Sepsis, which is characterized by life threatening organ dysfunction caused by an uncontrolled host response to
infection, is the leading cause of death in hospitals and is responsible for >250,000 deaths per year in the U.S.
at the cost of over $20 billion in patient care. Sepsis has a mortality rate nearing 50% at 2 years, which has
profound implications for patients recovering from severe COVID-19, a form of viral sepsis. Because sepsis is a
multi-organ disease, the quantification of circulating proteins and metabolites is central to the profiling of sepsis
and of its long-term effects, and some such assays (lactate, procalcitonin) have become the standard of care.
To date, the majority of studies, and particularly the large number in COVID-19 sepsis, have profiled plasma and
serum, which, compared to whole blood is fraught with variability and belies the important roles of the erythrocyte
and other cell-types. To this end, biorepositories at Duke University, which are banking samples from COVID-
19 sepsis patients in the intensive care unit (ICU) and in a post-COVID outpatient clinic, have been utilizing
volumetric absorptive microsampling (VAMS) on Neoteryx Mitra tips to collect and store whole blood specimens
for future ‘omic analyses. The overarching goals of this application are to develop and validate multiomic methods
for mass spectrometry-based quantification of proteins and metabolites from Mitra tips that comprehensively
profile sepsis pathobiology. In the R21 phase, we will develop and validate mass spectrometry-based methods
for the non-targeted quantification of proteins and post-translational modifications (phosphorylation,
glycosylation) in whole blood, and for targeted quantification of numerous metabolite classes. We will develop
reference materials and standard operating procedures for inter-laboratory translation of these approaches. In
the R33 phase, we will analyze proteins, PTMs and metabolites from over 600 patient timepoints from
biorepositories containing longitudinal samples in critically ill sepsis and post-COVID-19 cohorts, and we will
integrate these data with well-curated clinical datasets. Completion of these aims will establish the utility of blood
sampling by VAMS for future sepsis studies and will create a highly curated clinical and deep multiomic dataset
for future hypothesis generation.