Yale Psychiatry Grand Rounds: April 22, 2022

April 22, 2022

Ribicoff Lecture: "Translating Addiction: Molecular Insights to Risk and Therapeutic"

Yasmin Hurd, PhD, Professor, Icahn School of Medicine at Mount Sinai

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00:03Marina, thank you so much for that
00:06really great introduction and I'm let
00:08me share my screen and I do that.
00:10You know, it's it's quite an honor to
00:14be given this opportunity to be the.
00:18Ribicoff speaker for for this year and
00:21especially that great list of of previous
00:24speakers and just for me, I think.
00:27And obviously everybody.
00:28I think Yale has a great reputation
00:30of understanding the importance of
00:32evidence based medicine and the only
00:35way for us to really get there and
00:38move the field forward is having
00:39this close interaction between basic
00:41scientists and clinical and the
00:43and the clinical and clinicians.
00:46So again, thanks for this opportunity.
00:49So I'm going to move all the lovely
00:53pictures of everyone and the first
00:55thing I will say is that you know
00:58I don't have anything to dispose
01:00in terms of the the work that I'll
01:03be talking to you about today.
01:05And basically you know for me,
01:07the Addiction Institute of Mount
01:09Sinai that I direct is.
01:13It brought home the critical nature of
01:15of of research and where we are today,
01:18and the reason is in large part
01:21when we think about a lot of the
01:23substance use disorders at Mount Sinai,
01:25we treat over 6000 people with
01:27an opiate use disorder.
01:28So you can imagine the the challenges
01:30there and it really reflects the
01:32opioid crisis that we are still in.
01:35And as many of you know,
01:36you know the economic burden
01:38of the opioid crisis.
01:40The healthcare system under
01:42siege because it's.
01:43It does cost a lot,
01:45is it's costing nearly three times more
01:47than other medical disorders to trade,
01:49and the treatments that are that
01:51are available are often not used,
01:52and some of them not suitable and we
01:56still have so many overdose deaths
01:59today and that has been the crisis
02:02have been exacerbated by covid's.
02:04COVID has exacerbated so many things,
02:07but the stress and social isolation
02:09has increased drug use.
02:10And of course,
02:11you see that a lot with opioids,
02:12and especially unfortunately.
02:14With fentanyl,
02:15that 4th wave of opioids that
02:19contribute significantly to to dread
02:21to drug overdose and so that you know.
02:24During COVID you've seen so many
02:27more people dying of opioid overdose,
02:29so you know.
02:31So for me, the question has always been,
02:33you know, how can we improve?
02:35What's what we're doing and a part
02:37of that comes back to treatments.
02:39And when you look at treatments
02:41for substance use disorders,
02:42you know there is actually a long history.
02:44And it's interesting that there were
02:47morphine maintenance clinics already in 1919,
02:49and as you look across the years of the
02:52medications that have been developed.
02:55There've been a number.
02:56I'm going to focus on the opioids
02:58because most of the the treatments
03:00that have been developed and hear
03:02from 1964 with methadone to today.
03:05They are all focused on the opioid system.
03:08So opioid agonist in large part,
03:11not only methadone, buprenorphine,
03:13and of course,
03:14we have naltrexone,
03:16especially for trying to reduce overdose.
03:20And there are, of course
03:22behavioral therapies as well,
03:24but as I mentioned earlier,
03:26the problem that we have with a lot
03:27of the treatments is that only about
03:2920% of the people who need opioid
03:32use treatment actually receive it,
03:33and the the reasons are numerous
03:35things start with even the stigma,
03:38because unfortunately,
03:39even though these medications do work,
03:41they've saved millions of lives.
03:43The stigma associated with with opioid
03:47agonist and many programs not wanting
03:49to quote UN quote their their their
03:52clients to be on any medication.
03:54That might be have an addictive
03:57potential itself,
03:58so there are these challenges as well.
04:00And ironically,
04:01the governmental regulations
04:03of using these opioid.
04:05Treatments make it very challenging,
04:08clinically and for patients to
04:09even access these treatments,
04:11sometimes in places in the US
04:13where people have to drive or find
04:15a way to get to them for hours.
04:17And as I said, from 19 six to four,
04:20we really have had this one size fit
04:23all approach to treating opiate use disorder.
04:26So for me, when you look at
04:28the neurobiology of addiction,
04:29you know the question is what
04:31have we learned?
04:32We have learned a lot really a lot.
04:34We have learned a lot about.
04:36Different brain regions and
04:38neural circuits relevant to
04:40phenotypes important for addiction.
04:42For example,
04:43the ventral steroidal area that which is
04:45a nucleus accumbens important for reward,
04:47expectation, goal directed behavior.
04:49The dorsal striatum.
04:51Important for habit, habitual formation,
04:54the prefrontal cortex,
04:55and the number of the it's sub regions
04:58such as the orbital frontal region.
05:00Cognitive control goal, directed behavior.
05:04Cognitive flexibility.
05:06Emotional regulation and so on.
05:08And we do know that for the most
05:11drugs of abuse that the use acute
05:13use of the drug does lead to,
05:15for example,
05:16this increasing dopamine that's
05:18associated with business phoria and
05:21so many research has gone into this.
05:24This acute changes that occurs with drug use.
05:28So for me,
05:29a quick question that I asked many
05:31many years ago that then became the
05:33foundation for my research was really
05:35what have we learned about the human brain,
05:37especially the molecular level.
05:39Because it was so challenging
05:41to get that kind of insights.
05:43And so I thought,
05:45OK,
05:45let's start studying the
05:47postmortem human brain.
05:48And I mean clearly it's very complicated.
05:51But you know,
05:53we know that you know studying
05:55gene expression looking at aspects
05:57of genetics of individuals,
05:58and I'll come back to epigenetic
06:01mechanisms that Marina mentioned that
06:03altogether this leads to changing
06:05proteins in the function and the
06:07phenotype of events of the disease so.
06:09That's where I'm going to start and
06:11and spend like the first few part.
06:13The first half I've been going
06:15north of my talk in terms of what
06:17have we learned about the human
06:19brain that guides our animal
06:20models to then see if we can start
06:23developing new treatments.
06:24So when we looked for
06:26example here in the striatum
06:28of human heroin users.
06:30Using approaches that were agnostic,
06:35Welsh and unbiased in a way of
06:38looking at thousands of genes,
06:40whether it initially was microarray
06:42strategies or more recently RNA
06:45sequencing of the transcriptome,
06:47we could see clearly that the gene
06:50expression signature inhering users
06:52differed from normal controls and
06:55where it differed was interesting
06:58because we saw much greater.
07:00This regulation of glutamatergic genes
07:03and perhaps not really surprising of
07:06the synaptic plasticity related genes.
07:08This the striatum receives really
07:10strong innovations from the prefrontal
07:13cortex and we know from a number of
07:16animal studies that that is really
07:18critical for especially aspects
07:20of even of drug seeking behavior.
07:22But we were surprised when we saw these
07:25really profound changes of epigenetics.
07:28And, importantly,
07:29these epigenetic marks were were.
07:33Opened up a new line of research for us,
07:35also in some aspects of our
07:37developmental cannabis studies.
07:38Because we focused a lot initially
07:41in trying to understand the
07:44individual vulnerability that.
07:46Why do some people?
07:48Many people may take a drug,
07:50but only a certain percentage
07:51may develop that disorder and
07:53we focus initially on genetics.
07:55But the environment of this complex
07:58disorder addiction plays a critical
08:01role and it plays a critical role in
08:03being able to change gene expression.
08:06And and in fact sometimes can
08:08override these genetic blueprints.
08:10So genes that should be closed are
08:12now turned on and genes that are that
08:14should be turned on and now turned off.
08:16And there are numerous epigenetic
08:20Marks and epigenetic mechanisms.
08:21Trust me,
08:22we've only touching the surface
08:24of it right now,
08:26but we had a fundamental understanding
08:28of what some of these epigenetic
08:30marks may mean on a functional level.
08:33So, for example, if you have methylation.
08:36DNA often that was a repressive
08:39mark and would reduce transcription,
08:42while for example,
08:44assimilation of the histones
08:46around that wrap around the DNA,
08:49the DNA wrapped around these histone
08:52proteins that regulate transcription
08:54and assimilation would open up
08:56those the those regions of the
08:58gene and turn on transcription.
09:00And depending on where methylation
09:01of histones occur,
09:02you could also have transcriptional
09:05repression.
09:06And what we saw in the brains of heroin
09:08users in Australia and predicted in
09:11particular of these epigenetic remodelers,
09:13it was predictive of this
09:15enhanced transcriptional state.
09:17But it was specific to certain gene.
09:21Gene regulators and often came
09:24back to the synaptic plasticity,
09:26so these epigenetic marks
09:28would correlate very strongly.
09:30Inherent users with glutamatergic
09:32genes or synaptic plasticity genes
09:35and it was very specific where these
09:39epigenetic tags were predicted to occur,
09:42and a lot of the the genetic
09:44changes related to, for example,
09:47a civilation and assimilation
09:49of the the lysine.
09:52In a region,
09:53and I'm not going to get too detail
09:55on the molecular related to enhancer
09:57regulation of gene transcription and it
10:01related to their years of heroin use.
10:04We were able to look at this in many
10:07different using many different techniques,
10:09and we saw consistently this.
10:11This opening of the transcriptome in in
10:15relation to the years of heroin news
10:17and especially cassette around these
10:20synaptic plasticity related genes.
10:22But because humans have such a very
10:24life and we don't know that much about
10:26all their lives just about their
10:27toxicology and often like I said,
10:29some of the people we knew their
10:31history of of hearing news,
10:32we use our animal models where animals
10:35will self administer heroin and we
10:37could actually replicate where in the
10:39genome or in the in the transcription.
10:41Say that we saw this gene expression
10:43changes and when we looked at the
10:45epigenetic tags in these regions,
10:47they correlated exactly to what we saw in,
10:49for example,
10:50their civilation of these these
10:53synaptic plasticity related genes and
10:55this acetylation of lysine 27 and I'm
10:59I'm not getting as as detail I know
11:01because even more broad clinical audience.
11:04But what is?
11:06What does that epigenetic change really mean?
11:10So for for the the histones in terms
11:13of we have tags that are put on these
11:17histones and they're they're called writers.
11:20These enzymes and we have others
11:22that take the these tags off,
11:24and we then have a readers that must
11:27give information to the system as to
11:29what these signals mean, and and they're.
11:31As I said, they're called readers.
11:33And for assimilation the bromodomain
11:35are the readers.
11:37They bind these assimilated lysine residues,
11:39and there are.
11:41About four families of these BT.
11:43These bromodomain proteins 32,
11:45three and four.
11:47And there's also one in the testes,
11:48but we only have 3 at least so far
11:51that was identified in the brain,
11:53and these are expressed in numerous
11:56brain regions.
11:57BRD 2-3 and four.
11:58But when we look in the brains both in in
12:00human hair and users and and animal models,
12:03we didn't see changes in BRD two or three.
12:05What we saw were changes in beer D4 and
12:09in different cohorts that we study.
12:12So.
12:12The thing that was also fascinating
12:15for us was that the again,
12:17the changes in in BRD 4 correlated
12:21very strongly with synaptic markers
12:23of markers of synaptic plasticity,
12:27such as DLG 4,
12:28which is the gene that encodes
12:30PSD 95 this postsynaptic density.
12:34Proteins.
12:34So again,
12:35every single thing told us that there
12:38was something really interesting about
12:40the BRD 4, the Bromodomain readers.
12:43So the thing that's very important
12:46about epigenetics and and at
12:48least at the time when we started
12:50studying this was although now in
12:53neuroscience more and more focus is
12:55looking at epigenetic mechanisms.
12:58The greatest information that we know
13:00really comes from the cancer field.
13:02And there they've been able to identify many,
13:05many different.
13:10Chemicals that can inhibit specific
13:13epigenetic mechanisms or promote
13:16certain epigenetic mechanisms.
13:18Because the cancer is definitely a
13:20disorder of epigenetic gone awry,
13:22so we were able to leverage what was
13:25being developed at that time in terms
13:27of some of the the chemicals to see
13:29whether or not if we could inhibit BRD,
13:31we had hoped to inhibit bird for,
13:34specifically.
13:35Could that itself decrease or heroin self?
13:40Administration and at the time,
13:42the prototypical BRD inhibitor,
13:44they would promote it as BRD.
13:474 inhibitor.
13:47It really was not selected because
13:50it also could bind to the R.
13:52D2 and D3,
13:54so we nevertheless looked at the JQ
13:57one was approachable the inhibitor
13:59at a time in our animal models,
14:01and interestingly,
14:02when we gave it into the striatum,
14:06we could reduce heroin self administration
14:09and heroin seeking behavior.
14:10But the goal long term goal is to
14:13be able to develop medications.
14:15So we we know we're not going to
14:17infuse it into the brain of of people,
14:19and so even giving it systemically,
14:21we could significantly reduce heroin
14:24self administration behavior in the animals.
14:27So.
14:29The the leveraging you know the
14:32looking at the postmortems brains
14:34of heroin users you were able to
14:37see that these epigenetic changes,
14:38especially those that related to
14:41genes as part of synaptic plasticity,
14:44especially these little mergent genes,
14:46they correlated significantly
14:47with the years of heroin use and
14:50inhibiting it we could inhibit
14:52harrowing self administration
14:53behavior and more importantly,
14:55heroin seeking behavior.
14:59Unfortunately we have tried for many
15:01years to try to develop a or to obtain
15:05even a specific beauty for inhibitor and
15:08we still have not been able to do that.
15:11So you know, I I feel every time I
15:15give these this talk of what our our,
15:18you know these BT family of inhibitors,
15:22how they're being developed
15:23in the cancer field.
15:24But we still have not been able to
15:26find some that actually penetrate
15:27the brain on these the BRD.
15:29What specific purity for inhibitors that
15:33passes the blood brain barrier effectively?
15:35And so we're still trying,
15:38but it's important to also emphasize
15:40that it's not only our group that
15:42has seen these changes in beard for
15:44relevant to substance use disorders,
15:46other groups have seen that.
15:48In fact you cocaine self administration will
15:52also increase variety for and inhibiting.
15:56Giving the JQ one inhibitor the B
15:58inhibitor also will decrease cocaine,
16:00place preference and cocaine self
16:02administration and I think that that's
16:03really important because when we
16:05think about substance use disorders
16:06often we think OK we must get the
16:08most selective thing or I think in
16:11psychiatry in general I think you know
16:13the the strongest antagonists of the
16:16dopamine receptor and and selectivity,
16:18but for substance use disorders we know
16:21that many people are polysubstance users.
16:24So for me the fact that
16:25we see some commonality.
16:26Between the different substances of abuse,
16:29I think it's important so you know
16:32we're still optimistic about BRD 4.
16:36I'm still saying on this this,
16:39you know, in developing medications
16:40based on what we've seen,
16:42the human brain and how
16:43animal models can help us,
16:45I'm still going to focus on aspects of
16:47epigenetics and synaptic plasticity,
16:49and the reason is once again.
16:52When we look at the the brains
16:55of heroin users, as I said,
16:57you know when we we looked at the
17:00gene expression that transcriptome
17:02we were able to see these epigenetic
17:05genes related to epigenetic mechanisms
17:07and the synaptic plasticity.
17:08But you can also look at the epigenome
17:11itself in an agnostic manner.
17:12You can actually sequence the the
17:15epigenome and you can do that with
17:17a technique called a taxi which is
17:20assay for transposase accessible.
17:22Something basically,
17:23you're looking at chromatin state looking
17:26at where in the where in the epigenome,
17:29maybe in the chromatin is open,
17:31and therefore would be associated with gene
17:34transactivation and where it may be closed.
17:37And importantly, it's agnostic,
17:39because normally when you look at
17:42the epigenetic tags you actually
17:45choose a specific epigenetic mark.
17:48So for example,
17:49we might have chosen,
17:50like acetylation of lysine.
17:5227 that we showed that was changed
17:54in the brains of heroin users,
17:56but we know it's the combination of a
17:59number of epigenetic marks that will lead
18:01to these changes in gene transcription
18:03and therefore obviously downstream
18:05changes in in protein and function.
18:08So if we look agnostically using this,
18:11a taxi that has actually been extremely
18:14informative and finding out where which
18:16loci where in the in the epigenome is
18:19most significantly changed with hearing news.
18:22And importantly,
18:23this technique allows a something
18:25for me that's important for
18:26studying the human brain.
18:27It uses very little tissue,
18:31and this way we can look at different cells.
18:34Cell types in the human brain.
18:36Here in this particular study we
18:37look mainly at neurons and glia,
18:39not looking at specific neurons and
18:41specific non glia non neuronal subtypes.
18:44But even in doing that it was able to help
18:47us a lot and it was able to help us in here.
18:49A previous student who's now.
18:52Going for his first independent position,
18:56America Avari in in using the taxi we could
19:00see that indeed we were able to dissociate.
19:04Which epigenetic signatures that
19:07dissociated neurons from glia.
19:09But we could also see those that dissociated.
19:14Up here with disorders heroin use this.
19:17The dissociated heroin use in neurons
19:19and what we saw specifically was that
19:21the gene ontology of where these
19:24epigenetic changes were occurring again
19:26shows really strong dysregulation on an
19:28epigenetic level of synaptic plasticity.
19:32Again,
19:32these little meteorologic postsynaptic
19:35density dendritic changes and
19:37what were the genes changed?
19:40The top gene was Finn.
19:42And that was fascinating for us,
19:45because we had never studied fan and
19:47obviously it yellow come to that.
19:48You guys have been studying it for
19:50a while and the thing also was
19:53that the this this.
19:55The epigenetic changes that we saw for Finn.
19:58It actually explained Signal 6 to 8%
20:01of the variance for identifying heroin users.
20:05We were able to to show that my student Tony
20:07Roman was able to show that it's functional.
20:10The change this this enhancer region that
20:12we saw had the greatest chromatin change
20:15in Harry Newsers in the stratum and we were
20:18able to see also that it was self specific,
20:21at least that we didn't see these changes
20:23in glia, but it was mainly in neurons.
20:27And So what is Finn?
20:28As I said, you know.
20:29For Yale, I know many of you Vandyke and
20:34and me had been studying fit in relation
20:38to Alzheimer's and alcohol use disorders.
20:41For me, even though we had been getting
20:43more and more interested in the postsynaptic
20:46density in the glutamatergic signaling.
20:48Based on our postmortem results,
20:50it was interesting because we've never looked
20:52at fit and finish a stark tyrosine kinase,
20:55and it is a member of this group.
20:57Detergent, postsynaptic density and it
21:00regulates the cytoarchitecture dynamics.
21:02And we've found this in the
21:04nucleus accumbens,
21:05but my student run the Ellis also in
21:07using machine learning strategies,
21:09and I'll come to that a little later
21:10in the orbital frontal cortex.
21:12Another brain region critical in substance
21:14use disorders also identified as being
21:17part of a network predictive of heroin users.
21:21So in looking at Finn,
21:23we not only saw fin changes on the
21:26epigenetic level.
21:27We also saw it on the gene expression
21:29level in in the striatum.
21:31We saw it also change in animals
21:33that self administered heroin and
21:35we also even saw it in animals that
21:37self administered heroin.
21:38It correlated with their harrowing
21:40intake because here and at least
21:42the animals we know exactly how much
21:44hearing they take over their lives.
21:46We also saw it in cell cultures
21:50in with morphine.
21:51So.
21:53Thin as a kinase,
21:55it's activity in regulating
21:57downstream signaling relates to
21:59it being an phosphorylated and
22:01its phosphorylated active form.
22:03Inherent users was significantly
22:06increased as compared to the
22:08inactive form of first decreased.
22:10And indeed spin correlated with
22:12the years of heroin use in humans.
22:15So altogether,
22:16Finn was became very interesting for us and.
22:22As I mentioned, many people as I said,
22:25including you know a lot of the colleagues,
22:28your colleagues you know had shown
22:31Finn related to aspects of Alzheimer's
22:33because Finn phosphorylates the important
22:36downstream target of Finn is Tau,
22:39and hyperphosphorylated Tau is a
22:41pathological feature of Alzheimer's
22:43and another among other tauopathies,
22:46these neurodegenerative disorders.
22:48And years before we had seen
22:51these epigenetic changes,
22:52we had actually seen increase phosphorylate.
22:55Towel in the brains of heroin.
22:56Users in streaming, especially in Cortex.
23:00So the question is, you know,
23:03can you see the same things in animal models?
23:05So because humans,
23:06and especially I'd said at the time when
23:09we had found the increased phosphorylated
23:11Tau in the brains of heroin users
23:13before we started studying epigenetics,
23:15I thought that they had just hit their hats
23:17because you know when they're toxicated,
23:19maybe they had fallen.
23:20But we when animals self administered heroin,
23:23in addition to seeing increased fan it also.
23:26They also had hyperphosphorylated
23:28Tau that we could also replicate.
23:31You are in our cell culture model.
23:33Importantly,
23:34thin phosphorylated Tau phosphorylates
23:36toward a specific sites on Tau,
23:40and when you look at other sites that are
23:43phosphorylated by other kinases and Tau,
23:45we did not see those changes,
23:46so there was for Tau.
23:49There were five specific changes in regard
23:51to its phosphorylation and function.
23:56So. When we see these changes and said,
24:00you know, many, many years ago we
24:03see these hyperphosphorylated towel
24:04and like I said, I brushed it away.
24:07We see these changes in regard
24:09now to these epigenetic changes
24:11that really is not due to hitting.
24:14You know people hitting their heads
24:16or you know taking other substances
24:19but might be very, you know,
24:21newer toxic because we could also see
24:23that as I said in our animal models,
24:25so is opioid use predictive?
24:28Often your Commissioner type
24:30and so my student Brandy.
24:32We've been looking at this in
24:33a number of ways and I'm only
24:35going to show like one thing.
24:36So if we look at electronic health
24:40records and track opioid exposure.
24:43And your cognitive diagnosis later in life.
24:46Indeed, we actually see that if
24:48we look at people and look at we
24:50and if anybody wants to know,
24:52we can talk about it and the
24:54Q eight time period.
24:56But in looking at retrospectively
24:58and looking at 5 and 10 year
25:01follow-up following opioid exposure,
25:03we could see that indeed those individuals
25:05that had substance use disorder,
25:07especially opiate use disorder.
25:10It increased their diagnosis
25:12for neurocognitive their
25:14neurocognitive diagnosis.
25:15Later on in life.
25:17So, but back to our final story so.
25:22In animals, such self administered heroin,
25:24just like our heroin users.
25:26Like I said,
25:26we saw this increase in Finn and
25:28it made us interested to think of
25:30whether or not Finn may be important,
25:33for indeed directly causally regulating
25:35heroin self administration behavior.
25:37So we,
25:37we changed the expression of Finn
25:39by using viral media to minute
25:41manipulations and the animals,
25:43and we could see that indeed,
25:44if you're not down Finn,
25:46you could actually reduce heroin
25:48seeking behavior in these animals.
25:51But since our long term goal
25:54is medication development,
25:56we wanted to see whether or not
25:58a pharmacological inhibition
25:59of fan could also work,
26:01and there now this time we are lucky
26:05because then there was a inhibitor
26:07offence or ketamine that was being
26:10tested in Alzheimer's disease,
26:11and so we could use it in our animal
26:13models and we really made sure
26:15that animals were quite motivated.
26:17We increased increasing the work effort
26:19for them to self administer heroin.
26:21And then gave them sarcasm.
26:23And during those days that
26:24they have received sarcastic,
26:25it had reduced their currency and
26:29self administration behavior.
26:30In your when you're trying to
26:32develop medications for substance
26:34use disorders is really critical.
26:36Not to impact all aspects of
26:39their reward because, you know,
26:41we need to still have a regular
26:43hypnotic state.
26:44And when we gave a sarcastic name
26:46while their self administering food,
26:49for example, it didn't change that behavior.
26:52So when we look at the the human brain
26:54and our translational animal models,
26:57we see that there are specific
26:59epidemic and synaptic dysregulation,
27:01and a lot of that synaptic
27:03Goodman Turkic pathology.
27:05Really,
27:06the some of the targets that we
27:08have identified may be important
27:10for treatment development.
27:11But just as the epigenetic,
27:13the BRD the the Bromo domain family that we
27:16think is important for developing medication,
27:19others have and we saw and they saw
27:22other substances being impacted.
27:23It's like the the intake of other drugs.
27:26Other groups have been looking at then here
27:30dorid Bronze group at UCSF was able to see
27:33that in animals that are consumed alcohol,
27:36it also increased fat in the striatum and
27:39if they use their cabinet it could decrease
27:43the animals alcohol intake behavior.
27:45But at Yale, you guys have?
27:49They've been clinical studies,
27:51carried out with individuals,
27:52alcohol use disorder,
27:54and have not seen any significant
27:56changes with alcohol consumption.
27:58But it was really nice because it
28:00was a translational study and they
28:02also had a mouse model where they
28:03could see that perhaps perhaps the
28:05habitual responded for for ethanol,
28:08maybe what may be affected by
28:11our cabinet and there's and those
28:13of course may be important.
28:15So here we did not see, at least for alcohol.
28:18Just sort of carry that by.
28:21Work, you guys,
28:22that circadian pattern effect?
28:24But I think that there's still a
28:26long way because we still think then
28:28see on so many levels that thing is
28:31really important for a number of at
28:34least opiate use changes that we saw.
28:36So I'm gonna go back to the orbital
28:39frontal cortex where if you remember
28:41we had seen this changes in fan
28:43and the overall frontal cortex.
28:44As I said earlier,
28:45is is a critical brain region
28:47also for substance use disorder,
28:49especially in terms of guiding
28:52decision making.
28:53The values of reward,
28:55goal directed behavior and even
28:57in reversal learning aspects so.
28:59Although we saw Finn,
29:01Finn was not the primary.
29:03Significantly changed gene hub
29:06using machine learning approaches.
29:09It was shesa 7. And she's just seven.
29:13We solved that all models that Randy
29:16used and our colleagues in terms of
29:19machine learning models predicted
29:21so predicted that she's a seven
29:24was really key in dissociating.
29:28Differentiating the gene expression
29:31pattern of parent users from control.
29:33So basically that's what you're asking.
29:35You know, these machine learning algorithms,
29:38how? How well do these these?
29:41Transcriptional signatures tell you who's a.
29:45Maybe a heroin user and who is
29:47a control subject.
29:48And as I said,
29:49she's a 17 up all the time,
29:51so once she's a 7.
29:54It's usually 7 is an auxiliary
29:57subunit and the the.
29:59Unfortunate thing is that actually a
30:01good thing and an unfortunate thing.
30:03The good thing is that for us is that
30:05it's something novel because very,
30:07very few people have studied Chester 7.
30:10It's considered an auxiliary subunit,
30:13not only at the Ampang
30:15glutamatergic receptor,
30:16but even at the GABA a receptor
30:18and different researchers
30:20have evidence on both sides,
30:22so that's why we are working with the
30:25Yale your proteomics core and being
30:27able to see where is Jesus 7 binding,
30:31at least in our models,
30:32and so hopefully we'll be
30:34able to get some insights
30:36for that, but in the meantime,
30:37what we've done is to try to
30:39see is sheets of seven, really.
30:41Critical for parents,
30:42self administration and parents seeking
30:45behavior and here Randy and Jackie,
30:47Jackie Fuller and postdoc in a group.
30:49You could see that indeed it
30:52replicated that her she's a 70
30:54and the cohorts of heroin users
30:56is reduced in rats is reduced.
30:58But actually it correlates
31:00significantly with the the the
31:02rest heroin seeking behavior.
31:05So we could we wanted to see if
31:08overexpressing she's a seven could change
31:11was relevant to Q induced behavior
31:14so we had animals self administer
31:16heroin and also saline as comparison
31:20groups and then we overexpressed.
31:21She's a seven in the animals and we
31:24could see that in those animals that
31:25had heroin and she's the seven we can
31:28push that heroin seeking behavior.
31:29So she's a 7 augments human
31:32induced parents seeking behavior.
31:35As I mentioned,
31:36the orbital frontal cortex is
31:38important for reversal learning and
31:40indeed overexpressing she's a 7 in.
31:42Also change sucrose reversal
31:44learning so animals that first
31:47learn to self administer heroin,
31:50we would switch and give
31:51them a different reward.
31:53They're the conditions under which the
31:54levers and we could see that it was a.
31:56She's a 7 overexpression could
31:59promote reversal learning more
32:01versus the heroin animals.
32:02So the question is what is happening?
32:05In the brain, on a transcriptional level.
32:08With she's a 7.
32:09And so here what we did was to
32:13look at what gene expression
32:15patterns is similar in heroin,
32:18self administration and under
32:20conditions of no heroin,
32:22self administration.
32:23Just she's a 7 overexpression of
32:25the orbital frontal cortex and
32:27this is a strategy using rank rank.
32:29It's called hypergeometric overlap.
32:32In comparing 2 gene expression sets,
32:35it's a little convoluted.
32:37It's not,
32:37I mean that it's just the opposite
32:39in terms of.
32:40Where we see significant overlaps
32:43in genes that are downregulated
32:47in one comparison group.
32:49And over here these are genes
32:51that are upregulated,
32:52but there was a complete concordance
32:54between whether or not complete a
32:57very strong concordance between the
32:59gene genes that are upregulated in
33:02hiring users and those that are
33:04are also upregulated by she's a 7
33:06overexpression and the same thing.
33:08Those that are downregulated so there.
33:10This there is this.
33:11She's a 7 overexpression mimics a
33:13lot of the patterns in the in the
33:15orbital frontal cortex that we see
33:17with parents often ministration.
33:19So and not in the discordant genes.
33:22This is just the the odds ratio
33:24and this this significance.
33:26So what are the biological features
33:29that are that are changed in relation
33:33to the genes that are that she's
33:36a 7 regulates?
33:37Once again not surprising we see
33:40changes related to postsynaptic density,
33:42the synaptic plasticity,
33:44the cytoskeletal organization.
33:46Also we see a number of of
33:50of disorders related to,
33:52for example, Alzheimer's disease,
33:54Huntington's disease,
33:56these neurodegenerative disorders
33:57come into the gene transcriptional
34:01profile that she's a seven is inducing.
34:04And even going back to the machine learning,
34:06I'm not going to show a lot of it.
34:07We can see that other genes that have
34:09been identified even though they were
34:10not as strong as sheets of seven,
34:12for example, here this product could here,
34:14is it also in she's a 7.
34:17And when you change she's a 7 expression.
34:19It also changes a number of these
34:21genes that the machine learning
34:23strategies had shown to relate to heroin
34:27seeking behavior in in particular.
34:30So using these unbiased computational
34:33strategies, we were able to
34:36identify molecular alterations,
34:39again emphasizing synaptic
34:41dysregulation and also this aspect
34:44of heightened neurocognitive risk.
34:52So the last part of you know the
34:56strategies that we've been using to try
34:59to develop new treatments or potential
35:02new treatments for opiate use disorder.
35:05The first strategies, as I mentioned,
35:08started with our human subjects and
35:10looking at postmortem tissue and then
35:13going to our preclinical animal models
35:15and trying to manipulate them and
35:17moving them into the clinical studies.
35:21What we are. Also did was using our
35:26animal models in general to see what
35:29they may help to identify and here this
35:32was an unusual start because we've
35:35been looking at the developmental
35:36effects of cannabis for many years,
35:38both from the prenatal and analysing
35:41exposure and we had looked in in humans
35:45in terms of fetal samples for example,
35:47but even our animal models because
35:49our animal models we could allow
35:51them to grow into adults and really
35:52see doesn't impact on behavior.
35:54And one behavior that we we spend a lot
35:56of time looking at because initially,
35:58especially with the adolescent exposure,
35:59was this gateway hypothesis of cannabis
36:03exposure increasing addiction risk even
36:05to other substances later in life.
36:07And here we looked at opioids and.
36:11And then we can go into QA as to
36:13why we looked at opioids,
36:14but there is a they share a number
36:17of signaling mechanisms with the
36:20endogenous cannabinoid receptors.
36:22And we could see in animals that
36:24had been exposed to THC prenatally,
36:27that they would self administer
36:29heroin more than controls,
36:31and even on their conditions,
36:32when they we had them,
36:34they would take the same amount of heroin.
36:36If you just looked at even the first,
36:38they're running to the first letter lever
36:41to get that first hit of of of heroin.
36:44In those adult animals, with penalty, etc.
36:47It was much faster,
36:48and they had a greater drug seeking behavior,
36:50especially under stressful.
36:52Conditions similarly,
36:54when we gave adult animals have
36:56had adolescent exposure to to TFC.
36:59They also self administered this
37:02black line here that's missing.
37:04They also self administered more heroin.
37:07But when we?
37:09When we have our human studies,
37:11we're talking about cannabis and
37:13when we have our animal studies
37:15we're talking about THC.
37:17And we know that the cannabis
37:18plant is very complex,
37:19containing over 500 chemicals,
37:22many of them over 140, are cannabinoids.
37:25So THC yes is the primary psychoactive
37:29cannabinoid in cannabis plant,
37:31but other cannabinoids such as CBD,
37:33cannabidiol is.
37:35You know,
37:36also have from ecological and
37:39and and psychoactive properties,
37:42so one of the things we wanted to
37:44look at was that we said let's look
37:46at at least another cannabinoid.
37:47CBD, it's used to be well.
37:49It still technically is the second
37:52highest cannabinoid in the cannabis plant.
37:55The you know the normal cannabis plant.
37:57Today it's decreased dramatically
37:59as compared to the concentrations
38:01of THC that have gotten higher.
38:03And when we looked in our animal
38:06models that had been given CBD,
38:08we saw a different pattern to
38:10what we saw with THC.
38:11As I said earlier,
38:12with THC animals would invariably
38:14self administer more heroin is
38:15given to you to earlier and what we
38:18saw with CBD was that it decreased
38:20heroin seeking behavior,
38:21and it was very specific it
38:23was decreasing Q and juice.
38:25Currently seeking behavior so animals
38:27when they self administer heroin,
38:29just like humans,
38:31the environmental context becomes important.
38:34And for example if you showed
38:36them a cue or
38:37an odor when they get the drug then
38:39they will start to associate those cues.
38:41And if you only show them those
38:43cues they will press the lever.
38:44A lot of that's what we we call it.
38:46Seeking behavior and CBD was
38:48able to reduce that. So.
38:52Is CBD you know relevant?
38:54Could CBD be an A potential treatment?
38:58So as I emphasized when we look in
38:59the brains of human hearing users,
39:01we see all these changes in regard to
39:03synaptic plasticity in glutamatergic genes.
39:05We also see those in animals
39:08that self administer heroin and
39:09when we gave those animals CBD to
39:12actually reverse those changes,
39:14we could also see changes,
39:15for example with with heroin supply,
39:17administration of the cannabinoid receptor,
39:19and the cannabinoid receptor is
39:21very critical for regulating.
39:22About the transmission and
39:25CBD normalized those changes.
39:27So the question was,
39:30could it be effective in clinically?
39:32And so we ran a number of pilots
39:35here as a double.
39:36All of them were double blinded
39:38and randomized placebo controls,
39:39and those are things I think that
39:41are really critical when you're
39:42talking about cannabinoids,
39:43and especially today where everybody
39:45thinks that cannabis in general
39:47may be treatment for everything but
39:49those cannabis studies are difficult
39:51because especially if they're THC,
39:53people know when they're getting THC
39:55with CBD, it doesn't have intoxicating.
39:57Properties so at least we can have you know,
40:00good placebo control studies.
40:02So here we looked at individuals who
40:05had a heroin use disorder and when
40:08the individuals had been shown in
40:10heroin Q and then received placebo,
40:13they crave and CBD reduced that.
40:15So replicating what we saw in the
40:17animals in the in terms of their
40:20drug seeking behavior.
40:21One of the things that animals
40:23study had also showed us was that
40:25even weeks after the last CBD
40:27administration to the animals,
40:28it was still reducing their
40:30heroin seeking behavior.
40:31So when we brought people back
40:33into the lab about a week later,
40:35we could see that it was still
40:37reducing their their craving.
40:40One thing that we also saw,
40:41at least the our study participant would
40:44clear was CBD was also impacting on anxiety,
40:48so when they had gotten the heroin,
40:50Hugh and and been given placebo,
40:52they they were anxious and when
40:54they got to CBD it reduced the
40:56anxiety that cue induced again.
40:58Cue induced angularity and a week later
41:01it's still reduced their anxiety.
41:04We hadn't studied anxiety in
41:05our animal models,
41:06but we did try to look at other
41:08aspects of in terms of, you know,
41:10in our human studies in not just the
41:13self reports of craving and anxiety,
41:15but also these physiological
41:17measures of stress.
41:18For example, cortisol levels.
41:19So when when people had been given
41:23the drug queue and had gotten placebo,
41:26their cortisol levels went up
41:28and CBD reduced that.
41:29Similarly,
41:30their heart rate went up when given the
41:33drug queue and placebo and CBE reduce that.
41:36As I said,
41:37we hadn't really looked at anxiety in
41:39our animal models, but based on what
41:41we we were we saw in our humans,
41:43we've now gone back to look at that
41:46and to also try to understand what's
41:48the mechanism of action by which
41:50CD may be working here in my post
41:53that Jackie Ferlin we looked at,
41:55we tried to induce anxiety in our
41:58animals plastic with in terms of shocking
42:00animals and when the animals are shocked,
42:03they're given a queue.
42:04Here we give them this lemon odor
42:06and then we assess their anxiety.
42:08Related behavior here.
42:09This is using the light dark spots
42:12and we could see that in those
42:14animals that when they've been given
42:16vehicle and they had were exposed
42:18to the lemon queue and they showed
42:22increased anxiety related behavior.
42:24But given CBD CBD completely reduced,
42:28that Q induced limit.
42:31Behavioral response.
42:32We're now, as I said,
42:34trying to understand how CBD
42:36might be having its effects,
42:37so that we can perhaps identify
42:41even non CBD related development.
42:44Non CBD related medications based on
42:46the the biology of what's happening.
42:49Not going to tell you some of the
42:51the specific mechanisms that we see,
42:54but one of the things that's clear is
42:56that there are disturbances in these
42:58circuits related to the nucleus accumbens.
43:00The basolateral amygdala and prelimbic.
43:03Facts, not surprisingly,
43:05but interestingly,
43:06if we just, for example,
43:07within the nucleus accumbens,
43:08here is just a shell.
43:09We can see that there is a
43:11significant changes where some genes
43:12are downregulated in other genes,
43:13upregulated when in animals when
43:16they're exposed again to this,
43:18this queuing juice stressor and
43:20they show this anxiety behavior,
43:22and when you look at the animals
43:24that had gotten CBD and their
43:26behaviors were normalized.
43:28It's just it shows that CBD
43:30reverses or even eliminates these
43:32particular genes that are changed.
43:35So. Does cannabidiol hold promise?
43:38At least here,
43:39we've been able to do clinical trials.
43:41We're still working on one of
43:43the effective doses,
43:44and the formulations and
43:45the delivery systems,
43:47but similar to what we you know
43:50I mentioned in terms of the other
43:53strategies that we're looking at in
43:56translating to developing potential
43:57medications for opiate use disorder.
44:00It may not be specific CBD for just opioids.
44:03Other groups have shown,
44:04for example, with alcohol again.
44:06And here weeks after their
44:09last alcohol intake animals,
44:11CBD still reduce their alcohol
44:13seeking behavior.
44:14And even when they're the stress induced
44:17against here, it's just a shock.
44:19Again.
44:19You can also see that CBD still
44:22reduce their alcohol seeking behavior.
44:24But there may be some things that would sex.
44:27For example,
44:28a group looking at binge drinking
44:31in in the mouse model found that
44:34although CBD did reduce alcohol
44:36intake in the mail animals,
44:38it did not do that in the female until
44:41they increased the dose of CBD significantly.
44:44So there's still a lot that we have
44:47to learn and doses are important,
44:50but at least now we're able to
44:53expand our our small studies.
44:55And now we're also looking at in terms of a
44:58big clinical trial with CANNABIDOL to see.
45:01Indeed again, you know, placebo,
45:03randomized placebo control,
45:04and to see also doing your imaging
45:07studies to see if we can start in
45:09humans like our animal models to
45:11understand what are the neural
45:13systems that are changed with CBD.
45:16So clearly in looking at the human brain,
45:21it has taught us a lot that you know,
45:24although initially we have focused
45:26on dopamine and and even the opioid
45:29system in targeting for medication,
45:31but it's about epigenetics and
45:34synaptic plasticity and these to
45:37me and also those those neural.
45:41Networks and those systems that really
45:44relate as well to cognition in part,
45:47and those are things that we're
45:49definitely trying to develop,
45:50and the important thing about
45:52all of these epigenetic changes
45:54is that they are reversible.
45:56So yes,
45:57the epigenetic mechanisms maintain these
45:59long term sensitivity of the brain,
46:02but it's because we really haven't found
46:03the right targets to reverse them,
46:05because these tags are like genetics,
46:07are reversible.
46:08So I had mentioned a lot of the people.
46:11Who had you know,
46:12contributed to a lot of the
46:14work that we're doing?
46:16I didn't show like Alex **** where
46:18working on CBD and or heroin users,
46:21and a lot of our clinical
46:22team and Karen backing,
46:24and in large part in running the
46:26operations of our clinical trials
46:28and doctor and soul systems of
46:31Amazing Addiction medicine physician
46:33who's been great to work with.
46:35So with that I will take any
46:37questions that you might have.
46:38Thank you.