Temple University Researching CRISPR/Cas9 for HSV Cure

[Evaluate]

When it comes to CRISPR/Cas9, Drs. Cullen/Bloom and Editas are likely first to come to mind.  It turns out that Temple University is also using CRISPR/Cas9 to look into a HSV cure.

http://temple-news.com/news/researchers-aim-to-find-cure-for-herpes/

Below are a few snippets.

Quote

Lifan He, a first-year graduate student in the Health Informatics program, said CRISPR/Cas9 contains the instructions and the tool to cut out a specific protein in the herpes gene sequence, ICP0.

While eliminating ICP0 means the infection and its effects are greatly reduced, there are other parts of the herpes’ DNA without which herpes will not survive. The team has not eradicated any of these genes yet; but they are working on different targets for Cas9. He added the team has successfully eliminated the protein ICP0 from herpes’ gene sequence in the simplest platform of research: human cell lines.

“Once CRISPR/ Cas 9 is in the cell, it works like scissors,” He said. “We cut out ICP0 and once we cut it out, the whole viral infection is reduced.”

Team members said as this research progresses, if other proteins respond similarly to ICP0, an eventual cure could be found for herpes.

“The target I am working on now, which is not ICP0, can potentially beat Type 2 and Type 1 at the same time,” He added.

It's great that they're right now able to show promise for HSV type 1 AND 2 using the same sequence, as I think this will move clinical testing along as the safety profile will only need to be confirmed once.

Quote

Initial research involved testing ICP0 on the cell lines—and because the work has been successful at this level, the team is moving on to deceased rats.

The team is now working with primary neuron cell culture, which means they are now working with neurons in the brain. This level of research is much more complicated than the singular cell line, as they are working with actual subjects instead of home grown cells. Additionally, once the team figures out how to insert Cas9 into the rat’s brain, it still means work to come as a living brain functions differently than a dead one.

 

[dont quit!]

Good find, they are targeting the protein which enables HSV go into latency. I like that they are attempting to kill two birds with one stone. But I was hoping their primary focus would be HSV2 being that Editas and Cullen are focusing on type 1 already. Also, Cullen has already figured out how to get into the neurons which the Temple staff has yet to do. This research is pretty good news!

[Sanguine108]

Well this is cool.  I thought  the ICP# is a gene encoding for a surface protein on the virus.   While I think this is a nice lil' snippit of progress, I'd rather them snip out the promoter region for HSV replication.   Organisms, in general, have a lot of junk DNA floating around and I wouldn't mind if HSV was just some more disabled, junk DNA floating in the cytoplasm.  Deleted ICP# would not make it junk DNA.  The virus would still replicate but without a 'stealthy' component to itself.

A cure to me means that my body doesn't have to deal with the energetic weight/demand/burden of the virus.   clipping the ICP# region would still allow the virus to replicate but, since it's easily identified by the immune system, it gets picked up quick and on the surface there's no apparent symptoms.   I hope this isn't the route people go with this.  I'm all for progress and pushing forward in science and general research so to get as many angles as possible. So go Temple! 

 

So now that I've read the article...

"Initial research involved testing ICP0 on the cell lines—and because the work has been successful at this level, the team is moving on to deceased rats.

The team is now working with primary neuron cell culture, which means they are now working with neurons in the brain. This level of research is much more complicated than the singular cell line, as they are working with actual subjects instead of home grown cells. Additionally, once the team figures out how to insert Cas9 into the rat’s brain, it still means work to come as a living brain functions differently than a dead one.

“Because it is a tumor cell line, it is not as natural as primary cell culture,” He said. “And primary cell culture is not as natural as a live animal, which is not as close as human.”"

I'm just glad there's another lab focusing on this.  The more minds working on it the better.

Edited January 26, 2016 by Sanguine108

[JustHanginginThere]

Interesting news but Cullen and Bloom team are already into animals with CRISPR-Cas9 and they have spend their entire careers with Herpes. However, the part about beating both HSV-1 and 2 with the same target is quite interesting.   The good thing about University research is they all learn from each other.

Edited January 27, 2016 by JustHanginginThere

[Evaluate]

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Edited January 27, 2016 by Evaluate

[JustHanginginThere]

I read the article several times trying to translate it.  There is a lot of talk about ICP0 in the beginning.  Then Lifan states “The team has successfully eliminated the protein ICP0 from herpes’ gene sequence in the simplest platform of research: human cell lines.”   I start thinking the entire article will be about ICP0.  Then  (to my suprise) he states “The target I am working on now, which is not ICP0, can potentially beat Type 2 and Type 1 at the same time.”  This target which IS NOT ICP0 is not further described at all.  I am curious to know what target he is talking about.  They are very much in pre-clinical discovery and again not as far as other researchers.  However, additional minds on the same topic are always helpful.

Edited January 27, 2016 by JustHanginginThere

[Evaluate]

3 hours ago, dont quit! said:

Also, Cullen has already figured out how to get into the neurons which the Temple staff has yet to do.

Is there a reason Temple University hasn't obtained this information from Cullen/Editas?  I'm trying to figure out to what extent Cullen works for Editas and to what extent he continues to research for a university.  I get it if Editas doesn't want to share information, but if Cullen is still under his university doesn't it make sense to share these scientific lessons learned?

2 hours ago, Sanguine108 said:

Well this is cool.  I thought  the ICP# is a gene encoding for a surface protein on the virus.   While I think this is a nice lil' snippit of progress, I'd rather them snip out the promoter region for HSV replication.   Organisms, in general, have a lot of junk DNA floating around and I wouldn't mind if HSV was just some more disabled, junk DNA floating in the cytoplasm.  Deleted ICP# would not make it junk DNA.  The virus would still replicate but without a 'stealthy' component to itself.

So now that I've read the article...

"Initial research involved testing ICP0 on the cell lines—and because the work has been successful at this level, the team is moving on to deceased rats.

With CRISPR being reportedly as powerful as I hear it is, why would Temple pursue a novel approach as weakening the virus instead of disabling it?  The article itself says there are 78 essential proteins.  I would think they should keep trying other proteins that would actually disable the virus before moving the ICP0 to further testing.  Maybe I'm missing something.

[JustHanginginThere]

The NIH gives lots of different grants for Herpes. Cullen does NOT work for Editas.  They are on a joint Small Business Grant that Editas won.  Small Business Grants require you use some of the funding for Universities so they are collaborating with Editas on the grant.  It is specifically for HSV-1 Herpes Keratitis so while it seems similar it is different research.  It appears Temple is not researching HSV-1 Herpes Keratitis.  Researchers don't just get on the phone and start calling and say "I have this or that"---they typically end up reading about others work from scientific journals (they are all proud people).  I woudn't focus much on Editas--they are focusing on HSV-1 Herpes Keratitis as it is an Orphan Disease.  In addition, HSV-1 Herpes Keratitis will not be their first trial in 2017.

Edited January 27, 2016 by JustHanginginThere

[Sanguine108]

It's all about painting a picture.   This is just the beginning of working with the gene editing tech.  AND VERY MUCH the beginning of using it on HSV.  It's not like anyone really knows how anything is going to turn out.  For that matter, most of the discoveries out there happened by accident and were not on their track of thinking.  So I think the more push in a direction, even if it seems like a waste of time, will contribute to progress and better understanding of a "cure" or at least how to work with HSV DNA in the human body.

Like people want to poo poo on Dr. Ryder's DRACOs research.  And while I'm not entirely confident that it'd do anything for HSV as it would be tagging nerve cells for destruction. ...I think the medical community would gain so much insight from just exploring the use of that technology.  

Not to get too deep into this, but when I was working in a lab, I benefitted from the progression of the Human genome project, and genome projects on various birds, and the progress of human molecular research enhanced my research tools and abilities.  Science should grow like that.   But human biotech is a bit different.

I do think it's just a matter of time until the gene editing tech starts getting billions of dollars for research and development instead of millions or not very much.  I'm looking forward to Editas producing these proofs that this tech is worthy, it's powerful and it's safer than drugs/chemical therapies.   But we shall see!

Edited January 27, 2016 by Sanguine108

[dont quit!]

6 hours ago, Evaluate said:

 Is there a reason Temple University hasn't obtained this information from Cullen/Editas?  I'm trying to figure out to what extent Cullen works for Editas and to what extent he continues to research for a university.  I get it if Editas doesn't want to share information, but if Cullen is still under his university doesn't it make sense to share these scientific lessons learned?

I don't think Cullen/Editas would help out a competitor because that is how they are viewed realistically. I also know that Cullen has patents so IMO he is very unlikely to give a helping hand. 

[dont quit!]

6 hours ago, JustHanginginThere said:

The NIH gives lots of different grants for Herpes. Cullen does NOT work for Editas.  They are on a joint Small Business Grant that Editas won.  Small Business Grants require you use some of the funding for Universities so they are collaborating with Editas on the grant.  It is specifically for HSV-1 Herpes Keratitis so while it seems similar it is different research.  It appears Temple is not researching HSV-1 Herpes Keratitis.  Researchers don't just get on the phone and start calling and say "I have this or that"---they typically end up reading about others work from scientific journals (they are all proud people).  I woudn't focus much on Editas--they are focusing on HSV-1 Herpes Keratitis as it is an Orphan Disease.  In addition, HSV-1 Herpes Keratitis will not be their first trial in 2017.

If you cure HSV keratitis, you cure HSV 1. That is a big deal. I'm definitely tracking their progress.

[dont quit!]

9 hours ago, Sanguine108 said:

Well this is cool.  I thought  the ICP# is a gene encoding for a surface protein on the virus.   While I think this is a nice lil' snippit of progress, I'd rather them snip out the promoter region for HSV replication.   Organisms, in general, have a lot of junk DNA floating around and I wouldn't mind if HSV was just some more disabled, junk DNA floating in the cytoplasm.  Deleted ICP# would not make it junk DNA.  The virus would still replicate but without a 'stealthy' component to itself.

A cure to me means that my body doesn't have to deal with the energetic weight/demand/burden of the virus.   clipping the ICP# region would still allow the virus to replicate but, since it's easily identified by the immune system, it gets picked up quick and on the surface there's no apparent symptoms.   I hope this isn't the route people go with this.  I'm all for progress and pushing forward in science and general research so to get as many angles as possible. So go Temple! 

If they clip the ICP 0 gene in the neurons, once reactivated, it would then be unable to go back into latency. The virus would then be destroyed by the immune system. If I understand their method correctly this may be able to work. 

[fixme1]

The world needs its eyes opening to the damage hsv does instead of dismissing it, hopefully this editas etc cracks something. will it just take willing people to risk the gene editing or is it people like the fda stopping it all?

[Evaluate]

7 hours ago, dont quit! said:

If they clip the ICP 0 gene in the neurons, once reactivated, it would then be unable to go back into latency. The virus would then be destroyed by the immune system. If I understand their method correctly this may be able to work. 

From what the article says, it doesn't stop it entirely:

Quote

While eliminating ICP0 means the infection and its effects are greatly reduced, there are other parts of the herpes’ DNA without which herpes will not survive.

This is from the article.  Why I asked the previous question was they point out themselves that there are other parts of the DNA which would rid someone of herpes completely and not just weaken the virus as they say ICP0 would do.

@dont quit! and @JustHanginginThere, are either of you in the biotech field?  The reason I ask is I would like to get a handle on how quickly CRISPR/Cas9 research can progress versus vaccine testing.  From what I've read CRISPR/Cas9 is easy and cheap to work with.  I wonder if this means CRISPR/Cas9 research can progress quicker than typical vaccine research.

[StayingUpbeat]

This is miles away from the clinic.  It's really only someone's pet project at the moment so I wouldn't hold my breath that it is actually going to make it out of a University lab.  It's interesting, however, that they are clipping ICP0.  It doesn't seem like anyone mentioned it but ICP0 is the same gene that is deleted from Dr. Halford's vaccine strain.  From everything I've read about ICP0 its probably the best target you can go for as a single gene to disable HSV (with ICP10 being a close second).  There have been dozens of papers written on how important ICP0 is to the life cycle of HSV and its totally conserved across strains.  From an evolutionary genetics perspective that implies it removing it would dramatically reduce virulence if not totally disable it's ability to replicate.  Sign me up!

Edited January 28, 2016 by StayingUpbeat

[fixme1]

Cany anyone remove it yet or is it notnworked out yet

[JustHanginginThere]

12 hours ago, Evaluate said:

From what the article says, it doesn't stop it entirely:

This is from the article.  Why I asked the previous question was they point out themselves that there are other parts of the DNA which would rid someone of herpes completely and not just weaken the virus as they say ICP0 would do.

@dont quit! and @JustHanginginThere, are either of you in the biotech field?  The reason I ask is I would like to get a handle on how quickly CRISPR/Cas9 research can progress versus vaccine testing.  From what I've read CRISPR/Cas9 is easy and cheap to work with.  I wonder if this means CRISPR/Cas9 research can progress quicker than typical vaccine research.

I personally don't get involved in the vaccines discussion--they are a long way off and take a long time to go through trials......and then they only provide benefit for some.......

A Cure > Vaccines  (regardless of the technique)

Edited January 28, 2016 by JustHanginginThere

[Evaluate]

Very interesting observation, @StayingUpbeat.  Perhaps the article did not do it justice when citing that there are other, perhaps more effective, proteins to target.  It seems their decision to focus on ICP0 was a decision justified by previous research available. 

I think your point regarding the reality of the lab's ability to bring something to market is valid.  Still, the more the merrier when it comes to herpes research.  Hopefully there is a really intelligent grad student somewhere in there who can contribute something big to the field.

Edited January 28, 2016 by Evaluate

[dont quit!]

I agree with Stayingupbeat that this is light years away from the clinic. And probably doesn't have any funding. Maybe Dr. Jerome has something in the works? His description of his HSV therapy is solid and much more advanced than Temples at the moment. I think he just needs funding, boooo!!

[JustHanginginThere]

10 minutes ago, dont quit! said:

I agree with Stayingupbeat that this is light years away from the clinic. And probably doesn't have any funding. Maybe Dr. Jerome has something in the works? His description of his HSV therapy is solid and much more advanced than Temples at the moment. I think he just needs funding, boooo!!

Jerome doesn't work with CRISPR-Cas9--he works with Meganuclease gene editing  (older technology).  Also, he does not work with any support groups.  He gets funded through Pharma and they do not openly talk about what they are doing as much as some of the other researchers with updated blogs.

[JustHanginginThere]

Gene editing therparies are NOT "miles away" or "light years away" from the clinic.  Those are pessimistic views based on opinions that are not the reailty of the science or the companies involved with this technology.

Don't worry be happy (just like the song)--Hope is coming.

Edited January 28, 2016 by JustHanginginThere

[dont quit!]

Just now, JustHanginginThere said:

Jerome doesn't work with CRISPR-Cas9--he works with Meganuclease gene editing  (older technology).  Also, he does not work with any support groups.  He gets funded through Pharma and they do not openly talk about what they are doing as much as some of the other researchers with updated blogs.

Its still gene editing and a solid well thought of concept. So who cares if he isn't using Crispr Cas9. Just cure me!

[dont quit!]

Just now, JustHanginginThere said:

Gene editing therparies are NOT "miles away" or "light years away" from the clinic.  Those are pessimistic views based on opinions that are not the reailty of the science or the companies involved with this technology.

Don't worry be happy (just like the song)--Hope is coming.

Its not being negative. Truth is, is that they haven't even penetrated the neurons and they don't have funding. It is, what it is. 

[JustHanginginThere]

From Cullen's Blog----

"The big advantage of AAV is that you can get very high levels of virus—up to 10 billion infectious units per milliliter—and the Bloom lab has clearly shown, using an AAV that expresses green fluorescent protein (gfp), that he can infect essentially every single neuron in the trigeminal ganglia where HSV-1 establishes latency."

 

 

[Evaluate]

As a side note, I emailed Dr. Roehm (the lead researcher) to enquire about the lab.  She responded straight away.  I probably didn't ask the right questions (not being scientifically inclined) but she told me that there are full time researchers in the lab and they are using a similar approach to the Cullen Lab and Editas.  Are there any other questions I could ask her about Temple's research?  I can send them altogether later tonight if posted here.