Childbirth experiences of women with a history of physical, sexual, and child abuse: a cross-sectional study of 2,575 Russian women.

BACKGROUND: A substantial number of women who subsequently become pregnant and give birth have a history of physical, sexual, and/or child abuse. This study investigated the associations of these types of traumas and their cumulative effect with childbirth experiences, namely, mode of birth, maternal and child complications during pregnancy/childbirth, preterm birth, medical procedures, and obstetric violence during labour. METHODS: A group of Russian women (n = 2,575) who gave birth within the previous 12 months, completed a web-based survey, where they provided demographic information, details about their childbirth experiences, and a history of trauma. RESULTS: Women with any type of past abuse were at higher risk for maternal complications during pregnancy/childbirth (exp(ß) < 0.73, p < 0.010 for all). More specific to the type of trauma were associations of physical abuse with caesarean birth, child abuse with complications during pregnancy/childbirth for the baby, and physical and child abuse with obstetric violence (exp(ß) < 0.54, p < 0.022 for all). There was a cumulative effect of trauma for all the outcomes except for medical procedures during childbirth and preterm birth. CONCLUSION: This study provides insights into potential different individual effects of physical, sexual, and/or child abuse as well as their cumulative impact on the childbirth experiences. The robust findings about maternal complications during pregnancy/childbirth and obstetric violence highlight the importance of trauma-informed care, supportive policies, and interventions to create safe and empowering birthing environments that prioritise patient autonomy, dignity, and respectful communication.

Long-Term Effects of Maternal Depression during Postpartum and Early Parenthood Period on Child Socioemotional Development.

Most research on the impact of maternal depressive symptoms on child development is conducted during the first year postpartum. Findings on long-term effects of maternal depression are still contradictory and underexplored. The present study investigates the long-term impact of maternal depression during the postpartum and early parenthood periods on child behavioral problems at the mean age of 2.25 years. The Edinburgh postnatal depression scale and Beck depression Inventory were used to assess postpartum and early parenthood depression, respectively. The Child Behavior Checklist was used to examine child's behavioral problems. The regression analysis showed statistically significant associations between child behavioral problems and maternal depression during early parenthood and no significant associations with postpartum depression. Four maternal depressive symptoms' trajectories were identified: consistently low, consistently high, decreased, and increased. The children of mothers with consistently high depressive symptoms at both research stages had the most significant total, internalizing, and externalizing problems. Children of mothers whose depressive symptoms decreased over time had the lowest scores in all three domains of behavioral problems. It is extremely important to implement programs for screening and early intervention for maternal mental health problems that could greatly influence the well-being of women and their children's development.

Parental burnout, depression and emotional development of the preschoolers.

Introduction: Parental burnout is becoming more and more prevalent in the world, mainly incultures with high demands towards parents. Parental burnout is distinctive from depression and might have its unique influence on child development, which isunder current international research. This work contributes to the understanding of parental burnout, maternal depression and child emotional development(specifically emotion comprehension) interrelations. Additionally, we explored whether there are differences in the effects of parental burnout and depressionon boys and girls. Methods: To analyse the emotional development of the preschoolers, the Russian version of the Test of Emotional Comprehension (TEC) was used. We used the Russian version of the Parental Burnout Inventory (PBI) to analyse the level of PB and the Russian version of Beck depression Inventory (BDI) to assess participants' depression level. Results: Parental burnout positively correlates with child emotional comprehension skills, specifically understanding of external causes (B = 0.20, CI: 0.03; 0.37) and mental causes of emotions (B = 0.22, CI: 0.05; 0.40). This effect is gender dependent and is significantlyhigher for girls (B = 0.54, CI: 0.09; 0.98). The effect of maternal depression on emotion comprehension skills is also gender dependent: total scores on emotioncomprehension tasks are significantly higher for daughters of mothers with depression (B = 0.59, CI: 0.001; 1.18). Discussion: Maternal depression and parental burnout might provoke development of extra sensitivity and self-regulation strategies in girls.

Past Traumatic Life Events, Postpartum PTSD, and the Role of Labor Support.

The aim of this study was to investigate the association of postpartum post-traumatic stress disorder (PP-PTSD) symptoms and subjective rates of traumatic birth experience with past traumatic life events (physical and sexual assault, child abuse, perinatal loss, previous traumatic birth experience, and the cumulative traumatic experience). A sample of Russian women (n = 2579) who gave birth within the previous 12 months, filled in a web-based survey, where they reported demographic and obstetric characteristics and past traumatic experiences, evaluated their birth experience (0 = not traumatic, 10 = extremely traumatic), and completed the City Birth Trauma Scale (CBiTS). We found that PP-PTSD symptoms were higher among women who previously experienced physical (F = 22.02, p < 0.001) and sexual (F = 15.98, p < 0.001) assault and child abuse (F = 69.25, p < 0.001), with only associations with child abuse (F = 21.14, p < 0.001) remaining significant for subjective rates of traumatic birth experience. Perinatal loss and previous traumatic birth showed moderate but inconsistent effects. Support during labor did not have a buffering effect for participants with past traumatic experiences but showed a universally protective effect against PP-PTSD. Trauma-informed practices and allowing women to have a supportive birth team of choice during childbirth are promising avenues to minimize the incidence of PP-PTSD and improve the childbirth experience for all women.

Evidence of SARS-CoV-2 infection in postmortem lung, kidney, and liver samples, revealing cellular targets involved in COVID-19 pathogenesis.

There is an urgent need to understand severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-host interactions involved in virus spread and pathogenesis, which might contribute to the identification of new therapeutic targets. In this study, we investigated the presence of SARS-CoV-2 in postmortem lung, kidney, and liver samples of patients who died with coronavirus disease (COVID-19) and its relationship with host factors involved in virus spread and pathogenesis, using microscopy-based methods. The cases analyzed showed advanced stages of diffuse acute alveolar damage and fibrosis. We identified the SARS-CoV-2 nucleocapsid (NC) in a variety of cells, colocalizing with mitochondrial proteins, lipid droplets (LDs), and key host proteins that have been implicated in inflammation, tissue repair, and the SARS-CoV-2 life cycle (vimentin, NLRP3, fibronectin, LC3B, DDX3X, and PPARγ), pointing to vimentin and LDs as platforms involved not only in the viral life cycle but also in inflammation and pathogenesis. SARS-CoV-2 isolated from a patient´s nasal swab was grown in cell culture and used to infect hamsters. Target cells identified in human tissue samples included lung epithelial and endothelial cells; lipogenic fibroblast-like cells (FLCs) showing features of lipofibroblasts such as activated PPARγ signaling and LDs; lung FLCs expressing fibronectin and vimentin and macrophages, both with evidence of NLRP3- and IL1ß-induced responses; regulatory cells expressing immune-checkpoint proteins involved in lung repair responses and contributing to inflammatory responses in the lung; CD34+ liver endothelial cells and hepatocytes expressing vimentin; renal interstitial cells; and the juxtaglomerular apparatus. This suggests that SARS-CoV-2 may directly interfere with critical lung, renal, and liver functions involved in COVID-19-pathogenesis.

A genome-wide association study of total child psychiatric problems scores.

Substantial genetic correlations have been reported across psychiatric disorders and numerous cross-disorder genetic variants have been detected. To identify the genetic variants underlying general psychopathology in childhood, we performed a genome-wide association study using a total psychiatric problem score. We analyzed 6,844,199 common SNPs in 38,418 school-aged children from 20 population-based cohorts participating in the EAGLE consortium. The SNP heritability of total psychiatric problems was 5.4% (SE = 0.01) and two loci reached genome-wide significance: rs10767094 and rs202005905. We also observed an association of SBF2, a gene associated with neuroticism in previous GWAS, with total psychiatric problems. The genetic effects underlying the total score were shared with common psychiatric disorders only (attention-deficit/hyperactivity disorder, anxiety, depression, insomnia) (rG > 0.49), but not with autism or the less common adult disorders (schizophrenia, bipolar disorder, or eating disorders) (rG < 0.01). Importantly, the total psychiatric problem score also showed at least a moderate genetic correlation with intelligence, educational attainment, wellbeing, smoking, and body fat (rG > 0.29). The results suggest that many common genetic variants are associated with childhood psychiatric symptoms and related phenotypes in general instead of with specific symptoms. Further research is needed to establish causality and pleiotropic mechanisms between related traits.

Postpartum PTSD and birth experience in Russian-speaking women.

OBJECTIVE: The principal goal of this study was to examine risk factors for postpartum posttraumatic stress disorder (PTSD), such as mode of birth, obstetric violence and medical interventions during labor, in Russia. This study is also the first attempt to investigate the association between the support during labor and postpartum PTSD in a Russian sample. RESEARCH DESIGN: 611 Russian-speaking mothers aged 18 to 45 years (M = 31.2 + 4.5) filled in the online form, which included Edinburgh Postnatal Depression Scale, the City Birth Trauma Scale and survey on demographic characteristics and childbirth experience within one year after birth. FINDINGS: Among the participants 45.7% had clinically significant depressive symptoms and 15% fulfilled all the diagnostic criteria for PTSD. Postpartum PTSD symptoms were higher among women who had a cesarean vs vaginal birth (p = 0.009), women with medical interventions (p = 0.043) and women who experienced obstetric violence (p < 0.001) during childbirth. The more interventions they had and the more instances of obstetric violence they experienced, the higher were the PTSD symptoms (B = 1.03, 95% CI 0.23; 1.84, p = 0.012 and B = 5.08, 95% CI 3.80; 6.37, p < 0.001, respectively). The presence of a partner or a personal midwife/doula at birth was associated with lower rates of cesarean birth, fewer medical interventions and less obstetric violence (p <  0.017 for all). KEY CONCLUSIONS: Postpartum PTSD is widespread in Russia, and the problem of obstetric violence and overuse of medical interventions has a systematic nature. Increased availability of individual continuous support during labor and the development of professional midwife and doula communities might be a potential avenue to address these problems. IMPLICATIONS FOR PRACTICE: Our findings emphasize the importance of ethics in patient-doctor communication in the maternal healthcare system and the role of support during labor, which is still not widely available in Russia.

Preliminary safety assessment of CIGB-210, an investigational peptide for HIV infection.

Current human immunodeficiency virus treatments need to be periodically administered lifelong. In this study we assess the effect of repeated doses of an anti-HIV peptide drug candidate in C57BL6 strain. Two schemes of up to 15 administrations and one of 30, daily dosing for 5 days per week, all by the subcutaneous route were evaluated. Different dose concentrations of the peptide were assayed. CIGB-210 treated animals showed no symptoms or abnormal behavior as compared with placebo. All the animals gained weight during the study. Macroscopic evaluation showed no alterations in any of the organs studied. Microscopic analysis of the tissues did not show morphological changes in thymus, stomach, small and large intestines, kidney, brain, or cerebellum. The proliferative response of splenocytes and their capacity to secrete gamma interferon were not compromised by the repeated administration of CIGB-210. There were not statistically significant differences for any of the parameters evaluated during the study among treated and non-treated groups. We can conclude that CIGB-210 is well tolerated in C57BL6 mice in the dose concentration range explored and merits subsequent toxicological studies.

The impacts of the COVID-19 pandemic on birth satisfaction and birth experiences in Russian women.

Background: Women's satisfaction with their childbirth experiences has significant impacts on their health and the health of their children. Recently, childbirth and maternity care systems have been disrupted by the COVID-19 pandemic. This study aimed to investigate the association of birth satisfaction with mode of birth, medical interventions, support during labour, type of childbirth healthcare plan and antenatal education in the context of the COVID-19 pandemic in Russia. Methods: 1,645 Russian women who gave birth during the first year of the COVID-19 pandemic and 611 matched controls who gave birth in the previous year participated in an anonymous Internet survey about their childbirth experience. The survey included questions regarding women's demographic and obstetric characteristics as well as their childbirth experiences. Birth satisfaction was measured using the Birth Satisfaction Scale Revised Indicator (BSS-RI). Results: Birth satisfaction scores did not show notable changes before and during the pandemic (Pearson Chi-square = 19.7, p = 0.22). Women had lower BSS-RI scores if they tested positive for COVID-19 during labour (F = 9.18, p = 0.002), but not during pregnancy or postpartum (p > 0.32). In both cohorts women who had vaginal births rated birth satisfaction higher than those who had caesarean births. The more medical interventions there were, the lower were the BSS-RI scores (B = -0.234, 95% CI: -0.760; -0.506, p < 0.001), but only during the pandemic. Birth satisfaction was higher if women had a support person present during labour (F > 7.44, p < 0.001), which was not possible for over 70% of participants during the pandemic. In both cohorts birth satisfaction was associated with the childbirth healthcare plan (F > 5.27, p < 0.001), but not with antenatal education (F < 0.15, p > 0.43). Conclusions: Our study highlights the significant impacts of the COVID-19 pandemic on the birth experiences of Russian women. Sustaining the rights of women to informed decisions during labour, respect for their preferred childbirth healthcare plan, presence of the birth team of choice and professional support for home birth are essential for higher birth satisfaction and better health outcomes for mothers and their infants.

Postpartum Depression and Birth Experience in Russia.

Background: In European countries, postpartum depression (PPD) occurs in 13-19% of women. The statistics indicate that postpartum depressive disorders affect up to 300,000 women in Russia annually. There is still an extremely acute lack of psychological comfort provided to women during labor in Russia. Objective: To our knowledge, ours is the first study that examines the association between childbirth experience and the risk of PPD in Russia. Design: We collected data from 190 Russian-speaking mothers, ages 19 to 46, (M = 32 ± 4.3) two months after their delivery. Results: Birth satisfaction and physical well-being two months after delivery were significantly inversely associated with PPD. Birth satisfaction negatively correlated with the perceived severity and unpredictability of labor, and positively correlated with physical well-being two months after delivery. The presence of a partner and a personal midwife or doula at birth was associated with higher birth satisfaction. Conclusions: Our results emphasize the significance of childbirth satisfaction in the context of PPD and suggest the importance of individual professional support during labor.

Birth Experience, Postpartum PTSD and Depression before and during the Pandemic of COVID-19 in Russia.

The aim of the study is to investigate the changes in the maternal healthcare system during the pandemic and their associations with maternal mental health in Russia. A sample of Russian women who gave birth during the first year of the COVID-19 pandemic (n = 1645) and matched controls, i.e., women who gave birth before the COVID-19 pandemic (n = 611), completed an anonymous Internet survey about recent childbirth. They were assessed for childbirth-related posttraumatic stress disorder (PTSD) and postpartum depression (PPD). Clinically relevant symptoms of PPD and PTSD were high before the pandemic and showed no significant change during the pandemic (p = 0.48 and p = 0.64, respectively). We found a notable increase in the frequency of obstetric violence (p = 0.015) during the pandemic, which, in turn, has a strong correlation with birth-related PTSD and PPD. The problem of ethical communication with patients among maternal healthcare professionals is acute in Russia, and it has been exacerbated by the pandemic. Family and doula support during labor can be a potential protective factor against obstetric violence.

A polyepigenetic glucocorticoid exposure score at birth and childhood mental and behavioral disorders.

BACKGROUND: Maternal depression and anxiety during pregnancy may enhance fetal exposure to glucocorticoids (GCs) and harm neurodevelopment. We tested whether a novel cross-tissue polyepigenetic biomarker indicative of in utero exposure to GC is associated with mental and behavioral disorders and their severity in children, possibly mediating the associations between maternal prenatal depressive and anxiety symptoms and these child outcomes. METHODS: Children (n = 814) from the Prediction and Prevention of Preeclampsia and Intrauterine Growth Restriction (PREDO) study were followed-up from birth to age 7.1-10.7 years. A weighted polyepigenetic GC exposure score was calculated based on the methylation profile of 24 CpGs from umbilical cord blood. Child diagnosis of mental and behavioral disorder (n = 99) and its severity, defined as the number of days the child had received treatment (all 99 had received outpatient treatment and 8 had been additionally in inpatient treatment) for mental or behavioral disorder as the primary diagnosis, came from the Care Register for Health Care. Mothers (n = 408) reported on child total behavior problems at child's age of 2.3-5.8 years and their own depressive and anxiety symptoms during pregnancy (n = 583). RESULTS: The fetal polyepigenetic GC exposure score at birth was not associated with child hazard of mental and behavioral disorder (HR = 0.82, 95% CI 0.54; 1.24, p = 0.35) or total behavior problems (unstandardized beta = -0.10, 95% CI -0.31; 0.10, p = 0.33). However, for one standard deviation decrease in the polyepigenetic score, the child had spent 2.94 (95%CI 1.59; 5.45, p < 0.001) more days in inpatient or outpatient treatment with any mental and behavioral disorder as the primary diagnosis. Criteria for mediation tests were not met. CONCLUSIONS: These findings suggest that fetal polyepigenetic GC exposure score at birth was not associated with any mental or behavioral disorder diagnosis or mother-rated total behavior problems, but it may contribute to identifying children at birth who are at risk for more severe mental or behavioral disorders.

Zonation of Ribosomal DNA Transcription Defines a Stem Cell Hierarchy in Colorectal Cancer.

Colorectal cancers (CRCs) are composed of an amalgam of cells with distinct genotypes and phenotypes. Here, we reveal a previously unappreciated heterogeneity in the biosynthetic capacities of CRC cells. We discover that the majority of ribosomal DNA transcription and protein synthesis in CRCs occurs in a limited subset of tumor cells that localize in defined niches. The rest of the tumor cells undergo an irreversible loss of their biosynthetic capacities as a consequence of differentiation. Cancer cells within the biosynthetic domains are characterized by elevated levels of the RNA polymerase I subunit A (POLR1A). Genetic ablation of POLR1A-high cell population imposes an irreversible growth arrest on CRCs. We show that elevated biosynthesis defines stemness in both LGR5+ and LGR5- tumor cells. Therefore, a common architecture in CRCs is a simple cell hierarchy based on the differential capacity to transcribe ribosomal DNA and synthesize proteins.

Impact of school-based nutrition and physical activity intervention on body mass index eight years after cessation of randomized controlled trial (AVall study).

BACKGROUND & AIMS: The prevalence of obesity and overweight among children is increasing, representing a new and pressing societal problem. Excess weight in childhood is an independent risk factor for adult obesity. Although school-based interventions are generally considered effective in the short term, few studies analyze their longterm impact. The aim of study was to assess if the effect of the intervention on body mass Index (BMI) immediately after the intervention remained eight years later (AVall Study). METHODS: In 2006-2008, an intervention for primary school children promoted healthy eating habits and physical activity in the school setting using the Investigation, Vision, Action and Change (IVAC) educational methodology, which involves children as active participants in healthy change. At baseline, parents reported their weight, height and educational level by questionnaire. Children's weight and height were measured in situ in 2006, 2008, 2010, 2012 and 2016, and levels of physical activity were assessed by questionnaire. Multilevel mixed effects linear regression was used to assess changes in BMI over time. RESULTS: Increases in BMI from 2006 to 2016 were 0.68 kg/m2 (95% CI, 0.02, 1.34; P = 0.045) higher in the control group than in the intervention group. The prevalence of obesity and overweight combined in 2006 and 2016 was 22.2% and 27.9%, respectively, in the control group and 25.6% and 21.2% respectively, in the intervention group. Changes in BMI in the intervention group were maintained from 2006 on: at the end of the intervention in 2008, -0.4 kg/m2 (P = 0.001); in 2010, -0.23 g/m2 (P = 0.012); in 2012, -0.63 kg/m2 (P < 0.001) and in 2016, -0.27 kg/m2 (P = 0.025). The child's BMI increased by 0.52 kg/m2 (P = 0.046) if the father was obese, by 1.26 kg/m2 (p = 0.011) if the mother was obese, and by 2.37 kg/m2 (P < 0.001) if both parents were obese. Parental education levels were not associated with childhood obesity. CONCLUSIONS: A school-based healthy eating habits and physical activity intervention using IVAC methodology contributed to lower increases in BMI among children in primary school. Although parental BMI also influenced children's BMI, the intervention was effective. TRIAL REGISTRATION: ClinicalTrials.gov NCT01156805.

The epigenetic clock and pubertal, neuroendocrine, psychiatric, and cognitive outcomes in adolescents.

BACKGROUND: Molecular aging biomarkers, such as epigenetic age predictors, predict risk factors of premature aging, and morbidity/mortality more accurately than chronological age in middle-aged and elderly populations. Yet, it remains elusive if such biomarkers are associated with aging-related outcomes earlier in life when individuals begin to diverge in aging trajectories. We tested if the Horvath epigenetic age predictor is associated with pubertal, neuroendocrine, psychiatric, and cognitive aging-related outcomes in a sample of 239 adolescents, 11.0-13.2 years-old. RESULTS: Each year increase in epigenetic age acceleration (AA) was associated with 0.06 SD units higher weight-for-age, 0.08 SD units taller height-for-age, -0.09 SD units less missed from the expected adult height, 13 and 16% higher odds, respectively, for each stage increase in breast/genitals development on the Tanner Staging Questionnaire and pubertal stage on the Pubertal Development Scale, 4.2% higher salivary cortisol upon awakening, and 18 to 34% higher odds for internalizing and thought problems on the Child Behavior Checklist (p values < 0.045). AA was not significantly associated with cognition. CONCLUSIONS: Our findings suggest that already in adolescence, AA is associated with physiological age acceleration, which may index risk of earlier aging. AA may identify individuals for preventive interventions decades before aging-related diseases become manifest.

The Epigenetic Clock at Birth: Associations With Maternal Antenatal Depression and Child Psychiatric Problems.

OBJECTIVE: Maternal antenatal depression may compromise the fetal developmental milieu and contribute to individual differences in aging and disease trajectories in later life. We evaluated the association between maternal antenatal depression and a novel biomarker of aging at birth, namely epigenetic gestational age (GA) based on fetal cord blood methylation data. We also examined whether this biomarker prospectively predicts and mediates maternal effects on early childhood psychiatric problems. METHOD: A total of 694 mothers from the Prediction and Prevention of Preeclampsia and Intrauterine Growth Restriction (PREDO) Study provided information on history of depression diagnosed before pregnancy; 581 completed the Center for Epidemiological Studies Depression Scale throughout pregnancy, and 407 completed the Child Behavior Checklist at child's age 3.7 years (SD = 0.75 year). DNA methylation (DNAm) GA of fetal cord blood DNA was based on the methylation profile of 148 selected cytosine linked to guanine by phosphate (CpG) sites. Epigenetic GA was calculated as the arithmetic difference between DNAm GA and chronological GA and adjusted for chronological GA. RESULTS: Maternal history of depression diagnosed before pregnancy (mean difference = -0.25 SD units, 95% CI = -0.46 to -0.03) and greater antenatal depressive symptoms (-0.08 SD unit per 1-SD unit increase, 95% CI = -0.16 to -0.004) were associated with child's lower epigenetic GA. Child's lower epigenetic GA, in turn, prospectively predicted total and internalizing problems and partially mediated the effects of maternal antenatal depression on internalizing problems in boys. CONCLUSION: Maternal antenatal depression is associated with lower epigenetic GA in offspring. This lower epigenetic GA seems to be associated with a developmental disadvantage for boys, who, in early childhood, show greater psychiatric problems.

FoxO1, A2M, and TGF-ß1: three novel genes predicting depression in gene X environment interactions are identified using cross-species and cross-tissues transcriptomic and miRNomic analyses.

To date, gene-environment (GxE) interaction studies in depression have been limited to hypothesis-based candidate genes, since genome-wide (GWAS)-based GxE interaction studies would require enormous datasets with genetics, environmental, and clinical variables. We used a novel, cross-species and cross-tissues "omics" approach to identify genes predicting depression in response to stress in GxE interactions. We integrated the transcriptome and miRNome profiles from the hippocampus of adult rats exposed to prenatal stress (PNS) with transcriptome data obtained from blood mRNA of adult humans exposed to early life trauma, using a stringent statistical analyses pathway. Network analysis of the integrated gene lists identified the Forkhead box protein O1 (FoxO1), Alpha-2-Macroglobulin (A2M), and Transforming Growth Factor Beta 1 (TGF-ß1) as candidates to be tested for GxE interactions, in two GWAS samples of adults either with a range of childhood traumatic experiences (Grady Study Project, Atlanta, USA) or with separation from parents in childhood only (Helsinki Birth Cohort Study, Finland). After correction for multiple testing, a meta-analysis across both samples confirmed six FoxO1 SNPs showing significant GxE interactions with early life emotional stress in predicting depressive symptoms. Moreover, in vitro experiments in a human hippocampal progenitor cell line confirmed a functional role of FoxO1 in stress responsivity. In secondary analyses, A2M and TGF-ß1 showed significant GxE interactions with emotional, physical, and sexual abuse in the Grady Study. We therefore provide a successful 'hypothesis-free' approach for the identification and prioritization of candidate genes for GxE interaction studies that can be investigated in GWAS datasets.

A genome editing approach to study cancer stem cells in human tumors.

The analysis of stem cell hierarchies in human cancers has been hampered by the impossibility of identifying or tracking tumor cell populations in an intact environment. To overcome this limitation, we devised a strategy based on editing the genomes of patient-derived tumor organoids using CRISPR/Cas9 technology to integrate reporter cassettes at desired marker genes. As proof of concept, we engineered human colorectal cancer (CRC) organoids that carry EGFP and lineage-tracing cassettes knocked in the LGR5 locus. Analysis of LGR5-EGFP+ cells isolated from organoid-derived xenografts demonstrated that these cells express a gene program similar to that of normal intestinal stem cells and that they propagate the disease to recipient mice very efficiently. Lineage-tracing experiments showed that LGR5+ CRC cells self-renew and generate progeny over long time periods that undergo differentiation toward mucosecreting- and absorptive-like phenotypes. These genetic experiments confirm that human CRCs adopt a hierarchical organization reminiscent of that of the normal colonic epithelium. The strategy described herein may have broad applications to study cell heterogeneity in human tumors.

Associations between maternal risk factors of adverse pregnancy and birth outcomes and the offspring epigenetic clock of gestational age at birth.

BACKGROUND: A recent study has shown that it is possible to accurately estimate gestational age (GA) at birth from the DNA methylation (DNAm) of fetal umbilical cord blood/newborn blood spots. This DNAm GA predictor may provide additional information relevant to developmental stage. In 814 mother-neonate pairs, we evaluated the associations between DNAm GA and a number of maternal and offspring characteristics. These characteristics reflect prenatal environmental adversity and are expected to influence newborn developmental stage. RESULTS: DNAm GA acceleration (GAA; i.e., older DNAm GA than chronological GA) of the offspring at birth was associated with maternal age of over 40 years at delivery, pre-eclampsia and fetal demise in a previous pregnancy, maternal pre-eclampsia and treatment with antenatal betamethasone in the index pregnancy, lower neonatal birth size, lower 1-min Apgar score, and female sex. DNAm GA deceleration (GAD; i.e., younger DNAm GA than chronological GA) of the offspring at birth was associated with insulin-treated gestational diabetes mellitus (GDM) in a previous pregnancy and Sjögren's syndrome. These findings were more accentuated when the DNAm GA calculation was based on the raw difference between DNAm GA and GA than on the residual from the linear regression of DNAm GA on GA. CONCLUSIONS: Our findings show that variations in the DNAm GA of the offspring at birth are associated with a number of maternal and offspring characteristics known to reflect exposure to prenatal environmental adversity. Future studies should be aimed at determining if this biological variation is predictive of developmental adversity.

Early Life Stress, FKBP5 Polymorphisms, and Quantitative Glycemic Traits.

OBJECTIVE: Early life stress (ELS) has been shown to influence health later in life. Functioning of the hypothalamic-pituitary-adrenal axis, regulated partly by FKBP5 gene, may moderate these effects. We examined whether FKBP5 single-nucleotide polymorphisms (SNPs) interact with ELS on Type 2 diabetes, cardiovascular disease, and quantitative glycemic traits. METHODS: A total of 1728 Helsinki Birth Cohort Study participants born from 1934 to 1944 were genotyped for FKBP5 SNPs (rs1360780, rs9394309, rs9470080) and were administered a 2-hour (75 g) oral glucose tolerance test and a questionnaire on physician-diagnosed and medication use for chronic diseases at a mean age of 61.5 years. Of the participants, 273 had been exposed to ELS, operationalized as separation from their parents, at a mean age of 4.7 years due to evacuations during World War II. RESULTS: ELS interacted with FKBP5 SNPs in the analyses of fasting (rs1360780, p = .015), 30-minute (rs1360780, p = .031; rs9394309, p = .041) and incremental insulin (rs1360780, p = .032; rs9394309, p = .028; rs9470080, p = .043), insulin area under the curve (rs1360780, p = .044), and impaired fasting glucose (rs9470080, p = .049); among carriers of at least one copy of minor allele, but not among major allele homozygotes, insulin values were higher, as were the odds for impaired fasting glucose if they had been separated compared with if they had not. Corresponding associations were found with a haplotype formed by minor alleles in all three SNPs for fasting, 30-minute, and incremental insulin (p < .05). CONCLUSIONS: FKBP5 polymorphisms in combination with ELS exposure predict higher insulin and glucose values in midlife. Our findings support the role for hypothalamic-pituitary-adrenal axis dysregulation in health-related metabolic outcomes.