Pubertal Stress Recalibration in Postinstitutionalized Children PDF

Correction PSYCHOLOGICAL AND COGNITIVE SCIENCES, PHYSIOLOGY Correction for “Pubertal stress recalibration reverses the ef- fects of early life stress in postinstitutionalized c...

Correction PSYCHOLOGICAL AND COGNITIVE SCIENCES, PHYSIOLOGY Correction for “Pubertal stress recalibration reverses the ef- fects of early life stress in postinstitutionalized children,” by Megan R. Gunnar, Carrie E. DePasquale, Brie M. Reid, and Bonny Donzella, which was first published November 11, 2019; 10.1073/pnas.1909699116 (Proc. Natl. Acad. Sci. U.S.A. 116, 23984– 23988). The authors note that Bradley S. Miller should be added to the author list following Bonny Donzella. Bradley S. Miller should be credited with performing research. The corrected author line, affiliation line, and author contributions appear below. The online version has been corrected. Megan R. Gunnara, Carrie E. DePasqualea, Brie M. Reida, Bonny Donzellaa, and Bradley S. Millerb CORRECTION a Institute of Child Development, University of Minnesota–Twin Cities, Minneapolis, MN 55455; and bDivision of Pediatric Endocrinology, Department of Pediatrics, University of Minnesota, Minneapolis, MN 55454 Author contributions: M.R.G. and B.D. designed research; C.E.D., B.M.R., B.D., and B.S.M. performed research; C.E.D. analyzed data; M.R.G. and C.E.D. wrote the paper; and B.M.R. edited paper and assisted with data analysis. Published under the PNAS license. First published January 13, 2020. www.pnas.org/cgi/doi/10.1073/pnas.1922473117 Downloaded by guest on January 2, 2021 www.pnas.org PNAS | January 28, 2020 | vol. 117 | no. 4 | 2225 Pubertal stress recalibration reverses the effects of early life stress in postinstitutionalized children Megan R. Gunnara,1, Carrie E. DePasqualea, Brie M. Reida, Bonny Donzellaa, and Bradley S. Millerb a Institute of Child Development, University of Minnesota–Twin Cities, Minneapolis, MN 55455; and bDivision of Pediatric Endocrinology, Department of Pediatrics, University of Minnesota, Minneapolis, MN 55454 Edited by Ian H. Gotlib, Stanford University, Stanford, CA, and accepted by Editorial Board Member Renée Baillargeon October 8, 2019 (received for review June 10, 2019) Nonhuman animal models reveal that the hypothalamic–pituitary– Glucocorticoids have an inverted U shape relation to cognitive and adrenocortical (HPA) axis calibrates to the harshness of the envi- behavioral functioning due to differential activation of mineralo- ronment during a sensitive period in infancy. Humans exposed to corticoid and glucocorticoid receptors in the hippocampus and depriving institutional care in infancy show reduced HPA axis amygdala, among other areas (12). Thus, underactivity of the HPA responsivity, even years after they are placed in supportive, axis has been associated with various physical and mental health well-resourced families. This study examined whether puberty problems (13–15). The question is, can this effect be reversed? opens a window of opportunity to recalibrate the HPA axis toward Simply removing children from deprivation and placing them more typical reactivity when children shift from harsh deprived in supportive, well-resourced homes does not appear to be suf- conditions in infancy into supportive conditions in childhood and ficient to allow the HPA axis to recalibrate. This is the case even adolescence. Participants (n = 129 postinstitutionalized, 68.2% fe- when the parents in those homes score very high on observa- male; n = 170 comparison, 52.4% female) completed 3 annual tional measures of parenting quality (16) and children report sessions beginning at ages 7 to 15 (M = 11.28, SD = 2.31). Each high levels of perceived support (8). However, development may session assessed pubertal stage via nurse examination and cortisol allow windows of opportunity for recalibration. In particular, for reactivity to the Trier social stress test for children. The linear the HPA axis, puberty may reopen the system and functionally mixed-effects model controlling for sex and between-individual create a second sensitive period. Two cross-sectional studies differences in pubertal stage showed a significant group by pu- suggest this may be the case. Both involved the cortisol awak- bertal stage interaction: within-individual increases in pubertal ening response (CAR) and thus were not actually studies of the stage were associated with increases in cortisol stress reactivity stress response (17, 18). Both found a blunted CAR for early life for postinstitutionalized youth but not nonadopted comparison stress children at earlier stages of pubertal development but an youth. This study indicates that pubertal development reopens a increased CAR for those at later stages. The increased CAR was window of opportunity for the HPA axis to recalibrate based on more similar to that shown by children and adolescents who had significant improvements in the supportiveness of the environ- not experienced adverse care. ment relative to that in infancy. The peripubertal period may be The possibility that puberty may open a window of plasticity an important time in development where the caregiving environ- allowing the HPA axis to recalibrate is supported by studies in ment has a substantial impact on the HPA axis and, perhaps, other nonhuman animal models. Romeo (19, 20) has argued that the stress-mediating systems. Future research is needed to examine the peripubertal period is another sensitive period for programming mechanisms of recalibration and whether HPA recalibration impacts of the HPA axis. He has shown that stressors during that period physical and psychological health. have longer-term impacts on the axis than the same stressors HPA axis | puberty | recalibration | early life stress | institutional care Significance I nstitutional care during infancy impairs the responsiveness of the HPA axis to psychosocial stressors. It also is associated with reductions in risk-taking and sensation-seeking (1, 2), increased This longitudinal study demonstrates within-individual puber- tal recalibration of the HPA axis in humans. Findings provide empirical support for an adolescent window of plasticity dur- amygdala responses to threat stimuli (3, 4), and increases in ing which the brain resamples the environment and alters HPA anxiety symptoms (5) that are associated with alterations in brain functioning if the current caregiving environment is sufficiently connectivity (6) and white matter pathways (7). Taken together, different from the early caregiving environment in which the these effects suggest that the defensive system was calibrated system was originally organized. This suggests that interven- to the harsh, unsupportive environment of the institution (or- tion efforts to improve outcomes for children who have ex- phanage) and that even years after adoption into supportive, perienced early life adversity should include a focus on the well-resourced homes, the defensive system maintains its original prepubertal and peripubertal period in order to maximize their calibration. Studies of the adrenocortical response to stressors impact on recalibrating systems like the HPA axis. have shown that early institutional deprivation results in a blunted Author contributions: M.R.G. and B.D. designed research; C.E.D., B.M.R., B.D., and B.S.M. cortisol response to stressors (8–10). This is seen within the first performed research; C.E.D. analyzed data; M.R.G. and C.E.D. wrote the paper; and B.M.R. months after removal from institutional care (9) and lasts at least edited paper and assisted with data analysis. until middle childhood (8). The effect is causal, as shown by the The authors declare no competing interest. Bucharest Early Intervention Study. They found that the cortisol This article is a PNAS Direct Submission. I.H.G. is a guest editor invited by the and autonomic reactivity were both blunted in response to the Editorial Board. Trier social stress test in children randomly assigned to institu- Published under the PNAS license. tional care as usual and those randomly assigned to removal from Data deposition: The data from this manuscript are available at https://github.com/ institutional care. However, this was only true for those removed cdepasquale/Gunnar-DePasquale-Reid-Donzella-2019-PNAS. from the institution after 2 y of age but not for those removed 1 To whom correspondence may be addressed. Email: [email protected]. before age 2 (10). The causal nature of this blunting of the HPA This article contains supporting information online at https://www.pnas.org/lookup/suppl/ axis is also demonstrated in studies of nonhuman primates ran- doi:10.1073/pnas.1909699116/-/DCSupplemental. domly assigned to nursery rearing instead of maternal rearing (11). First published November 11, 2019. 23984–23988 | PNAS | November 26, 2019 | vol. 116 | no. 48 www.pnas.org/cgi/doi/10.1073/pnas.1909699116 imposed on adult animals. It is also the case that positive expe- change by group interaction was significant (t = 2.74, P = 0.01), riences in adolescent nonhuman animals functionally reverse the but the mean pubertal stage by group interaction was not (t = effects of prenatal stress (21) and postnatal repeated maternal −0.77, P = 0.44), suggesting that exposure to early institutional separations (22). These studies used environmental enrichment care moderated the association between within-individual changes to reverse the hyperresponsiveness of the axis induced by pre- in pubertal stage and cortisol reactivity but not between-individual natal and postnatal stress. differences in pubertal stage and cortisol reactivity (Fig. 1). Least- While studies using the CAR suggest that there may be a squares mean contrasts indicated that cortisol reactivity at pubertal recalibration of the HPA axis with puberty, the CAR is not a stages 1 and 5 were significantly different for postinstitutionalized measure of stress reactivity. To answer the question of whether youth (95% CI [−1.40, −1.15] and [−1.14, −0.92], respectively). the HPA axis stress response is recalibrated with puberty, it is There were no significant differences in cortisol reactivity for necessary to assess the cortisol response during a stressor task. We nonadopted youth at any pubertal stage. There was no evidence used the Trier social stress test for children (TSST-C), a social that sex moderated the recalibration effect. evaluative stressor (23, 24) and one of the most reliable stressor tasks in the literature, to examine whether youth adopted as infants Discussion from institutions would become increasingly more stress responsive Research suggests that there is an initial sensitive period in in- and would respond more similarly to low-risk, comparison youth fancy during which the HPA axis calibrates to the harshness of the with increasing pubertal stage. An accelerated longitudinal design environmental conditions which, for humans, tends to result in was used (25) in which children began the first session at 7 to 15 y reduced HPA responsivity (9, 10). This study sought to determine old and then were assessed annually over 2 y, totaling up to 3 whether puberty provides a second window of plasticity during sessions per participant. See SI Appendix, SI Materials and Methods, which the human HPA axis could recalibrate if conditions are for details of our method for maintaining responses across re- sufficiently different from those in infancy. In the case of the pre- peated trials of the TSST. We previously reported preliminary sent study, conditions were harsh and unsupportive in infancy, and evidence of recalibration using a cross-sectional examination of the once adopted, the children entered well-resourced and generally PSYCHOLOGICAL AND COGNITIVE SCIENCES data from the first session (26). The present analysis focused on the supportive homes. Thus, our prediction was that HPA axis reac- cortisol response to the TSST-C as a function of within individual tivity, with advancing pubertal stage, would increasingly resemble increases in pubertal stage. We hypothesized that youth exposed to the typical responsiveness of youth reared in well-resourced, sup- institutional care in infancy would exhibit increasing cortisol re- portive homes. If postinstitutionalized children’s HPA activity can activity with increasing pubertal stage, gradually becoming more be restored to moderate, but not prolonged, levels of responding to similar to the cortisol responses seen in typically developing youth, environmental challenges, this may signal other downstream im- who would show no or comparatively little change in cortisol re- provements in physical and mental health as well (13, 15). Because activity across puberty. Importantly, this framework begins to tease glucocorticoids are known to be critical regulators of plasticity apart the role of age- and puberty-related change, as within- (14), recalibration of the system with puberty may help support individual increases in pubertal stage do not differ if, for exam- the plasticity of this period of brain development. PHYSIOLOGY ple, an individual moves from pubertal stage 1 to 2 or stage 4 to 5. Indeed, the results of this study provide within-individual ev- To compare biological and psychological responses to the TSST-C idence of pubertal recalibration of the HPA axis in humans. across puberty, see SI Appendix, Pubertal Changes in Self-Reported Using the first year of data collection in this study, we previously Stress, for additional analyses examining whether increasing pu- reported that cross-sectional differences in puberty predict bertal stage predicts within-individual changes in self-reported greater cortisol reactivity. The present analysis allows us to subjective stress during the TSST-C. confirm that this was, in fact, a developmental process occurring within individuals. For postinstitutionalized youth, within-individual Results increases in pubertal stage significantly predicted increased (i.e., Salivary cortisol reactivity and pubertal status were characterized more typical) cortisol stress reactivity. Nonadopted comparison in 299 children and adolescents across 3 annual sessions beginning youth who did not experience institutional deprivation in infancy when participants were 7 to 15 y old. Participants were either pre- did not show a similar pattern of pubertal recalibration. These viously institutionalized as infants and toddlers (postinstitutionalized findings corroborate nonhuman animal models (19, 22) and ex- [PI]) or born and raised in their natal families (nonadopted [NA]). tend cross-sectional human research (17, 18, 28) which demon- At each session, saliva samples were collected across 2 h, capturing strated between-individual differences in HPA functioning across cortisol reactivity and recovery in response to a modified version the peripubertal period, particularly for those who experienced of the TSST-C (see SI Appendix, Fig. S1 and SI Materials and early life adversity. Methods, for detail on sampling protocol). A linear mixed-effects Importantly, this change in cortisol reactivity does not occur model was fit to examine whether within-individual changes in immediately upon removal from the depriving environment. The pubertal status would predict changes in salivary cortisol reactivity, majority of the youth in this study were adopted by age 2, and yet moderated by group. Data were modeled hierarchically with cor- even in the earlier stages of pubertal development (at least 5 to 7 y tisol sample nested within session, nested within participant. postadoption) the axis still appears relatively blunted. It ap- Between-individual differences in pubertal stage were modeled pears to take the physiological changes associated with pubertal by calculating each individual’s mean pubertal stage across all development to open the axis for recalibration. From a life his- sessions (hereon referred to as “mean pubertal stage”). The focal tory perspective (29), the axis is recalibrating prior to the onset predictor, within-individual changes in pubertal stage, was cal- of the reproductive period to match the conditions likely to be culated as the difference from an individual’s mean at each separate present during that life stage. However, the mechanisms of this session (hereon “pubertal change”). Because puberty may also in- recalibration are still unclear. There was no particular pubertal crease differences in HPA axis functioning between males and fe- stage that was associated with changes in cortisol reactivity in this males (27), we also explored the possibility that sex would modify study. For this reason, it is unlikely that the activity of any in- the pubertal stress recalibration effect. Participant information is dividual hormone is responsible for driving recalibration. In- described in Table 1. Descriptive statistics and correlations between stead, it may be that more global changes in neural plasticity and focal variables are included in SI Appendix, Table S1. the associated enhanced processing of environmental stimuli occurring during puberty (30) allow the caregiving environment Pubertal Recalibration of Salivary Cortisol Reactivity. Full model to have a disproportionately large influence on HPA axis func- results are displayed in SI Appendix, Table S2. The pubertal tioning during this period. Gunnar et al. PNAS | November 26, 2019 | vol. 116 | no. 48 | 23985 Table 1. Demographic information for each group Postinstitutionalized (n = 129) Nonadopted (n = 170) Test of difference Session 1 age in years, M (SD) 11.38 (2.39) 11.20 (2.24) t(279) = 0.63, P = 0.53 Sex, n (%) female 88 (68.22%) 89 (52.35%) χ2(1, n = 299) = 4.84, P = 0.03 Child race, n (%) χ2(4, n = 276) = 103.36, P < 0.001 American Indian/Alaskan Native 14 (11.29%) 1 (0.62%) Asian 48 (38.71%) 2 (1.23%) Black/African American/African 7 (5.65%) 4 (2.47%) White 50 (40.32%) 142 (87.65%) Multiracial 5 (4.03%) 13 (8.02%) Child region of origin, n (%) — Russia/Eastern Europe 63 (48.84%) — China/Southeast Asia 45 (34.89%) — Latin America 16 (12.40%) — Africa/Haiti 5 (3.88%) — Session 1 puberty, n (%)* t(86) = −1.15, P = 0.25 Stage 1 41 (33.61%) 60 (37.97%) Stage 2 16 (13.11%) 39 (24.68%) Stage 3 28 (22.95%) 14 (8.86%) Stage 4 19 (15.57%) 14 (8.86%) Stage 5 18 (14.75%) 31 (19.62%) Session 1 medication index, M (% 0)† 2.76 (74.24%) 2.26 (86.71%) t(279) = 3.69, P < 0.001 Annual family income, n (%) χ2(8, n = 276) = 9.73, P = 0.28 $200,000 15 (12.61%) 22 (14.01%) Primary caregiver education, n (%) χ2(9, n = 280) = 7.39, P = 0.60 High school degree/GED or less 2 (1.64%) 3 (1.90%) Some college 16 (13.11%) 25 (15.82%) Undergraduate degree 42 (34.43%) 68 (43.04%) Graduate/professional degree 62 (50.82%) 62 (39.24%) Significant group differences are indicated in bold. *Statistic tests group difference in age at pubertal stage 3 (central puberty) in order to examine group differences in pubertal timing. † Mean calculated excluding zeroes. Further, this study measured cortisol, which is a distal end comparable enough to be confidently treated as equivalent. In any product of the HPA axis that involves complex neural inputs and case, repeated exposure to the TSST-C is ecologically valid be- influences. It is unclear whether the change in HPA reactivity cause youth commonly encounter multiple experiences of speaking occurred at the level of the adrenal, pituitary gland, hypothala- in front of the class or engaging in other public social evaluations. mus, or limbic inputs to the hypothalamus. This knowledge could Thus, this study provides the best approximation of an adolescent’s have functional implications for our understanding of pubertal cortisol response to multiple exposures of social evaluative stress. development and the recalibration of the HPA axis following Of particular note, we did not find that age of adoption altered substantial changes in environmental conditions. Future studies are the impact of puberty on recalibration of the cortisol response. needed to determine the mechanisms through which pubertal We had a wide range of adoption age (5.5 to 59 mo, M = 19.43) recalibration of the stress response takes place in humans. It will and so would have expected to have identified an impact of also be important to examine the extent to which variations in adoption age if it was there. This means that even those adopted current environmental conditions impact the magnitude of pubertal later than age 2 were exhibiting changes in reactivity, consistent recalibration and whether recalibration is associated with physical with the idea that the increased plasticity of the peripubertal and psychological functioning in adolescence into adulthood. period allowed them to recalibrate. We might have expected less Repeated administration of the same stressor, as was done in recalibration among those adopted younger because previous this study, may change the individual’s psychological interpre- work would suggest that they would have a less blunted response tation and/or physiological response to the stressor. Indeed, (10); however, we did not see evidence of this. It should also be participants reported lower self-reported subjective stress across noted that other studies have found persistently reduced cortisol puberty (SI Appendix, Table S3). However, this would typically be reactivity into adulthood in individuals who experienced both associated with a reduced cortisol stress response over time. The early life stress and repeated, ongoing adversity (31, 32), sug- fact that this study found increasing cortisol reactivity despite re- gesting that puberty alone does not increase axis reactivity fol- duced novelty of the stressor and decreasing subjective stress fur- lowing early adversity in humans. Indeed, it seems likely that ther strengthens the evidence for pubertal recalibration. To test the conditions need to change markedly for recalibration to be evi- pubertal recalibration hypothesis, one could hypothetically use dent. One study examining postinstitutionalized children who equivalent but different stressors at each time point in order to were adopted prior to adolescence found continued HPA axis maintain novelty of the stressor. Unfortunately, there is not enough dysregulation in adulthood (33), although this was measured via evidence in the literature that any 2 laboratory stressors are the CAR and not reactivity to a stressor. It is yet unclear whether 23986 | www.pnas.org/cgi/doi/10.1073/pnas.1909699116 Gunnar et al.  cortisol stress reactivity and pubertal stage at 3 consecutive, annual sessions. We used an accelerated longitudinal design (25), with multiple age cohorts (7 to 15 y) that we followed once per year for 3 assessment periods. This design thus captured a wider developmental period (∼7 to 17 y of age, from the youngest participants at session 1 to the oldest participants at session 3) in a shorter time span than a traditional longitudinal design. Consent was received from 1 par- ent for each participant, and written and verbal assent were received from all participants. This study was approved by the University of Minnesota In- stitutional Review Board. Pubertal Staging. Trained nurses conducted pubertal staging, scored according to Marshall and Tanner criteria (35, 36). Breast and genital scores, for girls and boys, respectively, were used as a measure of central puberty ranging from 1, indicating pubertal development not yet begun, to 5, in- dicating pubertal development is complete (see ref. 37 and SI Appendix, SI Materials and Methods, for additional details on pubertal assessment pro- cedures, quality control analyses, and data processing). Salivary Cortisol Reactivity. At every annual session, participants completed a modified version of the TSST-C (see refs. 8, 24, and 26 for additional details on Fig. 1. Line graph illustrating model-implied results of a linear mixed-effects this procedure) while being filmed by an obvious camera in front of a 2-way model examining the association between pubertal stage and cortisol stress mirror. Seven whole, unstimulated saliva samples were collected throughout reactivity by group (postinstitutionalized, Left; nonadopted, Right). Saliva the 2-h laboratory visit, as shown in SI Appendix, Fig. S1 (26), to later be assayed samples were collected at time 0, +5, +20, +40, +60, and +80 min shown on for cortisol concentration. The samples were stored in a laboratory freezer at the x axis above, with time 0 indicating beginning of TSST-C prep. For clarity, −20 °C until being shipped to the University of Trier, Germany. All samples were PSYCHOLOGICAL AND the data shown are collapsed across multiple data points per participant; COGNITIVE SCIENCES assayed in duplicate using a time-resolved fluorescence immunoassay (DELFIA). however, model results in SI Appendix, Table S2, indicate that cortisol re- Additional details on laboratory measures, procedures, quality control analyses, activity is associated with within-individual increases in pubertal stage, not just and data processing are provided in SI Appendix, SI Materials and Methods. between-individual differences across puberty (see SI Appendix, Table S2, for model results and SI Appendix, Fig. S2, for 95% confidence envelopes). Data Analysis. A hierarchical linear mixed-effects model (38) was used to ex- amine change in cortisol reactivity over time. Time since start of the TSST-C preparatory period and the quadratic effect of time were used as predictors to evidence of recalibration will generalize beyond situations of model cortisol reactivity and recovery, with random intercepts and random early institutional deprivation followed by adoption into well- effects of time and quadratic time nested within session and within partici- resourced homes. It will be important to determine whether pant. The quadratic effect of time captures the expected inverted U pattern of other conditions of adversity (e.g., child abuse and neglect) that cortisol production across the TSST-C: an initial increase (reactivity) that PHYSIOLOGY remit prior to the peripubertal period also exhibit evidence of eventually peaks and starts to decline again (recovery). To account for the recalibration of the HPA axis. accelerated longitudinal design and disentangle between- and within-person In conclusion, pubertal development seems to open a window of effects of pubertal stage, and capture both an individual’s change in puberty opportunity for changes in the HPA axis of postinstitutionalized over time as well as an individual’s mean pubertal score compared with other children that tend not to be seen prior to puberty when they are individuals in the study, within- and between-individual pubertal change scores were calculated. Within-individual change in puberty, or the longitu- adopted out of institutions into well-resourced homes. There is dinal change in pubertal score for each individual (pubertal change), was also a steep increase in the incidence of mental health disorders modeled using pubertal stage centered around each participant’s mean across during the pubertal period (34). The ability to recalibrate the HPA all sessions. Between-individual change in puberty was modeled as each par- axis toward more typical functioning during puberty could pro- ticipant’s mean pubertal stage across all sessions (mean pubertal stage). Each mote resilience in high-risk adolescents who experienced early life pubertal stage variable was then moderated by group (PI vs. NA) to examine adversity, protecting against the emergence of mental health whether changes in pubertal stage predicted changes in cortisol reactivity symptoms, although this remains to be tested and should be a differently for the 2 groups. Thus, the focal predictor was the quadratic time focus of future research. To take advantage of the apparent by pubertal change by group moderation term, representing a change in heightened plasticity seen during pubertal development, the pre- cortisol reactivity and recovery associated with within-individual increases in puberty that differs by group. All models adjusted for sex, as well as sex by sent findings argue that interventions to improve the supportive- group and sex by pubertal stage interactions (additional details on model ness of children’s environments should include a focus on the construction are provided in SI Appendix, SI Materials and Methods). peripubertal period to maximize their effectiveness for children’s stress-mediating systems like the HPA axis. Data Availability. The data from this manuscript are available on GitHub (39). Materials and Methods ACKNOWLEDGMENTS. This study was funded by National Institute of Child Participants and Study Design. Participants included 299 youths aged 7 to 15 y at Health and Human Development Grant R01 HD075349 (to M.R.G.), National In- the start of the study (M = 11.28, SD = 2.31). Of those participants, 129 (88 stitute of Mental Health Grant T32 MH015755 (to C.E.D.), and NSF Graduate Re- female) were previously institutionalized as infants and toddlers (PI) and 170 search Fellowship 00039202 (to B.M.R.). The content is solely the responsibility of (89 female) were born and raised in their natal families (NA). PI youth were the authors and does not represent the views of the National Institutes of Health selected to have been adopted prior to age 5 (M = 19.43 mo, SD = 12.64) and or the National Science Foundation. The authors wish to thank the families who participated, the International Adoption Project, and the Center for Neurobehavioral spent at least 50% of their preadoptive life in the institution (M = 95.19%, SD = Development at the University of Minnesota. We also thank Tori Simenec, 9.54). PI and NA families were roughly matched on socioeconomic status; see Bao Moua, Lea Neumann, Heather Taylor, and Dr. Chris Desjardins for their Table 1 for demographic information and analyses of group differences in assistance with the study; our nurses Janet Goodwalt, Terri Jones, and Melissa demographics (see SI Appendix, SI Materials and Methods, for additional in- Stoll for Tanner staging; and Drs. Brad Miller and Lorah Dorn for providing formation on identification of study participants). We assessed participants’ training in pubertal assessment. 1. M. M. Loman, A. E. Johnson, K. Quevedo, T. L. Lafavor, M. R. Gunnar, Risk-taking and 3. F. S. Maheu et al., A preliminary study of medial temporal lobe function in youths sensation-seeking propensity in postinstitutionalized early adolescents. J. Child Psychol. with a history of caregiver deprivation and emotional neglect. Cogn. Affect. Behav. Psychiatry 55, 1145–1152 (2014). Neurosci. 10, 34–49 (2010). 2. C. Kopetz et al., Early psychosocial deprivation and adolescent risk-taking: The role of 4. N. Tottenham et al., Elevated amygdala response to faces following early deprivation. motivation and executive control. J. Exp. Psychol. Gen. 148, 388–399 (2019). Dev. Sci. 14, 190–204 (2011). Gunnar et al. PNAS | November 26, 2019 | vol. 116 | no. 48 | 23987  5. K. L. Wiik et al., Behavioral and emotional symptoms of post-institutionalized children 23. A. P. Allen et al., The Trier Social Stress Test: Principles and practice. Neurobiol. Stress in middle childhood. J. Child Psychol. Psychiatry 52, 56–63 (2011). 6, 113–126 (2016). 6. J. A. Silvers et al., Vigilance, the amygdala, and anxiety in youths with a history of 24. I. S. Yim, J. A. Quas, L. Cahill, C. M. Hayakawa, Children’s and adults’ salivary cortisol institutional care. Biol. Psychiatry Cogn. Neurosci. Neuroimaging 2, 493–501 (2017). responses to an identical psychosocial laboratory stressor. Psychoneuroendocrinology 7. J. Bick, N. Fox, C. Zeanah, C. A. Nelson, Early deprivation, atypical brain development, 35, 241–248 (2010). and internalizing symptoms in late childhood. Neuroscience 342, 140–153 (2017). 25. S. Galbraith, J. Bowden, A. Mander, Accelerated longitudinal designs: An overview of 8. C. E. Hostinar, A. E. Johnson, M. R. Gunnar, Early social deprivation and the social modelling, power, costs and handling missing data. Stat. Methods Med. Res. 26, 374– buffering of cortisol stress responses in late childhood: An experimental study. Dev. 398 (2017). Psychol. 51, 1597–1608 (2015). 26. C. E. DePasquale, B. Donzella, M. R. Gunnar, Pubertal recalibration of cortisol re- 9. K. J. Koss, S. B. Mliner, B. Donzella, M. R. Gunnar, Early adversity, hypocortisolism, activity following early life stress: A cross-sectional analysis. J. Child Psychol. Psychiatry and behavior problems at school entry: A study of internationally adopted children. 60, 566–575 (2019). Psychoneuroendocrinology 66, 31–38 (2016). 27. L. R. Stroud, G. D. Papandonatos, D. E. Williamson, R. E. Dahl, Sex differences in 10. K. A. McLaughlin et al., Causal effects of the early caregiving environment on de- cortisol response to corticotropin releasing hormone challenge over puberty: Pitts- velopment of stress response systems in children. Proc. Natl. Acad. Sci. U.S.A. 112, burgh Pediatric Neurobehavioral Studies. Psychoneuroendocrinology 36, 1226–1238 5637–5642 (2015). (2011). 11. J. P. Capitanio, S. P. Mendoza, W. A. Mason, N. Maninger, Rearing environment and 28. H. Blumenthal, E. W. Leen-Feldner, C. L. Badour, C. D. Trainor, K. A. Babson, Pubertal hypothalamic-pituitary-adrenal regulation in young rhesus monkeys (Macaca mulatta). maturation and cortisol level in response to a novel social environment among female Dev. Psychobiol. 46, 318–330 (2005). adolescents. J. Adolesc. 37, 893–900 (2014). 12. J. Herbert et al., Do corticosteroids damage the brain? J. Neuroendocrinol. 18, 393– 29. M. Del Giudice, B. J. Ellis, E. A. Shirtcliff, The adaptive calibration model of stress 411 (2006). responsivity. Neurosci. Biobehav. Rev. 35, 1562–1592 (2011). 13. E. Fries, J. Hesse, J. Hellhammer, D. H. Hellhammer, A new view on hypocortisolism. 30. C. Aoki, R. D. Romeo, S. S. Smith, Adolescence as a critical period for developmental Psychoneuroendocrinology 30, 1010–1016 (2005). plasticity. Brain Res. 1654, 85–86 (2017). 14. C. Liston, W. B. Gan, Glucocorticoids are critical regulators of dendritic spine devel- 31. S. Goldman-Mellor, M. Hamer, A. Steptoe, Early-life stress and recurrent psychological opment and plasticity in vivo. Proc. Natl. Acad. Sci. U.S.A. 108, 16074–16079 (2011). distress over the lifecourse predict divergent cortisol reactivity patterns in adulthood. 15. B. S. McEwen, What is the confusion with cortisol? Chronic Stress (Thousand Oaks) 3, Psychoneuroendocrinology 37, 1755–1768 (2012). 1–3 (2019). 32. E. S. Young et al., The dual impact of early and concurrent life stress on adults’ diurnal 16. J. M. Lawler, K. J. Koss, M. R. Gunnar, Bidirectional effects of parenting and child cortisol patterns: A prospective study. Psychol. Sci. 30, 739–747 (2019). behavior in internationally adopting families. J. Fam. Psychol. 31, 563–573 (2017). 33. R. Kumsta et al., HPA axis dysregulation in adult adoptees twenty years after severe 17. L. S. King et al., The impact of the severity of early life stress on diurnal cortisol: The institutional deprivation in childhood. Psychoneuroendocrinology 86, 196–202 (2017). role of puberty. Psychoneuroendocrinology 77, 68–74 (2017). 34. J. Ormel et al., Mental health in Dutch adolescents: A TRAILS report on prevalence, 18. K. Quevedo, A. Johnson, M. Loman, T. Lafavor, M. R. Gunnar, The confluence of early severity, age of onset, continuity and co-morbidity of DSM disorders. Psychol. Med. deprivation and puberty on the cortisol awakening response: A study of post- 45, 345–360 (2015). institutionalized children. Int. J. Behav. Dev. 36, 19–28 (2012). 35. W. A. Marshall, J. M. Tanner, Variations in pattern of pubertal changes in girls. Arch. 19. R. D. Romeo, Pubertal maturation and programming of hypothalamic-pituitary- Dis. Child. 44, 291–303 (1969). adrenal reactivity. Front. Neuroendocrinol. 31, 232–240 (2010). 36. W. A. Marshall, J. M. Tanner, Variations in the pattern of pubertal changes in boys. 20. R. D. Romeo, The metamorphosis of adolescent hormonal stress reactivity: A focus on Arch. Dis. Child. 45, 13–23 (1970). animal models. Front. Neuroendocrinol. 49, 43–51 (2018). 37. B. M. Reid et al., Early growth faltering in post-institutionalized youth and later an- 21. S. Morley-Fletcher, M. Rea, S. Maccari, G. Laviola, Environmental enrichment during thropometric and pubertal development. Pediatr. Res. 82, 278–284 (2017). adolescence reverses the effects of prenatal stress on play behaviour and HPA axis 38. J. Pinheiro, D. Bates, Mixed-Effects Models in S and S-PLUS (Springer Science & Busi- reactivity in rats. Eur. J. Neurosci. 18, 3367–3374 (2003). ness Media, 2006). 22. D. D. Francis, J. Diorio, P. M. Plotsky, M. J. Meaney, Environmental enrichment re- 39. M. R. Gunnar, Gunnar, DePasquale, Reid, & Donzella 2019 PNAS. GitHub. https://github. verses the effects of maternal separation on stress reactivity. J. Neurosci. 22, 7840– com/cdepasquale/Gunnar-DePasquale-Reid-Donzella-2019-PNAS. Deposited 8 October 7843 (2002). 2019. 23988 | www.pnas.org/cgi/doi/10.1073/pnas.1909699116 Gunnar et al.