Nestler_011020.mp4

January 14, 2020

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Eric J. Nestler, MD, PhD, Dean for Academic and Scientific Affairs, Nash Family Professor of Neuroscience, Professor of Pharmacological Services, Professor of Psychiatry, Icahn School of Medicine at Mount Sinai; Director, Friedman Brain Institute: "Inaugural Heninger Lecture: 'Transcriptional & Epigenetic Mechanisms of Addiction'"

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00:00Welcome everybody to the inaugural George are Henninger lecture.
00:06And this is about as good as it gets in science because we have the opportunity to both celebrate a beloved member of our faculty and 2:00 and then here from an extraordinary son of our Department before I get into that. Let me just let you know about the upcoming grand round lectures next week. We have Simon icon from the University of Dusseldorf will be talking about some very exciting.
00:37Brain imaging findings where artificial intelligence techniques are applied to the analysis of functional brain imaging scans so that should be in exciting grand rounds and then the following week, the 24th of January. We have Justin Baker from Harvard Medical School in McLean, Hospital, who's going to be talking about digital or computational phenotypes of psychosis. The use of passive data collection to try to identify people who are.
01:08At risk for psychosis and then and then at the end of the month. We have Darren Lattimore, who's our deputy Dean for diversity who's going to be talking about microaggressions in the health learning environment.
01:22So I'm going to make a few comments.
01:25About George Henegar, who needs no words to recognize his contributions to the Department and then introduce Eric Nestler, who needs no introduction in order to to celebrate the work that he's going to tell us about in his career. So I just wanted to say a few words about George for those of you who may not know know him as as well as as we all do I see people in the back of the room.
01:57We have, I would call them aisle seats available up here. There are certainly uh maybe a stairwells stare seat or or 2 we should. Look up in the right front. If you'd like to to make use of them You know this. Eric was reminding me that that when we renovated the CMHC Auditorium years ago. George was one of the people who bemoan the fact that we were going to lose seats.
02:31For grand rounds, but I would say that that there is.
02:37We should have anticipated this and had it in a bigger setting, but there's something about the communality of being altogether. In this very this room, which is meant so much for the Department in its history that that that makes the setting seemed very appropriate.
02:54But so not too long ago as you know, we had the celebration for the 2 George is George Henry Jerin George Agaj Anian. The two of them are seminal figures in our Department who have mentored so many people and equally important set the kind of scientific vision that everybody since them has followed in terms of the values of.
03:25Of Science and the possibilities of Science and the need for rigor in both thinking an end in experimental design.
03:35That that we had this wonderful celebration to celebrate both of their careers.
03:42Subsequent to that Eric and Dennis Charney had the idea that we should really do something more that we should.
03:54That we should create an Dau Electro ship that recognizes the contributions of these 2 incredible people for the Department.
04:03And once we heard about it once they share the idea with me the the IT was such an obvious thing for us to do and it took all of about 10 minutes to find all of the donors who wanted to participate to endow this lectureship, so this lectureship is really the work of a whole group of people who contributed to to endowing this lectureship. The other part of the lectureship. They agaj anian lectureship will be taking place.
04:34A little bit later this year and Doctor Charney will be the agagianian lectureship.
04:41Eric nestler the Henegar lecture.
04:44So uhm.
04:46So let me tell you a little bit about George in case in case you don't know him all that well.
04:52So so this is where George grew up Utah State Hospital. The mental hospital in Provo. UT he was not. It wasn't a patient in the in the in the hospital actually his father was the psychiatrist. The Superintendent of the state hospital and his brother also became a psychiatrist.
05:18And uh.
05:20George George, his who's been an athlete is his entire life tells the story of being one of the early people who?
05:30Discovered jogging or discovered running you know was running around in the farmers would shout at him as he was running by if you've got so much enerji you can help me on the farm here, but but he he came from this background. He developed a lifelong appreciation of the suffering of people with mental illness and it profoundly shaped his decision to pursue occur in psychiatry and 2.
06:00Conduct research.
06:02George was born in 1934 and attended the University of Utah and then had a seminal experience at Mass. Mental Health Center, which was probably one of the very Premier places to train in psychiatry in that era and then went to do a research Fellowship at NIH with Dick quiet on the biology of schizophrenia and came to Yale in 1966 with the opening of the Connecticut mental Health Center.
06:35And the creation of the clinical research unit.
06:39The establishment of that clinical research unit was really a partnership between George and the late Malcolm Bowers, who was the inpatient unit chief.
06:49Uh and they work together on that unit for a long time. Malcolm ultimately became the chief of psychiatry at Yale, New Haven Hospital and for a period of time head of the psychiatry residency program here. George ran the research enterprise, becoming first head of the research unit, then director of the Ribicoff research facilities.
07:15Then associate chair for research and ultimately professor emeritus.
07:22George.
07:24Throughout this period of leadership was the most fierce and the most generous advocate for research in the Department of protecting the research enterprise from from all sorts of challenges at over over the period of time of his leadership and more importantly, inspiring creative innovation in research in the Department of psychiatry and fostering their development.
07:55Of the careers of young psychiatrist who have gone on to to be seminal leaders.
08:03Probably uh the one of the very exciting, most important collaborations in the history of this Department was the collaboration between George and Dennis Charney.
08:20George the reflective introspective force and Dennis the volcanic creative energetic in the merger of that, too was of those two was extraordinarily creative in terms of research and but also incredibly creative in terms of mentor ship.
08:54I can say for myself.
08:56There was no better place to train in the world than on the clinical neuroscience research in the era. When Dennis and George were leading the enterprise.
09:09Um their work.
09:11Uh set the tone for psychiatry at Yale of being a serious neuroscience enterprise and that sounds almost obvious like how could it be.
09:23Not a serious enterprise and how could it not be a scientific enterprise, but but clinical research in the error that this was that this work was going on.
09:36Um was not well founded in neuroscience and in fact, Yale was one of the very few very few places in the world where the clinical psychiatry research and the basic neuroscience research. People got together and wrestled with common scientific problems, so that that we felt very much that we were in a common enterprise and and and had common values.
10:07And common names.
10:10And that was extraordinarily energizing extraordinarily inspiring both for the basic and the research communities here here at yeah.
10:22The other innovation where these 2 kids.
10:27Really that the foundation of molecular psychiatry.
10:31Um.
10:32Again, hard to believe but there was an era where neuroscience research in psychiatry meant measuring whether the dopamine level was up or the dopamine level was down.
10:45In the clinical research enterprise, we got to the level of trying to understand how receptors were regulated. It wasn't until the advent of molecular psychiatry at Yale that psychiatry as an enterprise.
10:59Seriously wrestled with what happened once a neurotransmitter bound to its receptor target an that was made possible really through the through the work of Eric, who brought the signal transduction focus from from his work with Paul Green Garden. I'll talk about that. A little bit later as well as Ron's work ultimately on on many complementary aspects of signal transduction and.
11:30Neurobiology but this this.
11:35Enterprise was so inspiring.
11:39That, it inspired Julio Licinio to steal that title and create a Journal called molecular psychiatry.
11:50OK, so George George George honors do not are great and there are many fold, the animatic apprised one of the highest most valued prizes and depression. Steven Fleck Award in the LPR a best teacher award. The Axelrod Julius Axelrod Mentorship Award from from a CNP.
12:15Um.
12:17In some ways, they don't, they don't adequately capture. Georges impact on science am much better way to get that feeling are the many famous productive scientists around the world. Some of whom are here in the room today. They have a car. I see and others. Eric myself, but Nelson many others.
12:46Um, who have been inspired by George and Ann.
12:55It's hard to capture what it is about George that is so inspiring. But maybe you'll get a feel for it. If I read to, you, the holiday card that I got this year from George.
13:08You know, let us welcome a new year blah blah blah blah blah and then.
13:15Dear John this is the holiday card right.
13:19One possibility is to measure oxidative stress in molecular changes in brain derived exosomes in schizophrenia in order to guide treatment. I will stop by to explain our plan now. Maybe that's not the kind of holiday card, usually right, but it says a lot about about Georges.
13:47Georges embrace how George embraces the science at the at the edge.
13:55At the at the forefront and is always thinking about how to bring that Horizon of neuroscience into clinical research and that.
14:06His unchanging throughout his life so George on behalf of everybody here today. We salute you an we express our gratitude and I hope everyone will stand and join join us in the round of class.
14:43Georges is reserving his comments until after Eric speaks.
14:48So.
14:52So.
14:54Eric Nestler.
14:57Eric is and is a person who has improved.
15:02Every group, he has been in probably since kindergarten.
15:09That's what his mother told me on the way in today.
15:15He is the Nash family professor of neuroscience director of the Freedmen Brain Institute. Dean for academic affairs at the Icahn School of Madison at Mount Sinai.
15:26Um so, so Eric's roots go very deep here at Yale.
15:33This in the back of the room, you probably can't see this, this is a tattoo of of the Yale sign.
15:42I'm not saying that Eric has a tattoo of Yale sign logo on him.
15:50But I'm not saying he doesn't have it have a tattoo the tattoo.
15:57Yell was Eric was a Phi Beta Kappa graduate of Yale College, publisher of the Yale Daily News. He stayed in New Haven for medical school and for pursuing his PhD with Paul Greengard, who I think in many ways, certainly scientifically and perhaps more broadly was a spiritual father for Eric. In some ways. He was class president of Yale Medical school I didn't even know we had a class president, but that's how accomplishment.
16:29They created a role for Ericas to be class president. He was Alpha Omega Alpha, which is the honors. Medical Honor Society and stayed here for psychiatry residency after a brief trip to Boston and received the Lessman toward the highest honor for psychiatry resident research.
16:53He has had by any standard a meteoric career with with Ron Doom and they established the laboratory of molecular psychiatry very soon. He was the Elizabeth mirrors in-house Jameson first assistant professor then associate professor then professor.
17:11He went on to lead the Abraham Ribicoff research facilities here and in so doing really transformed the science, bringing many new investigators into the labs inspiring new directions in molecular cellular in translation or research.
17:33And then he ran around 1999, two thousand he left for become chair of psychiatry at Utah, southwestern he had a transformative effect on on that Department and its research mission, and then was in a stroke of inspiration and brilliance was recruited by Dennis Charney to become chair of neuroscience and head of the Freemen Brain Institute at the Icahn School of Madison.
18:05Um.
18:06Along the way he became president of a CMP president of the Society for neuroscience and spine off company. Psycho genic's and has been an advisor to all sorts of groups and is now Dean for academic affairs.
18:30Eric has had a transformative impact on science, beginning with his work with Paul Green Garden. Synaptic fossil proteins, continuing with establishing his own line of research initially around signal transduction than transcriptional mechanisms around substance abuse stress an antidepressant.
18:48And and overtime essentially pushing the boundaries of what could be explored using animal models to study the biology of these kinds of approaches leading him to study aspects of synaptic construct brain structural Ramada Ling Epigenetic regulation.
19:11With and more recently, including the pathophysiology of psychiatric disorders using things like postmortem brain tissue in other other relevant paradigms. An Eric has had an inspired sense of where the important questions lies so for example, in studying the biology of depression. In a really groundbreaking paper to find out that the biology of depression.
19:42In in women and men as well as in male and female animals is really quite different a profound profound insight that has yet to be translated into a novel sex specific Therapeutics, but but is an example of the kind of of of really seminal work that he has done.
20:08Again, uh Eric is replete with honors member of the National Academy of Madison, a fellow in the American Academy of Arts and Sciences are recipient of the highest prize in neuroscience from the National Academy of Medicine called the Sarnak Prize recipient of the mood disorders, and cognitive neuroscience prizes from the brain and behavior research foundations.
20:33Um Deanna Monica Prize.
20:35Uh uh.
20:37The Society of biological psychiatry got a large gift and created a prize, which is only given every several years to people who have made really extraordinary indistinctive contributions to science and psychiatry called the Red Alzheimer Rentals Heimer Award in Eric was the first recipient of that award is received awards both for his science, Efron toward an mentorship the Axelrod Award from a CMP.
21:07And recently received something called the Wilbur Cross metal from Yale University, which is the highest honor given to a graduate of the graduate an alumnus or of the graduate department's so.
21:20Uh these these awards are signposts of Eric's extraordinary impact on science is extraordinary impact on people. His extraordinary impact on the institutions.
21:37And groups within an organizations within which he works. These impacts are lasting and meaningful so we are honored pleased to welcome my dear friend.
21:53Eric Nestler.
22:05Thank you so much John just give me a second while I set up my PowerPoint here. It's a blast to be back.
22:12And a tremendous honor to be here today to give the inaugural George Henninger Lecture John gave you.
22:21Background of Mycareer, but in looking back, I've been very fortunate to have so many senior faculty. Help me along the way.
22:31But I can identify 2 primary mentors. John mentioned Paul Greengard, who was my scientific mentor. He taught me how to do science had to run a lab.
22:42And George Henninger, who was my administrative mentor.
22:46More than anyone else, George taught me how to navigate the political intrigue of life in academia.
22:55But more importantly than that George helped me grow beyond.
23:00The inherent selfishness of running ones own lab.
23:04To build and manage a group of people help helping each to thrive and flourish. I'm going to come back to that in just a few minutes.
23:14George also influenced me and many other sitting in the audience here enormously through his truly unique way.
23:24Of framing a problem with provocative metaphors.
23:30I first met George Henninger formally in July 1984. I had just returned to Yale to start my PG 2 year in psychiatry had an appointment with George.
23:41The next week and a paper from National Geographic appeared in my mailbox focused on smelting copper and iron.
23:50In Preindustrial Africa with a note for discussion with Doctor Henninger next week and I remember my fellow PG 2 resident and I looking at one another saying.
24:00OK, who we had no clue what to expect.
24:06We read the article, which basically described a weeks long process involving dancing singing religious rituals.
24:13And ultimately heating the orc to thousands of degrees when we met with George, he made the point that this is just like psychotherapy.
24:23We know it works, but most of it is a series of superstitious rituals without knowledge of the active required step.
24:31Amazing insight and you know, I still use this metaphor with our residents today.
24:362 years later in this auditorium, George gave grand rounds to psychiatry and he began in my own memory with I think a 10 minute exposition.
24:47Of a storybook character called Flat Stanley, a children's book series.
24:52Flat Stanley is flat 2 dimensional, he can slip under a door.
24:58He can put himself in an envelope and Mail himself across country and all sorts of cool things.
25:04And I remember sitting where you are today thinking to myself, Oh Lord Where is this going.
25:11What were George brought it was to the bio cycle social theory of psychiatry? Which was
25:19Dominant in the field in those days.
25:23And Georges Point was is that it's the flat Stanley view of psychiatry flat 2 dimensional obvious trivial.
25:32Ultimately, unhelpful and explicating mechanisms of psychiatric disease.
25:37What was all the more remarkable to me is a PG for resident was that one of the major proponents of this theory was more riser who is chair of the Department at the time, meaning Georges Boss.
25:52As I said George was provocative.
25:56Then there are what we all.
25:58Lovingly call Henninger isms.
26:01Decorum prevents me from sharing these with you today lest I be cancelled.
26:10But I will share one.
26:12Being an administrator is like walking behind people and picking up their shit as it falls from them.
26:22I can tell you the number of times that I think of this to this very day and smile.
26:30In the mid 1990s, we held a 60th birthday party for George where we celebrated and roasted him.
26:37A week later in my mailbox appear to photograph of that from that party with Georges scribbles on the Top so there's Georgian Julie at the right. Denison in the middle Larry Price, Chris McDougall, myself in the foreground.
26:53About a week later.
26:56This appeared in my homes Mail.
27:00You know you may not recognize me with her, but that's me upside down in a 2 two I guess.
27:06This is a picture of what George showed at the party to roast Maine.
27:11And similar pictures of everyone else here roasting them certainly out roasting us compared to what we did with him, but was particularly remarkable is this was not addressed to me, this was dressed to our son, David, who had unbeknownst to us had sent. Georgia picture of me that he could use in this picture along with this note, which I'll read dear. David thanks for the note in picture for the party. I thought you might like it picture of what goes on. In these parties. These are the kind of adult ideas that get passed around.
27:42Sometimes dad's or not as big as they seem at least with trick pictures. You can make them smaller George George, new David because he would take David and my other son. Madden me on an annual fishing trip canoe trip and the like and really wonderful times, that we still reminisce about to this day.
28:05I loved Georges Irreverence.
28:09It gave me license to embrace my own.
28:14Along the way I learned from George that the worst kind of person in academia probably every.
28:21Path of life are those who kiss up and kick down.
28:26George of course was always the exact inverse.
28:31Providing maximal support to junior people, while challenging authority that resonated with me very much and helped shape the way I've tried to lead.
28:43According to George Fighting Convention.
28:47And bureaucracy in the interest of innovation and driving scientific and medical discoveries.
28:53Today we call that type of person, a disruptor.
28:56George was the consummate disruptor.
29:03In the early 1990s, George stepped down from his two administrative posts. Vice chair of research in psychiatry and director of the Ribicoff research facilities, so that Dennis Charney and I could have those career opportunities.
29:19What incredible an unprecedented generosity?
29:26I have worked very hard to pay forward that generosity.
29:31Ever since George thank you so much for being such a wonderful mentor to Maine.
29:37And to so many other people who have passed through this great Department of psychiatry.
29:43And congratulations on establishment of the George hanging or lecture at Yale University in perpetuity, let's give George another round of applause.
30:03So now from the sublime to the ridiculous.
30:06I'll spend the rest of the time that we have telling you a little bit about the work that we've done in the area of drug addiction.
30:14Back when I was still at Yale couple of decades ago. We proposed along with many other groups that addiction could be seen as a form of drug induced neural plasticity mediated in large part through changes in gene expression as depicted in this cartoon so we know that drugs of abuse.
30:34Find initially to proteins at the extracellular level of synapses receptors transporters channels.
30:41But the diction requires repeated exposure to a drug.
30:45And that ultimately produces changes in a wide array of intracellular signaling pathways, which ultimately lead, according to this notion to regulation of transcription factors proteins that bind to DNA in a sequence specific fashion to regulate expression of Target Jeans, so the hypothesis was that drugs of abuse over repeated exposures with cause induced neural plasticity associated with addiction by producing long lasting changes in gene expression.
31:13Now parents erikli all current treatments and all currently research treatments for addiction still remain focused on extracellular proteins without taking advantage of thousands of intracellular signaling proteins. I'll come back to that toward the end of the talk.
31:30In order to explore these drug induced changes in transcriptional mechanism. We had to know where in the brain to look for that. We've relied on a large literature over several decades, identifying so-called brain reward regions.
31:43Beginning with dopamine containing nerve cells in the ventral tegmental area of the midbrain and the many for brain regions to which those neurons project. Most importantly, the nucleus accumbens but also amygdala hippocampus regions of prefrontal cortex among others.
32:02And in the early days for Maine when I was still back at Yale and will work focused on 2 transcription factors.
32:09We identified Delta Fosby because it was a unique Foss June. EPI one family protein induced after chronic drug exposure.
32:17And we focused on crib because it's a downstream effector of the cycle campy pathway and we had shown along with George Agagianian colleagues that the cycle game. P pathway was very important in the actions of drugs of abuse on the brain.
32:34The panels to the right show the very different temporal properties of these 2 proteins and highlighting the unique temporal properties of Delta Fosby, which one modified by phosphorylation at specific residues becomes very stable, so that even though it is induced for a slightly in response to an initial drug exposure. These small amounts of Delta Fosby gradually accumulate feedback suppress these other Foss proteins. And so that faucet Delta Fosby becomes the dominant. AP one transcription factor present in nucleus comments after chronic drug exposure.
33:07And over the years at Yale in Dallas. More recently at Mount Sinai. We've learned a great deal about these 2 proteins. We believe that Delta Fosby, which is induced mostly in D1. Type medium spiny neurons. I'll come back and define that in just a moment.
33:23Increases an animals sensitivity to drugs and natural rewards represents a form of positive reinforcement.
33:31Crab does the opposite it's really a classical mediator of drug tolerance and dependance induced in both major subtypes of striatal medium spiny neurons.
33:41Opposes drug and natural reward mechanisms, but it to drive self administration through negative reinforcement and I want to credit.
33:50Bill Carl's on and David Self, 2, former postdocs of mine here at Yale. Bill is now at Harvard and David is at Utah southwestern.
33:59When we moved to Dallas. We decided to move beyond transcription per se to epigenetics Chrome and in biology.
34:06Let me just first define what I mean by that. We know that a mailing an Organism has about 3 billion nucleotides in its genome is stretched out linearly those nucleotides would be about 2 meters in length. That is compacted in every cell into a microscopic cell nucleus. One of the extraordinary achievements of evolution. We've learned a lot about the biological basis of that compaction over the last couple of decades. We know that the DNA double. Helix is wrapped around Oct rumors of histone proteins to form the unit of chromatin called a nucleosome.
34:38Which is then further organized in many ways in the most condensed form? What would be recognizable as a chromosome?
34:45In this organization is very important because spans of DNA that are in compacted regions of chromatin or can cannot be used to transcribe RNA or serve other functions, whereas DNA and open regions are biologically active, which means that we can use this information to help identify the jeans that are affected by drugs.
35:04Really providing the 1st.
35:07Insight into transcriptional mechanisms in the brain in Vivo.
35:11And then by analogy with the developmental biology and cancer biology fields, where certain types of epigenetic modifications that control. This opening and closing a chromatin once they occur permanent. We hypothesize that perhaps the same occurs.
35:25With behavioral experience that life experience produces similar types of permanent epigenetic changes in certain cells in the brain that drive these forms of emotional learning and memory.
35:37Let me tell you a little bit more about these mechanisms looking at this level of organization. Now the DNA or these gold stripes wrapped around these fears the histone octamer's forming a nucleosome looking at one end of the spectrum where Chrome and is open or active. The other end, where it's condensed an inactive and we've learned a great deal about the biological mechanisms, controlling that opening and closing histones proteins themselves or modified in hundreds of ways. I'll give you examples a bit later through acetylation phosphorylation metalation and many others.
36:11Transcription factors like Delta Fosby and crab combine 2 areas of DNA that are relatively open where nucleosomes are not deposited.
36:20Where they then recruit other ends other proteins to form the transcriptional complex mediating transcription of RNA?
36:29Mechanisms of repression are equally complicated and all told, and simpler systems where it's been studied in stem cells and yeast cells for example.
36:38Hundreds of proteins are involved in coming together to activate 1 gene or to suppress a different Genomic region.
36:47So we decided little over a decade now to take advantage of open ended next. Generation sequencing technologies to back up and take a more open ended view of the molecular changes that drugs induce in the reward circuitry with the idea that we pluck Delta Fosb, Ian crib based on reasonable hypothesis. But they were candidate jeans. They were not based on truly open ended unbiased data and we felt it was essential to do the latter.
37:18So we've used RNA sequencing for example, to identify rnas that are up or downregulated.
37:24We would overlay that with Chrome and and measures by chip sequencing. Another approach is that I'll discuss looking at different types of histone another modifications.
37:34Doing chip sequencing or related approaches to measure binding sites of Crab Delta Fosby. Other transcription factors and so on. This is an enormous bioinformatic challenge generating terabytes of data but we feel that it's absolutely essential to have biology teach us? What jeans are the most important to study and I'll give you some examples of how we've used this approach.
37:58So about a year ago, a little over a year ago. Idina Walker, who's on her way to Oregon, an error in Calipari, who's at Vanderbilt were in my lab and published. This study where they had my self administer cocaine or Saline for a period of time.
38:14We took one cohort of animals the day after this last self administration session.
38:19We left the other animals sit in their home cages and live normally for 30 days.
38:24At that 30 daytime point we then gave animals in acute dose of cocaine or Saline and analyze the animals an hour later.
38:32I said there were 6 experimental conditions and we isolated six brain regions. All the major brain reward all the major components of the brain's reward circuitry.
38:43This 30 day withdrawal point is an important one because we know and animals and to a certain extent. It's been shown in humans.
38:50That incubation of craving occurs during this type of withdrawal so if one takes a mouse and puts it back in the Self Administration Cage. At that 24 hour. Timepoint turns off the Lever. My mouse compressed the liver, but it no longer gives cocaine the mouse still presses. The lever in presumably anthropomorphizing searching for the cocaine, but after 30 days, putting that mouse back in the chamber.
39:17There is a much greater response called incubation of cocaine Cravings.
39:21So this is an enormous data set RNA sequencing of six time points across six brain regions. I'm just going to show you a very small sliver of the data focusing only on nucleus incumbents where we've used machine learning to identify jeans, particularly regulated at that 30 day withdrawal time point.
39:39So we ask the computer to tell us what jeans are uniquely regulated up or down up as in yellow blue is.
39:46Down at the 30 daytime point withdrawal cocaine Self Administration Saline Challenge or cocaine self administration with an acute cocaine challenge for those of you who are not used to looking at these heat Maps each vertical line represents a single RNA.
40:06So here at the left, I can show you jeans that are similarly uniquely altered up or down at either of these 230 day withdrawal conditions.
40:16And on the right those jeans in particular that are uniquely induced or suppressed by that.
40:23Challenge dose of cocaine, we would call these jeans that are in yellow to be primed. Most of them You see are not regulated by the first dose of cocaine the acute dose of cocaine that the animal receives.
40:35Or desensitized with the opposite phenomenon.
40:40Then we dug in to look at the behavior of individual mice and now in this heat map. I'm not showing you jeans. I'm showing you behaviors. We took on animals different aspects of pressing a lever and taking and seeking cocaine. Many dozens of behavioral measures an came up with what we call the addiction index with a big caveat that we don't know when a mouse is addicted, but it is showing a range of addiction related behaviors.
41:07And then we plotted here, then the behavior index diction index of the individual animals that self administered Saline or self administered cocaine. You could see the self animals itself administered sailing show low addiction index, which is what one would expect.
41:24And you see that the animals that self administered cocaine all meeting criteria for significantly self administering the drug showing actually a wide variation in that addiction response, even though these are genetically inbred mice. So this is looking at non genetic based differences, which we could talk about during the question period if you're interested.
41:43So Idina did next is then to ask.
41:46What jeans at that 30 day withdrawal time point?
41:51Or related to the addiction index, and I'm showing you examples here for 2 brain regions nucleus comments and ventral hippocampus.
41:59Showing jeans that are up regulated or down regulated by in these 30 day withdrawal time points that are significantly positively regulated in red or negatively regulated in correlated in Gray with the addiction index. Let me just highlight 1 high level conclusion or hypothesis that we made from these data note? How a large portion of the jeans that are primed or desensitized at that 30 daytime point by a challenge dose of cocaine.
42:30We're regulated oppositely.
42:32In response to an animal's first ever exposure to cocaine.
42:37Very highly significantly regulated, too, and animals inherent addictiveness Interestingly, suggesting that an individual's addictiveness actually could be derived from their response to initial drug exposure, even though the changes that are induced by chronic drug exposure are very different opposite in some brain regions to what's seen acutely this pattern, it was most apparent in nucleus accumbens for example, not seen in Hippocampus and then we can go into these data and ask what transcription factors are.
43:08Would be expected to regulate these subsets of jeans?
43:12And you know, sometimes one is lucky in life because among the highest rank transcription. Factors are crab and AP one representing Fosby's family that was reassuring for our older older data but we also found many transcription factors.
43:30Most of which have never been studied in addiction models previously and if I have time I'll come back to one called the E2F family.
43:40Now everything that I've been talking to you about so far is showing you heat Maps of jeans that are significantly up or downregulated so-called differential gene expression analysis, but because of the magnitude of these datasets. We've been able to do much more complicated. Bioinformatics for example, different forms of Co expression network analysis working very closely with bin Zhang at Mount Sinai and one of his postdoc John show.
44:06This is depicting the entire.
44:09Transcriptome all rnas within the nucleus accumbens under control conditions how each RNA is related to one another just by correlations within our data set notice how cocaine dramatically alters the structure of that transcriptome really driving the jeans to be much more tightly correlated. One can use cercos plots as shown here to look at different segments or branches of these trees called gene modules.
44:34To identify the modules that are most significantly altered by cocaine treatments and it provides ways to begin to digest enormous experiment so for example, Dina Walker in the lab is has a data set where she analyzed animals responses to acute and chronic cocaine, but comparing males and females. With those responses and also comparing normal animals to animals that have been stressed earlier in life.
45:05An enormous data set and I'm illustrating one aspect of that here showing that the ability of chronic cocaine to alter rnas.
45:14In the nucleus of comments of a male and female mouse under control conditions.
45:19Is quite different between the sexes notice the very different effects of cocaine under control conditions. But if one does this experiment in animals exposed to stress earlier in life. It obliterates most of these sex differences just to give you 1 example that we're now pursuing to understand.
45:39Now all the work that I've talked about thus far has been taking both dissections of nucleus accumbens and doing these molecular analysis, which is good as a start but we know that the nucleus accumbens.
45:51Contains about half neurons half glea of the neurons. Maybe 95% are called medium spiny neurons. The major projection neurons and there are 2 main types of projection neurons based on named by the dopamine receptor that they predominantly expressed. The D1 medium spiny neurons in the D2 Dopamine D2 medium spiny neurons and they've been shown to exert opposite effects on reward so for example, activation of D1 medium spiny neurons.
46:23Medically, or chemo genetically promotes drug reward activation of the two cells does the opposite.
46:29And using methods like fiber photometry. It's been possible to have an animal in. That's awake and behaving and responding to drug an be able to show that acute drug exposure activates D1 MSN's does the opposite to D2 MSN's?
46:45And that chronic drug exposure exaggerates these responses that are cell type different. These are just two of the papers done by Mary Kay Lobo and Aaron called party. My lab this work represents work by the field, many laboratories.
47:00So it becomes crucial to replicate these transcriptomic and molecular analysis on a more cell type specific basis. So more recently, Phillip Muse in the lab has begun to do this and this is an example of the data, he's accumulated.
47:171st I'll show you data that's called a tax sequencing. I'm not going to define it. It basically reflects a way to identify regions of the genome that are open that don't have nucleosomes deposited so these are presumed active regions of chromatin.
47:34And one can identify that in control animals. The pink lines that the D2 mediums, finding neurons have many more open regions of chromatin than D1 neurons. That's consistent with the fact by the way that there are many more transcripts. Rnas expressed in D2 neurons and D1 neurons at rest.
47:56On centering these plots around this transcription start site and then looking genome wide.
48:01One can see then in the D1 medium spiny neurons in acute dose of cocaine really causes dramatic opening of chromatin within the genome wide that is maintained through 30 days of withdrawal and opened still further in response to that priming dose of cocaine.
48:20Notice the responses in D2 mediums, finding neurons are much smaller.
48:26This is showing the data the same exact data. But in a different way. Each horizontal line is now a different Genomic region showing you. The dramatic opening a chromatin that occurs in D1 medium spiny neurons across these acute and chronic conditions. Much smaller responses seen in D2 medium spiny neurons and it's possible to validate these data in the following way, so for example, I can show that we can show that the gene encoding. The D1 Receptor is active open has a lot of a taxi peaks.
48:56In D1 medium spiny neurons.
48:58Whereas the gene encoding the D2 Dopamine Receptor shows evidence of being active in D2 medium spiny neurons and not vice versa.
49:08These are genome wide data so one can go in and take a look at what any gene is how any gene is behaving so let's look at Fosby.
49:17Not surprisingly, fosby is induced it shows an opening by a taxi can response to acute cocaine. This opening persists. After 30 days of withdrawal consistent with the fact that Fosby is drive is one of these primed jeans.
49:35More recently, we've obtained RNA sequencing data separately for D1 and D2 mediums, finding neurons and consistent with the attack seek data showing that it is the D1 meaning spiny neurons that show a truly dramatic number of prime jeans in the chronic cocaine state that 30 day withdrawal followed by a cocaine challenge much weaker response is seen in the D2 medium spiny neurons. And when we ask? What are the upstream regulators of these prime jeans in D1 medium spiny neurons?
50:09Again, the E2F family of transcription factors comes up as highly regulated.
50:15The Top most prime gene the gene that's prime to the greatest magnitude.
50:21Is phosphate?
50:22As I said sometimes one is just lucky.
50:28And one can even go beyond doing these analysis on single types of cells to doing the analysis on single cells. So so called single cell RNA sequencing.
50:40The interesting Lee one can take dissection of nucleus accumbens sequence every cell that you isolate a nucleus accumbens have a computer identify the different clusters of cells color coded and then based on the jeans that are expressed in those cells we can label them neurons.
51:01Astrocytes microglia and so on.
51:04But notice how the computer cannot differentiate D1 and D2 neurons.
51:10So the whole literature on striatum has focused on differences between D1 and D2 medium spiny neurons, which is not evident based on their transcriptomes. In fact, if one looks at expression levels of D1 and D2 receptors across these mediums, finding neurons. You can see rather broad distributions, suggesting that the main difference in this cell type is due to their circuitry, not inherent molecular features.
51:37One can dig even deeper we can isolate the D1 medium spiny neurons. The cell types that showing these dramatic prime responses to chronic cocaine.
51:46And ask the computer are there specific clusters embedded within and there are the computer identified over 20 clusters shown here. One cluster also expresses D2 receptors that's known a small subset of cells expressed both.
52:02And we identify the cluster of cells that are in Richton Fosby, another immediate early jeans.
52:08But I don't think that single cell sequencing is yet ready for prime time for the kind of work that we're doing where we want to understand a stimulus induced change in transcription for 2 main reasons first single cell sequencing can only identify 10% of the transcriptome. It's not deep enough to look at all transcripts.
52:28And I think this is just too much variability. I don't think there are 20 different types of D1 neurons. I think we're capturing neurons at different phases of activation or some other functioning.
52:39So that remains a work in progress.
52:44So what is driving this very long lived?
52:49Opening of chromatin in D1 medium spiny neurons after 30 days withdrawal from cocaine self administration in the old days, meaning up until 5 years ago.
53:02We would do chip sequencing for histone modifications based on what the field told us.
53:09Were important?
53:12But it turns out that there are hundreds of different histone modifications, which makes it impossible to really do a comprehensive analysis.
53:20So more recently, we've collaborated with Ben Garcia at Penn and one of his postdocs who's now at Albert Einstein Simones Adoli.
53:28To do a proteomic analysis on nuclei to identify which histone modifications are most dramatically altered under these different experimental conditions.
53:40Some of those data are shown to the right again. I mentioned several 100, so this is just showing you a very high altitude. Viewers subset of these findings. Let me show you the code, there are 4 known types of histone proteins labeled here by number.
53:54This refers to Histone H3 lysine 4, Mono Metalation and so on. Blue is down yellow is up and it's possible to identify several types of chromatin marks histone marks that are altered quite differently in the acute cocaine state versus the 30 day withdrawal state.
54:18And among the marks that are most statistically significantly highlighted.
54:25Is a mark called H3K79 Dian Trimethylation?
54:31Anna histone variant called H2, a point Z.
54:35And if one digs into H2, one eight point Z it's particularly H28.
54:40Point Z acetylation that is dramatically altered differently.
54:44Acutely versus chronically.
54:47Now it would have taken us a really long time to get to K 79 demethylation in H2, AZ. They were way low on our list. I think it again underscores the absolute essential feature of using of letting biology teach you what's important, and look at.
55:07So we're now doing film, using the lab is now doing chip sequencing for these various histone modifications in an effort to identify what we call chromatin scars. The idea being that chronic drug exposure scars chromatin. It creates changes in the chromatin architecture at specific genomic loci that are then near permanent.
55:29OneDrive these long lasting changes in gene expression. Let me just show you one example of some of these data.
55:35This is focusing on H2, AZ and H2, AZ acetylation.
55:39Doing chip sequencing on nucleus accumbens tissue showing that after acute cocaine. There is increased deposition of H2, AZ and H2, AZ acetylation. The acute being the green versus the pink control.
55:57But notice the dramatic depletion of these marks.
56:01After at the 30 day withdrawal time point barely detectable signals so again really highlighting and Validating. The findings from this open ended exploration, which we would not have gotten too without an open ended process.
56:23Everything that I've told you about today, so far has been correlations.
56:28One of the approaches that my lab has used over the years is to is the ability to establish causality and I want to demonstrate that with one example. This E2F family of transcription factors.
56:42Each have family has been widely implicated in the regulation of gene expression mostly in peripheral tissues. There have been very few studies in brain. Most studies in brain have focused on Olga Dendra sites on Mylan producing cells.
56:56However, based on these repeated datasets, we where we continue with lease. Find the E2F family is being among the most highly predicted upstream regulators of primed and desensitized jeans. We decided to take a closer look.
57:14It turns out that about 1/4 of all prime desensitized jeans in nucleus that companies have deduced E2F sequences within their upstream gene promoters.
57:26And this induction of E2F3 is selective for D1 mediums, binding neurons in response to cocaine.
57:33Not seen in D2 mediums, binding neurons.
57:36We're focusing on each have 3 because it's really the only family member out of 7 or so that expressed that appreciable levels in nucleus accumbens.
57:45Which expressed mainly in nerve cells?
57:48And there are 2 isoforms of this gene each have 3A which is the one that's regulated each have 3B which is not regulated and really barely expressed even in D2 nerve cells?
58:01So the approach that my lab has used over the years is one of viral mediated gene transfer in collaboration with Rachel Navy and colleagues at Massachusetts General Hospital.
58:11We would make viral vectors that either overexpressed each have 3A.
58:15Whorwood expresa micro RNA targeting the E2 of 38 M RNA and suppress that rnas translation.
58:25What we can show is that when we overexpress each have 38 in all nucleus accumbens nerve cells we can increase the rewarding response to cocaine. This is using a place conditioning assay for example.
58:40This is an effect.
58:42Exhibited by E2F38 with no effect seen when we overexpress the splice form that's not regulated E2F3B.
58:51Knockdown of E2F38 produces the converse effect now with a higher dose of cocaine that produces a significant place conditioning by itself. We can suppress that place conditioning by reducing it to have 3 levels again no effective to have 3B?
59:08And a similar effect of E 2 of 3 knockdown on decreasing cocaine self administration.
59:14This is work done by hand in case in the lab who is a graduate student and more recently for Edison Martinez has gone on to do this in a cell type specific basis now using viral vectors that expressed their trans jeans in a creative Pennant. Fashion express them in mice that expressed. Creina specific cell type showing that the ability of each have 3 overexpression to increase place conditioning responses.
59:38Is due to its effect in D1 medium spiny neurons not D2 medium spiny neurons?
59:44We can go beyond this for example, we can do RNA sequencing on the.
59:50Nucleus accumbens tissue where each have 3A is overexpressed.
59:54By heat map comparing the effect of chronic cocaine on up or down regulation of Rnas in nucleus accumbens and just asking a normal animal no drug.
01:00:05How are the same jeans affected by E2F3 over expression and you can see, there's an appreciable overlap statistics statistically shown with this RRHO rank right hypergeometric overlay approach highly significant coincidence of these 2 manipulations.
01:00:28One can then show that each have 3 overexpression also recapitulates a large fraction of the ability of cocaine to produce changes in alternative splicing of Rnas. Just very quickly. There are 4 major forms of RNA splicing each have 38 overexpression nucleus, Cummins re capitulating.
01:00:49A significant portion of each.
01:00:53In the interest of time, I'm going to skip over some slides here 'cause. I do want to leave some time for questions and we have to leave some time for George.
01:01:01To come back, I hope he has no more slides of Maine.
01:01:08To conclude that what I've shown you today is a?
01:01:12Very reduction istic approach to a problem like drug addiction that we know has very strong psychological social and cultural effects.
01:01:22Yet at its core we believe that addiction has a very strong biological component. After all, it reflects the ability of a physical substance certain type of drug to effect.
01:01:34A vulnerable brain.
01:01:36So we believe that the data sets that we're generating.
01:01:42Provide almost like a periodic table a template of all the jeans that are being affected in the chronic cocaine treated state.
01:01:52Which then could be exploited for therapeutics remember I said at the outset that all of today's treatments focus on the very narrow range of proteins expressed at the synapse that where are smart enough now to study like a dopamine receptor or transporter?
01:02:08Not thousands, of other proteins that are expressed throughout the neuron, including hundreds of other proteins at the synapse, which could be viable drug targets.
01:02:20So with that let me just make a few conclusions. I showed you how the degree to which chronic cocaine exposure, particularly after long periods of withdrawal dramatically reorganized the transcriptome of specific cell types within the nucleus accumbens.
01:02:38I focused today on the nucleus accumbens but the work again underscores the importance of unbiased open ended investigation. We have data and data are needed for many regions within the brain's reward circuitry.
01:02:54We need to look well beyond D1 and D2 mediums, finding neurons to interneurons and several types of glial cells, which we also know are very important.
01:03:04And we need to look well beyond cocaine to other drugs of abuse and Kayla Brown, in my lab is now generating essentially parallel data sets for opioids.
01:03:15Scientifically my lab is most interested today in defining what we term croman and scars. The ability of experience in this case drug exposure to produce near permanent changes in the chromatin structure at particular. Genomic sites that mediate these long lasting changes in gene expression that are so important to driving a state of addiction and I gave you 1 example of how we establish that causality I'd skipped over.
01:03:46The more recently developed tools where we can manipulate chromatin states in an animal. In a single cell type at a single gene to really establish these causal mechanisms and the hope is that this would provide an unprecedented template for addiction treatments, so with that. Let me stop. I'll show you where I'm from.
01:04:06This is Mount Sinai on the Upper East Side right off Central Park.
01:04:12And that's where we live. Thank you very much.
01:04:24So.
01:04:26Sure, Yeah.
01:04:33Jane.
01:04:48Yes.
01:04:54But yeah,
01:05:01One yeah, yeah.
01:05:06Right yeah, so well So what I would say is that what makes.
01:05:11Foss be induced in D1 sells uniquely might not be anything inherent in D1 sells although there are differences, obviously, but it may be the circuitry and in fact, if we take a new quote. We've done this for years, taking their nucleus. Komban slice, adding cocaine are dopamine. Receptor agonist to a slice we don't get fosby induction.
01:05:35So we think that fosby induction and regulation of these other intracellular pathways requires ongoing glutamatergic innervation of the nucleus common cells.
01:05:44Like to establish these patterns so that you'd get differences between the cell types, even if the molecular constituents are largely the same. And they're not fully the same, so I do think there are intrinsic autonomous differences as well.
01:05:58But maybe not so prominently.
01:06:02Yeah, and.
01:06:09Right.
01:06:27Yes.
01:06:29I use the word permanent provocatively. We haven't done that experiment. These are all adult animals. Looking 30 days later. So you can assess and put an adjective on how Long live that is.
01:06:42But I guess the idea is that a discrete experience in adulthood can produce changes in chromatin that persist for a fairly long time. Even though you and I know that a large majority of the chromatin changes are very label.
01:07:01And that's where this open ended approach is so useful because it helps us focus in on those changes that really do persist and as one example during development. We know that do do signals that remain poorly understood. There are changes in a cell that produce its differentiation that then it becomes truly permanent and so our hypothesis is that perhaps behavioral experience produces similar Long live changes.
01:07:27Maybe not permanent.
01:07:30Yeah, in the back.
01:07:38Work with patients with drug use is a lot of.
01:07:52What they need?
01:08:02Anybody.
01:08:10Yeah, no, I agree.
01:08:15Absolutely.
01:08:18Runt.
01:08:21Right.
01:08:24Thanks Ron,
01:08:26So.
01:08:30Yes.
01:08:35Yes, absolutely so notice here is the idea that perhaps some of these epigenetic enzymes or other proteins that are mediating these long lasting changes could themselves be drug targets like a Jack inhibitors?
01:08:51The specific methyltransferase is I'm not going to go back and slides they specific methyltransferase is 4H3K79 demethylation for example.
01:09:01And there are small molecule inhibitors developed for most of these enzymes for the treatment of cancer.
01:09:07So it becomes feasible to begin to test that in patients with drug abuse. The challenges that the molecules that have made so far act throughout the body and are quite toxic and so we'll have to wait for molecules that are safe enough innocuous enough in normal people and then try them in a drug abuse population. But something we're very interested in doing manipulating these epigenetic. Enzymes has a dramatic effect on animals behavior and gene expression.
01:09:35Should we stop or?
01:09:37Thank you all very much.
01:09:52Uh it would only be fair now to invite George Henniger up to.
01:09:57Lecturing to make a few comments.
01:10:14Well, thank you. The committee that set this up. It's a real reward. Most of all I would like to thank my wife Julie.
01:10:3663 years when I made associate professor, he real my mother took me aside and said George.
01:10:45When you came home at 2:00 AM with the car smelling of Baron's tobacco smoke. We thought you'd never amount to much and I tried to explain the other guys roll and she interrupted and said.
01:10:58You know, George everything you've achieved is because of Julie Ann into mothers know their children might need.
01:11:10So thank you Julie.
01:11:13The I didn't wanna take a lot of time because even here, you've heard a lot.
01:11:20I just had to thoughts and.
01:11:23Having been here for 53 years there.
01:11:28The whole life of the mental Health Center. I wanted to say 2 things about the past.
01:11:36In the past is.
01:11:39Maybe summarized by.
01:11:41Um locks it Vera Tos.
01:11:45The University.
01:11:47The medical school in the Department and I've been here through 7 presidents who the University 8. Deans of the medical school and 7 chairman of the Department of psychiatry, but that.
01:12:00That model of light and truth.
01:12:03His prevailed.
01:12:05And that's that's what's carried his foreign so that's why this is a great place to study in its been fertile for main the main thing with the establishment of the mental Health Center by Fritz Redlich, an then governor Abraham Ribicoff.
01:12:23Um an I had a couple of quotes I won't go along on that, but
01:12:29This is a quote from Abraham Ribicoff.
01:12:34Let us evaluate are thinking about mental illness and how we have dealt with it. Let us ask ourselves are even the modest advances. We have made in recent years on the right track or do we need an entirely new point of departure? Are we exploring every possibility in innovation psychological social and biological and so.
01:12:56That that was the charter of the mental Health Center Ann.
01:13:01Few years later, Malcolm Bowers interviewed for its red look for the Department and then that that.
01:13:07That video is available in their Department website and Malcolm eyes doctor ever.
01:13:14What's your advice to us and Relic said.
01:13:18Science.
01:13:19Use science to be helpful at summarize the whole thing so.
01:13:25Between those 2 this center, which is the cornerstone of the Department it spawned.
01:13:32Overtime growth throughout until the modern Department.
01:13:38It's been my home an but but the ideal of of what Doc Governor Ribkoff said is still there.
01:13:51And so I won't belabor that and let me just say.
01:13:55Quickly, about the future, I do that. An.
01:14:02And thinking about it, you get old.
01:14:05And so.
01:14:07People here have had cancer in this auditorium.
01:14:12And people in this auditorium are going to get cancer and people in the Department have died of cancer.
01:14:20But but look across the street look at what they're doing and.
01:14:25That hospital OK there's progress and so.
01:14:30On TV last night cancer rates are dropping.
01:14:35Why not because of detection?
01:14:39Uh or prevention.
01:14:41It's for treatment and if you look at the trim, and what they've done.
01:14:46It is opened up a new field immuno therapy OK.
01:14:52Car car T cells.
01:14:54Checkpoint inhibitors there's a whole. These guys are good and so that's our that's our older sibling.
01:15:05About how to go about it so.
01:15:09Uh.
01:15:10Let me just say an example that illustrates that of where we are, and where we need need to go because I'm I really want to talk to the 30 year olds here.
01:15:21Who have their 53 years in the Department OK? I won't? But you will recent report of 22. Q 11 syndrome degeorge syndrome. The most prevalent genetic cause of schizophrenia got an animal model for that.
01:15:40They found
01:15:41Reduce gene expression of a factor that's an anti oxidant.
01:15:47Yeah, that was good.
01:15:49But the interesting thing is that they gave.
01:15:53And an anti oxidant.
01:15:58They given antioxidant to the developing animals an actually prevented the condition.
01:16:04There's another paper where they been able to prevent.
01:16:08Animal of Down syndrome.
01:16:10So it so if the Sciences that powerful.
01:16:17We just need to apply it here and so.
01:16:21I just say to the young people.
01:16:24This is extremely rich environment.
01:16:28So seize the opportunity and and let me start stand in your way because in your 53 years. You'll be able to make.
01:16:41Miraculous advances on Curzan Prevention, so thank you.