Welcome back everybody to another episode of You Make Me Sick.
Today we're joined by a very special guest, this is Dr. Shawn Owen.
Dr. Owen is a microbiologist.
She is a or was a research fellow in the Department of Biomedical Informatics and
Laboratory of Systems Pharmacology at the BAME Laboratory at Harvard Medical School.
She's also completed research at the Jay Hinton Laboratory in Liverpool, England.
She's an expert in the field of bacteriophages.
Will be soon starting her own lab in Albany, New York at the Wadsworth Center, correct?
In conjunction with New York Department of Health.
She's actually been using experimental evolution and computational approaches to study the
phenomenon of antibiotic resistant bacteria, among other things and just really innovating
the way phages are looked at.
So I'm very happy to have her here today.
Thank you for joining us Shawn.
Thanks a lot Andrew, very excited to be here.
Shawn has also been published in a variety of journals including Nature, Nature Microbiology,
Science and the International Journal of Molecular Sciences, just to name a few.
She just completed her post doctorate here at the Harvard Medical School in the BAME
Laboratory.
As I said she's going to be heading up her own lab very soon and I'm actually really
excited to talk to her today about a, it's a subject that a lot of people have no idea
exists, bacteriophages.
So I guess question is Shawn to start with, what can you tell us about bacteriophages?
If I was someone off the street who had no background in biology, had no idea, could
you explain to me what a bacteriophage is?
Yeah, so a bacteriophage is a, or a phage.
I like to go for phage but it's a potato potato, right?
It's actually, it's your call.
It's a virus and so I usually like to explain it to people as it's like the coronavirus
for bacteria.
So these are exactly the same way as you and I get viral infections, bacteria are susceptible
to the exact same problem and frequently encounter these viruses in their everyday lives which
will infect them and often kill them.
And so more specifically in terms of the biology, a bacteriophage is debatably alive when it's
outside of a living cell.
A phage is just a protein shell surrounding some nucleic acids and that's either DNA or
RNA.
But that, those nucleic acids code for the instructions that the virus uses once it gets
inside a cell to repurpose that cell into a factory to make more of itself.
Exactly the same as when you and I get viruses ourselves.
How long have you had this great appreciation for phages?
Is this something, is this something that, I know you were telling me you grew up in
the south of England, a really big forested area, lots of nature around you.
Have you always been a biology fanatic?
Yeah, I think I've always been really interested in the natural world.
I've also always had a morbid fascination with infectious disease.
I think it's that my mum was also a nurse and so I think she instilled in me a kind
of interested disease and gross things.
And so yeah, I always remember being interested in microbiology.
I remember the first time I learned about bacteria was in high school science class
and we would go around the school swabbing surfaces and culturing them on bacterial plates.
And I just remember being amazed that there was this whole world that we couldn't see
going on.
And yeah, I think microbiology is so fascinating in that respect.
It's just a window into, we are titans, you know, in this like world that we cannot even
see.
Yeah, I have a, I had a microbiology professor that said it's their world.
We're just living in it because they're everywhere.
They're ubiquitous.
Exactly.
With regard to phages, so do you know anything about their history as far as when they were
first discovered, their first uses, especially in the field of medicine?
Yeah, so phage has a very, the field of phage has a really interesting starting point.
So phage were first kind of discovered in the late 1800s when scientists, I think the
first accepted kind of people who identified these were scientists in India actually who'd
noticed that the cholera bacteria they were growing would sometimes just kind of die when
they added water from the Ganges River.
And it was kind of known to be this sort of principle, this kind of lytic principle that
would occur sometimes in bacteria.
But that was kind of as far as that got.
And then it wasn't until the early 1900s and the First World War, a British microbiologist,
very proud of this, Fred Twart published a paper describing what he found when he was
growing some micro-cockas, I think it was, bacterial colonies.
And he noticed that on top of these colonies occasionally, he'd see these little glassy
spots of kind of like transparent circles in the bacterial colonies.
And crucially, he found that he could transpose those little areas of lysis from one colony
to the next.
So if you like, he could infect one of these glassy colonies into a healthy colony and
cause infection.
And so he, and secondly, so at that point we weren't really sure what this like agent
causing this lytic morphology was, but he showed that if he filtered this diseased part
of the colony through what was known as a chamberland filter, so basically a filter
that had such small holes that bacteria cannot pass through, you could still infect healthy
colonies, suggesting that whatever this thing was was smaller than bacterial cells.
And at that point, it would come known as like an ultramicrobe, they were calling it.
And I kind of, yeah, it's a really fascinating story involving many characters.
There's another, there's a lot of actually debate about who it was who first identified
the bacteriophage.
And another kind of famous name that often comes up is Felix Durell.
That's the name I'm familiar with.
Yeah.
So he's like often, I think, often was like first credited with studying the bacteriophage
for a long time, certainly because he was much more flamboyant personality.
But technically his paper came out a little bit later, but he was working on bacteria
causing dysentery in France, a chagella bacteria, and discovered like very much the same thing.
But I think Felix Durell is also often celebrated because he was the first person to really
think about therapy and what we could do with these ultramicrobes that could kill bacteria.
And he was very confident and kind of a bit of a maverick.
And he very early started pioneering the use of actually using these bacteriophages to
treat soldiers and patients who are suffering from enteric disease.
And it was successful, right?
They actually they had success using phages to treat disease early on.
This is pre-antibiotics.
So this is the turn of the century.
1900, 1920s, 1930s, correct?
Correct.
And obviously like a caveat here is nobody's reporting when it doesn't work.
That's like a you're going to realize that's a long standing issue with the phage therapy
literature.
But yes, it had early success.
Studies won't let you do that anymore, will they?
And so phages were actually acceptable in science.
And even here in the United States, I'm pretty sure Eli Lilly was the first manufacturer of
laboratory produced phages to treat just different forms of diseases.
Ankinteric diseases, you said, were one that were used quite commonly for.
I know that they were also used topically.
I'm not sure how early that was.
But I don't know if it was staff or what they were treating with that.
So there was over 100 years ago, I guess.
So then antibiotics came into play.
And this is kind of this will play into a lot of what I'd like to talk about today, just
with regarding phages and phage therapy.
So antibiotics, when they were first created, they've been used now for about 100 years.
And we're really starting to see just kind of the repercussions of their misuse and their
overuse and just a misunderstanding of how bacteria mutate to survive.
So from your perspective, what advantages do you think phages can provide over antibiotics
or in conjunction with antibiotics?
And can you kind of explain the difference of how phages destroy bacteria as opposed to
how antibiotics might kill bacteria?
Yeah.
And first, let's go back a little bit and talk a bit about what happened when antibiotics
were discovered, because that's kind of an important part of the history of phage research.
So you're right.
There was a lot of interest, I think, after Dorell was showing that phages could help
patients.
But shortly after that, phage therapy was particularly taken up in Eastern Europe.
And so there were some Georgian scientists who happened to be studying at the Pasteur
Institute in France with Dorell and kind of took this technology back to Georgia and Eastern
Europe with them and did some really good work and a lot of pioneering work into bacteriophages.
And then the 1930s comes, or late 1920s, and penicillin is discovered.
And so I think at this point, in the West, bacteriophages in comparison were seen as
very complicated things.
So a feature of bacteriophages is they tend to be very strain specific.
They are not great at curing every kind of E. coli infection you might have.
Like one phage might just hit one single strain of E. coli.
And so to actually formulate a therapy with these things is quite challenging and can
be very inconsistent.
Versus antibiotics, which were easy to mass produce.
You just grow fungus, you extract the antibiotics.
They tended to kill super broad spectrum kinds of bacteria, so you didn't need to have loads
of individual kinds of phages.
You could just have one antibiotic and hit everything.
And so pretty much the Western world stopped working on phages and really focused their
efforts towards antibiotics and very successfully, right?
Like antibiotics really hugely improved our human lives in the 21st century.
So yeah, it's kind of an interesting story.
But then obviously now we're kind of reaching a point where antibiotics have been so successful.
We are using them in really every part of life.
And so I think 66%, for example, of all antimicrobials are used in agriculture now.
They're not even used directly to treat human disease.
And the result of that is that we are just really putting a high degree of selection
on bacteria to evolve resistance to antibiotics in areas that aren't even the human body.
And hence we're having this huge problem with antibiotic resistance.
But back in Georgia and Eastern Europe, they've actually been using still phages this whole
time.
So you could go to Georgia and I have colleagues who were there for conferences and you can
go into a pharmacy and buy vials of phages for ear infections and gastric upset.
So we're now finally going back to our colleagues in Eastern Europe in Georgia to try and really
understand what these therapies are that they've been using and see if we can learn from them
as we become re-interested in phage therapy.
So there was an online article that was published from the pharmaceutical processing world.
That was the name of the public online publications in 2023.
From that article, they said that quote, the global antibiotics market size was valued
at approximately $42.33 billion in 2023 and is expected to grow because of due to increase
to antibiotic resistant infection and rising demand for novel antibiotics.
Do you feel that with phage therapy, you know, it's kind of an afterthought due to the monster
of some amount of revenue that these pharmaceutical companies bring in?
Do you think that, you know, I know that, and I saw again, this was Eli Lilly just pledged
another just large amount of money towards more antibiotic research and I don't even
know if they've dipped their finger in phage research or not.
But do you think that's something that's being overlooked that should be more closely
paid attention to?
Yes.
Well, I think the issue is very complicated.
The problem is we are way behind in terms of phage research compared to what we know
about antibiotics.
So because we abandoned the field for so long, we are really in the very early days of understanding
phages and using them to treat patients.
So you can kind of imagine we are back in terms of, in using phage, we are back in the
1930s still as far as our understanding goes.
And so I would say more than maybe pharmaceutical companies needing to invest in phage, I think
we do need more basic science research to be done.
And then I think later, once we're at the point where it does seem like we have technology
to really push these things into therapeutics, it would be great.
But I would say in general, we just need more government level support and reward for antimicrobial
development.
To that point, so last year the pharmaceutical health and beauty industry spent about $378
million in lobbying in the US.
Knowing how much profit these pharmaceutical companies can make from antibiotics, do you
feel that lobbyists are kind of, they're slowing things down governmentally with regulations
because they know that phage technology, even though it might be more successful, they're
making more money with antibiotics.
And at least in the United States, the dollar rules everything, even our political system,
which is how it goes, I guess.
But do you think that if there were more lobbying groups or proponents of phages and phage therapy,
that this would be more research and more funding would actually be, I don't know, they'd
be compelled to do more research and more funding?
Yeah, perhaps.
I mean, I think we also, as researchers need, there is some onus on us to really demonstrate
the therapeutic potential of these things.
And certainly that is not something that I'm 100% convinced of.
Cards on the table, I think like it's still not, I would still not bet that in 50 years
we'll be using phages widely to treat bacteria.
The jury's still out a little bit.
So I think we, yeah, a lot more basic science research needs to be done and time will tell
whether phages are really going to save us.
So it's something that we encounter a lot in the healthcare system, our multi-drug resistant
organisms.
Phages and especially some of the ones you've been studying have shown potential to actually
treat a lot of these that antibiotics can't kill.
And these are different strains of bacteria that just have gained resistance to some of
them multiple antibiotics and become really, really hard to kill.
There was a recent study in nature microbiology, it was the BT100 study, it was pretty small,
but it found that phage therapy used in combination with antibiotic treatment proved to be highly
effective in treating MDROs both in vivo and in vitro, so in the lab and in humans.
It was small, the sample size, but it did provide data just to show that phages are still effective
and can be very effective in conjunction with antibiotics.
Do you think that there may be a day when phages themselves could be as potent or even
more effective at treating and killing these multi-drug resistant organisms?
Yeah, so that study was hugely celebrated in the phage field.
It was a real labor of love primarily pushed by these researchers in Belgium and really
answers a lot of very important questions with regard to the success of phage therapy,
which has been something that's been very hard to answer for a long time because in
general the way phages have been used for therapy is very unsystematic.
So typically as there is no official regulation of phages as therapeutics, the only way in
which clinicians are able to use them is in these compassionate use cases.
And therefore every trial is a little different, the formulations are slightly different,
patients don't really have comparable illnesses.
And so it's been up until now extremely difficult to really know where the phage therapy is
working and of course there's quite a few case studies in the literature about successful
cases, but as we mentioned before how many failed cases were there for every successful
case.
And so this particular study, the VTE 100 study is really exciting because they didn't
cherry pick the data at all.
They really just systematically took the first 100 cases that they had done and were super
transparent about the data and it's actually a kind of optimistic picture.
So what it shows is that interestingly phages when they were delivered on their own weren't
particularly good at clearing infections.
The clinical prognosis didn't tend to improve too much.
But more happily when you combined phages with antibiotics there was a pretty convincing
increase in patient infection prognosis.
So it was pretty promising data that combined with antibiotics phages might be a pretty useful
clinical strategy.
So kind of staying on the subject of these MDROs.
So this year, this past year, or this spring or the end of last year, the CDC they released
data from a 2019 antibiotic threats report and in the United States here they stated
that more than 2.8 million antimicrobial resistant infections occur every year and 35,000 people
die as a result.
That's just here in the United States.
And then even beyond that the WHO, actually this is actually an article from the Lancet,
I'm sorry, from 2022 that again kind of confirmed and this is using, it might be even the same
data from 2019.
It's estimated that worldwide there were about 4.5 million deaths associated with antimicrobial
resistance.
A lot of death from this, you know, a lot of these, obviously they can't be treated
with antibiotics, they lead to a lot of bloodstream infections, sepsis and subsequent death.
The numbers are pretty staggering and with such an urgency that we have right now and
especially in the last few years there's something called the antimicrobial stewardship
and this is seen in hospitals where you're not prescribing antibiotics unless you are
100% certain someone has a bloodstream infection or if you're suspicious you stop them as soon
as if they don't have one you stop the antibiotics as soon as they're shown not to have one and
then you get a specificity as soon as possible and you narrow down the antibiotic and this
is to try and control some of that resistance in these microbes.
But with such an urgency do you think that like phage therapy there could be more attention
paid to this because it is a problem that's going to continue to grow and increase especially
over the next 25 years or so?
Absolutely.
And so actually the reason why phage therapy has been kind of neglected, interestingly
all goes back to that history we were talking about.
So because phages were kind of abandoned in favour of antibiotics they were kind of seen
as too complicated I think.
But the cool thing is that even though they were completely abandoned by people interested
in therapy they were taken up by molecular biologists and physicists who decided that
bacteriophages were a really fascinating way to study.
They considered them to be the simplest organisms so if we were going to understand life we
could start with a bacteriophage.
And so there was a huge effort to understand phages at the molecular level and that led
to all kinds of really important scientific breakthroughs in the 20th century including
the discovery of DNA as heritable material and tons of other interesting things.
The enzymes we use in molecular biology most of them come from bacteriophages.
And so it's kind of a double-edged sword here because even though we did kind of abandon
phages in terms of therapy we got a huge, we learned a huge amount from them in terms
of molecular biology.
And so hopefully we're now at a point where we need phages again perhaps therapeutically
but we have all of these tools in our pocket from almost kind of 80 years of studying them
at the molecular level and we can maybe now kind of combine our knowledge and take a kind
of synthetic biology approach to improve phages.
And it's very analogous really to what happened with antibiotics.
So when antibiotics were first discovered they are a natural product of fungi, right?
And originally people were just using the raw, the real natural product of fungi to treat
patients and it wasn't that great.
It had a lot of problems so occasionally these antibiotics were toxic to humans, they were
hard to produce.
And very quickly with research people moved from using these natural product antibiotics
to modifying them and making synthetic versions of those original antibiotics.
And now most of the antibiotics we're using don't look quite like the original natural
products that fungi are producing.
They're things that we have expanded upon and elaborated.
And I really think the phage field is going down the very much very similar trajectory.
So at the moment we're still in the early days where we have these like natural bacteria
phages and what we need to do is start expanding them and editing them and turning them more
towards our purposes.
So my personal feeling and it's controversial perhaps and not everyone in the field might
agree with me but my personal feeling is phage therapy as we know it today is never going
to be widely uptaken.
But as we move towards kind of custom therapies or edited or even machine designed bacteria
phages these things I think have the power to really influence the way we treat infectious
disease.
I'll touch on that a little bit later especially using artificial intelligence to help kind
of create or guide the production of these.
So the WHO recently released their list of priority pathogens.
This includes 24 pathogens 15 different families of antibiotic resistant bacteria included in
the list are Salmonella, Shijella, Nasiria gonorrhea, Pseudomonas, Pseudomonas aeruginosa
and Staph aureus.
I kind of wanted to talk a little bit you've done quite a bit of research actually with
Salmonella correct and phages and how phages can be used to treat MDI Rho Salmonella.
And I also wanted to talk a little bit about Pseudomonas and then MRSA the Methacillin
resistant Staph aureus which are two of the most common hospital acquired multi drug resistant
organisms.
So with your research with Salmonella exactly like how were you able to use phages to actually
treat that MDI Rho Salmonella strain that you guys were working on?
Yeah so it's an interesting story.
So I was originally working on Salmonella during my PhD and that's how I first got
interested in bacteriophages.
The way I stumbled upon them is I was working on two strains of Salmonella.
One was kind of a European classic gastrointestinal strain, Salmonella typhimurium and the other
was a derivative of Salmonella typhimurium from sub-Saharan Africa which had emerged
to cause bloodstream infection in the kind of immune compromised HIV positive population.
And but at the genomic level so when you looked at the DNA sequences of these two strains
of Salmonella they're actually very related like very similar.
So in Salmonella genome there's around 5 million base pairs of DNA and you could align end
to end these two genomes from the European and the African Salmonella.
So my task was kind of like look at these two genomes and figure out what is it in this
African Salmonella that made it more virulent, made it able to invade and cause these bloodstream
infections.
And the most obvious thing that struck me as soon as I looked at these genome sequences
was in the African strain there was these large insertions of new genetic material that
weren't present in the European strain.
And those it transpired were phages.
So one thing we haven't talked about yet is that phages can have this kind of dual lifestyle.
So they can behave very much like your classic virus where they'll infect a cell, make it
a virus factory and then explode the cell and go on and infect more.
Or they can do this more kind of covert lifestyle that's kind of analogous to what HIV does
right in humans.
They can infect the cell but instead of making it into a virus factory they will integrate
themselves into the genome, into the DNA of the bacteria and they'll just like hang
out there until you can imagine like some conditions get particularly good and then
the phage might emerge.
But the kind of really interesting thing about this in Salmonella and in some other very
notorious bacterial pathogens is that because they're spending a lot of time inside the
bacterial genome it's kind of in their interest to increase the fitness of the bacterium
itself.
And a lot of virulence factors in Salmonella and in many other bacteria, so cholera, staph
aureus, dip, the pathogen that causes diphtheria, many of these like famous infectious diseases,
the actual virulence genes that are making you sick are encoded on one of these viruses
that's integrated into the bacterial cell.
Crazy.
Yeah, it's really cool.
So it's worth thinking about in terms of therapy because phages aren't always bad to bacteria,
sometimes they're very good for bacteria.
So in this particular case in Salmonella I had an immediate hypothesis that these new
phage sequences that had integrated into the chromosome must be bringing some new virulence
factors that we hadn't found yet.
So I spent most of my PhD studying these phages, I got sent to an amazing phage conference
called Viruses of Micropes which happens every two years and just focusing on the viruses
and not just bacteria but also fungi and archaea.
And I just became very quickly completely fascinated by bacterium phages.
And in the end though we didn't find any virulence factors encoded on these new prophages.
Instead we found something else that the prophages were doing to help the Salmonella and that
was phage resistance.
So another thing that is good for bacteria is to be able to defend against viruses, right?
So it turns out that often these phages will integrate into a bacterial genome and then
make that cell resistant to other phages.
So it's kind of like get off my lawn kind of thing.
I'm here and no one else can come in and take my host.
And so actually now in the last five or so years this whole field of phage defense and
understanding how bacteria defend themselves against phages is huge.
And it turns out all of these genes that bacteria had that for a long time we didn't understand
what they did.
We now know a lot of them are actually involved in defending bacteria against phages.
And sometimes those are just on the bacterial chromosome and sometimes they're brought in
by other mobile genetic elements like phages and plasmids.
And that's kind of one of the fears regarding the use of phages when I was doing a little
research on them was just like similar to antibiotics.
Bacteria, they've been here forever and they know how to mutate.
There are no free lunches in nature like there's no silver bullet.
And bacterial infections, phage resistance is something that is one of the, I don't
know if it's a stumbling block, but it's something that has to be definitely brought
to light when trying to research phage therapy and create phages that will actually treat
these bacteria.
Because bacteria for even just being very small organisms are very, very crafty.
Very crafty.
And don't forget that bacteria have been defending themselves against phages for a few billion
years now.
The phages have probably existed for as long as bacteria have.
And so bacteria are exquisitely good at finding ways to evolve resistance to bacteria phages.
And certainly the first thing you see in the lab when you start working with phages is
how quickly bacteria can mutate to become resistant.
And I would argue probably it's even easier to become phage resistant than it is to become
resistant to antibiotics.
With gene editing, you mentioned gene editing prior to this.
Is there a way that you might be able to prevent phage resistance in the bacteria just kind
of manipulating gene editing?
And is that something that every so often would have to be recalculated for every batch
of new phages you brought out to make sure the resistance didn't last?
Yeah, that's a really interesting area that's starting to be explored.
So there's a lot of interest in kind of delivering some of these gene editing technologies within
bacteria phage particles.
And that's a really exciting idea because the phage, we're really like repurposing the
phage here.
The phage would no longer contain its own native genome, so it wouldn't be able to escape
and just like start replicating as a phage.
Instead it would be just used as a delivery vector.
So we are just tracking down the bacteria that we want to be, for example, a sensitive
to antibiotics again.
And we would deliver our gene editing technology and just get rid of those resistance genes
from a bacterial pathogen.
And there's quite a lot of interesting synthetic biology companies now developing those kinds
of products.
But it's still the early days, but I'm pretty excited about that direction.
It's so crazy.
It's like something out of a sci-fi novel.
Right.
It's really neat.
So you were recently published in Nature Communications.
In your article there were significant findings that actually might be relevant to treating
and stopping antimicrobial resistance.
I don't know if you can kind of go into that a little bit and then kind of explain how
these findings could help aiding against the antimicrobial resistance.
And maybe help explain there was one in particular, FT Midnight, which is a great name.
How did you guys come up with that name?
Who got to name it?
That's actually a cool story because we had one postdoc in the lab who was characterizing
those particular groups of phages.
She was goth, really into gothic stuff.
So she named all of her phages like dark colors because we have a general color theme for
our phage names.
And so all of Lucy's high-Lucy phages are dark goth colors.
It's awesome.
But yeah, so I can talk a little bit more about that project.
So essentially, so we've already talked about phages and how they enter bacterial cells.
And they typically, I told you, have a very narrow host range.
And that's usually dependent on some proteins on the surface of bacterial cells.
So phages just like human viruses, they use these receptors, molecules on the surface
of cells to know which cell they want to enter.
Similar to influenza, you'll have the H in the end.
So hemoglobin in, nernimides.
So it's like the H1N1, H5N1.
These are binding receptors essentially for viruses or just protein binding receptors
in general.
Exactly, exactly.
And so bacteria have an array of proteins and other structures on their surface that
phages can recognize and enter with.
And the one interesting thing about antibiotic resistance plasmids, so these are like these
mobile pieces of DNA that are frequently transmitting resistance genes, is they themselves produce
a structure on the bacterial cell surface.
And that's this like tube-like apparatus.
We call it conjugative pilus that's essentially there to mobilize the plasmid from cells to
cells.
And that's part of why antibiotic resistance is such a problem right now is these genes
can move themselves between bacteria.
They encode their own vehicles to transmit across bacterial populations.
So but the interesting thing when looking at this from a phage perspective is that kind
of that conjugative pilus, so that tube that the plasmid has made on the bacterial cell
surface becomes visible to phages because anything on the cell surface can be targeted
as a phage receptor.
So kind of interesting thing that was discovered in the kind of 70s and 80s is that there's
a whole populations of phages that have adapted to use these plasmid-encoded proteins as receptors
to infect cells.
So we call these plasmid-dependent phages.
And it's really cool because essentially now you have a way to specifically infect cells
that have resistance plasmids in them.
So we could, these phages won't do anything to a salmonella that isn't resistant to antibiotics,
but once that salmonella like gains a antibiotic resistance plasmid, it becomes sensitive to
these phages.
So I think they have like immense therapeutic potential because of that.
So back in the 70s and 80s, these plasmid-dependent phages were pretty widely reported, so people
were finding them all across the world for many different kinds of plasmids.
But then for some reason that I still don't quite understand, the field kind of shut down.
I think the labs who were working on it just happened to close and nobody took up that
research.
And phage research for a while kind of went out of fashion.
And then so recently, and yes, the other sad thing about that kind of research stopping
is it was pre-the genome sequencing era.
So we don't have any genome sequencing memory of what these phages were.
We don't, phages don't last all that long in the lab, so they've subsequently kind of
like died off.
We don't have any record of them physically.
We just have these reports that plasmid-dependent phages exist and they are widespread and
can target many different kinds of resistance plasmids.
So in our work, we kind of set out to rediscover these lost plasmid-dependent phages to see
whether actually we could start to go towards translation of these phages to healthcare.
So we designed an assay to systematically detect them from the environment, which was
previously kind of challenging.
But now with our assay, it's very easy and we were super surprised to find that these
things are extremely abundant.
So in the early days of this project, it was happening really during the COVID times and
we weren't allowed too much time inside.
So my co-first author and me on the project, we were biking around Boston and Cambridge
going to community gardens and sampling the compost and the soil and any kind of like
gross samples we could find and testing for these phages.
And we were amazed to find like every single sample we tested contained these plasmid-dependent
phages.
They were really just absolutely everywhere.
And so we went on to characterize the phages, we sequenced them, we showed that there's
a lot of genetic diversity in the collection that we've found.
And so FT Midnight, you mentioned, is one of these plasmid-dependent phages that is dependent
on a plasmid that's very commonly found in E. coli bacteria, the F. plasmid.
And the reason why FT Midnight kind of features more prominently in the paper and has maybe
a special name is because it's a unique amongst our collection.
And the FT Midnight looks like the classic bacteria phage that you might have in your
mind.
So this kind of spaceship, moonland-like structure I'm wearing earrings of this structure.
And that's maybe might not sound that interesting, but a weird thing about plasmid-dependent phages
is they never have that structure.
They actually tend to be very unusual in their morphology.
They have like very non-canonical structures.
And so we were very excited to find FT Midnight because it's a suggestion that there is much
more diversity in these plasmid-dependent phages than we currently know.
And also that many types of phages might be able to evolve to target these antibiotic
resistance plasmid-encoded receptors.
So I'm really excited going forward to just find more of these things and really see like
how broadly can we target antibiotic resistance plasmids with phages.
And would that kind of present a solution to some of the resistance that the bacteria
would gain over time if you could have these phages that are specific that do not affect
essentially resistance genes for being created?
Yeah.
So you can really cure bacteria of antibiotic resistance with these phages.
The only way bacteria can become resistant is by completely losing these plasmids that
encode the resistance.
Or the second way is they can mutate the genes that encode that protein tube.
But if they do that, the resistance plasmid can no longer move through bacterial populations.
So we're targeting dissemination of these genes and we're targeting the resistance
itself.
So it's kind of an exciting, like win-win evolutionary outcome that has given me a lot
of optimism in terms of therapy.
It's pretty amazing pride when you think about it.
So that's so cool.
And this is the disco technique that you guys used?
Disco, yeah.
And can you tell me how you guys came up with that name?
Yeah, well there's so much pressure inside us to come up with catchy names and show your
stuff.
So it took us a long time actually.
We went through many iterations of Bacronyms until we landed on disco.
But it's very simple.
The disco means discovery by co-culture.
And essentially our assay relies on there being two different types of bacteria co-cultured
together so we can find these plasmid-dependent phages.
Catchy name.
It's memorable.
People seem to like it.
It's going well.
I know that I think I read the samples from your research, a lot of them for this project
actually came from a wastewater treatment plant.
So who gets to actually procure that wastewater?
You guys get to travel there.
We get to go.
How much fun was it?
Us ourselves.
It's the highlight of our week going down.
No, and I'm not even joking here.
I'm a big fan of Deer Island wastewater treatment plant.
It's down in East Boston.
It's the second largest wastewater treatment plant in the mainland US.
I think the largest is in Illinois.
But Deer Island is incredibly futuristic.
You wouldn't believe this, but people from all over the world come on infrastructure
tourism to visit Deer Island because it's so famous amongst wastewater treatment plants
in terms of the technology it uses.
It's a fantastic place to find phages because it's just a mix of all of these effluents
from so many different local constituencies.
And actually I would really encourage the listeners of this podcast if you're local
to Boston, a little known fact is that actually the MWRA offers free tours to the public of
Deer Island Waste Waste Treatment Plant and they let you go all the way up the top of
the eggs.
And it's a fantastic experience that everyone should do.
How bad does it smell?
You know, people always ask that wastewater isn't that gross to work with.
It really is just kind of like brown, cloudy liquid with the large particulates have been
filtered out.
It smells bad, but the bacteria smell worse, trust me.
Yeah.
I get to deal with the fresh poop in my job, so at least you get some of the filtration.
This is much better, yeah.
So I can imagine it.
We kind of mentioned earlier some of the barriers to phages and phage therapy.
At the moment, and you had mentioned this earlier as well, at least in the United States,
it's only allowed as expanded access, which is otherwise known as compassionate use.
So compassionate use, we actually saw this during COVID with a number of drugs that were
just trialed just to try and see if they worked, kind of throw the spaghetti at the wall and
see if it sticks.
So it means that quote, and this is from, I think it's a USDA, treatment outside of
clinical trials when no comparable or satisfactory alternative therapy options are available.
And right now, that's the only way phages can be used to actually treat illness here
in the United States.
What do you see as the current barriers to actually being able to provide phage therapy
as an everyday treatment to kind of kill some of these hard to treat bacterial infections?
Yeah.
So I think the biggest issue is there are so many kinds of infections that people want
to treat.
And with developing phage therapies, it really has to be on A, a pathogen by pathogen basis,
and you need a therapy that's specific to each kind of infection.
And the biggest challenge tends to be delivery of these things.
So there are certain sites in the body that seem to be slightly easier to get to.
So there's a lot of promising research that respiratory diseases, a lot of people interested
in cystic fibrosis treatments, are using phage because phage are very easily nebulized.
So it's quite easy to deliver them into areas like the lungs.
On the other hand, areas of the human body that are harder to reach or are challenging,
phages are proteinaceous, so they elicit an immune response sometimes.
So delivery, intravenous delivery is something that's kind of not well explored because there's
a lot of, you have to get an extremely pure product to not elicit a dangerous immune response.
And even then, some phages seem to elicit high responses and will just be cleared by
the immune system as though they're a foreign virus or something.
So yeah, so it really needs to be kind of pathogen specific and infection specific.
But there's certainly some hope.
And in terms of compassionate use, that's like one of the other issues here is because
phage is such a new and unheard of therapy, there have been very few actual clinical trials
using phage therapy and therefore we only have these case studies, these compassionate
use cases.
And in some ways, those are really interesting data because they really allow researchers
and clinicians to do things that you wouldn't be allowed to do in other ways.
So an example of this was there was a quite high profile case of a mycobacteria obsessus
case.
I think it was in the UK actually and it was like a really awful systemic, I think, mycobacteria
obsessus case infection that was like pretty nasty and bad and they were eventually given
compassionate use grounds to use a phage.
And one of the interesting things about mycobacteria obsessus is there's no known phages that don't
enter this integrative lifestyle.
So we only have phages that we know can just integrate into the genome.
But Graham Hapful, who's a fantastic researcher in Pittsburgh, has kind of been working on
mycobacterium phages for a long time and knows how to make mutants so he genetically engineered
a version of these mycobacterium obsessus phages, preventing them from going down this integrative
life cycle, which means you essentially now have a GM phage that's much more idealized
for phage therapy purposes.
And in normal terms, right, you'd never get permission to use a GM bacteriophage.
It's so untested and unheard of, but they were given permission to use this in this case
and it was a successful outcome to the treatment.
So in that way, these kinds of like compassionate use cases can really push the field forward
because you can be given the chance to do something kind of wacky, something that you
might not get permission to do in a much broader clinical trial.
And it could be very successful and we can get amazing data.
So it's not all bad, but I think we definitely need kind of a combination.
So we can only go so far with compassionate use.
There are also difficult cases to get good data from because they tend to be cases where
patients are in pretty bad diastrates and the actual like improvements tend to be fairly
short term anyway.
So yeah, getting better clinical trials in other kinds of patients is something that's
really needed.
Yeah, it's even when looking to try and find cases, a lot of them, I wouldn't say they're
anecdotal, but there's definitely not a lot of concrete data to support like where they've
been used.
As far as funding goes for phage therapy, it's significantly lower than antibiotic research
by many, many, many, many millions of dollars.
What do you feel needs to happen to actually help to grow and expand funding for research?
Do you feel that there needs to be just a louder voice in the biology or even the medical
community to try and gain more funding for this?
Yeah, so change in science tends to be slow and I'm actually optimistic about that.
It does seem like most funding bodies do have an appetite for phage research now and certainly
funding for phage research is increasing.
So I think things are moving in the right direction.
I think funding for science in general needs to increase.
That's something that tends to be fluctuate a little bit depending on the political climate.
Certainly I think I'm optimistic that phage research is going to be funded, but we just
need general funding for basic science research.
So you had mentioned earlier just with the use of antibiotics and how a majority of that
is actually not for treating illness, but it's actually prevention and agriculture and
other areas.
I want to talk about some of the additional uses for phages just beyond right now treating
for MDIROS and such.
I know that there's a fellow phage researcher, Hanyi Nani, and he's a pretty big proponent
of using phages just to improve food safety.
Beyond food safety improvement, what other possibilities do you think phages could be
used for just in everyday life or maybe antibiotics are used right now?
Yeah, it was.
The food safety field for phages is very successful.
If people don't realise, but if you buy sliced meat and cheese in the US, it's probably
being sprayed with phages.
I love your consuming them every day.
Those kinds of products are outside of the human body.
Phages are much easier to make products for.
They're not really regulated in the same way.
So there's a lot of interest in therapy for, not therapy, but treatment for acne, things
like this.
If you go to San Francisco, I think you can buy from a startup there, tubes of phage that
you can put on your face to help reduce your acne.
There's a lot of mounting evidence from the microbiome world now that actually bacteria
in your body can affect all kinds of diseases beyond what we traditionally think of as infectious
disease.
So there's pretty good evidence now that your microbiome might be playing a role in your
susceptibility to things like Alzheimer's or type 1 diabetes.
And so as research like that progresses, it's possible that we might even start to be able
to use phages to reduce our risk of other kinds of diseases that we don't think of as
infectious.
Another interesting area is mitochondrial diseases.
So these are very rare diseases in humans, but they often have really terrible symptoms
and can be like really life-altering.
And so far in the gene therapy world, it's very difficult to edit the mitochondria because
it's a part of the human cell that's like enclosed.
And mitochondria themselves were once bacteria.
So mitochondria were engulfed by eukaryotic cells way, way, way, way back when eukaryotic
cells first evolved.
And so there's a potential there.
So mitochondria still look like bacteria.
So there's some interest in potentially using phage proteins or some strategies inspired
by phages to get gene editing technology inside human and animal mitochondria, which is exciting.
It should be pretty crazy, especially there have been links to long COVID and mitochondrial
disorders.
And that's one of the biggest complaints of people with long COVID is just lack of energy
and lethargy, which mitochondrial disorders, they call it the powerhouse of the cell and
it essentially creates, it drives the energy for cells to work.
So if you have a mitochondrial disorder, it's you, a lot of people just have no energy
to do anything.
Your body's just not producing it.
So that'd be pretty crazy.
It's pretty significant.
And it's amazing the avenues that can be discovered that we can go down with phages.
More research.
So in a recent research article that was published, I guess it was in nature May of 2023, you
and some of your fellow researchers, you developed a way to use genetic modification to help
cells avoid viral infection.
Can you actually explain, and this isn't, this is, you know, with viruses, how phages
might be able to help combat viral infections and possible limitations of this and how it
might actually change the way we tend to look at and treat viruses?
Yeah.
So that was a really exciting project I got to be involved with, led by the church lab
here at Harvard Medical School and the VISA Institute.
So essentially the church lab is really interested in reprogramming cells to be virus resistant
because obviously that would be fantastic, right?
But it's a very long-term goal here.
So the first step in this process is start with bacteria.
And the strategy that they're taking is essentially recoding the language that bacteria use to
translate these protein messages.
So what the church lab did, so they took a strain, the original strain had actually been
produced in the lab in Cambridge in the UK.
And in this strain, researchers had removed three of the codons that code for amino acids.
So we have, there are 60 codons, so these are like triplets, three letters, that code
for only 20 amino acids.
So essentially there's a redundancy in the code that we all use to code for our proteins
and that allowed them to take some of these redundant codons and make change what they
code for.
And that means that if a virus, which hasn't been recoded, it's a natural virus, comes
and tries to infect one of these recoded bacteria, it tries to translate its protein messages
and they get, they don't make sense.
They're translated into something that they wouldn't, they're not meant to be.
It's just like in a language, it's like changing a language and now your translation is wrong.
So the virus cannot build its proteins, it cannot replicate.
And so this original strain that was produced in Cambridge in the UK was claimed to be virus
resistant because of this recoding that had been done.
And so the church lab here at Harvard Medical School, though, were a little suspicious that
this is the case and so they set me in my lab about trying to find viruses that would
affect this recoded strain of E. coli and we were actually able to find them pretty easily
in Boston, dear island, wastewater.
And so the way that phages were getting around this was actually pretty simple.
They encoded for their own systems to translate their own protein messages.
And so in the end, so the kind of conclusion of that work was we made further edits to the
E. coli, further recoding that made it much harder for viruses to kind of escape this
process and also added in a kind of containment method.
So we made that E. coli strain dependent on an amino acid that doesn't exist in nature.
So essentially we've locked that E. coli strain into laboratory or special culture so it cannot
escape and there's no fears of a virus resistant E. coli ever escaping the lab.
And so even though this is a long way away from this very far off goal of making humans
virus resistant, it's an exciting development in terms of industry and industrial production
of a lot of compounds and enzymes.
So a lot of the industry produced things are done in bacteria, so enzymes that we use every
day and some medicines.
And actually there's a huge amount of revenue lost and also just wastage from viral phage
contamination into industrial like enzyme preps.
So having this virus is an E. coli strain is actually very exciting, not from a human
health perspective yet.
From a research perspective though, I mean just think about how many lost samples you
won't have if you have something that's...
Exactly, exactly.
And commercially it could be hugely profitable.
So it's a kind of exciting step forward.
And yeah, but who knows, but after one day we'll be able to use the same technology on
animals.
Once again, I don't know.
I'm always fearful.
And once again I said there's no free lunches in nature.
Like eventually a virus will find a way to get you in and it'll probably be really bad.
But when you tinker with science, it's for as much good sometimes you create bad.
But it's really cool.
So this brings me to my next question.
So with the growing use of artificial intelligence and all aspects of science and life in general,
I use it to create silly songs and weird pictures.
But you can use it for...
In science it's being used in so many different ways and it's unbelievable how fast it's growing.
And I can imagine it will be incredibly beneficial.
But how much do you think AI is going to improve phage research in the future?
Yeah, I'm cautious about that.
It's not something I have a ton of experience personally with.
One thing I have used it for though and I'm excited about is the potential for AI to help
us design proteins.
So AI has turned out to be extremely good for understanding the structure of proteins
that we already know about and potentially even designing proteins to have a specific
function that we need.
And so as we start to learn more about the proteins that bacteria phage is encode, I
am excited about the potential to maybe take one chassis of one phage and maybe adapt it
a different tail onto it so that it can infect a new host or many hosts.
And that kind of thing I think is going to start to be possible in the next decade with
advances of AI.
And also in terms of phage therapy, more traditionally, we have so much data about what phages infect
what strain of bacteria.
But as yet, that's not possible to do in the reverse.
So it's not possible to look at a strain of bacteria at the genome level and predict
what phages will infect it because it's just so complicated, like understanding what factors
the phage is using to infect that and what genes the bacteria might have that resist
the phage.
We can't yet do that.
But it's possible that as we start accumulating all this data into host range, there is speculation
that we might be able to start to train models, AI models, to just predict from a given bacterial
genome what phages we could use to treat it.
And that would certainly speed up phage therapy because one of the challenges right now is
these phages are so specific to a single strain.
So if you have a patient, you have to first isolate the strain of bacteria that they have
and then screen that bacteria across with all the phages that you have to find maybe
one or two that will work.
So we had a way to just pull from a library immediately.
It might really speed up the process.
And I was going to ask about phage libraries, if you could maybe explain what those are
and how they're used with phage therapy because it's almost like with antibiotics you have
a broad spectrum antibiotic.
It's one antibiotic that can do damage to a lot of different bacteria.
Whereas phage libraries contain numerous phages and can be pulled from to try and treat one
when you don't specifically know the susceptibility of the bacteria to phages, correct?
Exactly.
So it's like, yeah, it goes back to this problem or could be seen as a benefit depending on
how you look at it of phages and that they are very specific, they're narrow spectrum.
And so a phage library is essentially when you collect all the phages against a given
pathogen.
And that can be using one strain or it can be using many strains.
But having that large collection allows you to, when you have a new isolate that comes
in from a patient, you could very rapidly screen hundreds or thousands of phages that
might hit that strain until you have like a few good ones and then you can use them.
Because just putting one phage with one bacteria and hoping it's going to work is the probability
is very low here.
These things are exquisitely sensitive and specific.
And so the other kind of alternative to that, the solution that people have created is what's
called the phage cocktail.
So in this case, you would formulate a mixture of phages from your library that tend to have
a pretty good activity against most of the bacterial strains you're seeing.
So if it's like E. coli, in fact, UTI infections, you might have a cocktail of phages that hit
maybe 90% of the E. coli that people tend to get.
And then you can be fairly certain that your therapy will work and you can administer it
immediately without having to do this retroactive testing of the strain and the sensitivities.
Is that something that AI could help with as far as, because I mean large language models
just at their basic level is just gathering a multitude of information and finding the
best answers or answer for it.
With regard to phage libraries, because some of them are vast, is this something, if those
are all catalogued and you used AI to try and would it speed up possibly or help narrow
down specific phages or phage cocktails that could be used for treating specific bacteria?
Yeah, I know that people are definitely working on that and seeing if they can kind of train
AI models to predict which phages in our library will make the best cocktail for a given strain
or predict the outcomes.
Yeah, so to be continued, I guess, it's like ongoing research in the field.
So let's talk a little bit about the future of phages and then your own future as it pertains
to the lab that you'll be working in.
There seems to be a renewed interest in phage therapy, especially because of the antimicrobial
resistance and antimicrobial stewardship that's happening now.
What do you think the future will actually hold for phages?
You said earlier, you're optimistic, but at the same time with phage therapy, you're not
100% sure that there'll ever be a day where it's used regularly.
What do you hope for anyway with phages in the future?
I hope for more discovery.
So one of the things we haven't really talked about yet is the sheer diversity of phages
on this planet.
So phages are thought to be the most abundant biological entity on the planet, right?
Like it's very hard to understand what these numbers mean, but it's estimated that in your
body right now, there's probably a trillion bacteria phages, right?
For every grain of sand on the planet, there's a trillion bacteria phages, just like inconceivable
numbers.
And so the cool thing there is that there is so much biological diversity, and we only
have a tiny snapshot of it right now.
And so I'm really excited about further discovery and just mining, biological prospecting, if
you like, all of the functions that we might be able to get out of these diverse phages.
And certainly, another thing that I think is going to happen in the future is most of
the phage research has focused on a very small number of bacteria.
We've typically only are interested in human pathogens in microbiology.
There's so many neglected species of bacteria that are just over there infecting, I don't
know, amoeba or something, and we know nothing about their phages.
So yeah, I'm very excited about general discovery.
And yeah, looking into this vast unknown of bacteria phage diversity.
So could they be used?
Do you think I know you're talking about just abundantly different organisms beyond bacteria?
I know that there is malaria.
We're now having malaria resistant strains to antiparasitics.
Is that something that even with parasites that possibly we could find?
I mean, I don't know if they are phages specific just to bacteria.
Is it something that other microorganisms?
Yes.
So phages are the term phage specifically refers to a virus of a bacteria.
But indeed, any other any organism, I think, has a virus.
And yeah, we just don't always know about it.
So there's certain lifestyles that particularly parasites might have.
So malaria tends to live inside human cells.
So you can imagine it would be difficult for a virus to access it.
And therefore, viruses might not be there might not be immediate viruses of malaria.
But perhaps there are viruses of free living relatives of malaria that we could once we
discovered them, we could maybe repurpose them.
And yeah, I'm excited about the general diversity of viruses of microorganisms.
It's such an unexplored topic.
So with your loud that you're going to be heading up soon as you as you get out of Boston and
head to New York, we'll be continuing the same type of research, we'll be doing newer
research.
Do you do you even have an idea yet?
I'm sure you do.
Are you able to help just share with us what you're going to be working on over the coming
months, years and just into the future?
Yeah, so I will be heading to the Wadsworth Center in Albany.
And so the Wadsworth Center is affiliated with the New York State Department of Health.
So they have a remit to do basic science that is kind of adjacent to the public health surveillance
of New York State.
And so I'll be continuing my research into plasma-dependent phages.
I'm still very interested in antibiotic resistance and it's kind of a great grounding point for
me.
So my focus really will be on discovery of more plasma-dependent phages.
One of the exciting things for me about the Wadsworth Center is they are running the wastewater
surveillance for the whole state of New York.
So we're getting wastewater from over 200 wastewater treatment plants on a weekly basis.
It's a phage gold mine.
So yeah, so I'm very excited to do some phage disc going and discover a lot of new phages.
And I'm also interested in how bacteria become resistant to these phages.
So the phage defense field I mentioned earlier has really exploded in the last decade or
so.
But so far these plasma-dependent phages have remained kind of on the sidelines in terms
of defense.
We don't really know how bacteria, what specialized defense systems bacteria have against plasma-dependent
phages.
So that's going to be another focus of my research.
So cool.
With all your hard work, all the time and effort that you put in, the passion that you
have for it, which is obvious, and I love that, the enthusiasm, not only for just the
work you do, but it could help so much, it could change just kind of the way we treat
people and illnesses that are starting to get out of hand right now.
How exciting and do you actually find it really gratifying to now just be heading up your
own lab?
It must be great.
It's a dream.
It's really, really exciting.
I mean, I've been very lucky in my whole career and I've had mentors who've really let me
have a lot of freedom in science and really kind of do my own thing.
So I feel very ready for this.
I feel like I've been always leading a mini lab inside labs for a while now.
But it's very exciting to get to this final stage of total independence and being able
to look for any phages I want without asking for permission.
So it's extremely gratifying, yeah.
For anybody who might be interested in microbiology or phages or just science in general, what
advice would you have with regarding to just pursuing your passion?
I would say be tenacious.
Just keep going.
If it's interesting to you, it doesn't matter if it's interesting to other people.
Sometimes I think we feel pressure to do science that is exciting and is fun and will make
a nature paper.
And I think it's really important to try and let go of that and just follow your nose, follow
the topics that are really interesting to you.
And at the kind of more junior level or the public, I think just talk to everybody.
You'd be surprised at how willing scientists are to just talk to you.
We don't get out the lab that much.
So often scientists, if they're really interested in their research, they will talk your ear
off about it.
So yeah, if you're interested, reach out to your local scientist and talk to them about
it.
And this is proof of that because prior to just a couple of random emails, I had no prior
conversations with anybody at the BAME lab and I was so happy just to actually have you
here.
Thank you so much for coming on.
Any other advice, anything you want to talk about regarding phages or anything we didn't
touch on that you think might be important to discuss when it comes to like phages, phage
therapy, the future of phages?
One piece of advice I have for any undergrads or soon to be undergrads or parents of undergrads
who might be listening and are interested, have any interest in phages is to look for
colleges that run the C-phages program.
So this is an amazing undergraduate program that was spun out of Graham Hattfall's lab
at the Pittsburgh.
And essentially they allow undergrads across the country to find their own phages.
You get to sequence the genome, you get to name the phage.
It's an incredible program and it's a really great way to get into molecular microbiology.
How much lab equipment do you need to have to be able to discover phages?
Honestly, not a lot.
It's really amazing how the techniques we use in the lab today for phage research have
not changed in almost a century now.
Which is crazy.
You would think that with technology.
I mean, we're digital now.
Everything is digital.
Back then it was microscopes and light and that was about it.
Do you see the future of research being molded to, I don't know, being upgraded I guess.
With digital technology and artificial intelligence once again and being more computerized.
I mean, there will always be the lab work that will have to happen, the physical aspects
of it.
But just from computing and just from that aspect.
Absolutely.
And it's happening in all areas of biology.
I would say though that phage research has been a little late to kind of develop new
technologies.
So that is something I see coming and already is happening as we're starting to develop
more modern technologies for studying phages.
And yeah, that's again something I'm integrating into my research.
Sean, thank you so much for coming on.
This has been awesome.
I appreciate it so much.
Really happy to get you on.
Especially as you're heading off in just a few days to get out of this place.
I appreciate it.
I'll be following you with your new discoveries in your lab.
And I don't know if you want to share any of your social media, anything like that.
Or if that's something you want to keep private, it's up to you.
But if you want to feel free.
Yeah.
Thanks, Andrew, so much for having me.
And thanks everyone for listening.
You can follow me on Twitter at implosion.
Which is a great name.
Awesome.
Thanks, Sean.
I appreciate it again.
And for everybody else listening out there, thank you very much.
As always, emails at makemesickpod at gmail.com and you can find us on Twitter at makemesickpod.
Thank you all for listening.
And remember everybody to please wash your hands.