N E WS
FEATURES
A PAINFUL LEGACY
Mice hint at how people’s emotional trauma may
affect the biology of their children—and their
children’s children By Andrew Curry, in Zurich, Switzerland
T
he children living in SOS Chil-
dren’s Villages orphanages in Paki-
stan have had a rough start in life.
Many have lost their fathers, which
in conservative Pakistani society
often effectively means losing their
mothers, too: Destitute widows of-
ten struggle to find enough work to
support their families and may have
to give up their children.
The orphanages, in Multan, Lahore, and
Islamabad, provide shelter and health care
and send kids to local schools, trying to
provide “the best possible support,” says
University of Zurich (UZH) physician and
neuroscientist Ali Jawaid. “But despite that,
these children experience symptoms similar
to PTSD [post-traumatic stress disorder],” in-
cluding anxiety and depression.
Beyond these psychological burdens,
Jawaid wonders about a potential hidden
consequence of the children’s experience.
He has set up a study with the orphanages
to probe the disturbing possibility that the
emotional trauma of separation from their
parents also triggers subtle biological altera-
tions—changes so lasting that the children
might even pass them to their own offspring.
That idea would have been laughed at 20
years ago. But today the hypothesis that ex-
perience might alter the cells and behavior of
212 19 JULY 2019 • VOL 365 ISSUE 6450
an individual’s children and grandchildren
has become widely accepted. In animals, ex-
posure to stress, cold, or high-fat diets has
been shown to trigger metabolic changes
in later generations. And small studies in
humans exposed to traumatic conditions—
among them the children of Holocaust sur-
vivors—suggest subtle biological and health
changes in their children.
The implications are profound. If our
experiences can have consequences that re-
verberate to our children or our children’s
children, that’s a powerful argument against
everything from smoking to immigration
policies that split families. “This is really
scary stuff. If what your grandmother and
grandfather were exposed to is going to
change your disease risk, the things we’re do-
ing today that we thought were erased are af-
fecting our great-great-grandchildren,” says
Michael Skinner, a biologist at Washington
State University in Pullman.
Skinner’s own research in animals sug-
gests changes to the epigenome, a swirl of
biological factors that affect how genes are
expressed, can be passed down through mul-
tiple generations (Science, 24 January 2014,
p. 361). If trauma can trigger such epigen-
etic changes in people, the alterations could
serve as biomarkers to identify individuals at
greater risk for mental illness or other health
Isabelle Mansuy (shown in her lab at the University of Zurich) found that mice with traumatized
forebears took more risks when exploring open, elevated platforms (left).
problems—and as targets for interventions there in the public mind about epigenetics variations in traits such as kernel color last year at historical records showed the
that might reverse them. probably can never be proved.” that persisted for hundreds of generations. sons of Civil War soldiers who had spent
But proving that emotional trauma, as To investigate, Jawaid is collecting blood The work was initially controversial, as time as prisoners of war (POWs) were more
distinct from physical stress, can be passed and saliva samples from the Pakistani or- geneticists saw it as a revival of the non- likely to die early than the sons of their fel-
on to subsequent generations in people is phans and from classmates who live with Darwinian ideas of 19th century scientist low veterans. (The researchers controlled for
a challenge. “The difficulty … is being able parents. As a researcher in the lab of Isabelle Jean-Baptiste Lamarck. socioeconomic status and maternal health.)
to disentangle what comes through social Mansuy of UZH and ETH Zurich, he hopes to But experiments in many organisms But the human studies faced an obvi-
inheritance—which must be massive—and learn whether the trauma of loss and forced suggested epigenetic inheritance was real. ous objection: The trauma could have been
what doesn’t,” says neuroscientist Johannes separation has left identifiable marks at the In simple creatures like Caenorhabditis el- transmitted through parenting rather than
Bohacek of ETH Zurich university. “The jury cellular level. But to really prove transgen- egans worms, researchers found that genes epigenetics. Something about the POW ex-
is still out on humans.” erational inheritance, he’d have to study the turned off once by altering RNA remained perience, for example, might have made
Some of the field’s biggest names also orphans for years—until they have children silenced for 80 generations or more. Some those veterans poor fathers, to the detri-
worry that the idea could have dangerous of their own. That’s why Mansuy herself has examples were even more dramatic: Water ment of their sons’ lives. The psychological
consequences. Rachel Yehuda of the Icahn turned to mice. fleas exposed to the scent of a predator have impact of growing up with a parent who
School of Medicine at Mount Sinai in New offspring with spiky, armored heads. And in starved as a child or survived the Holocaust
York City studied the children of 40 Holo- ONE RECENT AFTERNOON, Mansuy donned a mice, researchers including Skinner found could itself be enough to shape a child’s be-
caust survivors and found lower baseline fresh lab coat and blue sanitary booties and that parents exposed to altered diets, low havior. Answering that objection is where
levels of the stress hormone cortisol as well gently cracked the door of a darkened room temperatures, or toxins had descendants mouse models come in.
as a distinctive pattern of DNA methyla- at her lab at UZH. A powerful smell—some- with behavioral changes and weight gain. Mansuy began in 2001 by designing a
tion, an epigenetic marker. But in a paper thing like dog chow mixed with animal Epidemiological studies of people have mouse intervention that re-creates some
last year, she said it would be “premature” musk—wafted out on a gust of warm air. revealed similar patterns. One of the best- aspects of childhood trauma. She separates
to conclude that trauma causes heritable Inside were hundreds of mice in 40 breed- known cases is the Dutch hunger winter, a mouse mothers from their pups at unpre-
changes, adding that hyped media cover- ing cages. “We keep it dark during the day famine that gripped the Netherlands in the dictable intervals and further disrupts par-
age could promote a misleading narrative to preserve their circadian rhythm when we closing months of World War II. The chil- enting by confining the mothers in tubes
of hopelessness, spreading the idea that work with them,” Mansuy says in a hushed dren of women pregnant during the food or dropping them in water, both stressful
PHOTO: PIOTR PIWOWARSKI
PHOTO: PIOTR PIWOWARSKI
one generation’s trauma permanently scars voice. “This is our 31st cohort.” shortages died earlier than peers born just experiences for mice. When the mothers
later generations. The idea Mansuy is exploring--that not before, and had higher rates of obesity, dia- return to the cage and their pups, they’re
“There’s a lot of overinterpretation of all inherited characteristics are rooted in betes, and schizophrenia. Studies of other frantic and distracted. They often ignore
initial results,” says Columbia University bi- DNA--dates back more than half a century. groups suggested the children of parents the pups, compounding the stress of the
ologist Katherine Crocker, who studies non- Tantalizing early results came from maize, who had starved early in life—even in the separation on their offspring.
genetic inheritance in crickets. “What is out in which plants with identical DNA had womb—had more heart disease. And a look Mansuy says the mice’s suffering has a
sciencemag.org SCIENCE SCIENCE sciencemag.org 19 JULY 2019 • VOL 365 ISSUE 6450 213

NE W S | F E AT U R E S
purpose. “We’re applying a paradigm that
is inspired by human conditions,” she says.
“We’re doing it to gain understanding for
better child health.”
Unsurprisingly, the pups of stressed
mothers displayed altered behavior as
adults. But to Mansuy’s surprise, the behav-
ioral changes persisted in the offspring’s
offspring. Initially, she thought this could
be a result of the offspring’s own behavior:
Mice traumatized as pups could have been
bad parents, replicating the neglect they
experienced in childhood. Thus they might
simply be passing on a behavioral legacy—
the same lasting psychological
effect that might explain such
findings in humans.
To rule out that possibil-
ity, Mansuy studied only the
male line (see graphic, p. 215),
breeding untraumatized, “na-
ïve” female mice with trauma-
tized males, and then removing
males from the mother’s cage
so that their behavior did not
impact their offspring. After
weaning, she raised the mice
in mixed groups to prevent lit-
termates from reinforcing each
other’s behaviors.
Her lab repeated the proce-
dure, sometimes going out six
generations. “It worked imme-
diately,” she says of the protocol.
“We could see that there were
symptoms [in descendants] that
were similar to the animals that
were themselves separated.”
Descendants of stressed fathers
displayed more risk-taking be-
havior, like exploring exposed
areas of a platform suspended
off the ground. When dropped
in water, they “gave up” and
stopped swimming sooner than Ali Jawaid (back right) works with children in a Pakistani orphanage. One boy has
control mice, an indicator of de- just given a blood sample for an epigenetic study.
pressivelike behavior in mice.
Mansuy is “definitely a pioneer,” says Ro-
main Barrès, a molecular biologist at the
University of Copenhagen. Other research-
ers have developed conceptually similar
models, for example giving male mice al-
tered diets or exposing them to nicotine and
tracing metabolic and behavioral changes
out for generations.
“If you’re asking, ‘Does the experience of
the parent influence the process of develop-
ment?’ the answer is yes,” says epigenetics
researcher Michael Meaney at McGill Uni-
versity in Montreal, Canada, whose own
studies have shown that differences in ma-
ternal care can have epigenetic effects on
brain development. “Isabelle and others
have documented the degree to which the
214 19 JULY 2019 • VOL 365 ISSUE 6450
no indent
experience of the parent can be passed on.
The question [is] how.”
THREE MASSIVE FREEZERS, down the hall
from Mansuy’s office, are filled with thou-
sands of samples of mouse blood, liver, milk,
microbiome, and other tissues. These serve
as a –80°C archive of more than 10 years of
data. Mansuy estimates she’s collected be-
havioral data and tissue samples from thou-
sands of mice over the past decade.
She hopes the biological markers of
trauma are hidden in those freezers, waiting
to be revealed. Many of the early mammalian
epigenetic studies focused on DNA methyla-
tion, which “tags” DNA with methyl groups
that switch genes off. But those changes
seemed unlikely to be directly inherited:
In mammals, methylation is mostly erased
when egg and sperm come together to form
an embryo.
Mansuy and others still think methylation
could have some role. But they are also ze-
roing in on tiny information-rich molecules
called small noncoding RNAs (sncRNAs).
Most RNA is copied from DNA, and then
acts as a messenger to instruct the cell’s ribo-
somes to produce specific proteins. But cells
also contain short strands of RNA that don’t
produce proteins. Instead, these noncoding
RNAs piggyback on the messenger RNAs, in-
terfering with or amplifying their function, saw the same behavioral changes in the Troubled offspring But this body of research hasn’t con-
thus causing more or less of certain proteins resulting mice. To explore how trauma affects generations of mice, such as the vinced everyone. Geneticist John Gre-
to be produced. But although the cause, in the form grandfather, father, and male pup below, researchers stressed ally at the Albert Einstein College of
Mansuy and others think stress may influ- of altered RNA, and the effect, in the mother mice. Their pups then exhibited both molecular and Medicine in New York City has been a
ence sncRNAs, along with the many other form of altered behavior and physiol- behavioral changes, such as taking more risks on an elevated vocal critic of the evidence for epigen-
biochemical changes it causes, from higher ogy, are identifiable in mouse experi- maze. These changes persisted for up to 5 generations. etic inheritance of trauma, pointing at
levels of hormones like cortisol to inflamma- ments, everything in between remains small sample sizes and an overreliance
tion. They have focused on the sncRNAs in maddeningly difficult to untangle, espe- on epidemiological studies. For now,
sperm, which may be especially vulnerable to cially in people. “The field has come a he says, “Mouse models are the way to
stress during the weeks that newly formed long way in the last 5 years,” Bale says. go.” He’s not yet seen definitive experi-
sperm spend maturing in a twisting tube on “But we don’t know what’s going on in ments even in mice, he says. “I’d like
top of the testes. Later, when sperm and egg humans because we don’t have a con- Elizabeth Culotta to see•us7/12/19
be more10:08
bold andAM brave and
come together, altered sncRNAs could mod- trolled environment.” move from preliminary associations to
ify the production of proteins at the very be- Still, mouse data in hand, Mansuy
Epigenetic changes, such as methylation
definitive studies—and be open to the
ginning of development in a way has been looking for similar epigenetic of DNA and alteration of RNA
idea that there may be nothing there.”
that ripples through the millions changes in people. She analyzed blood [changing "marks" to "changes". I can't
and millions of cell divisions that samples from Dutch soldiers, collected make this change IN Ain k4] ROOM down the hall
DARKENED
follow. “Hosts of signals happen before and after deployment to Afghan- from Mansuy’s office, just outside the
as those cells become a zygote,” istan between 2005 and 2008. And she’s mouse breeding area, two cages stand
says epigeneticist Tracy Bale at working with clinicians in Nice, France, side by side on a table. One is a stan-
the University of Maryland in to examine blood samples from survi- dard lab mouse enclosure, not much
Baltimore. “If dad brings small vors of a horrific 2015 terror attack. bigger than a shoebox. Wood chip–
noncoding RNAs that have an Other researchers had found altered strewn cages like this are where most
effect on mom’s RNAs, that can sncRNAs in the blood of the soldiers. lab mice, including most of Mansuy’s
Mother separated
change the trajectory of embryo In 2017, for example, Dutch research- animals, spend their lives.
from pups and
development.” ers showed soldiers exposed to combat traumatized. Next to it, black-furred, pink-tailed
Bale found evidence that trauma had recognizable differences Mother often mice scurry up and down in a luxury
trauma can affect sncRNAs in in dozens of sncRNA groups, some of ignores pups. two-story mouse house, equipped with
sperm—and that the effects them correlated with PTSD. But Man- three running wheels and a minimaze
might be transmitted to off- suy couldn’t find the same kinds of RNA upstairs. Their environment is designed
spring. She stressed mice during changes that appeared in her lab’s mice. to stimulate their senses and engage
adolescence by barraging them That could be because the soldiers’ sam- 3-month-old more of their brains in play and explo-
for weeks at unpredictable inter- ples were years old, or simply because male offspring ration (Science, 9 February 2018, p. 624).
vals, with things like fox odors, mice and people are different, showing mated with In 2016, Mansuy published evidence
loud noises, and bright light. the limits of mouse models. But Man- untraumatized that traumatized mice raised in this
females.
Then, she examined the sncRNAs suy hopes it means epigenetic changes enriched environment didn’t pass the
in their sperm and offspring. She are sensitive to the type of trauma and symptoms of trauma to their offspring.
Offspring show
found differences in nine types when it occurs in the life course. Mice The limited data—Mansuy says they’re
epigenetic and
of microRNAs--small noncoding can never perfectly replicate human suf- behavioral changes now working on an expanded study—
RNAs--including one that regu- fering, but, she says, “the best approach” without having suggests life experience can be healing
lates SIRT1, a gene that affects for research “is to select a population of experienced as well as hurtful at the molecular level.
metabolism and cell growth. humans who have gone through condi- trauma. “Environmental enrichment at the right
She then created RNA mol- tions which are as similar as possible to time could eventually help correct some
ecules with similar alterations our model.” Breeding of the alterations which are induced by
and injected them into early- That’s where the Pakistani orphans carried out for 6 trauma,” Mansuy says.
stage embryos. When those em- come in. The children’s chaotic early generations This and a few other studies sug-
bryos grew to adults, they carried years may have some similarities to gesting epigenetic change is reversible
RNA alterations like those seen in the sperm. what the mice in Mansuy’s lab experience, at Tufts University in Boston exposed male have the potential to change the narrative of
CREDITS: (GRAPHIC) V. ALTOUNIAN/SCIENCE; (PHOTO) PIOTR PIWOWARSKI
This second generation also had lower levels she says, including unpredictable separation mice to social stress by routinely changing doom around the topic, researchers say. “If
of corticosterone, the mouse equivalent of from their mothers. their cage mates. Their sperm had altered it’s epigenetic, it’s responsive to the environ-
cortisol, after a stressful spell inside a tight Early results are promising. “We have levels of specific sncRNA groups—albeit dif- ment,” says Feig, who found similar effects
tube. “If you do the same RNA changes, you overlapping findings with the mouse ferent ones from those altered in Mansuy’s on brain function across generations by giv-
produce offspring with the same phenotype,” model,” Jawaid says. In a preprint uploaded mice—and their offspring were more anx- ing mice play tubes, running wheels, toys,
Bale says. last month to bioRxiv, Mansuy and Jawaid ious and less sociable than the offspring of and larger cages more than a decade ago.
Mansuy found similar RNA changes in documented changes in the levels of fatty unstressed parents. “That means negative environmental effects
her male mice traumatized as pups. They acids in the orphans’ blood and saliva Working with a sperm bank, Feig then are likely reversible.”
had higher levels of specific sncRNAs, in- that mimicked changes in the traumatized looked for the same sncRNAs in human In public talks and interviews, Mansuy
cluding miR-375, which plays a role in mice—as well as similar sncRNA altera- sperm. He also asked donors to fill out the says she’s careful not to promise too much.
stress response. Mansuy is convinced those tions. The presence of similar biomarkers Adverse Childhood Experience (ACE) ques- As confident as she is in her mouse model,
molecular changes account for some of the “suggests that comparable pathways are op- tionnaire, which asks about abusive or dys- she says, there’s lots more work to be done.
PHOTO: ALI JAWAID
inherited behavioral traits she documented. erating after trauma in mice and children,” functional family history. The higher the “I don’t think the field is moving too fast,”
In one experiment, her team injected RNA Mansuy says. men’s ACE score, the more likely they were Mansuy says. “I think it’s moving too slow.” j
from traumatized male sperm into the fertil- In a conceptually similar effort to go to have sperm sncRNA profiles matching
ized eggs of untraumatized parents [ck] and from mice to people, biologist Larry Feig what Feig had seen in mice. Andrew Curry is a journalist in Berlin.
sciencemag.org SCIENCE SCIENCE sciencemag.org 19 JULY 2019 • VOL 365 ISSUE 6450 215
A PAINFUL
LEGACY
“Pullquote or liftout quote piece tops on baseline as shown a “synthesis of dummy type goes
here.”
Name Lastname, Affiliation
Headline box goes here ragged type
R
e1ici viti con rem doluptaqui numque seribus2am, aut quatia quatur, cuptio et harcit ani3aspi ditatiorem. Ariamus nobisita natius-
a4nt ditem desteserovid mo quatia volupta5tia non perum cum rem quia veliquas qu6e volorum exerest a corectas molup7ta inciend
ionsed esequatur, aut voloria non8esse lab in rernam ea sim alitis ea dest qua9tiun tionece ptionecum imagnimi, omni10gniet
maximet que dolor ma voluptat al11tur?
Velicid isquo12 deres di con poresen isinim13st liquiae des dolesciam, tes sequi rem r14 ne volorporerum culparum quae pla il
m15s ea dior am andelectibus dolorrorem idi co16equi nisciae voluptasi ipis ipis et dolum 17ndior epelessi ad unt exerempt

Sidehed here. Mostius aut officie 70cese vent es molorupitas re cus diaesti71, est quo magnate sanim ium eturi doll72um volesti struptat.
Upiciam, nosap73um volorum ut lant. –Writer Lastname