A Functional Variant of CXCL16 Is Associated With Predisposition to Sepsis and MODS in Trauma Patients: Genetic Association Studies

Jianhui Sun, Huacai Zhang, Di Liu, Li Cui, Qiang Wang, Lebin Gan, Dalin Wen, Jun Wang, Juan Du, Hong Huang, Anqiang Zhang, Jin Deng, Jianxin Jiang, Ling Zeng, Jianhui Sun, Huacai Zhang, Di Liu, Li Cui, Qiang Wang, Lebin Gan, Dalin Wen, Jun Wang, Juan Du, Hong Huang, Anqiang Zhang, Jin Deng, Jianxin Jiang, Ling Zeng

Abstract

Purpose: CXC chemokines are mediators which mediate immune cells migration to sites of inflammation and injury. Chemokine C-X-C motif ligand 16 (CXCL16) plays an important role in the occurrence and development of sepsis through leukocyte chemotaxis, leukocyte adhesion and endotoxin clearance. In this study, we selected a set of tagging single nucleotide polymorphisms (tag SNPs) in the CXCL16 gene and investigated their clinical relevance to the development of sepsis and multiple organ dysfunction syndrome (MODS) in patients with major trauma in three independent Chinese Han populations.

Methods: A total of 1,620 major trauma patients were enrolled in this study. Among these patients, 920 came from Chongqing in western China, 350 came from Zhejiang Province in eastern China, and 350 came from Guizhou Province in southwestern China. The improved multiplex ligation detection reaction (iMLDR) method was employed in the genotyping and genetic association analyses to determine the associations between CXCL16 haplotypes and sepsis morbidity rate and higher MOD scores in three cohorts.

Results: Only CXCL16 T123V181 haplotype was associated with an increased risk for sepsis morbidity and higher MOD scores in the three cohorts (OR = 1.89, P = 0.001 for the Chongqing cohort; OR = 1.76, P = 0.004 for the Zhejiang cohort; OR = 1.55, P = 0.012 for the Guizhou cohort). The effect of T123V181 haplotype on the chemotaxis, migration and endotoxin clearance of immune cells were further observed. Protein modeling analysis showed that T123 and V181 might alter the structure of the CXCL16 active center. Thus it enhanced the chemotaxis and adhesion ability of immunocytes.

Conclusion: We demonstrate the mechanism of CXCL16 T123V181 haplotype which regulates the sepsis morbidity rate and thus provide a new biomarker for early diagnosis of sepsis and MODS.

Clinical trial registration: www.ClinicalTrials.gov, identifier NCT01713205 (https://www.clinicaltrials.gov/ct2/results?cond=&term=+NCT01713205&cntry=&state=&city=&dist=).

Keywords: CXCL16; multiple organ dysfunction; sepsis; single nucleotide polymorphisms; trauma.

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2021 Sun, Zhang, Liu, Cui, Wang, Gan, Wen, Wang, Du, Huang, Zhang, Deng, Jiang and Zeng.

Figures

FIGURE 1
FIGURE 1
Overview of tagging single nucleotide polymorphisms (tag SNPs) in the CXCL16 gene. (A) CXCL16 gene organization and the location of two missense mutations, I123T (rs1050998) and A181V (rs2277680), on chromosome 17. (B) Location of SNPs in the CXCL16 gene with a minor allele frequency ≥5%. A linkage disequilibrium (LD) plot of these SNPs is displayed by a color scheme. Black represents very high LD (r2 = 1.0), and white indicates the absence of correlation (r2 = 0) between two SNPs. The r2 between I123T and A181V is 1.0.
FIGURE 2
FIGURE 2
T123V181 haplotype enhances the chemotaxis and adhesion of inflammatory cells. (A) Map of the CXCL16 expression plasmid. (B) Schematic diagram of the chemotactic assay. The percentage of THP-1 cells recruited to the lower chamber was calculated to represent chemotaxis. (C) The chemotaxis of the CXCL16-T123V181 protein was significantly higher than that of the other three proteins (**P < 0.01). (D) The percentage of THP-1 cells with green fluorescence expressing transmembrane forms of CXCL16-T123V181 protein attached to RAW264.7 cells was calculated to represent the adhesion ability. (E) The adhesion ability of cells expressing CXCL16-T123V181 protein was significantly higher than that of the other three proteins (**P < 0.01).
FIGURE 3
FIGURE 3
Conservative analysis and molecular modeling of I123T and A181V. (A) Conservative analysis shows that the two missense mutations are both located in the non-conserved region of the CXCL16 gene. (B) The structural diagram shows that the active center of the CXCL16 protein was changed by V → A substitution, which might make the active center of the CXCL16 protein smaller.

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