Androgen receptor pathway activity assay for sepsis diagnosis and prediction of favorable prognosis

Abstract

IntroductionSepsis is a life-threatening complication of a bacterial infection. It is hard to predict which patients with a bacterial infection will develop sepsis, and accurate and timely diagnosis, as well as assessment of prognosis, is difficult. Aside from antibiotics-based treatment of the causative infection and circulation-supportive measures, treatment options remain limited. Better understanding of the immuno-pathophysiology of sepsis may lead to improved diagnostic and therapeutic solutions.Functional activity of the innate (inflammatory) and adaptive immune response is controlled by a dedicated set of cellular signal transduction pathways in the various immune cell types. To develop an immune response-based diagnostic assay for sepsis and provide novel therapeutic targets, signal transduction pathway activities were analyzed in whole blood samples from patients with sepsis.MethodsA validated and previously published set of signal transduction pathway (STP) assays, determining immune cell function, was used to analyze public Affymetrix expression microarray data from clinical studies containing data from pediatric and adult patients with sepsis. STP assays enable quantitative measurement of STP activity on individual patient sample data, and were used to calculate activity of androgen receptor (AR), estrogen receptor (ER), JAK-STAT1/2, JAK-STAT3, Notch, Hedgehog, TGFβ, FOXO-PI3K, MAPK-AP1, and NFκB signal transduction pathways.ResultsActivity of AR and TGFβ pathways was increased in children and adults with sepsis. Using the mean plus two standard deviations of normal pathway activity (in healthy individuals) as threshold, diagnostic assay parameters were determined. For diagnosis of pediatric sepsis, the AR pathway assay showed high sensitivity (77%) and specificity (97%), with a PPV of 99% and NPV of 50%. For prediction of favorable prognosis (survival), PPV was 95%, NPV was 21%. The TGFβ pathway activity assay performed slightly less for diagnosing sepsis, with a sensitivity of 64% and specificity of 98% (PPV 99%, NPV 39%).ConclusionThe AR and TGFβ pathways have an immunosuppressive role, suggesting a causal relation between increased pathway activity and sepsis immunopathology. STP assays have been converted to qPCR assays for further evaluation of clinical utility for sepsis diagnosis and prediction of prognosis, as well as for prediction of risk at developing sepsis in patients with a bacterial infection. STPs may present novel therapeutic targets in sepsis.

Measuring host immune response status by simultaneous quantitative measurement of activity of signal transduction pathways that coordinate functional activity of immune cells from innate and adaptive immune system

“…AR pathway activity also increased in activated monocytes, in line with our earlier reporting on increased AR pathway activity in blood samples of patients with bacterial sepsis, in which monocytes play a dominant role (Bouwman et al, 2021), (Reyes et al, 2020). For the ER pathway low activity scores were consistently measured, with the only difference found between plasmacytoid and myeloid dendritic cells.…”

“…To enable future implementation, it is necessary to have STP assays with shorter time-to-result than the Affymetrix microarray; for this reason the assay platform has been converted to qPCR, enabling a result within a few hours (www.philips.com/oncosignal). Clinical implementation will also require establishment of normal ranges for STP PAS, which is likely to be possible given the similarity of I-PAPs for healthy individual samples, and earlier reported results (Bouwman et al, 2020), (Bouwman et al, 2021). Another challenge is the type of blood sample to analyze.…”

“…Another challenge is the type of blood sample to analyze. We have shown previously that STP analysis of mixed immune cell type samples, such as PBMCs or whole blood, may already provide clinically relevant results for a number of applications (Bouwman et al, 2021), (Bouwman et al, 2020). However, detailed analysis of the immune response of an individual will require isolation of specific immune cells of interest.…”

“…Development and validation of Affymetrix-based assays for measuring activity of the estrogen (ER) and androgen receptor (AR), PI3K-FOXO, MAPK, NFκB, TGFβ, Notch, JAK-STAT1/2, and JAK-STAT3 pathways has been described before, and can in principle be used to analyze pathway activity on any cell type (Bouwman et al, 2020), (Verhaegh et al, 2014), (Stolpe et al, 2019), (Canté-Barrett et al, 2020), (Wesseling-Rozendaal et al, 2021), (van Ooijen et al, 2018) including blood cells (Bouwman et al, 2020), (Wesseling-Rozendaal et al, 2020),(van de , (Bouwman et al, 2021). For the current study, quantitative pathway activity scores (PAS) were measured on Affymetrix expression microarray data.…”

“…Wesseling-Rozendaal et al, 2020)), (2) differential diagnosis of viral versus bacterial infection, (see our publication(Bouwman et al, 2020)), (3) assessment and monitoring of the severity of a viral infection(Bouwman et al, 2020), (4) prediction of risk at development of sepsis in a patient with a bacterial infection, sepsis diagnosis and prognosis and prediction of treatment response (see our publication(Bouwman et al, 2021)), (5) diagnosis and treatment response prediction for many immune-mediated diseases (unpublished results, e.g. for Crohn's Disease and Ulcerative Colitis, sarcoidosis, glomerulonephritis, psoriasis, pulmonary fibrosis); (6) prediction of late complications of kidney transplantation, such as fibrosis and glomerulonephritis (unpublished results).…”