AUTHOR=Corrêa Raquel da Silva , Leal-Calvo Thyago , Mafort Thiago Thomaz , Santos Ana Paula , Leung Janaína , Pinheiro Roberta Olmo , Rufino Rogério , Moraes Milton Ozório , Rodrigues Luciana Silva 

TITLE=Reanalysis and validation of the transcriptional pleural fluid signature in pleural tuberculosis

JOURNAL=Frontiers in Immunology

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2023.1256558

DOI=10.3389/fimmu.2023.1256558

ISSN=1664-3224

ABSTRACT=<sec><title>Introduction</title><p>Pleural tuberculosis (PlTB), the most common site of extrapulmonary TB, is characterized by a paucibacillary nature and a compartmentalized inflammatory response in the pleural cavity, both of which make diagnosis and management extremely challenging. Although transcriptional signatures for pulmonary TB have already been described, data obtained by using this approach for extrapulmonary tuberculosis and, specifically, for pleural tuberculosis are scarce and heterogeneous. In the present study, a set of candidate genes previously described in pulmonary TB was evaluated to identify and validate a transcriptional signature in clinical samples from a Brazilian cohort of PlTB patients and those with other exudative causes of pleural effusion.</p></sec><sec><title>Methods</title><p>As a first step, target genes were selected by a random forest algorithm with recursive feature elimination (RFE) from public microarray datasets. Then, peripheral blood (PB) and pleural fluid (PF) samples from recruited patients presenting exudative pleural effusion were collected during the thoracentesis procedure. Transcriptional analysis of the selected top 10 genes was performed by quantitative RT-PCR (RT-qPCR).</p></sec><sec><title>Results</title><p>Reanalysis of the public datasets identified a set of candidate genes (<italic>CARD17, BHLHE40, FCGR1A, BATF2, STAT1, BTN3A1, ANKRD22, C1QB, GBP2</italic>, and <italic>SEPTIN4</italic>) that demonstrated a global accuracy of 89.5% in discriminating pulmonary TB cases from other respiratory diseases. Our validation cohort consisted of PlTB (<italic>n</italic> = 35) patients and non-TB (<italic>n</italic> = 34) ones. The gene expressions of <italic>CARD17</italic>, <italic>GBP2</italic>, and <italic>C1QB</italic> in PF at diagnosis were significantly different between the two (PlTB and non-TB) groups (<italic>p</italic> &lt; 0.0001). It was observed that the gene expressions of <italic>CARD17</italic> and <italic>GBP2</italic> were higher in PlTB PF than in non-TB patients. <italic>C1QB</italic> showed the opposite behavior, being higher in the non-TB PF. After anti-TB therapy, however, <italic>GBP2</italic> gene expression was significantly reduced in PlTB patients (<italic>p</italic> &lt; 0.001). Finally, the accuracy of the three above-cited highlighted genes in the PF was analyzed, showing AUCs of 91%, 90%, and 85%, respectively. <italic>GBP2</italic> was above 80% (sensitivity = 0.89/specificity = 0.81), and <italic>CARD17</italic> showed significant specificity (Se = 0.69/Sp = 0.95) in its capacity to discriminate the groups.</p></sec><sec><title>Conclusion</title><p><italic>CARD17</italic>, <italic>GBP2</italic>, and <italic>C1QB</italic> showed promise in discriminating PlTB from other causes of exudative pleural effusion by providing accurate diagnoses, thus accelerating the initiation of anti-TB therapy.</p></sec>