Convergence of coronary artery disease genes onto endothelial cell programs.

Linking variants from genome-wide association studies (GWAS) to underlying mechanisms of disease remains a challenge1-3. For some diseases, a successful strategy has been to look for cases in which multiple GWAS loci contain genes that act in the same biological pathway1-6. However, our knowledge of which genes act in which pathways is incomplete, particularly for cell-type-specific pathways or understudied genes. Here we introduce a method to connect GWAS variants to functions. This method links variants to genes using epigenomics data, links genes to pathways de novo using Perturb-seq and integrates these data to identify convergence of GWAS loci onto pathways. We apply this approach to study the role of endothelial cells in genetic risk for coronary artery disease (CAD), and discover 43 CAD GWAS signals that converge on the cerebral cavernous malformation (CCM) signalling pathway. Two regulators of this pathway, CCM2 and TLNRD1, are each linked to a CAD risk variant, regulate other CAD risk genes and affect atheroprotective processes in endothelial cells. These results suggest a model whereby CAD risk is driven in part by the convergence of causal genes onto a particular transcriptional pathway in endothelial cells. They highlight shared genes between common and rare vascular diseases (CAD and CCM), and identify TLNRD1 as a new, previously uncharacterized member of the CCM signalling pathway. This approach will be widely useful for linking variants to functions for other common polygenic diseases.

Institutional leadership after Dobbs: a mixed methods analysis of US medical schools' public statements regarding abortion.

INTRODUCTION: Medical schools, as significant and influential organisations within their communities, have the potential and the capacity to impact abortion policy. Organisations often engage in advocacy by issuing public statements that clarify their stance on specific policies. This study analyses the quantity and quality of publicly discoverable statements that US medical schools issued regarding Dobbs v Jackson Women's Health Organization. METHODS: We conducted a mixed methods study using an explanatory sequential design. Using qualitative analysis, an inductive thematic approach was used to identify themes from public statements made within 6 months of 2 May 2022, Dobbs leak. Descriptive statistics and logistic regression analysis were used to assess the association between themes and institutional characteristics. RESULTS: Most institutions (n=124/188, 65.96%) did not issue public statements regarding Dobbs. Among all 188 US medical schools, allopathic institutions (OR=12.19, 95% CI (2.83 to 52.57), p=0.001), schools in protective states (OR=3.35, 95% CI (1.78 to 6.29), p<0.0001) and those with family planning divisions (OR=4.60, 95% CI (2.33 to 9.08), p<0.0001) were at increased odds of issuing statements. Of the 64 medical schools with statements, 64.06% (n=41/64) espoused pro-choice views, 34.37% (n=22) were neutral/non-committal and 1.56% (n=1) expressed antiabortion views. Those in protective states were at 3.35 times increased odds of issuing pro-choice statements (95% CI (1.16 to 9.72), p=0.03) compared with restrictive counterparts. CONCLUSION: Medical schools largely did not take a public stance on Dobbs. By refraining from actively engaging in this critical discourse, medical schools are foregoing a leadership opportunity to affect meaningful sociopolitical change, particularly in states with restrictive abortion laws.

Multiomic Analysis and CRISPR Perturbation Screens Identify Endothelial Cell Programs and Novel Therapeutic Targets for Coronary Artery Disease.

Endothelial cells (EC) are an important mediator of atherosclerosis and vascular disease. Their exposure to atherogenic risk factors such as hypertension and serum cholesterol leads to endothelial dysfunction and many disease-associated processes. Identifying which of these multiple EC functions is causally related to disease risk has been challenging. There is evidence from in vivo models and human sequencing studies that dysregulation of nitric oxide production directly affects risk of coronary artery disease. Human genetics can help prioritize the other EC functions with causal relationships because germline mutations are acquired at birth and serve as a randomized test of which pathways affect disease risk. Though several coronary artery disease risk variants have been linked to EC function, this process has been slow and laborious. Unbiased analyses of EC dysfunction using multiomic approaches promise to identify the causal genetic mechanisms responsible for vascular disease. Here, we review the data from genomic, epigenomic, and transcriptomic studies that prioritize EC-specific causal pathways. New methods that CRISPR (clustered regularly interspaced short palindromic repeats) perturbation technology with genomic, epigenomic, and transcriptomic analysis promise to speed up the characterization of disease-associated genetic variation. We summarize several recent studies in ECs which use high-throughput genetic perturbation to identify disease-relevant pathways and novel mechanisms of disease. These genetically validated pathways can accelerate the identification of drug targets for the prevention and treatment of atherosclerosis.

Bronchiectasis in children following kidney transplantation in New Zealand.

AIM: Bronchiectasis is an acquired chronic respiratory condition with a relatively high incidence in New Zealand children. Bronchiectasis following kidney transplant has been reported internationally. This study aimed to identify the incidence rate of bronchiectasis following paediatric kidney transplantation. Secondary aims were to assess the impact on kidney allograft function and identify risk factors that might prompt earlier diagnosis. METHODS: Case control study of children who developed bronchiectasis following kidney transplant in New Zealand. All children who were transplanted during the 16-year period from 2001 to 2016 were included. Each identified case was matched with two controls (children who did not develop bronchiectasis and received a kidney transplant within the closest time period to their matched case). Data were collected on baseline demographics, clinical variables, immunosuppression and allograft function. RESULTS: Of 95 children who had a kidney transplant during the specified time period, eight (8.4%) developed bronchiectasis at a median of 4 years post-transplant. The mean incidence rate of bronchiectasis was 526 cases per 100 000 paediatric kidney transplant population per year. The majority of children were Maori or Pasifika ethnicity and lived in areas of greater socio-economic deprivation. Immunosuppression burden and allograft function were not significantly different between groups. CONCLUSIONS: The incidence rate of bronchiectasis following paediatric kidney transplantation is substantially higher than the baseline paediatric incidence rate in New Zealand. A high index of suspicion for bronchiectasis and prompt investigation of children post kidney transplantation with a history of recurrent lower respiratory tract infection or chronic cough are advised.

CRISPR-Cas9 Genome Editing of Primary Human Vascular Cells In Vitro.

Genome editing of primary human cells with CRISPR-Cas9 is a powerful tool to study gene function. For many cell types, there are efficient protocols for editing with optimized plasmids for Cas9 and sgRNA expression. Vascular cells, however, remain refractory to plasmid-based delivery of CRISPR machinery for in vitro genome editing due to low transfection efficiency, poor expression of the Cas9 machinery, and toxic effects of the selection antibiotics. Here, we describe a method for high-efficiency editing of primary human vascular cells in vitro using nucleofection for direct delivery of sgRNA:Cas9-NLS ribonucleoprotein complexes. This method is more rapid and its high editing efficiency eliminates the need for additional selection steps. The edited cells can be employed in diverse applications, such as gene expression measurement or functional assays to assess various genetic perturbation effects in vitro. This method proves effective in vascular cells that are refractory to standard genome manipulation techniques using viral plasmid delivery. We anticipate that this technique will be applied to other non-vascular cell types that face similar barriers to efficient genome editing. © 2021 Wiley Periodicals LLC. Basic Protocol: CRISPR-Cas9 genome editing of primary human vascular cells in vitro.

Rare, Damaging DNA Variants in CORIN and Risk of Coronary Artery Disease: Insights From Functional Genomics and Large-Scale Sequencing Analyses.

BACKGROUND: Corin is a protease expressed in cardiomyocytes that plays a key role in salt handling and intravascular volume homeostasis via activation of natriuretic peptides. It is unknown if Corin loss-of-function (LOF) is causally associated with risk of coronary artery disease (CAD). METHODS: We analyzed all coding CORIN variants in an Italian case-control study of CAD. We functionally tested all 64 rare missense mutations in Western Blot and Mass Spectroscopy assays for proatrial natriuretic peptide cleavage. An expanded rare variant association analysis for Corin LOF mutations was conducted in whole exome sequencing data from 37 799 CAD cases and 212 184 controls. RESULTS: We observed LOF variants in CORIN in 8 of 1803 (0.4%) CAD cases versus 0 of 1725 controls (P, 0.007). Of 64 rare missense variants profiled, 21 (33%) demonstrated <30% of wild-type activity and were deemed damaging in the 2 functional assays for Corin activity. In a rare variant association study that aggregated rare LOF and functionally validated damaging missense variants from the Italian study, we observed no association with CAD-21 of 1803 CAD cases versus 12 of 1725 controls with adjusted odds ratio of 1.61 ([95% CI, 0.79-3.29]; P=0.17). In the expanded sequencing dataset, there was no relationship between rare LOF variants with CAD was also observed (odds ratio, 1.15 [95% CI, 0.89-1.49]; P=0.30). Consistent with the genetic analysis, we observed no relationship between circulating Corin concentrations with incident CAD events among 4744 participants of a prospective cohort study-sex-stratified hazard ratio per SD increment of 0.96 ([95% CI, 0.87-1.07], P=0.48). CONCLUSIONS: Functional testing of missense mutations improved the accuracy of rare variant association analysis. Despite compelling pathophysiology and a preliminary observation suggesting association, we observed no relationship between rare damaging variants in CORIN or circulating Corin concentrations with risk of CAD.