title: PREDICTING THE SEVERITY OF ACUTE PANCREATITIS AND CHARACTERISING THE IMPACT OF SEVERITY ON DISEASE OUTCOMES
Creators: name: Sud, Ajay
Publisher: name: University of Liverpool Repository
Year: 2025

description: Introduction: Established early stratification predictors and established outcomes measures in acute pancreatitis (AP). The aims of this body of work encompass the optimisation/calibration and then utilisation of mass cytometry conjunction with machine learning to characterize the early regularity T-cells changes that govern the severity of the disease process. Proteomics array analysis to novel identify candidate early biomarkers. Sarcopenia, hand grip strength, feeding nutrition and opioid parameters interrogated as potential outcome measures.

Methods: Helios is a Cytometry by time-of-flight system optimised and calibrated utilising cryopreserved frozen peripheral blood mononuclear cells, which were then thawed and assessed for biologically plausible Peripheral blood phenotyping panel and regulatory T-cells leukocytes. This protocol workflow was applied phenotype regulatory T-cell phenotyping from revised Atlanta criteria (RAC) stratified severity mild and non-mild acute pancreatitis patient blood samples from 24 and 48 hour from admission. Analysis of results undertaken manually and utilising artificial narrow intelligence machine learning algorithms. Proteomic analysis utilising Multi-Analyte Profile Multiplex, automated Luminex assay, workflow analysed manually and using supervised regularization machine learning. Retrospective Liverpool Acute Pancreatitis Biobank cohort data from 2010-2016 was collated to obtain nutritional, analgesic, and clinical particulars. The data was analysed using SPSS 22. Hand-grip strength patient clinical particulars were collected prospectively with grip strength measurement undertaken with 3 successive measures with a hydraulic dynamometer. Sarcopenia measurements were undertaken using 2 sets of axial CT images taken at the interval of at least 5-7 days. This sarcopenia data included specialist tertiary transfer patients rather being just primary index admissions. 

Results: A reduction in naive Tregs was demonstrated between 24 and 48 hours, this was a greater magnitude within the non-mild cohort. Memory regulatory Tregs were greater in severer AP patients within day of admission. Terminal and effector Tregs also demonstrated proportionally increased magnitude in day 1 non-mild AP patients. Cluster identification, characterization, and regression analysis demonstrated key cluster events pertaining to adaptive immune processes that differentiated RAC mild and non-mild AP severity at Day 1. These were naive inactive Tregs, T-cell suppression, some TH17 activated Tregs and some non-intrinsic CD127+ve Treg features. EN-RAGE proved to be an effective early research candidate biomarker. Least absolute shrinkage and selection operator (LASSO) at Day 1 of CRP &amp; EN-RAGE Area under the receiver operator curve of (AUROC) 0.95 and Day 2 EN-RAGE AUROC 0.88). Median days on opioids for RAC mild, moderate, and severe was 4 (2-7), moderate 7.5 (5.5-14) and severe (2-21). The median days to reestablish food/oral nutrition was 2(1-2), 2(1-4) and 7(3-13) rs=0.471, p&lt;0.001) for RAC mild, moderate, and severe disease respectively. Hand grip strength (HGS) spearman’s correlation with CRP (r2=0.521). At day 2 from admission HGS was 26.9% less than admission in the complicated AP in comparison to uncomplicated. Sarcopenia assessed in revised Atlanta non-mild patients proved to be more pronounced, in comparison to the non-intensive care cohort. Right psoas co-efficient variation of 4.31%. There was a significant (1 tail t-test p&lt;0.10) difference in % muscle loss between intensive care versus ward cohorts and between those invasively ventilated and those not, 0.52 and 0.74 respectively. There was a significant correlation between the percentage muscle loss correlated with CRP of 0.96 (2-tailed test significance level of &lt;0.001). 

Conclusion: Stratification and prediction can be achieved with these novel predictors and outcome measures. Distinct RAC severity-based adaptions to the regulatory Tregs-cell populations occur within the first 24 hours of admission with the disease. EN-RAGE +/- CRP at day 1 from admission is effective predictor of clinically significant mild and non-mild AP. Total morphine administration and time to eating drinking also demonstrated to differentiate more severe disease. RAC Severity based differentials in &gt;Day 2 HGS (mild and moderate) and CT-cross sectional L4 determined sarcopenia (moderate and severe) can be leveraged as outcome measures of the impact of RAC AP severity.