Deep learning, 3D ultrastructural analysis reveals quantitative differences in platelet and organelle packing in COVID-19/SARSCoV2 patient-derived platelets
Sagar S. Matharu,
Cassidy S. Nordmann,
Kurtis R. Ottman,
Rahul Akkem,
Douglas Palumbo,
Denzel R. D. Cruz,
Kenneth Campbell,
Gail Sievert,
Jamie Sturgill,
James Z. Porterfield,
Smita Joshi,
Hammodah R. Alfar,
Chi Peng,
Irina D. Pokrovskaya,
Jeffrey A. Kamykowski,
Jeremy P. Wood,
Beth Garvy,
Maria A. Aronova,
Sidney W. Whiteheart,
Richard D. Leapman,
Brian Storrie
Affiliations
Sagar S. Matharu
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Cassidy S. Nordmann
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Kurtis R. Ottman
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Rahul Akkem
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Douglas Palumbo
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Denzel R. D. Cruz
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Kenneth Campbell
Department of Internal Medicine, University of Kentucky College of Medicine, Lexington, KY, USA
Gail Sievert
Center for Clinical Translational Science, University of Kentucky College of Medicine, Lexington, KY, USA
Jamie Sturgill
Department of Internal Medicine, University of Kentucky College of Medicine, Lexington, KY, USA
James Z. Porterfield
Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, KY, USA
Smita Joshi
Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY, USA
Hammodah R. Alfar
Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY, USA
Chi Peng
Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY, USA
Irina D. Pokrovskaya
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA
Jeffrey A. Kamykowski
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA
Jeremy P. Wood
Department of Internal Medicine, University of Kentucky College of Medicine, Lexington, KY, USA
Beth Garvy
Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, KY, USA
Maria A. Aronova
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Sidney W. Whiteheart
Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY, USA
Richard D. Leapman
Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA
Brian Storrie
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA
AbstractPlatelets contribute to COVID-19 clinical manifestations, of which microclotting in the pulmonary vasculature has been a prominent symptom. To investigate the potential diagnostic contributions of overall platelet morphology and their α-granules and mitochondria to the understanding of platelet hyperactivation and micro-clotting, we undertook a 3D ultrastructural approach. Because differences might be small, we used the high-contrast, high-resolution technique of focused ion beam scanning EM (FIB-SEM) and employed deep learning computational methods to evaluate nearly 600 individual platelets and 30 000 included organelles within three healthy controls and three severely ill COVID-19 patients. Statistical analysis reveals that the α-granule/mitochondrion-to-plateletvolume ratio is significantly greater in COVID-19 patient platelets indicating a denser packing of organelles, and a more compact platelet. The COVID-19 patient platelets were significantly smaller –by 35% in volume – with most of the difference in organelle packing density being due to decreased platelet size. There was little to no 3D ultrastructural evidence for differential activation of the platelets from COVID-19 patients. Though limited by sample size, our studies suggest that factors outside of the platelets themselves are likely responsible for COVID-19 complications. Our studies show how deep learning 3D methodology can become the gold standard for 3D ultrastructural studies of platelets.
