Single-cell RNA sequencing reveals small extracellular vesicles derived from malignant cells that contribute to angiogenesis in human breast cancers.

BACKGROUND: Breast cancer is the most common cancer affecting women across the world. Tumor endothelial cells (TECs) and malignant cells are the major constituents of the tumor microenvironment (TME), but their origin and role in shaping disease initiation, progression, and treatment responses remain unclear due to significant heterogeneity. METHODS: Tissue samples were collected from eight patients presenting with breast cancer. Single-cell RNA sequencing (scRNA-seq) analysis was employed to investigate the presence of distinct cell subsets in the tumor microenvironment. InferCNV was used to identify cancer cells. Pseudotime trajectory analysis revealed the dynamic process of breast cancer angiogenesis. We validated the function of small extracellular vesicles (sEVs)-derived protein phosphatase 1 regulatory inhibitor subunit 1B (PPP1R1B) in vitro experiments. RESULTS: We performed single-cell transcriptomics analysis of the factors associated with breast cancer angiogenesis and identified twelve subclusters of endothelial cells involved in the tumor microenvironment. We also identified the role of TECs in tumor angiogenesis and confirmed their participation in different stages of angiogenesis, including communication with other cell types via sEVs. Overall, the research uncovered the TECs heterogeneity and the expression levels of genes at different stages of tumor angiogenesis. CONCLUSIONS: This study showed sEVs derived from breast cancer malignant cells promote blood vessel formation by activating endothelial cells through the transfer of PPP1R1B. This provides a new direction for the development of anti-angiogenic therapies for human breast cancer.

The influence of high worry on static and dynamic insular functional connectivity.

Worry is a form of repetitive negative thought. High worry-proneness is one risk factor leading to anxiety disorder. Several types of research indicated that anxiety disorder was highly associated with disrupted interoception. The insula is consistently considered to play a key role in interoception. However, the relationship between worry and the interoception network is poorly investigated in worry-prone individuals. Thus, it is essential to identify the neural characteristic of high worry-proneness subjects. A total of 32 high worry-proneness (HWP) subjects and 25 low worry-proneness (LWP) subjects were recruited and underwent magnetic resonance imaging scanning. Six subregions of insula were chosen as regions of interest. Then, seed-based static and dynamic functional connectivity were calculated. Increased static functional connectivity was observed between the ventral anterior insula and inferior parietal lobule in HWP compared to LWP. Decreased static functional connectivity was found between the left ventral anterior insula and the pregenual anterior cingulate cortex. Decreased dynamic functional connectivity was also shown between the right posterior insula and the inferior parietal lobule in HWP. Moreover, a post-hoc test exploring the effect of changed function within the insular region confirmed that a significant positive relationship between static functional connectivity (ventral anterior insula-inferior parietal lobule) and dynamic functional connectivity (posterior insula-inferior parietal lobule) in LWP but not in HWP. Our results might suggest that deficient insular function may be an essential factor related to high worry in healthy subjects.

Erratum: lncRNA LCPAT1 Upregulation Promotes Breast Cancer Progression via Enhancing MFAP2 Transcription.

[This corrects the article DOI: 10.1016/j.omtn.2020.07.015.].

The Neural Mechanisms of Cognitive Control in the Category Induction Task.

According to the conflict monitoring hypothesis, conflict monitoring and inhibitory control in cognitive control mainly cause activity in the anterior cingulate cortex (ACC) and control-related prefrontal cortex (PFC) in many cognitive tasks. However, the role of brain regions in the default mode network (DMN) in cognitive control during category induction tasks is unclear. To test the role of the ACC, PFC, and subregions of the DMN elicited by cognitive control during category induction, a modified category induction task was performed using simultaneous fMRI scanning. The results showed that the left middle frontal gyrus (BA9) and bilateral dorsal ACC/medial frontal gyrus (BA8/32) were sensitive to whether conflict information (with/without) appears, but not to the level of conflict. In addition, the bilateral ventral ACC (BA32), especially the right vACC, a part of the DMN, showed significant deactivation with an increase in cognitive effort depending on working memory. These findings not only offer further evidence for the important role of the dorsolateral PFC and dorsal ACC in cognitive control during categorization but also support the functional distinction of the dorsal/ventral ACC in the category induction task.

Frequency-specific alternations in the moment-to-moment BOLD signals variability in schizophrenia.

Variability of neuronal activity is considered as the fundamental mechanism for the flexible and optimal brain function. Moreover, different frequency neuro signal is related to specific function. While little is currently known regarding changes in spontaneous BOLD variability of schizophrenia. The current study used resting-state fMRI data from 53 chronic schizophrenic subjects and 67 healthy subjects to investigate this issue. The data-driven method was used to measure the BOLD variability (MSSD: mean square successive difference) in two different frequency bands respectively (slow-5: 0.01-0.027 Hz; slow-4:0.027-0.073 Hz). Schizophrenic subjects exhibited decreased BOLD variability in thalamus region, sensorimotor and visual networks, and increased BOLD variability in salience network compared to matched healthy controls. Moreover, the interaction effects between frequency and group were observed in thalamus and right dorsolateral prefrontal cortex (DLPFC). These findings identified that altered BOLD variability is frequency dependent in schizophrenia. Importantly, the severity of patients' negative symptom was related to the increased BOLD variability of DLPFC within slow-4 frequency band, highlighting the evidence that abnormal BOLD variability of frontal cortex is likely to have effects on the pathophysiology of negative symptom in schizophrenia.

Catalytic ozonation of norfloxacin using Co3O4/C composite derived from ZIF-67 as catalyst.

In this study, Co3O4-carbon composite was synthesized by calcined metal organic framework (MOF) ZIF-67 and used as efficient catalysts for ozonation of norfloxacin (NOF). The MOF-derived Co3O4-C composite remained similar polyhedrons structure of ZIF-67, suggesting that Co3O4 was well-dispersed in Co3O4-C composite. Furthermore, a larger amount of surface carbon-oxygen functional groups were distributed on Co3O4-C composite, which resulted in the diversification of active sites for catalytic ozonation reaction. NOF degradation and mineralization could be effectively enhanced in Co3O4-C/O3 process. Moreover, NOF mineralization by catalytic ozonation strongly depended on the solution pH, while other operational conditions, such as O3 concentration and catalyst dosage had not obvious influence on it. Co3O4-C composite could significantly accelerate O3 decomposition to produce active free radicals (such as •OH), which enhanced the mineralization of NOF. The possible catalytic mechanism of Co3O4-C composite was proposed. Additionally, after five consecutive use of Co3O4-C composite in catalytic ozonation process, there was no obvious decrease in TOC removal efficiency, indicating a stable performance of Co3O4-C composite, which was suitable for the catalytic ozonation for wastewater treatment.

lncRNA LCPAT1 Upregulation Promotes Breast Cancer Progression via Enhancing MFAP2 Transcription.

The importance of long noncoding RNA (lncRNA) in tumorigenesis has been supported by increasing evidence in recent years. However, the mechanism linking lncRNA function with cancer progression remains poorly understood. lncRNA LCPAT1 plays a role in lung cancer. However, how it works in breast cancer (BC) is largely unclear. In this study, we found that LCPAT1 was highly expressed in BC tissues and cell lines. High LCPAT1 expression predicted a low survival rate in BC patients. LCPAT1 promoted BC cell proliferation, migration, and invasion while inhibiting apoptosis in vitro. LCPAT1 knockdown suppressed BC growth in vivo and vice versa. LCPAT1 interacted with RBBP4 and recruited it to the MFAP2 (microfibril-associated protein 2) promoter and then activated MFAP2 transcription. Restoration of MFAP2 rescued the effects of LCPAT1 knockdown in BC cells. In sum, LCPAT1 promotes BC progression through recruiting RBBP4 to initiate MFAP2 transcription.

Increased Temporal Dynamics of Intrinsic Brain Activity in Sensory and Perceptual Network of Schizophrenia.

Schizophrenic subject is thought as a self-disorder patient related with abnormal brain functional network. It has been hypothesized that self-disorder is associated with the deficient functional integration of multisensory body signals in schizophrenic subjects. To further verify this assumption, 53 chronic schizophrenic subjects and 67 healthy subjects were included in this study and underwent resting-state functional magnetic resonance imaging. The data-driven methods, whole-brain temporal variability of fractional amplitude of low-frequency fluctuations and regional homogeneity (ReHo), were used to investigate dynamic local functional connectivity and dynamic local functional activity changes in schizophrenic subjects. Patients with schizophrenia exhibited increased temporal variability ReHo and fractional amplitude of low-frequency fluctuations across time windows within sensory and perception network (such as occipital gyrus, precentral and postcentral gyri, superior temporal gyrus, and thalamus). Critically, the increased dynamic ReHo of thalamus is significantly correlated with positive and total symptom of schizophrenic subjects. Our findings revealed that deficit in sensory and perception functional networks might contribute to neural physiopathology of self-disorder in schizophrenic subjects.

Frequency-specific alteration of functional connectivity density in antipsychotic-naive adolescents with early-onset schizophrenia.

Early-onset schizophrenia (EOS) is a severe mental illness associated with dysconnectivity that widespread in the brain. However, the functional dysconnectivity in EOS are still mixed. Recently, studies have identified that functional connectivity (FC) arises from a band-limited slow rhythmic mechanism and suggested that the dysconnectivity at specific frequency bands may provide more robust biomarkers for schizophrenia. The frequency-specific changes of FC pattern in EOS remain unclear. To address this issue, resting-state functional magnetic resonance imaging data scans from 39 EOS patients (drug-naive) and 31 healthy controls (HCs) were used to assess the FC density (FCD) across slow-4 (0.027-0.073 Hz) and slow-5 (0.01-0.027 Hz). Results revealed that a remarkable difference between the FCD of the two bands existed mainly in the default mode network (DMN) and subcortical areas. Compared with the HCs, EOS patients showed significantly altered FCD involved in audiovisual information processing, sensorimotor system, and social cognition. Importantly, a significant frequency-by-group interaction was observed in the left precuneus with significantly lower FCD in the slow-4 frequency band, but no significant effect in the slow-5 frequency band. In addition, decreased FC was found between the precuneus and other DMN regions in the slow-4 band. Furthermore, the change in FCD in precuneus was inversely proportional to the clinical symptom in slow-4 band, indicating the key role of precuneus in schizophrenia progress. Our findings demonstrated that the dysconnectivity pattern in EOS could be frequency-dependent.

Shared atypical default mode and salience network functional connectivity between autism and schizophrenia.

Schizophrenia and autism spectrum disorder (ASD) are two prevalent neurodevelopmental disorders sharing some similar genetic basis and clinical features. The extent to which they share common neural substrates remains unclear. Resting-state fMRI data were collected from 35 drug-naïve adolescent participants with first-episode schizophrenia (15.6 ± 1.8 years old) and 31 healthy controls (15.4 ± 1.6 years old). Data from 22 participants with ASD (13.1 ± 3.1 years old) and 21 healthy controls (12.9 ± 2.9 years old) were downloaded from the Autism Brain Imaging Data Exchange. Resting-state functional networks were constructed using predefined regions of interest. Multivariate pattern analysis combined with multi-task regression feature selection methods were conducted in two datasets separately. Classification between individuals with disorders and controls was achieved with high accuracy (schizophrenia dataset: accuracy = 83%; ASD dataset: accuracy = 80%). Shared atypical brain connections contributing to classification were mostly present in the default mode network (DMN) and salience network (SN). These functional connections were further related to severity of social deficits in ASD (p = 0.002). Distinct atypical connections were also more related to the DMN and SN, but showed different atypical connectivity patterns between the two disorders. These results suggest some common neural mechanisms contributing to schizophrenia and ASD, and may aid in understanding the pathology of these two neurodevelopmental disorders. Autism Res 2017, 10: 1776-1786. © 2017 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: Autism spectrum disorder (ASD) and schizophrenia are two common neurodevelopmental disorders which share several genetic and behavioral features. The present study identified common neural mechanisms contributing to ASD and schizophrenia using resting-state functional MRI data. The results may help to understand the pathology of these two neurodevelopmental disorders.

Atypical developmental trajectory of local spontaneous brain activity in autism spectrum disorder.

Autism spectrum disorder (ASD) is marked by atypical trajectory of brain maturation, yet the developmental abnormalities in brain function remain unclear. The current study examined the effect of age on amplitude of low-frequency fluctuations (ALFF) in ASD and typical controls (TC) using a cross-sectional design. We classified all the participants into three age cohorts: child (<11 years, 18ASD/20TC), adolescent (11-18 years, 28ASD/26TC) and adult (≥18 years, 18ASD/18TC). Two-way analysis of variance (ANOVA) was performed to ascertain main effects and interaction effects on whole brain ALFF maps. Results exhibited significant main effect of diagnosis in ASD with decreased ALFF in the right precuneus and left middle occipital gyrus during all developmental stages. Significant diagnosis-by-age interaction was observed in the medial prefrontal cortex (mPFC) with ALFF lowered in autistic children but highered in autistic adolescents and adults. Specifically, remarkable quadratic change of ALFF with increasing age in mPFC presented in TC group was absent in ASD. Additionally, abnormal ALFF values in diagnosis-related brain regions predicted the social deficits in ASD. Our findings indicated aberrant developmental patterns of spontaneous brain activity associated with social deficits in ASD and highlight the crucial role of the default mode network in the development of disease.

Decreased amygdala functional connectivity in adolescents with autism: A resting-state fMRI study.

The human brain undergoes dramatic changes in amygdala-related functional connectivity network during adolescence. Given that the amygdala is a vital component of the "social brain", the Amygdala Theory of Autism has been proposed to account for atypical patterns of socio-emotional behavior in autism. Most of the previous neuroimaging evidence has concentrated on local functional or structural abnormalities of the amygdala in relation to social deficits in autism, rather than on its integrated role as part of larger brain networks. To examine whether functional integration pattern of the amygdala is altered in autism, the current study examined sixty-five adolescent subjects (30 autism and 35 healthy controls, 12-18 years old) from two independent datasets (UCLA and Leuven) of the Autism Brain Imaging Data Exchange. Whole-brain resting-state functional connectivity maps seeded in the amygdala were calculated and compared between patient and control groups. Compared with healthy controls, adolescents with autism showed decreased functional connectivity between the amygdala and subcortical regions in both datasets, including the bilateral thalamus and right putamen. These findings support the Amygdala Theory of Autism, demonstrating altered functional connectivity pattern associated with the amygdala in autism, and provide new insights into the pathophysiology of autism.

Atypical effective connectivity of thalamo-cortical circuits in autism spectrum disorder.

Autism spectrum disorder (ASD) is a neurodevelopment disorder characterized by atypical connectivity within and across multiple brain systems. We aimed to explore information transmission from the sensory periphery to information processing centers of the brain across thalamo-cortical circuits in ASD. A large multicenter dataset from the autism brain imaging data exchange was utilized. A thalamus template derived from the Automatic Anatomic Labeling atlas was subdivided into six subregions corresponding to six cortical regions using a "winner-takes-all" strategy. Granger causality analysis (GCA) was then applied to calculate effective connectivity from subregions of the thalamus to the corresponding cortical regions. Results demonstrate reduced effective connectivity from the thalamus to left prefrontal cortex (P = 0.023), right posterior parietal cortex (P = 0.03), and bilateral temporal cortex (left: P = 0.014; right: P = 0.015) in ASD compared with healthy control (HC) participants. The GCA values of the thalamus-bilateral temporal cortex connections were significantly negatively correlated with communication scores as assessed by the autism diagnostic observation schedule in the ASD group (left: P = 0.037; right: P = 0.007). Age-related analyses showed that the strengths of the thalamus-bilateral temporal cortex connections were significantly positively correlated with age in the HC group (left: P = 0.013; right: P = 0.016), but not in the ASD group (left: P = 0.506; right: P = 0.219). These results demonstrate impaired thalamo-cortical information transmission in ASD and suggest that atypical development of thalamus-temporal cortex connections may relate to communication deficits in the disorder. Autism Res 2016, 9: 1183-1190. © 2016 International Society for Autism Research, Wiley Periodicals, Inc.

Transcriptome sequencing of HER2-positive breast cancer stem cells identifies potential prognostic marker.

In cancer stem cell theory, breast cancer stem cells (BCSCs) are postulated to be the root cause of recurrence and metastasis in breast cancer. Discovery of new biomarkers and development of BCSC-targeted therapy are practical issues that urgently need to be addressed in the clinic. However, few breast cancer stem cell targets are known. Given that there are few BCSCs, performing transcriptome sequencing on them thus far has not been possible. With the emergence of single-cell sequencing technology, we have now undertaken such a study. We prepared single-cell suspensions, which were sorted using flow cytometry from breast tumor tissue and adjacent normal breast tissue from two HER2-positive patients. We obtained BCSCs, breast cancer cells, mammary cells, and CD44+ mammary cells. Transcriptome sequencing was then performed on these four cell types. Using bioinformatics, we identified 404 differentially expressed BCSC genes from the HER2-positive tumors and preliminary explored transcriptome characteristics of BCSCs. Finally, by querying a public database, we found that CA12 was a novel prognostic biomarker in HER2-positive breast cancer, which also had prognostic value in all breast cancer types. In conclusion, our results suggest that CA12 may be associated with BCSCs, especially HER2-positive BCSCs, and is a potential novel therapeutic target and biomarker.

Disrupted resting-state insular subregions functional connectivity in post-traumatic stress disorder.

Post-traumatic stress disorder (PTSD) is suggested to be a structural and functional abnormality in the insula. The insula, which consists of distinct subregions with various patterns of connectivity, displays complex and diverse functions. However, whether these insular subregions have different patterns of connectivity in PTSD remains unclear. Investigating the abnormal functional connectivity of the insular subregions is crucial to reveal its potential effect on diseases specifically PTSD. This study uses a seed-based method to investigate the altered resting-state functional connectivity of insular subregions in PTSD. We found that patients with PTSD showed reduced functional connectivity compared with healthy controls (HCs) between the left ventral anterior insula and the anterior cingulate cortex. The patients with PTSD also exhibited decreased functional connectivity between the right posterior insula and left inferior parietal lobe, and the postcentral gyrus relative to HCs. These results suggest the involvement of altered functional connectivity of insular subregions in the abnormal regulation of emotion and processing of somatosensory information in patients with PTSD. Such impairments in functional connectivity patterns of the insular subregions may advance our understanding of the pathophysiological basis underlying PTSD.

Frequency-specific alterations in the fractional amplitude of low-frequency fluctuations in amyotrophic lateral sclerosis.

This study used resting-state functional magnetic resonance imaging and fractional amplitude of low-frequency fluctuations (fALFF) method to investigate low-frequency spontaneous neural activity at the bands of slow-5 (0.01-0.027 Hz) and slow-4 (0.027-0.073 Hz) in 20 patients with amyotrophic lateral sclerosis (ALS) and 20 healthy controls. We determined that, at slow-5 band, patients with ALS showed increased fALFF in the right middle frontal gyrus and decreased fALFF in the left middle occipital gyrus. However, compared with healthy controls, patients with ALS exhibited higher fALFF in the right caudate nucleus, left superior frontal gyrus, and right anterior cingulate cortex and lower fALFF in the right inferior occipital gyrus and bilateral middle occipital gyrus at slow-4 band. Furthermore, the fALFF value in the left superior frontal gyrus at slow-4 band was negatively correlated with functional rating scale-revised score. Our results demonstrated that the fALFF changes in ALS were widespread and frequency dependent. These findings may provide a novel way to look into the pathophysiology mechanisms underlying ALS.

Abnormal Functional Connectivity Density in Post-traumatic Stress Disorder.

Post-traumatic stress disorder (PTSD) is a psychiatric disorder that occurs in individuals who have experienced life-threatening mental traumas. Previous neuroimaging studies have indicated that the pathology of PTSD may be associated with the abnormal functional integration among brain regions. In the current study, we used functional connectivity density (FCD) mapping, a novel voxel-wise data-driven approach based on graph theory, to explore aberrant FC through the resting-state functional magnetic resonance imaging of the PTSD. We calculated both short- and long-range FCD in PTSD patients and healthy controls (HCs). Compared with HCs, PTSD patients showed significantly increased long-range FCD in the left dorsolateral prefrontal cortex (DLPFC), but no abnormal short-range FCD was found in PTSD. Furthermore, seed-based FC analysis of the left DLPFC showed increased connectivity in the left superior parietal lobe and visual cortex of PTSD patients. The results suggested that PTSD patients experienced a disruption of intrinsic long-range functional connections in the fronto-parietal network and visual cortex, which are associated with attention control and visual information processing.

Multivariate classification of autism spectrum disorder using frequency-specific resting-state functional connectivity--A multi-center study.

BACKGROUND: Resting-state functional magnetic resonance imaging studies examining low frequency fluctuations (0.01-0.08 Hz) have revealed atypical whole brain functional connectivity patterns in adolescents with autism spectrum disorder (ASD), and these atypical patterns can be used to discriminate individuals with ASD from controls. However, at present it is unknown whether functional connectivity at specific frequency bands can be used to discriminate individuals with ASD from controls, and whether relationships with symptom severity are stronger in specific frequency bands. METHODS: We selected 240 adolescent subjects (12-18 years old, 112 with autism spectrum disorder (101/11, males/females) and 128 healthy controls (104/24, males/females)) from 6 separate international sites in the Autism Brain Imaging Data Exchange database. Whole brain functional connectivity networks were constructed in the Slow-5 (0.01-0.027 Hz) and Slow-4 (0.027-0.073 Hz) frequency bands, which were then used as classification features. RESULTS: An accuracy of 79.17% (p<0.001) was obtained using support vector machine. Most of the discriminative features were concentrated on the Slow-4 band. In the Slow-4 band, atypical connections between the default mode network, fronto-parietal network and cingulo-opercular network were detected. A significant correlation was found between social and communication deficits as measured by the ADOS in individuals with ASD and the classification scores based on connectivity between the default mode network and the cingulo-opercular network. Connections of the thalamus were of the highest classification weight in the Slow-4 band. CONCLUSIONS: Our findings provide preliminary evidence for frequency-specific whole brain functional connectivity indices that may eventually be used to aid detection of ASD.

Intranetwork and internetwork functional connectivity alterations in post-traumatic stress disorder.

BACKGROUND: A large number of previous neuroimaging studies have explored the functional alterations of post-traumatic stress disorder (PTSD). However, abnormalities in the functional architecture of resting-state networks in PTSD were rarely elucidated. METHODS: This study used independent component analysis to explore the resting-state intranetwork and internetwork functional connectivity differences between 20 PTSD patients and 20 matched healthy controls (HCs). RESULTS: Selective alterations of intranetwork and internetwork intrinsic functional connectivities were found in the PTSD patients. Compared with HCs, the PTSD patients exhibited significantly decreased network connectivity within the anterior default mode network, posterior default mode network (pDMN), salience network (SN), sensory-motor network, and auditory network. Furthermore, the PTSD patients exhibited increased internetwork connectivity between SN and pDMN. LIMITATIONS: This study lacked recruitment of trauma-exposed HCs, which limits our ability to determine whether the alterations are caused by PTSD or trauma exposure. CONCLUSION: The findings suggested that the PTSD patients exhibited abnormal functional connectivity at the brain network level. Notably, the enhanced internetwork connectivity between SN and pDMN in the PTSD patients may be associated with hyperarousal and heightened anxiety in PTSD.

Altered cortical hubs in functional brain networks in amyotrophic lateral sclerosis.

Cortical hubs are highly connected nodes in functional brain networks that play vital roles in the efficient transfer of information across brain regions. Although altered functional connectivity has been found in amyotrophic lateral sclerosis (ALS), the changing pattern in functional network hubs in ALS remains unknown. In this study, we applied a voxel-wise method to investigate the changing pattern of cortical hubs in ALS. Through resting-state fMRI, we constructed whole-brain voxel-wise functional networks by measuring the temporal correlations of each pair of brain voxels and identified hubs using the graph theory method. Specifically, a functional connectivity strength (FCS) map was derived from the data on 20 patients with ALS and 20 healthy controls. The brain regions with high FCS values were regarded as functional network hubs. Functional hubs were found mainly in the bilateral precuneus, parietal cortex, medial prefrontal cortex, and in several visual regions and temporal areas in both groups. Within the hub regions, the ALS patients exhibited higher FCS in the prefrontal cortex compared with the healthy controls. The FCS value in the significantly abnormal hub regions was correlated with clinical variables. Results indicated the presence of altered cortical hubs in the ALS patients and could therefore shed light on the pathophysiology mechanisms underlying ALS.