A Novel Approach to Link Genetics and Human MRI Identifies AKAP7-Dependent Subicular/Prefrontal Functional Connectivity as Altered in Suicidality.

Background: Brain imaging and genetics are fields acquiring data at increasing speed, but more information does not always result in a better understanding of the underlying biology. We developed the ProcessGeneLists (PGL) approach to use genetics and mRNA gene expression data to generate regions of interest for imaging studies. Methods: We applied PGL to past suicide attempt (ATT): We averaged the mRNA expression levels of genes (n = 130) possibly associated with ATT (p ≤ 10-3 in a published genome-wide association study, GWAS) in each brain region studied in the Human Allen Brain Atlas (6 ex-vivo brains, 158 to 946 regions/brain have mRNA expression data) and compared that to the averaged mRNA expression levels of all other genes in each region in each brain in the atlas. Results: PGL revealed 8 regions where "attempt-related genes" were differentially expressed (Wilcoxon test with Bonferroni correction 8.88-11 =

Mega-analysis methods in ENIGMA: The experience of the generalized anxiety disorder working group.

The ENIGMA group on Generalized Anxiety Disorder (ENIGMA-Anxiety/GAD) is part of a broader effort to investigate anxiety disorders using imaging and genetic data across multiple sites worldwide. The group is actively conducting a mega-analysis of a large number of brain structural scans. In this process, the group was confronted with many methodological challenges related to study planning and implementation, between-country transfer of subject-level data, quality control of a considerable amount of imaging data, and choices related to statistical methods and efficient use of resources. This report summarizes the background information and rationale for the various methodological decisions, as well as the approach taken to implement them. The goal is to document the approach and help guide other research groups working with large brain imaging data sets as they develop their own analytic pipelines for mega-analyses.

Cortical and subcortical brain structure in generalized anxiety disorder: findings from 28 research sites in the ENIGMA-Anxiety Working Group.

The goal of this study was to compare brain structure between individuals with generalized anxiety disorder (GAD) and healthy controls. Previous studies have generated inconsistent findings, possibly due to small sample sizes, or clinical/analytic heterogeneity. To address these concerns, we combined data from 28 research sites worldwide through the ENIGMA-Anxiety Working Group, using a single, pre-registered mega-analysis. Structural magnetic resonance imaging data from children and adults (5-90 years) were processed using FreeSurfer. The main analysis included the regional and vertex-wise cortical thickness, cortical surface area, and subcortical volume as dependent variables, and GAD, age, age-squared, sex, and their interactions as independent variables. Nuisance variables included IQ, years of education, medication use, comorbidities, and global brain measures. The main analysis (1020 individuals with GAD and 2999 healthy controls) included random slopes per site and random intercepts per scanner. A secondary analysis (1112 individuals with GAD and 3282 healthy controls) included fixed slopes and random intercepts per scanner with the same variables. The main analysis showed no effect of GAD on brain structure, nor interactions involving GAD, age, or sex. The secondary analysis showed increased volume in the right ventral diencephalon in male individuals with GAD compared to male healthy controls, whereas female individuals with GAD did not differ from female healthy controls. This mega-analysis combining worldwide data showed that differences in brain structure related to GAD are small, possibly reflecting heterogeneity or those structural alterations are not a major component of its pathophysiology.

Cingulate Cortex Structural Alterations in Substance Use Disorder Psychiatric Inpatients.

BACKGROUND AND OBJECTIVES: Substance use disorder (SUD) includes maladaptive patterns of substance use despite negative consequences. Previous structural neuroimaging studies showed some structural alterations in SUD, but it remains unknown whether these alterations are specifically associated with SUD or common comorbidities. This study attempts to validate the findings of structural differences between SUD, healthy controls (HC), and psychiatric controls (PC). METHODS: We used HC (N = 86) matched for demographics, and PC (N = 86) matched for demographics and psychiatric diagnoses to a group of SUD patients (N = 86). We assessed the group differences of subcortical volumes, cortical volumes, thickness, and surface areas between SUD and HC. We then analyzed the group differences between SUD and PC within regions showing differences between SUD and HC. RESULTS: SUD had smaller left nucleus accumbens, right thalamus, right hippocampus, left caudal anterior cingulate cortex (ACC) volume, and larger right caudal ACC volume, and right caudal ACC, right caudal middle frontal gyrus (MFG), and right posterior cingulate cortex (PCC) surface than HC. Increased right caudal ACC volume and right PCC surface in SUD were the only findings when compared with PC. Several areas showed thickness alterations between SUD and HC, but none survived multiple comparisons vs PC. DISCUSSION AND CONCLUSIONS: Our findings suggest that cingulate structures may be altered in SUD compared with both HC and PC. SCIENTIFIC SIGNIFICANCE: These results are among the first to indicate that some structural alterations may be SUD-specific, and highlight a cautionary note about using HC in psychiatric biomarker research. (Am J Addict 2021;30:72-79).

Subcortical brain morphometry of avoidant personality disorder.

BACKGROUND: Avoidant personality disorder (AvPD) is a condition typified by social inhibition, feelings of inadequacy, and hypersensitivity to negative evaluation. AvPD has a high comorbidity rate with other personality disorders and other psychological diagnostic categories. There is very little research investigating subcortical volumetry in AvPD. We studied subcortical brain morphometry in AvPD as compared to both healthy controls and comorbidity-matched psychiatric controls (patients in the same clinic matched for age, sex and all psychiatric diagnoses except for AvPD). METHODS: We compared volumetric measures of 9 bilateral subcortical brain regions between AvPD patients, healthy controls, and psychiatric controls (n = 100 each group). The Bonferroni correction was used to control for multiple comparisons across regions (p < 0.0028). RESULTS: Compared to healthy controls, AvPD patients had lower volume of the left accumbens and left thalamus. However, no significant results were found when comparing AvPD patients and psychiatric controls. An exploratory study of cortical regions showed similar results: statistically significant differences between HC and AvPD (left lateral occipital, left and right pericalcarine smaller in AvPD) but no differences between AvPD and PC. LIMITATIONS: MRI and neuroimaging provides correlational information, and no causal claims can be made. CONCLUSIONS: These results suggest there may be no overt subcortical volumetric differences specific to AvPD, and provide strong cautionary advice when comparing patients to healthy controls, a common practice in psychiatry biomarker research.

Hippocampal Volume in Psychiatric Diagnoses: Should Psychiatry Biomarker Research Account for Comorbidities?

BACKGROUND: Many research papers claim that patients with specific psychiatric disorders (major depressive disorder, posttraumatic stress disorder, borderline personality disorder, alcohol use disorder, and others) have smaller hippocampi, but most of those reports compared patients to healthy controls. We hypothesized that if psychiatrically matched controls (psychiatric control, matched for demographics and psychiatric comorbidities) were used, much of the biomarker literature in psychiatric research would not replicate. We used hippocampus and amygdala volume only as examples, as these are very commonly replicated results in psychiatry biomarker research. We propose that psychiatry biomarker research could benefit from using psychiatric controls, as the use of healthy controls results in data that are not disorder-specific. METHOD: Hippocampus/amygdala volumes were compared between major depressive disorder, sex-/age-/race-matched healthy control, and psychiatric control (N = 126/group). Similar comparisons were performed for posttraumatic stress disorder (N = 67), borderline personality disorder (N = 111), and alcohol use disorder (N = 136). RESULTS: Major depressive disorder patients had smaller left (p = 8.79 × 10-3) and right (p = 3.13 × 10-3) hippocampal volumes than healthy control. Posttraumatic stress disorder had smaller left (p = 0.018) and right (p = 8.64 × 10-4) hippocampi than healthy control. Borderline personality disorder had smaller right hippocampus (p = 7.90 × 10-3) and amygdala (p = 1.49 × 10-3) than healthy control. Alcohol use disorder had smaller right hippocampus (p = 0.034) and amygdala (p = .024) than healthy control. No differences were found between any of the four diagnostic groups and psychiatric control. CONCLUSION: When psychiatric controls were used, there was no difference in hippocampal or amygdalar volume between any of the diagnoses studied and controls. This strategy (keeping all possible relevant variables matched between experimental groups) has been used to advance science for hundreds of years, and we propose should also be used in biomarker psychiatry research.

Orbitofrontal, dorsal striatum, and habenula functional connectivity in psychiatric patients with substance use problems.

Substance abuse is commonly defined as the persistence of drug use despite negative consequences. Recent preclinical work has shown that higher input from the orbitofrontal cortex (OFC) to the dorsal striatum was associated with compulsive reward-seeking behavior despite negative effects. It remains unknown whether drug use is associated with the connectivity between the OFC and dorsal striatum in humans. We studied the resting state functional connectivity (RSFC) between the OFC, dorsal striatum, and habenula (and the whole brain in a separate analysis) in psychiatric inpatients with high (PU, problem users) and low (LU, low users) substance use. We matched PU and LU for psychiatric comorbidities. We found that PU showed higher RSFC between the left OFC and the left dorsal striatum than LU. RSFC between the habenula and both OFC and dorsal striatum was also higher in PU, which suggests the habenula may be a part of the same circuit. Finally, higher RSFC between the OFC and insula was also observed in PU. Our data shows that OFC, habenula, dorsal striatum, and insula may play an important role in PU. Furthermore, we postulate that the habenula may link the mesolimbic and cortico-striatal systems, which are altered in PU.

Process genes list: An approach to link genetics and human brain imaging.

BACKGROUND: Linking human genetics and brain imaging data is extremely challenging, among other reasons because both fields suffer from multiple comparison problems. NEW METHOD: ProcessGeneLists (PGL) links genetics and human brain imaging by using genes associated with a disease and calculating a normalized mRNA expression average of those genes in each brain region. Brain regions in which those genes are most co-expressed become regions of interest (ROIs) to perform brain imaging in participants with and without the disease, decreasing multiple comparisons. Once a region is identified as "imaging-relevant", the genes most responsible for that ROI being highlighted can be genotyped in the imaged sample. This allows to re-analyze imaging data under the light of likely relevant genetics, to study possible brain imaging/gene variant interactions. RESULTS: As proof-of-concept, we created two lists of genes expressed in the habenula and the striatum, to verified that PGL would highlight those regions. Next, we used a list of genes likely important in alcohol abuse from the literature, which identified several brain regions previously associated with alcohol abuse such as the striatum, habenula, and hippocampus. COMPARISON WITH EXISTING METHODS: To our knowledge there is no current method to obtain brain regions of interest from genetics data. CONCLUSIONS: Genetics typically asks "which genes are associated with a disease?" while human brain imaging typically asks "which brain regions are associated with a disease?" PGL asks "which genes, via modulation within specific brain regions, are found to be associated with a disease?".

Right temporal pole volume reduction in PTSD.

Previous magnetic resonance imaging studies of post-traumatic stress disorder (PTSD) have reported cortical volume alterations in the parahippocampal, anterior cingulate cortex, and temporal pole. It is unclear, however, if these cortical regions are specifically associated with PTSD or associated with common comorbidities. Here, we present the result of cortical volume differences between PTSD and healthy and psychiatric controls. In this study, healthy controls (n = 67) were matched for demographic characteristics (age, sex, race) and psychiatric controls (n = 67) were matched for demographic characteristics plus all other psychiatric diagnoses (past and current) to a group of PTSD patients (N = 67). We assessed group differences of 34 bilateral cortical structure volumes using statistically defined brain regions-of-interest from FreeSurfer between PTSD patients and healthy controls. We found 10 regions to be significantly different between PTSD and healthy controls and analyzed the group differences between PTSD and psychiatric controls within these regions. The right temporal pole volume in PTSD was found to be significantly smaller than both healthy and psychiatry controls. Our finding suggests only right temporal pole volume reduction is specifically associated with PTSD, and also highlights the need for using appropriate controls in psychiatry research.

Altered habenula resting state functional connectivity in deprived veteran tobacco smokers: A pilot study.

This study aimed to examine habenular resting state functional connectivity (RSFC) abnormalities in tobacco-smoking veterans. The authors explored RSFC in sated smokers (n = 3D 18), overnight deprived smokers (n = 3D 13), and nonsmoker controls (n = 3D 26). Seed-to-voxel analysis was used to explore RSFC in the habenula. Compared to sated smokers, deprived smokers demonstrated higher RSFC between the right habenula and two clusters of voxels: one in the right fusiform gyrus, and one in the left lingual gyrus. To study nicotine withdrawal, the authors used the Shiffman-Jarvik Withdrawal Questionnaire (SJWQ) score as a regressor and found higher RSFC between the right habenula and the left frontal pole in deprived compared to sated smokers. Right habenula RSFC distinguished between sated and deprived smokers and differentiated between sated and deprived smokers when using SJWQ as a regressor, suggesting a habenular role in tobacco withdrawal.

Altered habenula to locus coeruleus functional connectivity in past anorexia nervosa suggests correlation with suicidality: a pilot study.

PURPOSE: Despite anorexia nervosa having the highest mortality rate of mental illnesses, little is known regarding the brain mechanisms involved. Given that lack of interest for food in anorexic patients is related to alterations in the reward system, we tested the hypothesis that patients with past anorexia nervosa (pAN) have altered resting state functional connectivity (RSFC) between the habenula (a major component of the reward system) and its targets. METHODS: RSFC between the habenula and major targets (locus coeruleus, median and dorsal raphe nuclei, substantia nigra, and ventral tegmental area) was studied in 14 psychiatric inpatients with pAN and 14 psychiatric inpatient controls (PC, never-anorexic patients in same clinic, matched for comorbidities). Next, we tested possible correlations between RSFC and suicidal ideation, depression, and anxiety as determined by self-report questionnaires. RESULTS: Left habenula/locus coeruleus RSFC was lower in pAN patients compared to PC. The left habenula/locus coeruleus RSFC was positively correlated with suicidal ideation (past 2 months) in pAN patients, but not in controls. CONCLUSIONS: pAN patients showed long lasting alterations in habenular connectivity. This may have clinical implications, possibly including future evaluation of the habenula as a therapeutic target and the need to carefully monitor suicidality in pAN patients. NO LEVEL OF EVIDENCE: Basic science.

Habenular connectivity may predict treatment response in depressed psychiatric inpatients.

INTRODUCTION: The habenula (Hb) is a small midbrain structure that signals negative events and may play a major role in the etiology of psychiatric disorders including depression. The lateral Hb has three major efferent connections: serotonergic raphe nuclei, noradrenergic locus coeruleus, and dopaminergic ventral tegmental area/substantia nigra compacta. We wanted to test whether Hb connectivity may be important to predict treatment outcomes in depression patients. METHODS: We studied whether habenular connectivity at admission into a psychiatric clinic can predict treatment response. We used an inpatient sample (N = 175) to assess habenular connectivity (diffusion tensor imaging and resting state functional connectivity (RSFC) between the Hb and its targets) close to admission. In addition, we obtained the Patient Health Questionnaire-depression module (PHQ-9) close to admission and at discharge. Inpatients in the study entered the clinic with at least moderately severe depression (score 15 and up). Inpatients considered treatment resistant had scores of 9 or more at discharge. RESULTS: Compared to responders, treatment non-responders had lower fractional anisotropy in the right Hb afferent fibers and lower RSFC between right Hb and median raphe, but higher RSFC between left Hb and locus coeruleus. A logistic regression model was significantly different from chance, and explained 27.7% of the variance in treatment resistance (sensitivity = 75%; specificity = 71.9%). DISCUSSION: The anatomical and functional connectivity of the Hb may be a predictor of treatment success in psychiatric populations. Limitations include the Hb small size and the limited time (5 min) of resting state data obtained.

Insular resting state functional connectivity is associated with gut microbiota diversity.

The gut microbiota has recently gained attention as a possible modulator of brain activity. A number of reports suggest that the microbiota may be associated with neuropsychiatric conditions such as major depressive disorder, autism and anxiety. The gut microbiota is thought to influence the brain via vagus nerve signalling, among other possible mechanisms. The insula processes and integrates these vagal signals. To determine if microbiota diversity and structure modulate brain activity, we collected faecal samples and examined insular function using resting state functional connectivity (RSFC). Thirty healthy participants (non-smokers, tobacco smokers and electronic cigarette users, n = 10 each) were studied. We found that the RSFC between the insula and several regions (frontal pole left, lateral occipital cortex right, lingual gyrus right and cerebellum 4, 5 and vermis 9) were associated with bacterial microbiota diversity and structure. In addition, two specific bacteria genera, Prevotella and Bacteroides, were specifically different in tobacco smokers and also associated with insular connectivity. In conclusion, we show that insular connectivity is associated with microbiome diversity, structure and at least two specific bateria genera. Furthemore, this association is potentially modulated by tobacco smoking, although the sample sizes for the different smoking groups were small and this result needs validation in a larger cohort. While replication is necessary, the microbiota is a readily accessible therapeutic target for modulating insular connectivity, which has previously been shown to be abnormal in anxiety and tobacco use disorders.

Prefrontal cortex, temporal cortex, and hippocampus volume are affected in suicidal psychiatric patients.

Suicide is a leading cause of death in America, with over 40,000 reported suicides per year. Mental illness is a major risk factor for suicidality. This study attempts to validate findings of volumetric differences from studies on suicidality. Psychiatric inpatients classified as having mildly severe or severe depression were separated into two groups: suicide attempted in the past two months (SA; n=20), non-suicidal control group (DA; n=20); these patients were all depressed and not significantly different for age, gender, race, marital status, education level, anxiety level, and substance abuse. Healthy controls (HC; n=20) were not significantly different from the suicidal groups for age and gender. Volunteers underwent MRI to assess volumes of cortical lobes, corpus callosum, and subcortical regions of interest, including the thalamus, insula, limbic structures, and basal ganglia. The right hippocampal volume of the SA group was significantly reduced compared to healthy controls. The frontal and temporal lobe volumes of the SA group were significantly decreased compared to the DA group. These volumetric reductions confirm previous findings and support the hypothesis that fronto-temporal function may be altered in suicidal patients.