ABSTRACT
Purpose of Study: COVID pneumonia caused by SARS-CoV-2 can result in a depletion of surfactant & lung injury, which resembles neonatal respiratory distress syndrome. Exogenous surfactant has shown promise as a therapeutic option in intubated hospitalized patients. Our preliminary data in human lung organoids (LOs) with a deficiency of surfactant protein B (SP-B) showed an increased viral load compared to normal LOs. Single cell RNA sequencing (scRNAseq) revealed that SP-B-deficient cells showed increased viral entry genes (ACE2 receptor) & dysregulated inflammatory markers emanating from the lung cells themselves. Our objective was to determine: (1) cell-specific transcriptional differences between normal & SP-B deficient human lung cells after infection with SARS-CoV-2 and (2) a therapeutic role of SP-B protein & surfactant in COVID-19 pneumonia. Methods Used: We used normal and SP-B mutant (homozygous, frameshift, loss of function mutation p.Pro133GlnfsTer95, previously known as 121ins2) human induced pluripotent stem cells (hiPSC) and differentiated them into 3D proximal lung organoids. The organoids were infected with the delta variant of SARS-CoV-2 for 24 hours at an MOI of 1. Infected and uninfected organoids were fixed in trizol in triplicate and underwent processing for bulk RNA sequencing. We tested for differentially expressed genes using the program DEseq. We also plated normal iPSC derived lung organoids as a monolayer and pre-treated them with 1mg/ml of Poractant alfa or 5 uM of recombinant SP-B protein. The delta strain of SARS-CoV-2 was added to the 96 wells at an MOI of 0.1 for one hour with shaking, then an overlay with DMEM/CMC/FBS was added and left on for 23 hours. The plate was fixed and stained for nucleocapsid (NC) protein. Summary of Results: Bioinformatic analysis of the bulk RNA sequencing data showed an increase in the multiple cytokines and chemokines in the SP-B mutant LOs compared to control. We also saw differential gene expression patterns in the SP-B mutant LOs including a reduction in SFTPC, FOXA2, and NKX2-1 and an increase in IL1A, VEGFA, PPARG and SMAD3. In the exogenous surfactant experiments, there was a decrease in total expression of viral NC in the Poractant alfa & rSP-B-treated cells compared to SARS-CoV-2 infection alone (p<0.001). Conclusion(s): Surfactant modulates the viral load of SARS-CoV-2 infection in the human lung. Deficiency in SP-B results in the dysregulation of the lung epithelial inflammatory signaling pathways resulting in worsening infections.
ABSTRACT
ROSSI'S PRINCIPLES OF TRANSFUSION MEDICINE: Transfusion Medicine impacts patients with hematologic, oncologic, and surgical conditions as well as all areas of critical care medicine and multiple areas of chronic care. This book aims to be the single best source for information related to any aspect or application of Transfusion Medicine. Contributors for the sixth edition have once again been drawn from various scientific, medical, and surgical disciplines. Thus, this book ranges from encouraging and managing donors, to collecting and preserving the blood, to matching it to the appropriate recipient, all the way to its clinical uses. It also extends these concepts to implantable tissue and regenerative medicine. Other sample topics covered within the work include: Contemporary issues in donation and transfusion: patient blood management, clinical and technical aspects of blood administration, and donor and patient Hemovigilance. Blood components and derivatives: red blood cell metabolism, preservation and oxygen delivery, blood groups, and composition of plasma. Apheresis, transplantation, and new therapies: hematopoietic growth factors, therapeutic phlebotomy and cellular apheresis, HLA antigens, alleles, and antibodies. How Transfusion Medicine has been affected by the coronavirus pandemic, the role of pathogen reduction and other modern trends. This book serves as a complete and comprehensive resource on Transfusion Medicine for clinicians who prescribe blood, students who expect to enter clinical practice, and for the scientists, physicians, nurses, technologists, and others who assure the quality and availability of blood services. © 2022 John Wiley & Sons Ltd. All rights reserved.