Yale Psychiatry Grand Rounds: October 16, 2020

Name: Yale Psychiatry Grand Rounds: October 16, 2020
Uploaded: 2020-10-16T16:21:57.293Z
Duration: 1 h 19 min 55 s
Description: Tracy L Bale, PhD, University of Maryland: " Extracellular vesicles as stress signals: Identifying novel systemic mechanisms of trauma programming "

October 16, 2020

Tracy L Bale, PhD, University of Maryland: "Extracellular vesicles as stress signals: Identifying novel systemic mechanisms of trauma programming"

Information

ID: 5758
To Cite: DCA Citation Guide

Download Transcript

00:00I first was introduced to Doctor Bale.
00:05Uh, maybe 5 six years ago at SFN I heard
00:08her give a talk at a women's symposium
00:11and I have to say I was mesmerized.
00:14There were a lot of strong women speakers,
00:17but I had an impression on me and so
00:21followed work for a few years and asked.
00:24To actually be able to come and give a
00:26grand rounds talk at Yale and between
00:28our schedules, this was over a year ago,
00:30so now she's finally. Here, Sortof.
00:35Seattle is a professor pharmacology and drug
00:37tour of the center of epigenetic research
00:40in child health and brain development.
00:43In the School of Medicine and yelling
00:46at University of Maryland in Baltimore.
00:49She completed her PhD University of
00:51Washington Department of pharmacology
00:53and her postdoctoral work at
00:55Salk Institute with doctor Vail.
00:58Doctor bail was Professor Neuroscience,
01:00an onion soup,
01:01and city in 15 years before moving.
01:05Her research focuses on understanding
01:07the role of stress dysregulation in your
01:10developmental and your psychiatric disease.
01:12And the sex differences in underlying
01:15disease vulnerability in humans in using
01:18the mouse model as it the Monster Model
01:21she's interested in developing models of
01:24Parenteau stress and the germ cell movement.
01:27An intergenerational programming
01:28of newer development,
01:30she serves on many internal and
01:32external advisory committees,
01:34panels and boards.
01:35She's been the recipient of many many
01:37awards including early career award
01:39achievements by Society for neuroscience
01:42and exceptionally promising an
01:44investigator through Endocrine Society.
01:46And others,
01:47and especially Metro Matronic Award
01:49from Society of Women's Health for
01:52Outstanding Research that has led
01:54to the improvement of women's help.
01:56She was recently elected president
01:59of the international brain research
02:01organization and it's my honor to welcome
02:04doctor bill to give grand rounds here.
02:07Thanks so much,
02:09Irina,
02:09and thank you again for the invitation
02:12and I guess one of the benefits of Kovid
02:16if there can be any is that we're all
02:19sort of trapped at home with no travel,
02:22which makes availability of
02:23scheduling a little bit easier.
02:25I think I really excited to
02:27talk about our data today,
02:29and I think the Yale Psychiatry audience
02:32is just such a tremendous group of broad,
02:35basic clinical translation.
02:36Are researchers that makes this.
02:39Even more exciting for me to be able
02:41to talk science and the translation of
02:44potential, I think of the of the science,
02:48and I'd love to hear your feedback.
02:51So with that,
02:52I also know that because there is such
02:55a broad background of people on the
02:57zoom today that rather than waiting
03:00until the very end with questions.
03:03If there's something that really
03:04clarification would make my talk
03:06more digestible or understandable,
03:08please don't hesitate.
03:09To interrupt just for clarification,
03:11points 'cause I'm happy I'm happy to do that,
03:14especially for a grand wrong talk.
03:16The topic I decided to put together
03:20today encompass is a kind of a wide
03:23area of the research in my lab and
03:26the common theme is going to be
03:28about extracellular vesicles and it's
03:30actually a timely topic, I think.
03:33But it's also to me personally
03:35relevant to meaningful to Yale
03:37because the first time I started
03:39talking about these extracellular
03:41vesicles was actually about five
03:43years ago at the ACM P the American
03:47College of Neuropsychopharmacology.
03:48Meeting and in that session
03:50actually was was organized by one
03:52of the organizers was Ron Duman,
03:55and we had a great discussion
03:57afterwards and he was really
03:58intrigued and excited by this idea of
04:01these extra set of their vesicles,
04:03and so to me it's very meaningful to
04:06be discussing this with the audience.
04:09Again thinking of Ron missing
04:10him so much so many I know it's
04:13a Yale and psychiatry,
04:15especially their interested in stress as
04:17it relates to nuro psychiatric disorders,
04:19so I hope that you find.
04:22This talk,
04:23even though sometimes it might get
04:25down in the weeds a little bit for
04:27clinicians to think about the translation
04:29of value of thinking about Biomarkers,
04:32and I think great conversation
04:33to have is is the word biomarker.
04:36What does biomarker mean in some cases?
04:38Biomarker means indicative
04:40or associated with.
04:41But a lot of times biomarkers can
04:43be pushed a little bit further to
04:45talk about mechanisms that those
04:47biomarkers that are identified
04:48in many clinical studies actually
04:50have mechanistic or causal value.
04:522,
04:52and I think that there's an incredible
04:54value in partnerships between clinical
04:56research and basic research that
04:58those biomarkers can cross cross that divide.
05:00Alright,
05:01here we go.
05:02Let's do this.
05:03OK,
05:03so before I get started then
05:05because I want to make sure
05:07everybody is on the same page of
05:09what in the world is she talking
05:11about with extracellular vesicles?
05:13I'm going to sprinkle this
05:15in throughout the talk today,
05:17but I wanted to just make sure
05:19everyone's on the same page as to
05:21what exercise or vesicles are and
05:23that they really do hold great
05:25translation of value but also
05:27reverse translation with potential
05:29toward understanding mechanisms
05:30and causal aspects of disease.
05:32So an extracellular vesicle is
05:33broadly termed as a small lipid
05:35and contain draft that is released.
05:38It is not a cell itself,
05:40but it is lipid contained.
05:42That contains many proteins in
05:45its membrane's structure and many
05:47small noncoding RNA's and other
05:50proteins that are its cargo inside.
05:52These exercise are vesicles you
05:54may have heard them called exomes.
05:56Exosomes are a small version of
05:59an extracellular vesicle,
06:00so exercise vesicle is a broad term that
06:02encompasses all types of these vesicles.
06:05Whereas eggs om is only one
06:07type of exercise or vesicle,
06:09I will abbreviate exercise in vesicles,
06:11often in the slides as Yves.
06:13Just you know what that is.
06:16And so in order for anybody
06:18to really say that something,
06:20for instance is an eggs ome versus just
06:23classifying it more generally as an EV,
06:25you have to actually go to great lengths.
06:28And there are actually societies
06:30and rules that determine the
06:32clarification and the rigor
06:33by which you have to validate
06:36that you're calling something and eggs om.
06:38So my lab we stay away from really
06:41defining these small types of
06:43vesicles versus just more grandly
06:45extracellular vesicles for that reason.
06:47The proteins have been characterized
06:49greatly in the content of these vesicles.
06:52All tissues in all mammals secrete
06:55exercise their vesicles into circulation
06:57and these vesicles travel in high
06:59concentration throughout the circulation.
07:02They travel in a somewhat specific manner.
07:05I like to use the analogy to the
07:08endocrine system whereby you have,
07:10for instance, gonadal release of
07:13steroid hormones that travel in
07:15circulation and act at distant sites.
07:18EV's are similar to that only in that
07:21they travel in circulation to distant
07:23sites or they can act locally as well.
07:25And I'll give you examples of both.
07:28But TV's also have an incredible
07:30specificity of where they act.
07:31Unlike things in the endocrine system
07:33that can act in many different issues.
07:36The way that that specificity happens is
07:38that if you actually look at the membranous
07:41structure of an extracellular vesicle,
07:43there are very specific protein
07:45combinations that determine both the
07:47tissue that the EV is released from,
07:49and the tissue in circulation that they
07:52will act upon that cargo that they deliver,
07:55both in interacting at the membrane
07:57at local cells, in a tissue.
07:59Oftentimes, the immune system will
08:01deliver cargo internally to a cell,
08:03and as you can imagine,
08:04if that cargo contains,
08:06for instance,
08:07small noncoding RNA.
08:08Lot of Micro RNA's is an example
08:10that those micro RNA can have an
08:12immediate and profound effect on the
08:15translation of Gene and transcription
08:17translation machinery such that
08:19the more of a given micro RNA
08:22delivered rapidly to a given cell,
08:24the more rapidly it can degrade a given
08:27targeted M RNA and prevent its translation.
08:30So it's really important I
08:31think is a biomarker,
08:33both Association with many disease states.
08:35Some of the greatest examples are that
08:38extracellular vesicles are being discovered
08:40as being communication within the brain.
08:42As well as released from the brain,
08:44but they also travel in circulation
08:46from many other tissues.
08:47The cancer field has really done the
08:49most work on exercise are vesicles,
08:51and they've done a lot of that work in
08:54relationship to signals to the immune
08:55system so will kind of come back to that.
08:58So that's what an exercise or vesicle is just
09:01to make sure we're all on the same page.
09:03There is,
09:04unfortunately,
09:04at this time alack of really
09:06rigorous tools and that is something
09:08that labs are working on.
09:09My lab is in the process now
09:11of making a mouse.
09:13That allows us to conditionally
09:14target and look at release of
09:17vesicles from specified tissues
09:18and follow them to other tissues,
09:20but those are tools that don't
09:23currently exist unfortunately,
09:24and that limits a lot of our
09:26interpretation of these Eves.
09:28These eaves have also been associated
09:30with many different disease States
09:32and will will come back to that.
09:34So as Irina introduced my
09:36lab covers a different areas
09:37around stress and neuro psychiatric disease.
09:40We focus a lot in my lab and
09:43understanding stress across the lifespan.
09:45And its impacts,
09:46especially in neuro development.
09:47I'm going to tell you a couple different
09:50stories from the lab today that focus
09:52a lot on understanding both male
09:54and female experiences with stress
09:56in their environment and adversity.
09:58One of the aspects, a spec.
10:00And now that I'm in Baltimore,
10:02which is a majority black city and
10:05engagement with the community that we're
10:07very interested in in doing service
10:09and understanding and appreciating the
10:11health disparities to this community is
10:14understanding the mental health aspects,
10:16especially.
10:16As you can imagine nowadays and
10:18understanding how it contributes to
10:20intergenerational changes in Nuro
10:22Development and risk for things like neuro,
10:25psychiatric neurodevelopmental disorders.
10:26So we know that there's disparities
10:28across mental health especially for.
10:30African Americans that now more than ever,
10:33that racism and discrimination
10:35produced profound effects and
10:37exacerbate mental health issues.
10:38But what I want to show you is
10:41models and translation of potential
10:43for understanding the long term
10:46consequences and intergenerational
10:47impacts of that discrimination stress.
10:51One of the key aspects that my lab
10:53is interested in is really modeling
10:55and understanding impacts on both
10:56maternal and infant mortality.
10:58In the long term,
10:59risks of even the survivors
11:00of discrimination,
11:01distress and how it's passed on.
11:03So as you can see from this graph here
11:05that that only currently goes up to 2013.
11:08If you compare a white mom versus a
11:10black mom in the United States that
11:12they have a four times increased
11:14risk for maternal and infant death,
11:16the bottom panel had just shows
11:18you within the state of Maryland
11:20that even though this green line,
11:22which is black babies has been decreasing.
11:24That infant mortality rates
11:26remain still four times higher,
11:27so a black mom with an advanced
11:29degree so it advanced education
11:31still has a four times increased
11:33risk for maternal and infant death
11:35within the first year than a white
11:38mom with a high school education.
11:40So it is not just related to social
11:42economic that there are many
11:44factors we need to understand in the
11:46environment and related to stress.
11:48We're very interested in trying to
11:50understand the long term influences
11:51of discrimination stress.
11:53So this term has been defined,
11:55brought more broadly in the.
11:56The social sphere is weathering
11:58and the toll that racism places
12:00as a social psychological tale,
12:01but also a physiological toll on the
12:03body and the germ cells as well that
12:06give rise to effects for the next generation.
12:08So Arlene Jeronimus,
12:09who's in Michigan,
12:10has studied this for many years and
12:12I give her credit for all of her
12:15are incredible work in this area.
12:17And an coining the term weather,
12:19rain,
12:19and the stresses that impact black
12:21individuals are chronic and repeated
12:23throughout their whole life course.
12:25And one of the aspects that we're
12:27particularly interested in trying to
12:29understand those influences in pregnancy.
12:31So many of you have heard the
12:33term aces or
12:34adverse childhood experiences,
12:36you know that Kaiser Permanente
12:37developed this ace protocol for
12:39understanding the effects throughout
12:40the lifespan on health outcomes.
12:42The original aces were
12:44really being the California.
12:46So can I ask that everybody try and
12:49mute themselves and getting lots
12:50of feedback for people in their
12:53offices were coming up on lunch time?
12:55Thank you so adverse childhood
12:56experiences as they relate to
12:58the Kaiser Permanente study,
12:59which was done in the state of
13:02California which many of you I'm
13:04sure are very aware of his was done
13:06in the largely white population.
13:08Many of those aces are now being
13:10redone and evaluated in inner city
13:12and especially in black populations,
13:14but there originally done to
13:16determine if adversity experienced,
13:17especially prior to adulthood,
13:18could predict risks for all kinds of
13:21health disparities in health outcomes.
13:22And so they were really qualified as 10.
13:25Adverse childhood experiences and those
13:27childhood experiences give rise to
13:29risk for many things that behavioral
13:31outcomes such as drug abuse and addictions,
13:34as well as physiological and mental risk.
13:37But there are also determined that
13:39the number of these aces prior to the
13:41age of 18 was also very productive,
13:44and in their study they determined
13:46that the accumulation of four or more
13:48of these adverse childhood events,
13:50which include things like divorce
13:51of your parents,
13:52incarceration of apparent drug
13:53addiction in the household,
13:55and violence,
13:55neglect,
13:56accumulation of four more of
13:57these aces in an individual,
13:59was a strong predictor of a
14:01lifetime of health consequences.
14:02So that's the number of the cloud software
14:05they determined was the highest risk.
14:07So going back to the interests of
14:09my lab and trying to understand
14:11so we can develop better models
14:13to look at the mechanisms.
14:14If you just look at the term of
14:17pregnancy itself and you were
14:18to look across the United States
14:20of America and ask all pregnant
14:21women the number of their adverse
14:23childhood events so those exposed
14:25to them before the age of 18,
14:27now that they're much older and
14:29pregnant themselves and you can look
14:30at the distribution across the US for
14:32those experiencing zero all the way
14:34up to four more highlighted in yellow,
14:36is that point of increased health risk?
14:38So across the US is about 12 1/2%
14:41of women who are currently pregnant
14:43that have experienced for more.
14:45But understanding the population
14:46that we are trying to serve and
14:48understand the risk factors for
14:50in the city of Baltimore.
14:52Arecent Baltimore mom study found
14:54an actual evaluation of inner city
14:56women here that that number is much
14:58greater and in fact that six more
15:00than 61% of pregnant women in inner
15:03city Baltimore have experienced four
15:05more aces and I think I want to take
15:07a second just to reflect on what that
15:10means in terms of trying to understand.
15:12The experiences and the risk factors
15:14for these women during their pregnancy
15:16for both themselves and as well as
15:18that of the health and the outcomes
15:20for their their baby,
15:22and I think that's incredibly
15:23important for us to think about
15:25in terms of the developing brain.
15:27So back to the translation
15:29of potential and biological
15:31signals that we want to study.
15:32So on the left is is a schematic
15:34from a recent review from Rachel
15:36Yehuda's group that I think highlights
15:39on the human side reflects the
15:41differences across adverse experiences
15:43that gives rise and influences on
15:45things like the gametes, the cell,
15:47the cells that come together at
15:49fertilization, egg and sperm.
15:51The Neo Nate itself in the developing
15:53infant and all of the experiences
15:55of mom during her pregnancy prior
15:57to her pregnancy and giving rise to
16:00health risks and resiliency as well
16:02for the 1st and next generations
16:04is important to think about.
16:06It is very difficult in a human and
16:08there really is relatively little
16:10evidence at the mechanistic or causal level.
16:13Especially,
16:13we're thinking about germ cells
16:15of what really contributes to the
16:17differences in brain development.
16:18We really have to look at models
16:21and largely we have to use again
16:23mice and rats and other other
16:25models to think about how we can
16:28mimic stress and adversity.
16:29While of course never being able to
16:31fully model that adverse environment that
16:33that someone of color might experience,
16:35especially in.
16:36In inner cities such as Baltimore,
16:39but in order for us to really
16:41get at these mechanisms,
16:43even going back to thinking
16:45about the extracellular vesicles,
16:46we really have to begin to understand
16:49both the risks and the changes in germ
16:52cells related to Dadan related to mom,
16:55mom's preconception,
16:56environment,
16:57pregnancy environment itself and
16:59postpartum environment are all
17:00encompassing in disease risk.
17:02So this is a very sensitive subject
17:04in getting into.
17:06How do we develop models in a
17:07mouse to begin to understand those
17:10mechanisms of something so important,
17:12such as discrimination,
17:13stress or high aces and adversity.
17:15So obviously we can't.
17:17Mice are not humans and they do
17:19not live in those same experiences,
17:21but what we can do is we can begin to
17:24appreciate the stress axis itself.
17:26The responses to stress in the environment,
17:28whether it be acute or chronic effects,
17:31and begin to understand things
17:32at the very cellular level,
17:34and I think that's an important topic,
17:36especially for a grand rounds.
17:37Is that what is stress?
17:39So it is psychiatric level
17:40are relative to the brain.
17:42The perception of stress can
17:44be psychological in nature,
17:45can be physical in nature,
17:46you're hungry, you're cold,
17:48those are stresses.
17:49But you're you're feeling chronic
17:50stress at work or at home,
17:52covid within the black lives
17:54matter community.
17:54Those are all stresses and
17:56they can be chronic in nature.
17:59But we also have to remember
18:01that the cell itself,
18:02whether it be in neuron or a germ cell,
18:05as somatic cell lining
18:07the reproductive track,
18:08is an example.
18:09We'll come back to that the
18:10cell itself doesn't understand
18:12it's not connected directly to
18:14the perceptions of the brain.
18:16So how do we model biologically
18:18the stresses in the environment?
18:20The minimal aspect of this and Jane
18:22Taylor's work has contributed tremendously
18:24to thinking just about glucocorticoids.
18:26You can't have stress without
18:28elevating glucocorticoids.
18:29But there are many other aspects.
18:31Of course with stress that
18:32are beyond glucocorticoids.
18:33But at the very cellular level,
18:35the perception of a cell,
18:37whether it be a liver cell,
18:39whether it be a reproductive
18:40cell along reproductive tract,
18:42or whether it be in neuron,
18:44the perception of stress in the
18:46environment minimally has to be
18:47an elevation in glucocorticoids
18:48that can be acute or chronic
18:50in nature and those will change
18:52your intracellular structures.
18:54And I'm going to give credit to
18:56Bruce McEwen as well for his
18:58incredible career work on the term.
19:00Allostasis, which will come back to.
19:03But the perception of the cell
19:05of stress of what stress is is a
19:08change in glucocorticoids minimally.
19:09And I'm also going to highlight that
19:11that change in glucocorticoids is
19:13really important for the cell when
19:16it's outside the normal circadian rhythm.
19:18So the Google Corticoids in all
19:20mammals rise and fall in a 24 hour
19:23cycle that happens every single day.
19:25The cells are aligned in their
19:28rhythm by those glucocorticoids.
19:29So really to model stress is really
19:32just elevating glucocorticoids.
19:33Outside that normal rhythm, right?
19:35That's where about so at the
19:37at the most cellular level,
19:38so I'm going to give you an example of
19:41some of our work of how we think about
19:44this at the in terms of pregnancy.
19:47So moms experiences,
19:48whether it be preconception or
19:49during pregnancy are manifested
19:51in intersections at very important
19:53issue that gives rise to signals
19:55to the developing embryo and
19:56fetus throughout gestation,
19:57and that tissue is the placenta,
19:59so the Internet.
20:00Action of maternal stress and that again
20:03that stress can be psychological in nature.
20:05That stress can be changes
20:07in her diabetic status.
20:08That stress can be many different
20:11factors that influence her perception
20:12and maintenance of homeostasis so we
20:15know I'm giving you example here of
20:17showing you that that intersection
20:19of the maternal endometrium and moms
20:21cells that have been experiencing
20:23moms life adversity at that first
20:25intersection where the blastocyst
20:27these purple cells outlined here are
20:29the trophectoderm that will give rise
20:31to trophoblast cells of the Placenta.
20:33And the inner cell mass here,
20:36which will give rise to the Amber
20:38developing embryo and eventual fetus.
20:40These cells are feeding information
20:42from moms environment and our
20:44programming themselves that will
20:46be remained throughout pregnancy.
20:48These this trophectoderm Here that
20:50gives rise to the trophoblast,
20:51will differentiate into all necessary
20:53cells of the Placenta as it invades moms,
20:56and Dmitri 'em and develop developing
20:58the decidua that gives rise to all
21:01the different cells of the Placenta.
21:03So why,
21:04in a psychiatric psychiatry grand rounds
21:06are we talking about the placenta?
21:08Well then to is an ender can tissue.
21:11It is critical,
21:12obviously for survival and
21:13development of the
21:14fetus, the trans placental signals
21:16coming from the placenta are
21:18intersecting with that developing fetus,
21:20including the developing brain.
21:21It's going to feed that developing
21:23fetus all the information about the
21:26environment it's about to be born into and
21:28throughout the course of pregnancy first,
21:30second trimesters especially.
21:31But even into the 3rd trimester.
21:34The Placenta provides all
21:35the necessary growth factors,
21:36oxygen support and information
21:38that the developing brain needs,
21:40but it also shapes the developing brain.
21:42So if there's changes in moms stress
21:44environment, her glucocorticoid
21:45levels are going to change.
21:47That's going to intersect at the
21:49level of the Placenta as an example,
21:52but also of her energy availability changes
21:54because of diabetes because of famine,
21:56those those.
21:57Those clues are also really important
22:00for the developing placenta.
22:02OK, so the way that we have
22:04thought about this in modeling
22:05over the last decade in the lab,
22:07this is just giving you an
22:08example of a rodent model.
22:10Here you have F0, which is mom.
22:12You can model this in
22:13terms of maternal stress,
22:14which we've studied in the lab
22:16and I'm going to talk about today.
22:18We've also manipulated mom's diet,
22:19which I'm not going to have
22:20time to talk about today,
22:22so that's the F zero generation.
22:23Just stating in mom is the F1 generation,
22:26so these are largely somatic
22:27cells as well as germ cells which
22:29give rise to the F2 generation.
22:30So those are the primordial germ cells.
22:32Those are also programmed during gestation,
22:34so you can imagine a scenario where
22:37moms stress in her environment,
22:39be it preconception or during pregnancy,
22:41changes in her diet,
22:42give rise to changes in a developing
22:44placenta that will guide and
22:46shape these somatic cells,
22:48putting within a range of
22:50risk and resiliency.
22:51Because of course you're on
22:52a genetic background here,
22:54so this is again goes back to
22:56RGB Y your genetic background.
22:58That's mom and dad contributes to
23:00as well as the environment this.
23:02Is a very specific timing window
23:05of a shaping environment,
23:06but it also shapes a lot about these
23:09primordial germ cells that will
23:11give rise to a next generation.
23:14This is intergenerational future
23:16generations make it trans generational.
23:19So I don't have time to go into
23:21how this model has developed over
23:23the last 10 to 15 years.
23:25All of this work has been published,
23:28but let me just quickly summarize,
23:30we developed a mouse model.
23:32We're really wanted to understand
23:33chronic stress in moms environment.
23:35During pregnancy,
23:36we identified a particular window
23:37which is basically early pregnancy,
23:39the equivalent in the human of the
23:411st trimester where moms experience
23:43with stress in a chronic manner out
23:45of sync with her normal circadian
23:47rhythm produced offspring.
23:49Where the male offspring because
23:51they were doing this in mice,
23:53so they have a litter,
23:55were able to study sex differences here.
23:57This in Utoro stress provided
23:59male offspring but not her female
24:01offspring with changes in their
24:03adult behavioral stress responses
24:05as they grow and develop changes
24:07in their physiological stress
24:09axis. Changes in their stress
24:11regulatory genes in their brain,
24:13cognitive learning and memory deficits,
24:15as well as some really interesting
24:17neuroendocrine phenotypes of
24:18their weight gain and changes.
24:20Post puberty, so again to summarize,
24:22this is all published work.
24:24It is in male offspring of the moms
24:26that were stressed, not female.
24:29Why is this relevant?
24:31Because we know that most neurodevelopmental
24:34disorders in humans also show a male bias.
24:37So autism, early onset,
24:39schizophrenia, oh oh, CD, etc.
24:41Most nuro psychiatric disorders
24:42related to neuro developmental
24:44timing tend to show a male bias.
24:47There seems to be something in utoro
24:50that is either extra at risk for
24:53males that are in development or
24:55resilient or preventative in females,
24:58or both.
24:58We know that if you go into a Nick
25:01you any necu nurse will tell you.
25:04They often see more males
25:06in the neque than females.
25:07Females tend to be more resilient
25:09going home earlier than males do,
25:11and that there's again seems to be
25:13this protective effect of in utero.
25:14Insults or females are protected
25:16in males are at risk.
25:18So in modeling that one of the genes
25:20that we have identified looking in
25:22the placenta is potentially this
25:24tissue that is protected for females,
25:26we identified 1 gene through many
25:28different transcriptomic approaches
25:29called OG tierro glycosyltransferase.
25:31This is an enzyme,
25:32it's an X linked gene.
25:34I'm not going to go into all the
25:36work of how we identified it,
25:38but it turned out to be what I
25:40defined as sort of a Canary in
25:42the coal mine gene that in the
25:45placenta in the trophoblast cells
25:47coming from the developing.
25:48Embryo that this X linked
25:50Gene Escapes X inactivation,
25:52meaning in female placental tissue.
25:54So if it's a female fetus,
25:56that placenta that is developed out of that
25:59embryo has more oh GT than a male placenta.
26:03So in the rodent world,
26:04because we have litters,
26:06we can look at the same
26:08intrauterine environment and
26:09look at sex specific outcomes.
26:11Why is this gene so interesting?
26:14Well,
26:14for many reasons,
26:15one of which is if you were to.
26:18Postulate what gene could produce hosts
26:20of effects very quickly that would
26:22be relevant evolutionarily to develop
26:24in terms of the developing brain.
26:26You would say, Well,
26:28something related to energy availability in
26:30the environment would be very important.
26:32Turns out oh,
26:33GT is an enzyme is regulated by
26:35glucose and mom's environment.
26:37It's also associated with hosts of
26:39effects that are really important
26:41for the developing embryo and fetus.
26:43It is strong epigenetic regulator,
26:45so it allows it to regulate at the
26:47level of the trophoblast cells
26:49in the Placenta.
26:51Very dynamically responses to moms
26:52environment and for those interested
26:54in sex differences in outcomes
26:56this gene is on the X chromosome,
26:58and in fact it's located very
26:59close to the long non coding RNA
27:02exist biochemically.
27:03There are interactions of these two
27:05genes that may provide again this
27:07extra resilience that will come back to for
27:09females in Utoro.
27:10This is all the boring biochemistry
27:12than required by law to show you
27:15that oh GT not only its RNA but its
27:17protein levels twice as high in a
27:20female placenta in a male placenta.
27:22The data I'm showing you here is
27:24all from mouse, but we've replicated
27:25this all in human placenta as well,
27:27so it escapes X inactivation twice as high
27:30in a female placenta compared to a male.
27:33And as an enzyme,
27:34what one of the things that Ogede
27:37Duzer by its effects on regulation
27:39is it places a sugar mark,
27:41an oblique inoculation mark
27:43on Syrian threonine residues.
27:44So you can think of any of your
27:47favorite enzymes or other proteins
27:48that are regulated by phosphorylation
27:51on Syrian training,
27:52and chances are it competes with old
27:55Lincoln alkylation and the take home message.
27:57Here is that typically Algonac olation
27:59is a break or phosphorylation is a gas,
28:02so the regulation of hundreds of proteins.
28:05Bio Glick Inoculation.
28:06Is what I'm showing you here.
28:08If you take out the placenta,
28:10either this is mouse,
28:12same results in a human placenta.
28:14There is way more protein,
28:15oblique inoculation in a female
28:17placenta compared to a male,
28:19and that's important because it says
28:20not only are its protein levels important,
28:23but it's what it's doing is similarly
28:25regulated and what that says is that
28:28the ability to break is much higher in
28:30a female placenta than a male placenta.
28:33By this biochemical mark.
28:34So our current model is is
28:36that there's a threshold.
28:38A vulnerability here where if you compare
28:40a control versus early prenatally
28:42stressed within females and males
28:44coming out of the same uterus so we
28:46can take out a male placenta in a
28:48female presenter from the same uterus,
28:51the same experience and show you that
28:53the levels protein and M RNA levels
28:55and Oakley can alkylation itself are
28:57much higher in the female and even
29:00though they are all affected by this
29:02stress that the male presented drops
29:04below some threshold of vulnerability
29:06where by its actions the break being
29:08placed on it is now released such that.
29:11The male placenta is going to
29:12dynamically respond to changes in
29:14moms environment where the female
29:16placenta is much more resilient,
29:17and that's important for functionality.
29:19One of the things locally if we dig
29:21way down in the weeds for those who
29:23don't spend a lot of time thinking
29:26about transcriptional regulation.
29:28One of the things that this important
29:30enzyme does broadly is it regulates
29:33dynamically the transcriptional
29:34and epigenetic state of a cell.
29:37So GT is a stabilizer of a
29:40methyltransferase called Easy H2.
29:42All you need to take home message
29:45from this is that easy.
29:47H2 is a predominant methyltransferase
29:49for histone three at Leising 27 so
29:52that take home messages if there
29:54is more oh GT around,
29:56it will stabilize more of
29:58this methyltransferase,
29:59resulting ultimately within the.
30:00Now of more histone three lysine 27,
30:03what's called Trimethylation,
30:04and that mark can be shown here
30:07by where if there is more oh GT in
30:10a female cell compared to a males
30:12male cell of a placenta, right?
30:14Female placenta?
30:15Male placenta comparison that you
30:17should see more of this trimethylation,
30:19and in fact we do again both.
30:22This is actually from mouse.
30:23We see the same effect within human
30:26placenta in a female placenta,
30:28you see way more of the H3K27
30:31trimethylation than you do of the Mail.
30:33And I'm going to go back to that
30:36that same take home message,
30:39which is more oblique,
30:40inoculation more trimethylation
30:42is more break.
30:43What that break allows is it allows
30:46the female placenta to really
30:48titrate its responses to an ever
30:50changing maternal environment.
30:52Mails missing that break are more
30:54vulnerable because they're going to
30:56respond there percent is going to
30:59constantly be responding to moms
31:01stress or dietary challenge, etc.
31:03Producing hosts of differences
31:04in the transplacental signals,
31:06making it to the developing brains,
31:08developing fetal and fetal brain.
31:11So I'm just going to show you a
31:13couple of quick pieces of evidence
31:15of why this is important.
31:16OK,
31:16so this is showing you if we
31:19just look at just broadly at
31:21mid gestation of a placenta.
31:23And we look at sex differences
31:25just broadly in transcription
31:27that are related to OGT.
31:28So what I'm showing you here for
31:30those again that don't do a lot of
31:33transcriptional regulation studies.
31:34This is from an RNA sequencing experiment.
31:36This is just asking all genes
31:38expressed in a female placenta at the
31:40same time point as a male placenta
31:42from the same uterus and looking at
31:44all genes widely being expressed.
31:46So this is plotted here on the Y
31:49axis is showing you at least in a
31:51log 2 fold change on the Y axis so.
31:54Increased gene expression
31:55decreased gene expression.
31:56The X axis is showing you
31:58the level of expression,
32:00so really highly expressed
32:01genes are out here.
32:03Lowly expressed genes are in here.
32:06A red dot is showing you a robust,
32:09significant difference
32:10between a male and female,
32:13both increased and decreased gene
32:15expression at this one time point.
32:18OK,
32:18if oh GT is important for titrating
32:21responses to the environment.
32:23If we are able to transcriptionally
32:26genetically alter oh GT in a female placenta,
32:30making her hemizygous meaning
32:32her level of oh GT,
32:34because now we're eliminating 1X.
32:36Of Oh GT expression,
32:37so her levels in a heavy zigas
32:40environment should be very similar to
32:42the levels of oh GT in a normal male.
32:45Now what happens to those differences
32:47between males and females?
32:49They are almost completely eliminated,
32:51and in fact the jeans that remain
32:53here that are significantly different
32:55between the two or either X or Y linked.
32:59Suggesting again,
32:59those are not going to be
33:01affected by the OG transcription.
33:03This evidence,
33:04the take home message here being
33:06this one gene that we identified
33:08shows you as a candidate approach
33:10that is widely important for
33:12what happens in the Placenta
33:14and then altering all of those
33:16differences in the transplacental
33:18signals that are going to happen being
33:20relayed to the developing fetal brain.
33:22Where's our evidence of that?
33:25What I'm showing you here is now
33:27an RNA sequencing experiment.
33:28This is these are heatmaps.
33:30These are the identical brains.
33:32From those placentas whoops,
33:33sorry from these placentas
33:34that I just showed you.
33:36So if we take out the fetal brain
33:38from the same midpoint of gestation
33:40and we now examine it for gene
33:43expression in the brain, OK,
33:45we already know in all mammals,
33:47mice, and humans that at any given
33:49time point in brain development
33:50or fetal development in general
33:52because males and females develop
33:54it slightly different rates.
33:56Likely accounting for differences that
33:57we're seeing here in gene transcription.
33:59OK, so this is the The Geno
34:01type of the placenta.
34:03That transcription I'm showing
34:04you here is of the brain,
34:06so a normal female brain had mid
34:09gestation compared to a normal male brain.
34:11Same pot, same time point.
34:13You can see that a lot of the jeans
34:15that are really elevated right?
34:17So each column here is a different brain.
34:20Each row is a different gene.
34:22OK so this is this is RNA sequencing.
34:24What this is showing you?
34:27Again,
34:27is that if we examine this by looking
34:29at the effects in a normal female
34:31versus a normal male jeans that
34:32are ultimately highly expressed at
34:34this time point in a female are
34:36not so highly expressed in a male
34:38and jeans that are lowly expressed
34:40in the female brain are.
34:41Again,
34:41this is the hypothalamus of the brain
34:43are more highly expressed in a male.
34:45The take home message here is
34:47that we already know this.
34:48We know that even before birth
34:50the male and female brain are
34:51in different trajectories.
34:52But if we now alter the level
34:54of oh GT in the placenta,
34:56leaving the brain totally normal.
34:57Trance. Altering transplacental signals?
35:00What does her brain look like?
35:02And I think you can see here that
35:05ultimately you're seeing is that
35:07this female brain were not her OG
35:09T levels in Placenta are similar
35:11to that of a male, not a female.
35:14Her brain becomes more male like
35:16and that tells us is that these,
35:18oh GTO,
35:19Glick inoculated proteins are really
35:20important for transplacental signals
35:22guiding the development of the fetus,
35:24and no doubt the rate of the fetal
35:26development, and that no doubt,
35:28is important for risk for
35:30neurodevelopmental disorders.
35:31Ultimate thing we've been working
35:33on for the past decade that we
35:36keep sort of running,
35:37beating our heads against the wall,
35:39trying to figure out is OK.
35:41What are these transplacental signals?
35:43We've identified many host of effects
35:45at the level of the placenta itself.
35:47We've identified even many of the
35:49transplacental potential signals
35:50from the placenta,
35:51but it's really difficult to get at.
35:54Are there steroid hormone differences
35:56that guide the developing fetal
35:58tissues and brain?
35:58Are there other proteins that are
36:01important information for relaying?
36:02Both the rate of development
36:04and fetal brain development.
36:06One of the signals that we found
36:08that I'm going to talk to you about
36:11is the extracellular vesicle.
36:13There is no time in million lifespan
36:16where if you were to take out through the
36:19blood and isolate exercise or vesicles,
36:22the highest concentration of extracellular
36:24vesicles in circulation is pregnancy
36:26because the placenta itself produces
36:28a ton of extracellular vesicles.
36:30Those both come from the trophoblast cells
36:32that interact with the developing fetus.
36:35But also the decidua side that
36:37interacts again with moms circulation.
36:38So trying to understand these extracellular
36:41vesicles and what signal that they
36:43might pose and how stress affects them
36:45could be a really a potential important
36:47biomarker that we're interested in.
36:49So just to remind you again,
36:50what are extracellular vesicles?
36:52There are many different types says from a
36:56review for you from your Sadowski's lab.
36:58He also said ASCII is at Pitt and he runs
37:01the Magee Women's center and is really done.
37:04The most work in this field and understanding
37:07placental extracellular vesicles.
37:08There important signals for fetal development
37:10and maintaining homeostasis for mom.
37:12So those exercise the vesicles produced
37:14by the Placenta Act both locally on all
37:17different cell types of the Placenta
37:19and also can be found in circulation.
37:22Larger exerciser vesicles are termed
37:23Micro Vesicles and as I told you small
37:26exercise vesicles are termed exomes.
37:28Again,
37:28there are many different types that you
37:31can detect that contain different cargo,
37:33both intracellular cargo protein
37:34cargo weather going and a lot of
37:37work is being done.
37:38Now to develop these tools to
37:40try and understand how they are
37:42released from intracellular stores.
37:43So that's a really important
37:45and interesting point.
37:46That way in which these vesicles are
37:48loaded up and are secreted from the
37:51cell differ from the type of vesicle
37:53that they are, how they released,
37:55and how they travel in circulation.
37:57So they're super super interesting
37:59and really important.
38:00Cellular signaling mechanisms for
38:01these extracellular vesicles.
38:03As I've said,
38:04there are many components of the
38:06protein content related to coming
38:08from the placenta itself,
38:09although none that are truly
38:11exclusively from the placenta,
38:13so there's not really any way to
38:15say that they're absolutely from
38:17the trophoblast cells,
38:19but the cargo has also been well
38:21characterized of many of these
38:23vesicles and their contribution for
38:25maternal millu and fetal development.
38:28So I'm just going to show you touch
38:30on a little bit of work just to
38:32Pique your interest in why these
38:34things are so incredibly important.
38:36Is that thinking about exercising
38:38vesicles and?
38:39Understanding them as a biomarker
38:40you can both sequence the exercise
38:42of vesicles that you can isolate.
38:44This is showing you some of this
38:46small noncoding RNA.
38:47The thing I want to draw your
38:48attention to is that we've sequenced
38:50by small non coding RNA sequencing
38:52of the exercise in vesicles,
38:53in circulation in mice.
38:55You can see that if you look at
38:57this heat map that is showing you
38:59here on the far end is non pregnant
39:01females versus pregnant
39:02that have been control or stressed.
39:05Obviously the biggest difference in
39:07that the micro RNA content I'm not
39:09even showing you all of the data here,
39:12I just want to show you
39:14that pregnancy itself,
39:15just to highlight again the presence
39:17of placenta dramatically shift the
39:19content of circulation of extracellular
39:21vesicle in the micro RNA content and
39:24that there are some really important
39:26known micro RNA that can be very
39:28specific clues as to what effect
39:30they're having on distal tissues,
39:32especially intersecting
39:33with the immune system.
39:35And to that point in our proteomics studies,
39:38this is work done by Brigid Nugent
39:40when she was a postdoc in the lab.
39:43The protein content of these
39:45extracellular vesicles really important,
39:46also because it tells you both
39:48the tissues it's being released
39:50from and where it's acting,
39:51but what's really important here is
39:54thinking about the intersection of
39:56where we found that with stress,
39:57the protein content dramatically shifts in A.
40:00Ocean of immune related protein
40:02signaling molecules in these vesicles
40:04to a huge increase in metabolic anan,
40:06proteolysis and complement activation etc.
40:08Proteins in these vesicles so the take
40:11home message here is the cargo both
40:13in terms of its actions at the distal
40:16site for the small noncoding RNA but
40:18also the tissues it's traveling to
40:21seem to significantly change with
40:23stress and moms environment and
40:24just one last slide on these 'cause
40:27I want to move on to other topics.
40:29Here is that when we.
40:31Take out these exercise in vesicles
40:33from a mouse who's pregnant.
40:35From both stress and controls,
40:37we fluorescently labeled these vesicles
40:38and inject him back into a pregnant mom.
40:41To ask the question of is there
40:43transplacental signaling going from the
40:44maternal mil you into the fetal compartment.
40:46We have no evidence that it
40:48actually makes it through.
40:49In fact,
40:50if you take out the uterus you can see
40:53here by this way this is lighting up here.
40:56This is only in the Placenta And if
40:58you actually dissect out the Placenta
41:00and fetus you can see that in fact,
41:02no crossing of maternal produced exercise
41:04vesicles makes it into the fetal compartment.
41:07Why that's important?
41:08Because it tells us that the source
41:11of communication then from mom to
41:13the fetus is likely the vesicles
41:15acting again at the placenta,
41:17producing changes that then changed
41:19what is targeted and communicated
41:20to the developing fetus.
41:22But I want to return back to the
41:24weathering question and the adversity
41:26as detailed by the Aces questionnaires
41:28and thinking about the effect of
41:31racism and discrimination stress.
41:32We've recently developed a collaboration
41:34with Tonya Ivanovic as part of
41:36the Grady trauma.
41:37Project that Carrie wrestler first
41:39started at when he was at Emory and
41:42developing the Grady Trauma project
41:44that when he left for Harvard,
41:45Tonya took over and now Tonya has
41:48actually moved herself to wings state
41:50where she is now starting a similar project.
41:53I'm at Wayne State is the greatest trauma.
41:55I'm sure many of you are very
41:57familiar with the
41:58Grady Trauma Project.
42:00It's produced some of the best
42:02data looking at discrimination,
42:03stress and effects of the environment.
42:05Long-term health outcomes in
42:06neuro psychiatric disease,
42:07especially PTS di.
42:09So in this study, we had developed
42:11a mouse model of understanding the
42:14timing specificity of when trauma or
42:16stress happens in development itself.
42:18Does it produce those lasting
42:20effects and so just quickly,
42:22the current collaboration that we're
42:24working on took that mouse model
42:27and asked in a translation away if
42:29we look at a very specific type of
42:32trauma abbreviated here as IPV.
42:34This is actually interpersonal,
42:36not enter certain, not intimate partner.
42:38This is interpersonal a type
42:40of interpersonal trauma.
42:41Which is the greatest predictor of PTS
42:44di development and lasting effects?
42:47We wanted to know if we asked a
42:49specific type of interpersonal trauma
42:51that being sexual trauma because it's
42:54very easily identified when that happened.
42:57If that when that timing of that happened
43:00related to brain developmental timing.
43:03So under 14.
43:04So basically entering into the
43:07pubertal Timepoint and before or during
43:09adolescence or into adulthood or.
43:12After that adolescent window,
43:13so these three different time points here,
43:15could we look at physiological outcomes
43:17that were specific to that timing,
43:19whether it be adversity in PTS,
43:21D development,
43:21metabolic outcomes in other health
43:23issues related to diabetes,
43:24and hypertension?
43:25Could we then look at these
43:27exercise or vesicles or cell free
43:29mitochondrial DNA which will come
43:31back to as biomarkers related to
43:33these physiological outcomes and
43:35we can again take these exercise or
43:37vesicles from these human subjects
43:39with this specific timing and ask
43:41related to no trauma or no sexual trauma.
43:44In this particular case,
43:45and even doing proteomics by mass
43:47spectrometry and smaller in a sequencing,
43:50can we begin to identify what we
43:52would classify as a biomarker to
43:55then go in and ask about causality
43:57in our mouse models?
43:59So related to the timing sense
44:01of sensitivity,
44:01I'm not going to go into all of this data.
44:04We're working on with Tonya,
44:06but I want to show you that if you
44:08look at sort of the metabolic and
44:10physiological outcomes related
44:11to obesity risk,
44:12these are now women who were recruited
44:14in through the Grady Trauma Project,
44:16whose sexual trauma experiences were
44:18noted were then captured both for
44:20the readout of their body weight,
44:21which then Maps directly on with
44:23their hip circumference as well
44:24as their waist circumference,
44:26which I'm not showing you on here
44:28similar data as well as their
44:30hypertension risk.
44:31And what you'll see consistently
44:32is that there is effect of trauma
44:35of sexual trauma in this case.
44:37But if you actually parse it
44:39out for when it happened,
44:41you'll see again and again.
44:42In this case that women experienced
44:44it prior to adolescence and
44:46adulthood is driving this data,
44:48so both but there by increased body weight,
44:50increased hip and waist circumference,
44:52which I'm not showing you that these women,
44:55also with hypertension risk,
44:56which is known associations of these
44:58outcomes compared to no
45:00sexual trauma divided.
45:01Into elevated hypertension also
45:02there's got cutoff hypertension,
45:04stage one and two on here.
45:06What you see is,
45:07I think is fascinating is that if your
45:10sexual trauma experience and this is again,
45:13these women are all in there and mid
45:15to late 30s now that their earlier
45:18traumatic experience related to
45:20interpersonal violence or trauma.
45:22Under 14, they start to escalate or enter
45:24earlier into hypertension stage two,
45:26which is this?
45:27This this green aspect here versus
45:29if it happened during adolescence?
45:32Versus if it happened after the age of 17,
45:35I think he's very clearly shown here
45:37that this risk related likely again,
45:39are these women showing a phenotype
45:41unique or just earlier onset?
45:42I think is a really important question.
45:45OK, so now if you move that into the
45:47nuro psychiatric and neurophysiological
45:48outcomes at Tonya's group is worked
45:51on in the Greater China Project.
45:53If you start to look at startle related
45:55to PTS di outcomes, it's interesting.
45:57This is distinct from whether it's
45:59group here was 14 and under that this
46:02group between 14 and 17 is actually
46:04showing you unique differences.
46:05And baseline of fear potentiated startle
46:08as well as skin conductance outcomes as well.
46:11So the take home message here that
46:13we're starting to see in this
46:15collaboration with Tonya's group
46:16is that when trauma happens,
46:18can be a predictor or an Association
46:20for unique and distinct health risks,
46:22and I think that's interesting when you
46:25start to look at our biomarker data.
46:27So I'm just going to quickly show you
46:29some of this data just to just to be
46:32provocative here is that we isolated
46:34extracellular vesicles from these women.
46:36These are all black.
46:3795% of the cohort is black.
46:39They all have a high background
46:41of trauma in the environment.
46:43But this is again related specifically
46:45on a non sexual trauma versus
46:47sexual trauma experience.
46:49OK,
46:49so this is showing you a heat
46:51map of the proteomics data.
46:53All of the proteomics was isolated at
46:55the same time is all blinded when we
46:58do the proteomics assessment and I'm
47:00telling you that because for having
47:02worked in this field for a decade now,
47:04this is some of the most provocative
47:06data that I've seen in human subjects
47:09related to the proteomics outcomes.
47:10So you can see if we categorize
47:13all proteins here in H,
47:14showing you the non sexual
47:16versus sexual trauma.
47:17Overall,
47:17we're seeing thousands of differences in
47:19the protein content of these vesicles.
47:21If you parse it out by when
47:24the trauma happened,
47:25you'll see that there's this
47:26group of proteins down here in
47:28pink which are largely reduced,
47:30which will come back to in a second.
47:33And there's this really interesting
47:35group of proteins that are really
47:37increased only in this 14 to 17 group.
47:39Many of the proteins that we're
47:41seeing that are dramatically changed,
47:43I mean dramatically by either
47:45being only present,
47:46such as these protein cereal,
47:47come back two or hundred
47:49fold changed are related to
47:51skin proteins. Even if globulins and
47:53other things related to EV function,
47:55and if you actually look at the
47:56three sexual trauma groups,
47:58there was some overlap where
47:59they all show these differences.
48:01Some of these proteins in here
48:02and related I'll talk about,
48:04but there are very unique proteins that
48:06categorized by when the trauma happened.
48:08OK, which ones what I'm showing
48:10you here in KR again log 2 fold,
48:12change in these proteins and
48:13these are dramatic differences.
48:15These are not subtle.
48:17What stands out about these
48:19proteins and why I'm showing them
48:21to you across the sexual trauma,
48:23and these are these proteins
48:25down here in pink.
48:27There is a robust reduction in a number
48:29of key proteins and specific types of
48:32extracellular vesicles secretion broadly.
48:34So what these proteins in XNA one and
48:36two and collected three and seven.
48:39They point to specific.
48:41Types of exercise,
48:42their vesicles,
48:43but also how the vesicles are released.
48:45So that's super interesting and very
48:48important that we're finding in this cohort,
48:50but the piece I want to leave you
48:53with from this collaboration that
48:55we're currently working on is
48:57that we found we examined this.
49:00This blue group here of proteins,
49:02some really interesting outcomes we
49:04found about 24 carat and related proteins,
49:06so carrot and type one and two
49:09of cuticular origin.
49:10Meaning of the hair and skin.
49:13And keratin associated proteins again
49:15meaning related to hair and skin.
49:18Only in the sexual trauma experienced
49:20in adolescence in this group.
49:21Now remember,
49:22these women are now in their mid to late 30s,
49:25and this is.
49:26These are log 2 fold change meaning
49:28there are dramatically increased
49:30in expression and likely only
49:32found in this one unique group,
49:33and that's something we're
49:35currently following up on.
49:36But I just want to be
49:38provocative here and remind you.
49:40That the data we found,
49:42which is largely driven by
49:43this group in fear,
49:45potentiated startle and skin conductance,
49:46was was driven a lot by this
49:49specific group as well.
49:50Now this comes back to the
49:52provocative biomarker question.
49:54Some biomarkers are just
49:55biomarkers that can predict a risk.
49:58Some biomarkers are predicted
49:59actually of Amecon.
50:00Sticker causal outcome,
50:01and that's something we're following up on,
50:04and this is really gets to the
50:06point of the translation and
50:07reverse translational potential,
50:09because we can now model and are
50:11doing these in keratinocytes in
50:12culture where we can ask from eaves
50:15from human subjects applied in
50:17culture with keratinocyte because
50:18these turns out all of these proteins
50:21predict a dramatic change in something
50:23that happened at the level of the
50:25skin cell called the keratinocyte,
50:27and also fit with some of this data as well,
50:31so that suggests.
50:32That there may be something
50:33unique to the adolescent window
50:35related to the skin and the skin
50:38being your greatest tissue.
50:39The largest organ in the biggest
50:41barrier and involved highly and
50:43Autonomic Regulation is something
50:44very important to think about.
50:46For those biomarkers.
50:47OK,
50:48so I'm going to jump from
50:50that provocative statement to my last topic
50:52that I'm going to peak your interest in,
50:55which is which is jumping from females,
50:57whether it be in the studies
50:59were doing Tonya of women,
51:01specially in the black community at risk and.
51:04And during pregnancy.
51:06To thinking about dad,
51:08we oftentimes in neurodevelopmental
51:10disorders forget about dad's contribution
51:12and so it's really within the last five
51:15going on 10 years that this has become a
51:18very hot topic of discussion about dads.
51:20Germ cells and his adversity
51:23in experiences as he passes on.
51:25So what about the preconception,
51:26male stress effects,
51:28and how this may be passed on,
51:30so Rachel, Yehuda,
51:31and others have started doing
51:33these asking these questions,
51:35especially within the community of
51:37veterans as well as Holocaust survivors.
51:39Another traumatic life events.
51:41I think it's really important to
51:44think about the data that goes all
51:46the way back to aspects of the
51:48Swedish famine and Chinese famines.
51:50Data has been really well mind for dads.
51:53Effects related to the stress and
51:56traumatic intersection of a famine
51:58itself with incredible records that
52:00have been kept in this overkalix region
52:02of Sweden for looking at birth and
52:05death rates and growth and development,
52:07there's been some data mined
52:09here with schizophrenia risk and
52:11adds experiences and the timing.
52:13Of those experiences and the Dutch hunger,
52:15winter has also been really well mind
52:18again for autism and schizophrenia
52:19risk for some of that data.
52:22So how do we mechanistically or
52:24causally ask questions about a
52:25germ cell as neuro scientists?
52:27This was a topic that has piqued a lot
52:30of interest and a lot of skepticism.
52:32Shall I say it actually reached
52:34the level of the New York Times
52:37in December of 2018,
52:38where I had a New York Times Reporter
52:40who is very interested in this area?
52:43Contact me and say,
52:44not just that they wanted to talk
52:46about the data and the potential
52:49importance of the data,
52:50but you really wanted to talk about that.
52:53Controversy in this field,
52:54and so it is a It is a difficult aspect
52:58to get at mechanistically in humans,
53:00so we have to use a lot of our
53:02animal models to understand
53:04mechanisms about germ cells.
53:05And of course,
53:06it is very difficult to model many of
53:09these these experiences in a mouse.
53:11Mice are not humans.
53:13They cannot experience stress or
53:15trauma in the way that a human can,
53:17but we can begin to understand
53:19some of the mechanisms,
53:20and I'm just quickly touch on a couple.
53:23So in our mouse model that we've
53:25been working on for the better
53:27part of the last decade,
53:29we can expose male mice
53:30across many different windows.
53:32It turns out it doesn't matter the
53:34window of development because the germ
53:36cells that are affected with about
53:38four weeks it requires of chronic stress,
53:40again getting dad's stress
53:41levels elevated enough,
53:42requires about four weeks of stress.
53:45This is a model that his
53:46offspring show a very
53:47hyper responsive HPA strikes
53:49acid accesses their phenotype.
53:50There is no sex difference here.
53:52Males and females show
53:53the exact same phenotype.
53:54We've been able to mechanistically
53:56replicate this data.
53:57I don't have time to talk about
53:59by micro injecting specific
54:00micro RNA at the Zagat Level.
54:02I'll kind of come back to
54:04that a little bit later.
54:05Later on, I want to jump to in this
54:08model sort of showing you the phenotype
54:10and why this phenotype is important.
54:13So mice, like humans,
54:14like all mammals,
54:15one of the reasons I like studying the
54:17stress axis is that it's translate rible.
54:20The jeans and involvement of the
54:21hypothalamus in the human brain
54:23are the exact same pathways.
54:25Circuitry in genes involved
54:26in the mouse brain.
54:27So it's the most translate Rible,
54:29and it's easy in a human error
54:31mouse to stimulate the stress axis.
54:33So this just shows you from our our paternal,
54:36so we stressed the male mice.
54:38We then gave them time off from stress,
54:41and that's important.
54:42We then bred them.
54:43This is the stress response of
54:45their male and female offspring,
54:47and this is just showing you
54:49the hypo responsive iti here.
54:50This is a normal in black when a
54:53mouse or a human will look like
54:55when you acutely stress them.
54:57Their glucocorticoids levels
54:58rise and fall in both males and
55:01females and if their dad had been
55:03previously stressed normally or if
55:05we really cranked up his stress
55:07during that same time window and
55:09then bread him doesn't matter.
55:10His male and female offspring show
55:12a hypo responsive stress axis.
55:14Why is that important?
55:15Because we know that in PTS di,
55:18oftentimes,
55:18in major depressive disorder that
55:20those individuals show a blunted stress axis.
55:22Blunted is not necessarily better,
55:23is just different,
55:24and I want to quickly highlight that
55:27this work from Jennifer Shannon,
55:28graduate student, which is all published.
55:30Now,
55:31that what's important here is that
55:32this is not an acute response,
55:34and in fact,
55:35if you breed the animals during
55:37the stress experience that they
55:39are exposed to or right at the
55:41end of their stress experience,
55:43you don't get this phenotype.
55:45In fact, if you breed, yeah,
55:46if you breed dad during or
55:48after immediately after.
55:49So one week after the stress is ended.
55:53Yet no phenotype, so it requires about
55:55four weeks to three months and it sorry it
55:58requires about four weeks of integration
56:00and lasts out to at least three months.
56:03As far as we've gone.
56:04So what that means is if we breed dad
56:07immediately, he doesn't pass on the effect.
56:09If we give him time for that post
56:12stress allostatic setpoint to happen,
56:13he passes on the effect and
56:15it's long lasting.
56:16It does not reverse,
56:18and that's really important,
56:19so we just quickly look at at at what we're
56:22currently thinking here as terms of how dad.
56:25Right, that's the mechanism
56:26we're looking for in the mice,
56:28so we can ask the question in humans
56:30if we compare at the end of stress
56:32here at this time point that the
56:34effect is not passed on to offspring.
56:36We see very few changes in the small
56:38non coding RNA content of dads sperm.
56:40If we wait three months and breed him.
56:44Where we see the effect passed on here
56:46that is required in three months of time.
56:49We in fact now see huge
56:50differences in these dads.
56:52So the control versus those that
56:54experience the stress but have
56:55now had three months to recover,
56:57which in the life of a
56:59mouse is a very long time.
57:00The take home message here is OK,
57:03so there's a signal.
57:04There's something in the sperm mice
57:06and I'll show you human data later.
57:08That is important for possibly transmitting
57:10the signal to the developing embryo.
57:12So I'm sure this is a little bit cloudy,
57:15but I just want to quickly say for
57:17those who think about the brain
57:19all the time and don't think about
57:21testes as much as my lab does that.
57:24Why is this important?
57:25The timing is important and the
57:26transcriptional inert activation
57:27of sperm is important,
57:29so sperm go through a course
57:30of spermatic Genesis.
57:31Everybody understands that that's
57:33about six weeks in most mammals.
57:34At the end of Schematic,
57:36Genesis sperm are not mature,
57:37they cannot swim, and they cannot fertilize.
57:39They are pushed by a fluid motion through.
57:42These DuckTales into the head
57:44of the epididymis.
57:45Epididymis is required portion of
57:47the reproductive track of dad.
57:49These tubules secrete all kinds of
57:52important factors to mature the
57:54sperm such that they can fertilize.
57:56They can swim,
57:57and they are fully mature into
57:59the lumen of many weeks that the
58:02sperm in all mammals go through.
58:04In this the capit region of the epididymis.
58:08There is another factor that is secreted,
58:10which is the extracellular vesicle.
58:12So in this tiny little piece of
58:14Anatomy of males is exercise are
58:16vesicles that are circulating
58:18interacting with the sperm.
58:19These extracellular vesicles we
58:21know are important for delivering
58:22the signals to the maturing sperm,
58:24so it's not transcription happening
58:25in this firm.
58:26It is a signal from cymatics cells
58:28and this is just quickly to show you.
58:31Why would this be?
58:32Because I know that as a neuro
58:34scientist I'm sure within psychiatry
58:36we often look at
58:37animal studies and say OK but why?
58:39I know you have an effect. Who cares?
58:41What does it matter and how does
58:44it happen and why would it happen?
58:46What I'm showing you here is the course
58:49of embryonic development in birth,
58:51and the germ cells that are
58:53important to what they go through.
58:55The take home message here is
58:57why would this mechanism happen?
58:59Well, because evolutionarily,
59:00you don't want Dad's germ cells.
59:02You don't want those primordial germ
59:04cells or those developing some additives.
59:07Where dad's DNA is vulnerable to
59:09be impacted by the environment.
59:11This is occurring in the testes,
59:13which is a privileged blood brain barrier.
59:16Sorry blood testis barrier,
59:17we can make some jokes about
59:19blood brain barrier here,
59:21but in the blood testis barrier
59:23that the environment itself it is
59:25privileged and these cells that are
59:27really required for preservation
59:29of the species evolutionarily are
59:31protected from dads environment right?
59:33This is protection happens through a
59:35course of proteins known as Pi RNA's.
59:37That are really highly expressed
59:39during these periods of time,
59:41but by the time the sperm leave the
59:43testes and enter into the Capital Region,
59:45there is still a slight protective barrier,
59:47but nothing like the testes.
59:49And the reason evolutionarily again,
59:51this is a hypothesis,
59:52is likely for those exercise vesicles
59:54to be interacting with the sperm.
59:56Here is,
59:57this is a way for dads germ cells
60:00to be impacted in a way that does
01:00:03not affect dad's DNA.
01:00:04OK, so sperm are transcriptionally inert,
01:00:06they are not responding to an
01:00:09active environment.
01:00:09Therefore they need to interact
01:00:11with in the lumen of the epididymis.
01:00:14These exercise are vesicles if
01:00:15they want to carry a message about
01:00:18dads environment and that is just
01:00:20simply modeled here,
01:00:21here is the lumen of the epididymis.
01:00:24Here are the sperm that are
01:00:26undergoing this maturation process.
01:00:27They are interacting with these somatic
01:00:30cell derived vesicles that contain
01:00:32all kinds of proteins and micro RNA
01:00:34that are transcriptionally actively
01:00:35changed by dads environment in a lasting way.
01:00:39So quickly,
01:00:40how do we look at this in terms
01:00:43of a mechanism?
01:00:44Because we cannot ask about a pure population
01:00:47of vesicles to test our hypothesis in
01:00:50vivo because as I told you before,
01:00:52every tissue produces vesicles.
01:00:54So there's many different cell
01:00:56types in this lumen besides just
01:00:58these epididymal epithelial cells,
01:01:00but also, as you can imagine,
01:01:02the capit region of the epididymis
01:01:04and a mouse you cannot easily or
01:01:07really at all isolate a unique.
01:01:10Extracellular environment here.
01:01:11So this brilliant graduate student
01:01:13was able to model this in a dish
01:01:15where she was able to model these
01:01:17DC 2 mouse epididymal epithelial
01:01:19cells that secrete extracellular
01:01:20vesicles into the environment.
01:01:21She was able to develop a model
01:01:24which I'm not going to go into
01:01:26because it's way more down the
01:01:28weeds than you need to be.
01:01:30She was able to model exactly what
01:01:33we see in vivo on dad's sperm
01:01:35environment with what she sees in
01:01:38vitro with the micro RNA extracellular
01:01:41vesicle content from stress.
01:01:43This very complicated figure is a way
01:01:45of telling you she pharmacologically
01:01:47worked out the stress conditions with the
01:01:50glucocorticoid treatment and recovery.
01:01:52This is showing you 11 days
01:01:55after stress recovery.
01:01:56The concentration of stress,
01:01:58relevant glucocorticoids
01:01:59isolating those vesicles.
01:02:00And looking at them by proteomics,
01:02:03she was able to model this
01:02:05beautiful heat map here,
01:02:06where she looked at the vesicles
01:02:08in culture versus previously
01:02:10vehicle treated versus court
01:02:11treated but now recovered right.
01:02:13This is 11 days and three
01:02:15media changes later.
01:02:17The same vesicle protein changes
01:02:18we see in vivo and in vitro,
01:02:21and this is just showing you is
01:02:24orthogonal report here that they are
01:02:26very different and even the size of
01:02:28these vesicles is changed long term.
01:02:31By that court treatment,
01:02:32same thing we see in vivo.
01:02:34This just shows you that if we
01:02:35label those vesicles and injected
01:02:37into mouse they again have a
01:02:38specificity of where they go.
01:02:40This is showing you your meat
01:02:41market sort of approach here.
01:02:43If you go and look at where all your
01:02:45meat comes from and in your cow or pig
01:02:47or whatever this is showing you the mouse.
01:02:50If we take out all of these tissues
01:02:52lay them out and look at the DIII
01:02:54where it's been transferred.
01:02:55You can see everything shows up in the liver.
01:02:58Ignore that but it's going
01:02:59to be immune system.
01:03:00The spleen here and really amazingly.
01:03:02Dad's reproductive track.
01:03:03The testes and the capital,
01:03:05the epididymis.
01:03:05Amazing, profound specificity.
01:03:06But for everybody cares about the brain.
01:03:09It turns out it also gets into
01:03:10dads brain that's a topic for
01:03:12another conversation of how these
01:03:14capit epididymal pacilio derived
01:03:16vesicles get into an effect.
01:03:17Adds brain, but we don't have time for that.
01:03:20What I do want to end with is
01:03:23the take home message of why
01:03:25these ev's are so important.
01:03:27'cause we're overtime if we actually
01:03:29take these eaves I was just telling
01:03:31you about and perform an experiment
01:03:33where we take out dads sperm.
01:03:35From a control mouse divided into 2 pools.
01:03:38Perform fertilization technique.
01:03:39Also used in humans where we can
01:03:42incubate the control vesicles with
01:03:44half of dad sperm and the formerly
01:03:46stress treated vesicles without sperm.
01:03:48Perform ixi where we're actually
01:03:51doing intracytoplasmic sperm injection
01:03:53so we can say in this pool versus
01:03:56this pool and transfer them into
01:03:58the left and right side of the same
01:04:00mom with eggs from a different mom.
01:04:03So all factors being controlled for.
01:04:06We can look at the outcome both
01:04:08during development at mid gestation
01:04:10for the developing brain and then
01:04:12the long term outcome and this is
01:04:15that capall amazing endpoint year
01:04:17to show you that if we look at the
01:04:19transcriptional mid gestational
01:04:21development of the brain we see
01:04:23huge differences only because and
01:04:25in partner to the vesicles that
01:04:27those the sperm are incubated with.
01:04:29So the developing brain around
01:04:31synaptic signaling etc likely changing
01:04:34again the rate of the
01:04:35brain but ultimately here.
01:04:36Those offspring showing you the same
01:04:39phenotype that stress signal coming from
01:04:41dads epidemie up at the other cells
01:04:43shaped the developing brain of his offspring.
01:04:45They show the same hypo responsive HPA axis.
01:04:48I don't have time because we're way over to
01:04:51tell you all about the human side of this,
01:04:54but in a collaboration longstanding
01:04:56collaboration that we have
01:04:57with Neil Epperson,
01:04:59who is now chair of psychiatry.
01:05:02At Denver, we're doing the
01:05:03human side of this question,
01:05:05and so we've just published a
01:05:07study just came out actually
01:05:08yesterday in scientific reports,
01:05:10so you can look at all this amazing
01:05:13data where we are able to look at the
01:05:15micro RNA and looking at the small
01:05:18noncoding RNA content with perceived
01:05:20stress in human subjects and found
01:05:22amazing data here for the micro RNA content.
01:05:25the T RNA fragment content and
01:05:27the Pi RNA content.
01:05:28And I'm just going to point
01:05:30out to Q Your interest.
01:05:32If you look within subject here,
01:05:34so each of these is a different
01:05:36Mail over time of collection,
01:05:38so they came in each month.
01:05:40You start to see patterns in both the
01:05:42Pi RNA's and you can also see the same
01:05:45pattern in the micro RNA content.
01:05:47So cyclicity of pattern has never
01:05:49been shown before in male germ
01:05:50cells so I think super interesting
01:05:52and this just highlights it.
01:05:54Again the data is all in the paper.
01:05:56I'm not going to go through it
01:05:58that we were with really rigorous
01:06:00bioinformatic modeling,
01:06:01able to identify those specific
01:06:02within and between subjects.
01:06:04So we had 20 subjects with
01:06:06six donations a month apart.
01:06:07We could identify actually patterns
01:06:09of specific RNAs T RNA's in my
01:06:11garnas that fit with their perceived
01:06:13stress outcomes,
01:06:14and this is where you taking this back in,
01:06:17then to the mouse where we can
01:06:19now ask if we take those specific
01:06:21targets and ask what do they do to
01:06:24the developing brain is a really
01:06:26reverse translation approach.
01:06:27So just to summarize again,
01:06:29there are many different ways for
01:06:30these exercise or vesicles to be
01:06:32really important for different populations,
01:06:34whether it be related to maternal
01:06:36health within the black community
01:06:38and discrimination stress.
01:06:39Within moms and dads and how the
01:06:41environment influences their signaling.
01:06:43And I'm not going to go through
01:06:45all the conclusions because I
01:06:47already highlighted them all,
01:06:49but I think as as an example
01:06:51of how biomarkers this case,
01:06:53extracellular vesicles can
01:06:54both serve as a biomarker,
01:06:56but also likely reverse reverse translate
01:06:58Aghbal causal mechanism to be explored,
01:07:01I think is very important.
01:07:03I do need to take the time to thank
01:07:05all the amazing people in the lab that
01:07:08have done all of this incredible work.
01:07:10Bridget Nugent worked on all of the
01:07:12placental studies and started looking
01:07:13at those exercise or vesicles early on.
01:07:15Chris Morgan has been doing the mouse
01:07:17work as well as the human studies
01:07:19that we're currently working on with
01:07:21Neal Epperson's group.
01:07:22Katie Morrison who is now an assistant
01:07:23professor at the West Virginia University,
01:07:25West Virginia, started the studies in
01:07:27collaboration with Tonya Jovanovitch
01:07:29is group who is now at Wayne State.
01:07:30We're right in the midst of of
01:07:32submitting a grant right now taking those
01:07:34extracellular vesicles and doing some.
01:07:36Happy Genomic Studies and again,
01:07:38of course Neil Epperson,
01:07:39who I haven't changed the slide.
01:07:41It looks like who's on here as pen,
01:07:43but now is moved.
01:07:44His chair of psychiatry again or please
01:07:46follow us as we really do do a lot
01:07:48of community engagement around these
01:07:50topics in Baltimore and around the world,
01:07:53especially as I'm now president of Ebro.
01:07:56And all of our funding agencies,
01:07:58including any MH on ahs and NICHD,
01:08:00and I'm happy to take questions first time.
01:08:03And I apologize for for going off on too
01:08:05many tangents and take questions now.
01:08:08Thank you.
01:08:15Hi. I'm Rafael Perez hanging.
01:08:21It was an excellent talk.
01:08:23I have a question about.
01:08:25The sequencing that you had
01:08:27done when patients overnight
01:08:29patients against your subjects,
01:08:31so one of the things that we see in
01:08:33the Miz win stresses this segregation
01:08:36of resiliency and susceptibility,
01:08:39and I do see somebody ability
01:08:41to data that you showed today.
01:08:44Have you guys began stratifying the
01:08:46data looking at kind like Arnold,
01:08:49different outcomes in terms of startle
01:08:51or like psychiatric condition?
01:08:53Yeah, so several, great question.
01:08:55It is across a lot of our
01:08:58human subject studies,
01:08:59both the collaboration with
01:09:00Neil as well as with Tanya,
01:09:02that we started looking at looking
01:09:05at the different responses,
01:09:06both satisfying it as well as looking
01:09:08at trying to find associations within
01:09:11our data set so I didn't have time to
01:09:14talk about the cell free mitochondrial DNA,
01:09:17which also is an incredibly
01:09:18interesting biomarker,
01:09:19especially as has been recently shown by
01:09:21looking at many outcomes related to suicide.
01:09:24Alice, suicidality risk,
01:09:25and major depressive disorder.
01:09:27And responsiveness to
01:09:28antidepressant treatment.
01:09:29Also, something really is easy to look at
01:09:31if you're interested in new biomarkers,
01:09:34so trying to look at cell
01:09:36free mitochondrial DNA,
01:09:37which you can get from urine,
01:09:40plasma, saliva, hair.
01:09:41There's lots of ways to do this.
01:09:43Martin Picard is doing tons of work on this,
01:09:47so it's something really.
01:09:48And Raffield your question about both
01:09:51looking at the risk and resilience piece,
01:09:53but also the associations within the data.
01:09:55So those that have high levels of
01:09:57changes in protein in Eves or high
01:10:00levels of cell free mitochondrial
01:10:01DNA does that also associate with
01:10:03any of our physiological outcomes
01:10:05in those different groups.
01:10:06So yes, we are.
01:10:08We have begun looking at those outcomes
01:10:10related to risk and resilience.
01:10:12Doctor Bell,
01:10:13there's a question in chat.
01:10:16Are the stress related changes
01:10:18in sperm alleviated with
01:10:20pharmacological manipulations?
01:10:21SSR eyes, cortisol suppression, etc.
01:10:23Great great questions,
01:10:25so we're currently both looking at
01:10:28this question in the rodent model
01:10:30and inhuman studies with Neil,
01:10:33so I'll grant that we just submitted
01:10:36last cycle with Neil is doing another
01:10:39recruitment of human subjects
01:10:41to test our bioinformatic model.
01:10:44And so, in one of that Grant is
01:10:46dude really asked the question.
01:10:48We've developed this really complex
01:10:49model if we tested on another cohort,
01:10:52can we replicate it?
01:10:53But aim two is actually looking if
01:10:55we start to add factors to our model.
01:10:57So one of the things we're adding is aces,
01:11:00but we'd like to also go into
01:11:02different cohorts of individuals.
01:11:04So for instance,
01:11:05working with the VA and recruiting
01:11:07individuals with and without PTS di,
01:11:08or with and without PTS di treatment.
01:11:10So that answers the question of the human
01:11:13subject studies and looking at associations.
01:11:15We are just now because we
01:11:17have a culture model as well.
01:11:19So culture model as well as
01:11:21mouse model on this starting
01:11:23to ask about reverse ability.
01:11:25I want to be careful with
01:11:27that because I don't
01:11:28want to suggest that the changes we're
01:11:31seeing are necessarily diseased.
01:11:32Predicting necessarily there just changes,
01:11:34right? So we see differences in the
01:11:36offspring and how they respond to stress.
01:11:39That may be a predictor of other
01:11:41nerve psychiatric disease risk,
01:11:43but it could be, as Rafael pointed out,
01:11:46a hallmark of resilience, right?
01:11:47We don't know, because there are mice.
01:11:50What we do know is that we can model
01:11:52that stress in the environment
01:11:55and resolution of that stress.
01:11:57Returning the cells in their reproductive
01:11:59track to a new allostatic setpoint alters
01:12:02the content of the sperm that I can say.
01:12:06But these questions of can we reverse
01:12:08them with drug treatment, etc.
01:12:11Something we're currently exploring we've.
01:12:13Tossed around the idea in the mice of if
01:12:16stress and resolution produces these changes.
01:12:19What kind of effector would
01:12:20reverse them at that.
01:12:22We were looking at already at the
01:12:24epigenetic the histone modifications
01:12:25that produce these changes long lasting
01:12:27So what would reverse that would
01:12:29it be the perception of something
01:12:31rewarding environment for example,
01:12:33is something we're looking at.
01:12:35It's a great question.
01:12:37OK, there's another question.
01:12:39There's evidence that in response
01:12:41to a maternal infection,
01:12:43placental inflammatory cytokines such
01:12:45as one L-1B can be detrimental to fetal
01:12:49brain development in your stress model.
01:12:51Do you see evidence of
01:12:53placental inflammation?
01:12:54An if so,
01:12:56are these responses also sex specific
01:12:59correlating with placental oh GT?
01:13:02'cause am I laughing because
01:13:03it's such a great question.
01:13:05It makes me wonder if someone like
01:13:06actually already know our data to ask that.
01:13:08But yes,
01:13:09great question and we've done all of
01:13:12those in the workers all published.
01:13:14Uh Stephanie Bronson is first author in
01:13:16a couple of those papers that we looked at.
01:13:20We've done both looking at the
01:13:22inflammasome and inflammatory responses
01:13:24in the placenta with her stress model.
01:13:26So the answer is yes,
01:13:28we do see increases and we do
01:13:30see it sex specifically.
01:13:32So while I showed you data on Strat How
01:13:36come we see this time and time again?
01:13:39Whether it's giving mom an inflammatory
01:13:42response, giving mom a stressor,
01:13:44giving mom a dietary challenge,
01:13:46that again,
01:13:47the male placenta shows thousands of genes
01:13:50that respond to that that challenge RE.
01:13:53Little, very selectively that happens in
01:13:55the female placenta in this same uterus,
01:13:57so it fits the hypothesis that oh
01:14:00GT is titrating that for females.
01:14:02So again yes we see a lot of
01:14:04those same outcomes.
01:14:06Anna manuscript that just came
01:14:08out in Placenta from Yasmine.
01:14:09See say here she's doing again.
01:14:11This is actually a picture
01:14:14right here of Yasmine.
01:14:15She's doing work in both the mouse
01:14:18and the human studies as well.
01:14:20In the black community,
01:14:21where we see very similar outcomes
01:14:23and that work just published in
01:14:25Placenta shows really dramatic
01:14:27changes in the Mail placenta as it
01:14:29relates to inflammatory responses
01:14:30and really very subtle differences
01:14:32in the female placental.
01:14:38Tracy, I have a question Hygiene
01:14:41High that was just amazing talk on
01:14:44all fronts and I have a I guess sort
01:14:49of basic biology question and that
01:14:52is in terms of your placental oh GT
01:14:55data in this sex specific effects.
01:14:59When you did you manipulation in terms of
01:15:03the header zygosity of the effects, why?
01:15:06I guess I was thinking.
01:15:09Why wouldn't X? Inactivation.
01:15:15Mitigate some of the sex specific
01:15:17differences or do not have inactivation
01:15:20at that part of development.
01:15:23Yep, so this is a great question
01:15:25and something that we're looking at
01:15:27from a very genetic and molecular
01:15:29point we've looked at.
01:15:31Oh GT, we have an OG T conditional
01:15:34mouse that allows us to look at it,
01:15:37and in lots of places in including the brain,
01:15:40the Placenta appears to be the only
01:15:43tissue that we've found where,
01:15:45oh GT escapes X inactivation.
01:15:47I don't have a justification or
01:15:50even a hypothesis as to why.
01:15:52So related to the point of why
01:15:54we see sex specific effects,
01:15:57or we know that oh GT and oblique
01:16:00inoculation intersects with exist.
01:16:02I don't know why oh GT escapes
01:16:05Axon activation of the placenta,
01:16:07but it clearly does in the
01:16:10trophoblast cells only.
01:16:12But oh GT and oblique inoculation
01:16:14as I showed you with the changes
01:16:18in H3K27 Trimethylation is a
01:16:20required event for X inactivation.
01:16:23So there is a lot of sex differences that
01:16:26end up resulting from the requirement
01:16:29of that increased repressive Mark.
01:16:32H3K27 Trimethylation necessity
01:16:33as part of X inactivation.
01:16:35So it's sort of a circular involvement.
01:16:38So Jane,
01:16:39I think that's a provocative question.
01:16:41Is there something evolutionarily
01:16:43as to why a female trophoblast cell
01:16:46escapes X inactivation for this gene?
01:16:48So if you look at our,
01:16:51there's a PNS paper from 2013, I think.
01:16:55From Chris Howerton.
01:16:57That that we did his initial
01:17:00screening looking for candidate genes.
01:17:01There was about seven or eight
01:17:04of them that came up,
01:17:06and they were all X or Y linked,
01:17:09so the placenta seems to
01:17:11be this interesting tissue.
01:17:12Evolutionarily,
01:17:12I don't know why that has very sex
01:17:15specific chromatin regulation that we
01:17:17have identified some of the players
01:17:20with no GT beans clearly important,
01:17:22but it's a great question.
01:17:24I wonder if the four core
01:17:26genotyped mice Mount.
01:17:27Might be a model where you could
01:17:30look at that right so S or Y for
01:17:32the four cores that James talking
01:17:34about this or why gene is what
01:17:37drives those four core that Jane
01:17:39your group is published.
01:17:40Some really intriguing brain data out of.
01:17:42For that those studies.
01:17:45The four core are based off of the
01:17:47SRY that's been placed AutoZone
01:17:49Moe so they can segregate testes
01:17:51development and testosterone production
01:17:52as part of those four cores.
01:17:54Yeah, it's an interesting question.
01:17:56If you if you made if you took oh
01:17:59GT out of its locus and made it
01:18:01over expressed in a way that it
01:18:04wasn't X inactivated or it's X
01:18:06levels increased his expression
01:18:07in the Mail trophoblast cell for
01:18:09instance, would you see the opposite
01:18:11sorts of effects and protective
01:18:13mechanism in males? Great question.
01:18:17There is another question.
01:18:19Will interventions at any specific age group,
01:18:22for example 13 to 18 year olds,
01:18:25reduce reverse stress related changes
01:18:27that could be passed on to the offspring?
01:18:31It's a great question.
01:18:33I assume that's in reference to
01:18:35the collaboration with Tonya,
01:18:36but there's lots of studies,
01:18:38certainly not just ours,
01:18:39like that field is pretty wide.
01:18:42That adolescence is a unique time
01:18:44window for specially for women,
01:18:45but not just women but a unique time
01:18:48window for vulnerability for the brain,
01:18:50for adversity in the environment.
01:18:52I I'm super excited about this data
01:18:54with Tonya, and this is really,
01:18:57really brand new data that Tony
01:18:59and I've been talking about.
01:19:01What does it mean?
01:19:02Why is this skin as part of your as part
01:19:06of your wide cast sort of stress network?
01:19:09This skin has really ultimately been ignored.
01:19:11How is that uniquely
01:19:12vulnerable during that window?
01:19:14I mean, it's there.
01:19:15Great questions,
01:19:16haven't really figured it out.
01:19:17I'm not sure that keratinocyte
01:19:19maturation or even the stem cells
01:19:21there have been that well documented
01:19:23changes in your skin over adolescence.
01:19:25I think we can all account for,
01:19:27but what it means for the stem cells?
01:19:30I don't know.
01:19:38That is it in chat. Unless
01:19:42anybody else has a question.
01:19:48Alright, thank. Q Doctor bail.
01:19:51Thank you so much, Tracy.
01:19:53Thank you. Nice to see everybody.