A characterization of the molecular phenotype and inflammatory response of schizophrenia patient-derived microglia-like cells

Different lines of evidence support a causal role for microglia in the pathogenesis of schizophrenia. However, how schizophrenia patient-derived microglia are affected at the phenotypic and functional level is still largely unknown. We used a recently described model to induce patient-derived microglia-like cells and used this to analyze changes in the molecular phenotype and function of myeloid cells in schizophrenia. We isolated monocytes from twenty recent-onset schizophrenia patients and twenty non-psychiatric controls. We cultured the cells towards an induced microglia-like phenotype (iMG), analyzed the phenotype of the cells by RNA sequencing and mass cytometry, and their response to LPS. Mass cytometry showed a high heterogeneity of iMG in cells derived from patients as well as controls. The prevalence of two iMG clusters was significantly higher in schizophrenia patients (adjusted p-value <0.001). These subsets are characterized by expression of ApoE, Ccr2, CD18, CD44, and CD95, as well as IRF8, P2Y(12), Cx3cr1 and HLA-DR. In addition, we found that patient derived iMG show an enhanced response to LPS, with increased secretion of TNF-alpha. Further studies are needed to replicate these findings, to determine whether similar subclusters are present in schizophrenia patients in vivo, and to address how these subclusters are related to the increased response to LPS, as well as other microglial functions

Generation and Characterization of a RACK1 Knockout Cell Line

The µ-opioid receptor (MOR) is an important regulator of nociception, bearing the largest role in analgesia of all opioid receptors. This function has made MOR an effective drug-target for the clinical treatment of pain. However, opioid drugs such as morphine and fentanyl are highly addictive, and prolonged abuse can be fatal due to respiratory depression. A key determinant in the cellular and systemic responses to opioids is MOR. At the transcriptional level, MOR is positively regulated by the transcription factors, including poly-C binding protein 1 (PCBP1). Using the two-hybrid screening, PCBP1 was found to interact with receptor for activated C kinase (RACK1). RACK1 expression was negatively correlated with MOR expression. Here, the CRISPR technique (CRISPR-Cas9n) was used to create the deletion of the RACK1gene using human NMB neuronal cells. Screening for the deletion of RACK1 in candidate clones detected only RACK1+/- cells, but no RACK1-/- cells. Preliminary data showed that partial deletion of the RACK1 gene successfully decreased RACK1 mRNA expression and increased MOR mRNA expression in RACK1+/- cells. The stability of RACK1+/- cells was further examined. Partial deletion of the RACK1 gene, as well as alterations in mRNA expression of RACK1 and MOR, were consistently detected across a period of about five months. These preliminary results show that RACK1 can negatively regulate MOR expression

Generation and Characterization of a RACK1 Knockout Cell Line

The µ-opioid receptor (MOR) is an important regulator of nociception, bearing the largest role in analgesia of all opioid receptors. This function has made MOR an effective drug-target for the clinical treatment of pain. However, opioid drugs such as morphine and fentanyl are highly addictive, and prolonged abuse can be fatal due to respiratory depression. A key determinant in the cellular and systemic responses to opioids is MOR. At the transcriptional level, MOR is positively regulated by the transcription factors, including poly-C binding protein 1 (PCBP1). Using the two-hybrid screening, PCBP1 was found to interact with receptor for activated C kinase (RACK1). RACK1 expression was negatively correlated with MOR expression. Here, the CRISPR technique (CRISPR-Cas9n) was used to create the deletion of the RACK1gene using human NMB neuronal cells. Screening for the deletion of RACK1 in candidate clones detected only RACK1+/- cells, but no RACK1-/- cells. Preliminary data showed that partial deletion of the RACK1 gene successfully decreased RACK1 mRNA expression and increased MOR mRNA expression in RACK1+/- cells. The stability of RACK1+/- cells was further examined. Partial deletion of the RACK1 gene, as well as alterations in mRNA expression of RACK1 and MOR, were consistently detected across a period of about five months. These preliminary results show that RACK1 can negatively regulate MOR expression

SARS-CoV-2 infection, inflammation and birth outcomes in a prospective NYC pregnancy cohort

Associations between antenatal SARS-CoV-2 infection and pregnancy outcomes have been conflicting and the role of the immune system is currently unclear. This prospective cohort study investigated the interaction of antenatal SARS-CoV-2 infection, changes in cytokine and HS-CRP levels, birthweight and gestational age at birth. 2352 pregnant participants from New York City (2020–2022) were included. Plasma levels of interleukin (IL)-1β, IL-6, IL-17A and high-sensitivity C-reactive protein (HS-CRP) were quantified in blood specimens obtained across pregnancy. Quantile and linear regression models were conducted to 1) assess the impact of antenatal SARS-CoV-2 infection, overall and by timing of detection of SARS-CoV-2 positivity (&lt; 20 weeks versus ≥ 20 weeks), on birthweight and gestational age at delivery; 2) examine the relationship between SARS-CoV-2 infection and maternal immune changes during pregnancy. All models were adjusted for maternal demographic and obstetric factors and pandemic timing. Birthweight models were additionally adjusted for gestational age at delivery and fetal sex. Immune marker models were also adjusted for gestational age at specimen collection and multiplex assay batch. 371 (15.8%) participants were infected with SARS-CoV-2 during pregnancy, of which 98 (26.4%) were infected at &lt; 20 weeks gestation. Neither SARS-CoV-2 infection in general nor in early or late pregnancy was associated with lower birthweight nor earlier gestational age at delivery. Further, we did not observe cytokine or HS-CRP changes in response to SARS-CoV-2 infection and thus found no evidence to support a potential association between immune dysregulation and the diversity in pregnancy outcomes following infection.</p

Data for: Transcriptome and single-cell protein expression landscape of schizophrenia patient-derived microglia-like cells

RNA sequencing data (raw counts and TPM) of monocyte-derived microglia-like cells, monocytes and primary microgli