key: cord-1000990-m43jd6nj
authors: Posadas-Sánchez, Rosalinda; Sánchez-Muñoz, Fausto; Martin, Carlos Alfonso Guzmán; Couder, Adrian Hernández-Díaz; Rojas-Velasco, Gustavo; Fragoso, José Manuel; Vargas-Alarcón, Gilberto
title: Dipeptidylpeptidase-4 levels and DPP4 gene polymorphisms in patients with COVID-19. Association with disease and with severity
date: 2021-03-24
journal: Life Sci
DOI: 10.1016/j.lfs.2021.119410
sha: b2b2273688d6df06b6480941d9655f44abb240a7
doc_id: 1000990
cord_uid: m43jd6nj

BACKGROUND: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes de COVID-19 disease use as a principal receptor the angiotensin-converting enzyme-2 (ACE2). It has been suggested that Dipeptidyl peptidase-4 (DPP4) can be another possible receptor for this virus. The present study aimed to establish if the DPP4 levels and DPP4 polymorphisms are associated with COVID-19 disease and its severity. METHODS: The study included 107 COVID-19 patients and 263 matched-healthy controls. Fifty patients required invasive mechanical ventilation. The DPP4 was quantified in serum using the Bioplex system. Based on the previous results and the functional prediction analysis, we select for the study 5 DPP4 polymorphisms (rs12617336, rs12617656, rs1558957, rs3788979, and rs17574) and these were determined using the 5´exonuclease TaqMan assays. RESULTS: Low levels of DPP4 were observed in COVID-19 patients (46.5 [33.1–57.7] ng/mL) when compared to healthy controls (125.3 [100.3–157.3] ng/mL) (P < 0.0001). Also, patients that required mechanical ventilation showed lower DPP4 levels (42.8 [29.8–56.9] ng/mL) than those that did not need this procedure (49.2 [39.9–65.6] ng/mL) (P = 0.012). DPP4 levels correlated negatively with age, fibrinogen, and platelet levels, and positively with albumin, alanine aminotransferase, and percentage of neutrophils. The DPP4 rs3788979 polymorphism was associated with a high risk of COVID-19 disease and, the TT genotype carriers had the lowest DPP4 levels. CONCLUSIONS: In summary, in the present study, an association of low levels of DDP4 with COVID-19 disease and severity was found. The association of the DPP4 rs3788979 polymorphism with COVID-19 is also reported.

sample. The patients were attended in the intensive care unit of our Institute from May 11 to July 31, 2020. Fifty patients required invasive mechanical ventilation. Demographic, clinical, and laboratory parameters of Covid-19 patients were obtained. All patients or their relatives signed the institutional consent letter.

To evaluate if DPP4 serum concentrations in COVID-19 patients were different from healthy controls, we used a case control study design. For each COVID-19 patient, we matched 3 controls by age, sex, and body mass index. Of the 107 COVID-19 patients, 83 patients were individually matched with 3 healthy controls, while 7 patients were matched with only 2 controls each. Therefore, in the present study, we examined circulating DPP4 serum concentrations in 90 COVID-19 patients and 263 healthy controls (172 men and 91 women). The healthy control individuals were selected among the participants of the Genetics of Atherosclerotic Disease (GEA) Mexican study control group [19] . The GEA control group included individuals without personal or family history of premature coronary disease. Anthropometric, biochemical, clinical, and demographic characteristics, as well as cardiovascular risk factors were determined in all participants as previously described [19] [20] [21] . Criteria definitions and methods have been reported previously [22] [23] [24] [25] . Obesity [19, 20] , current smoking [20] , hypertension [19, 20] , T2DM [19, 20] definitions have already been published.

The samples were obtained in the intensive care unit following the institutional security protocols. Competent trained personnel carried out the handling and processing of the patients' blood samples, which were later transported to the laboratory for processing.

Samples were centrifuged, and the serum was separated in a class II biological safety cabinet. Personnel handling the samples used personal protective equipment that included disposable gloves, a lab coat, and a surgical mask. Samples were collected and processed in a laboratory that adheres to the guidelines established in the Official Mexican Standards 

Bioplex system (R&D Systems, Minneapolis, USA) was used to quantified DPP4 serum concentration, according to manufacturer's instructions. A Bio-Plex Manager software was used for the data analyses. Results are expressed in ng/mL.

For the genomic DNA isolation from whole blood (containing EDTA) of the for all analyses.

One hundred-seven COVID-19 patients (68% male) were included in the study, with a median age of 55.76 years. (Table 4 ).

In three COVID-19 patients from the total of 107, DPP4 levels were measured several days after the diagnosis and their admission to the intensive care unit. As can be seen in Fig. 3 , in the two COVID-19 patients who recovered without IMV, serum SPP4 levels increased, whereas they decreased in one mechanically ventilated patient. 

Of the 5 SNPs studied, rs3788979 showed a different distribution of its genotypes in (Fig. 4) . On the other hand, in the total population (patients and controls), different DPP4 levels were observed in the rs3788979 genotypes; of note, the lowest levels were found in the TT genotype carriers (P = 0.003) (Fig. 5 ).

In a cohort of COVID-19 patients and controls, we found significantly lower levels of DPP4 in COVID-19 patients. Enzyme levels were also low in patients receiving IMV; by the same token, a low risk of requiring IMV was associated with a high DDP4

concentration. High enzyme levels correlated negatively with age, fibrinogen, and platelets, This effect could decrease the amount of biomolecule in the cell membrane and, consequently, in the serum of the patients. As mentioned, DPP4 is expressed in several blood cells, but predominantly in T lymphocytes [30] and specifically in CD4 T cells [41] .

Measurement of DPP4 in lymphocyte culture has suggested that these cells are an important source of soluble DPP4 [42] . As in other coronavirus infections (SARS and MERS), the individuals infected with SARS-CoV-2 present with a marked lymphopenia [43] [44] [45] ; this disorder is more severe in those individuals who do not survive the disease, [47] [48] [49] [50] [51] . Located in the 2p24.3 region, the gene that encodes DPP4 is polymorphic and some of its SNPs have been associated not only with the risk of developing some diseases, such as myocardial infarction [18] , rheumatoid arthritis [52] , and T2DM [17] , but also with variations in levels of DPP4 [17] and apolipoprotein B [53] . In our study, an association of the DPP4 rs3788979 polymorphism with risk of COVID-19 was established.

We also found that the lowest levels of this enzyme were present in TT genotype carriers;

this polymorphism was previously associated with risk of myocardial infarction in patients with atherosclerosis [18] . Data are shown as mean ± standard deviation or percentage.

J o u r n a l P r e -p r o o f J o u r n a l P r e -p r o o f J o u r n a l P r e -p r o o f 

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The authors declare that they have no conflict of interest.