Ontology highlight
ABSTRACT:
PROVIDER: PRJNA552157 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR9617930_1.fastq.gz | Fastqsanger.gz | |||
| SRR9617930_2.fastq.gz | Fastqsanger.gz | |||
| SRR9617940_1.fastq.gz | Fastqsanger.gz | |||
| SRR9617940_2.fastq.gz | Fastqsanger.gz | |||
| SRR9617931_1.fastq.gz | Fastqsanger.gz |
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Project description:The etiology of severe forms of COVID19, especially in young patients, remains a salient unanswered question. Here we build on a 3-tier cohort where all individuals/patients were strictly below 50 years of age and where a number of comorbidities were excluded at study onset. Besides healthy controls (N=22), these include patients in the intensive care unit with Acute Respiratory Distress Syndrome (ARDS) (“critical group”; N=47), and those in a non-critical care ward under supplemental oxygen (“non-critical group”, N=25). This highly curated cohort allowed us to perform a deep multi-omics approach which included whole genome sequencing, whole blood RNA-sequencing, plasma and peripheral-blood mononuclear cells proteomics, multiplex cytokine profiling, mass-cytometry-based immune cell profiling in conjunction with viral parameters i.e. anti-SARS-Cov-2 neutralizing antibodies and multi-target antiviral serology. Critical patients were characterized by an exacerbated inflammatory state, perturbed lymphoid and myeloid cell compartments, signatures of dysregulated blood coagulation and active regulation of viral entry into the cells. A unique gene signature that differentiates critical from non-critical patients was identified by an ensemble machine learning, deep learning and quantum computing approach. Within this gene network, Structural Causal Modeling identified several ARDS driver genes, among which the up-regulated metalloprotease ADAM9 seems to be a key driver. Inhibition of ADAM9 ex vivo interfered with SARS-Cov-2 uptake and replication in human epithelial cells. Hence we apply a machine learning approach to identify driver genes for severe forms of COVID-19 in a small, uncluttered cohort of patients.
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Project description:The molecular and functional characteristics of peripheral blood immune cells in childhood may contribute to the specialized immune system in children. At present, few researches have compared immune cells in childhood with other age groups, and there is a lack of understanding of immune cells in childhood from the whole lifespan. To display the molecular characteristics of peripheral blood immune cells in childhood at the single-cell level, we performed a comparative study by constructing a peripheral blood mononuclear cell (PBMC) atlas of multiple age groups, including children, young adults, middle-aged adults, aged adults and supercentenarians. The atlas revealed that monocytes in childhood were markedly different from those in other periods. B cells in childhood had a greater ability to respond to viruses and to activate mast cells. Remarkably, we identified a new subtype of CD8+ cytotoxic T cells expressing both naïve and cytotoxicity genes specifically in childhood. The marker genes of this subtypes were used as marker proteins for flow cytometry analysis, and the results confirmed the findings of the scRNA-seq. Moreover, there was more frequent crosstalk between PBMCs in childhood than in other periods, which may be one of the characteristics of the immune system in childhood. Finally, the differences in the expression of susceptibility genes in the PBMCs of different age groups may indicate differences in the disease spectrum of different age groups. In conclusion, this research systematically described the molecular characteristics of peripheral blood immune cells in childhood from multiple levels, and provided new evidence to elucidate the special immune environment of childhood.
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Project description:Although the nose, as a gateway for organism-environment interactions, may have a key role in asthmatic exacerbation, the rhinobiome of exacerbated children with asthma was widely neglected to date. Deep nasopharyngeal swab specimens, nasal epithelial spheroid cultures (NAEsp), and blood samples of acute exacerbated wheezers (WH), asthmatics (AB), and healthy controls (HC) were used for culture (n=146), 16 S-rRNA gene amplicon sequencing (n=64), proteomic and cytokine analyses. Interestingly, Proteobacteria were over-represented in WH (WH to AB: p=0.005; WH to HC: p=0.021), whereas Firmicutes and Bacterioidetes were associated with AB. In contrast, Actinobacteria commonly colonized HCs (PermANOVA p=0.005). Moreover, Staphylococcaceae (p<0.05), Enterobacteriaceae (p<0.05), Burkholderiaceae (p<0.05), Xanthobacteraceae (p<0.05), and Sphingomonadaceae (p<0.05) were significantly more abundant in AB compared to WH and HC. The α‐diversity analyses demonstrated an increase of bacterial abundance levels in atopic AB and a decrease in WH samples. Microbiome profiles of atopic WH differed significantly from atopic AB. The NAEsp bacterial exposure with M. catarrhalis, S. aureus and H. influenzae experiments provided a disrupted epithelial cell integrity, a cytokine release and cohort specific proteomic differences especially for M. catarrhalis cultures. Our comprehensive dataset contributes to a deeper insight into the poorly understood plasticity of the nasal microbiota, and, in particular, may enforce our understanding in the pathogenesis of asthma exacerbation in childhood.
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Project description:Background The timing and mechanisms of asthma inception remain imprecisely defined. Although epigenetic mechanisms likely contribute to asthma pathogenesis, little is known about their role in asthma inception. Objective To assess whether the trajectory to asthma begins already at birth and epigenetic mechanisms, specifically DNA methylation, contribute to asthma inception. Methods We used a combination of methyl binding protein-dependent DNA capture and microarrays to survey DNA methylation in cord blood mononuclear cells (CBMC) from 36 children (18 non-asthmatic, 18 asthmatic by age 9) from the Infant Immune Study (IIS), an unselected birth cohort closely monitored for asthma for a decade. SMAD3 methylation in IIS (n=60) and in two replication cohorts (The Manchester Asthma and Allergy Study, n=30, and the Childhood Origins of ASThma Study, n=28) was analyzed by bisulfite sequencing or Illumina 450K arrays. CBMC-derived IL-1b was measured by ELISA. Results Neonatal immune cells harbored 589 differentially methylated regions (DMRs) that distinguished IIS children who did and did not develop asthma by age 9. In all three cohorts, methylation in SMAD3, the most connected hub within the network of asthma-associated DMRs, was selectively increased in asthmatic children of asthmatic mothers and was associated with childhood asthma risk. Moreover, SMAD3 methylation in IIS neonates with maternal asthma was strongly and positively associated with neonatal production of IL-1b, an innate inflammatory mediator. Conclusions The trajectory to childhood asthma begins at birth and involves epigenetic modifications in immunoregulatory and pro-inflammatory pathways. Maternal asthma influences epigenetic mechanisms that contribute to the inception of this trajectory.
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