the top of the cage and see how they run,
or you can put them on a rotating rod called the rotarod and determine how long they stay
on top of the rod. Or have them swim and see if they can keep the head out of the water.
With this regeneration in Usher mouse models and others, the vestibular function was it
was like restored as well. However, in these mouse models the hair cells
kind of were still in there, and it was just a gene that was lacking. The gene was lacking
and reintroduced with this viral vector. If the hair cells are gone, the vestibular
hair cells are gone, it will be relatively hard at the moment to kind of grow them back.
However, also in the vestibular system you have supporting cells that could regrow into
the hair cells in the future. I mean, I'm not we're not talking about years here. It's
decades probably if I'd have to say anything about a timeline. So, yeah. Tricky.
>> NANCY MACKLIN: Okay. Katherine said, can you talk about the role of deteriorated brain
pathways in people with long term loss? Would they be eligible for gene therapy or regeneration?
>> LUKAS LANDEGGER: That's kind of question that I answered before where, yeah, it's kind
of the gain, et cetera. >> NANCY MACKLIN: Okay. Got it. Is effectiveness
of Anc80 versus AAVs specific to mouse inner ear, or are there similar studies on other
organs or organisms? >> Organs, yes. There are different studies,
so the initial paper I highlighted where they showed it in the liver, muscle and retina,
so everywhere there Anc80 seems to outperform the other AAVs. It's synthetic, so it's the
first of a class of synthetic AAVs, so in different organs it works. Regarding different
species, right now there's nothing published on that, whether it's translatable. That's
why we're so interested in having larger animal models not just for Anc80s but AAVs in general.
>> NANCY MACKLIN: Did male and female respond equal to the therapy? The females did much
better. >> LUKAS LANDEGGER: That's an excellent question,
and the NIH requires us to analyze male and female mice now because traditionally most
of the studies were carried out in male mice because they have fewer hormonal influences
that play a role in noise exposure in general. What we did for the in vivo studies is we
tested it in male and female mice. In vivo studies means the pups that were injected,
we tested the male and female mice for the so that was the same number, and pretty much
the same effect. For the in vitro studies, we don't know. At postnatal day four it's
really hard to differentiate the sex. We don't specifically look for that.
After a few weeks it's relatively easy to differentiate the sex in mice, but in the
very small mouse pups it's relatively hard. >> NANCY MACKLIN: Okay. Has a gene been identified
to explain cookie bite hearing loss? >> LUKAS LANDEGGER: I have to admit I'm not
familiar with cookie bite. >> NANCY MACKLIN: With that term? I'm not
either. >> LUKAS LANDEGGER: Like the shape of the
audiogram? >> NANCY MACKLIN: I believe so.
>> LUKAS LANDEGGER: Probably. Cookie bite hearing loss. Yes, that's the shape of the
audiogram. I'm not familiar with some of the clinical terms in English. This is not my
native language. That is a good question, and I'm, I mean,
I'm sure there are one of like one of the hundreds of genes or several of the hundreds
of genes have such a form, but I like it's hard to give you a diagnosis now just based
on this audiogram. So I don't think that that's possible.
>> NANCY MACKLIN: Okay. Would hearing loss due to meningitis be similar to that of age
related hearing loss discussed, or is that totally different?
>> LUKAS LANDEGGER: That is also a good question, and, yeah. So it depends what is like what
was inflamed. If there was actually a if the inflammation took part in the ear as well
or if it's just the not just, but if it's primarily the central parts that are affected.
If it is the central part, so the part of the brain where the processing of the signals
actually takes place, then I would say that it would be different than the well, I'm not
actually on this question, I'm not sure. I don't think I can answer this question.
I'm sorry. I have to pass on that. >> NANCY MACKLIN: Okay. All right. And last
question. Is the talk about hair cell regeneration in birds, mice, and fish applicable in nature
or just in research clinical trials? >> LUKAS LANDEGGER: No. For mice only in the
very, very neonatal mice. But for birds and fish, that's applicable in nature. So they
really regenerate their cells (their hair cells) constantly basically.
>> NANCY MACKLIN: Okay. Is there a reason for the lack of therapy trials? Is it strictly
because of funding, or you mentioned that in the very beginning.
>> LUKAS LANDEGGER: Right. That is a good question. I mean, the eye is just way more
accessible than the ear. The inner ear is kind of encapsulated in one of the hardest
bones in the human body and it's really tricky to deliver something there. In the eye you
can use a syringe and inject it in there. Yeah, it was just easier to kind of access
that, and then funding for blindness in general is or like hearing research is a relatively
small community in general, whereas the blindness foundations are definitely larger. So that
might have played a role as well. >> NANCY MACKLIN: Okay. And if you had to
guess, when would you think that there would be human clinical trials?
>> LUKAS LANDEGGER: There is already one human clinical trial.
>> NANCY MACKLIN: For the mass you know, for more people to get involved in the trials.
>> LUKAS LANDEGGER: So I don't think there will be clinical trials for like everybody
with deafness. It will be a clinical trial for a certain disease, for Usher syndrome
or that specific type for example with hearing data and so on. I'd really hope that we'd
have something to offer patients of like for at least like one specific syndrome within
the next two decades or so, but it's really hard to make these predictions. There's one
person from 20 years ago that said it would only take them five years to translate into
clinic. So you really have to be conservative there.
>> NANCY MACKLIN: It seems that the research is so promising and on the verge of great
discovery. I know everybody is anxious, and we definitely like to keep up on this topic.
So I welcome you to present again, even if it is from Europe or wherever you may land
from here. Thank you so much for doing the webinar. It was rather short notice, I know,
and you really did a great job. Thank you again to Darcy, who provided CART
tonight, and we'll look forward to seeing you next month when we talk about HLAA2018
in Minneapolis coming up in June. Good night, everybody, and thanks again.
>> LUKAS LANDEGGER: Good night. Thank you.
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