Method for predicting acute myocardial infarction in young patients with covid-19

Abstract

FIELD: cardiology.SUBSTANCE: used to predict acute myocardial infarction in young patients with COVID-19. The percentage of TRAIL and TGFβ1 is determined in peripheral blood monocytes using flow cytometry.β The obtained values are substituted into the formula for calculating the prognostic index (PI). If the PI value is equal to or greater than 12.90, a high risk of developing acute myocardial infarction is predicted. If the PI value is less than 12.90, the risk of developing acute myocardial infarction is predicted to be absent.EFFECT: method improves the accuracy of predicting acute myocardial infarction in young patients with COVID-19 by determining the PI taking into account the TRAIL and TGFβ1 values in peripheral blood monocytes.1 cl, 2 ex

Description

[0001] The invention relates to the field of medicine, in particular to cardiology, as well as to the field of scientific research - pathological physiology, and can be used to predict the risk of developing acute myocardial infarction in young patients with COVID-19.

[0002] According to the data available today, COVID-19 can cause damage not only to the respiratory system, but also to the cardiovascular system, which is due to the expression of angiotensin-converting enzyme type 2 [1].

[0003] According to the Russian registry “AKTIV”, in 31.8% of patients who had COVID-19, the predominant causes of death in the post-hospital period were acute coronary syndrome, stroke or acute heart failure [2].

[0004] Cardiovascular damage includes destabilization of existing pathology due to coronavirus infection, or the development of new COVID-associated complications: cardiomyopathy, pericarditis, thromboembolism, heart failure, arrhythmia, and acute myocardial injury [3]. The presence of pre-existing or newly developed cardiovascular pathology in patients predisposes to a more severe course and unfavorable prognosis [4].

[0005] Pathophysiological mechanisms of myocardial injury are diverse and may include both cell lysis associated with direct viral toxicity and inflammatory phenomena associated with hyperproduction of proinflammatory cytokines [5].

[0006] Among the most dangerous cardiovascular complications is acute myocardial infarction, which can develop both as a result of classic atherothrombotic mechanisms and as a result of direct myocardial damage by inflammatory mediators [6]. Cytokines and their receptors, including TNF-related apoptosis-inducing ligand - TRAIL (TNFSF10) and transforming growth factor beta 1 (TGFβ1), play a key role in this process. These molecules are involved in the regulation of apoptosis, inflammation, and fibrosis [7-13], which is of great importance for the prognosis and prevention of acute myocardial infarction in patients with COVID-19.

[0007] One of the mediators synthesized by monocytes during acute inflammation is TNFα, which regulates a wide range of signaling events within cells, including the NF-κB and MAP kinase pathways that mediate the inflammatory process, the expression of adhesion molecules, and cardiomyocyte apoptosis [9]. TNFα plays a key role in the development of the critical form of COVID-19 and the “cytokine storm” associated with this disease by stimulating the secretion of inflammatory cytokines [14, 15].

[0008] In patients with COVID-19, increased TNFα activity is associated with the development of myocarditis, myocardial ischemia, and cardiogenic shock

[10] . One of the mechanisms by which TNFα causes this damage is the activation of the apoptotic protein TRAIL. According to Bujak M., Frangogiannis NG (2007), excessive apoptosis contributes to the development of myocardial ischemia, post-ischemic cardiac remodeling, and coronary atherosclerosis

[13] . At the same time, TRAIL has anti-inflammatory and anti-atherogenic effects, especially in relation to the endothelium [7], and a disturbance in its content in the blood serum is associated with an increased risk of acute coronary syndrome and myocardial infarction [8].

[0009] Another cytokine involved in the development of cardiovascular complications in patients with acute myocardial infarction is TGFβ1, a multifunctional cytokine involved in the regulation of inflammation, apoptosis, and fibrogenesis. Although it has anti-inflammatory properties, its excessive activation can lead to fibrosis and myocardial remodeling

[11] . In COVID-19, an increase in TGFβ1 levels is observed in the late phase of the disease and is associated with the formation of myocardial fibrosis after inflammatory damage, including that caused by infarction

[12] . In addition, TGFβ1 is involved in the regulation of the coagulation system and can contribute to thrombus formation and secondary myocardial injury

[13] .

[0010] Therefore, the results of studies by both domestic and foreign authors prove that pro-inflammatory cytokines (TRAIL and TGFβ1) contribute to the development of acute myocardial infarction.

[0011] The above data highlight the need to develop diagnostic methods for cardiovascular disorders to identify factors leading to the development of acute myocardial infarction in young patients with COVID-19. According to the World Health Organization (WHO) classification, young patients include individuals aged 18-44 years, who are also at risk of developing severe cardiovascular complications from COVID-19.

[0012] Thus, predicting the risk of developing acute myocardial infarction in young patients with COVID-19 is a promising and optimal strategy for this pathology.

[0013] Based on the described information, we proposed a method for predicting the risk of developing acute myocardial infarction by assessing the percentage of TRAIL and TGFβ1 in peripheral blood monocytes in young patients with COVID-19 using flow cytometry.

[0014] The reviewed scientific, medical, and patent literature did not reveal a method for predicting acute myocardial infarction in young patients with COVID-19; therefore, the proposed method meets the criterion of novelty.

[0015] There are known studies in which the differential expression of 27 cytokine genes is studied to predict acute myocardial infarction, including proinflammatory mediators and their receptors: IFNα2, IFNαR1, IFNγR1, IL-1β, IL-16, IL-18, TNFR1, LTβR, CD137, TRAIL, Cxcl1, Cxcl2, CxcR2, CD137, CD40, CD27, etc.

[16] .

[0016] The main disadvantages of the above method are:

[0017] 1) the need to study the expression of a large number of cytokines and their receptors, which makes the procedure expensive;

[0018] 2) lack of integration of the studied indicators with the infectious pathology caused by SARS-CoV-2.

[0019] A well-known analogue of the claimed invention is also the study by del Rosario Espinoza Mora M. et al. (2014), who studied subpopulations of Treg cells (TGFβ and IL-10) using enzyme-linked immunosorbent assay in patients with cardiogenic shock

[17] .

[0020] The main drawback of this method is the lack of integration of the studied parameters with the infectious pathology caused by SARS-CoV-2. Furthermore, it does not take into account the role of TRAIL in the development of cardiovascular disorders.

[0021] There are known studies in which factor and cluster analysis of the profile of 10 biomarkers associated with fibrosis, inflammation and remodeling of cardiac tissue, such as MMP-2, MMP-3, MMP-8, MMP-9, periostin, N-terminal propeptide of procollagen I, osteopontin, TGFβ1, TIMP-1 and TIMP-4 are used to predict cardiovascular disorders

[18] .

[0022] The main disadvantage of this method is the multifactorial nature of the parameters studied, as well as the lack of integration of the studied indicators with the infectious pathology caused by SARS-CoV-2, which limits its application.

[0023] The closest analogue of the proposed invention is the method proposed by foreign researchers

[19] , which allows diagnosing the risk of developing acute myocardial infarction by determining the expression of TRAIL and its receptors, including DR4, DR5, and TNFR1, in peripheral blood mononuclear cells. We have chosen this method as the closest analogue (prototype).

[0024] A disadvantage of the prototype method is that it does not effectively predict the development of acute myocardial infarction in young patients, who may have weakly expressed classical risk factors, but have pathogenetically significant changes in the inflammatory mediator system, including TRAIL and TGFβ1, which makes the proposed method more specific and sensitive for this category of patients. Traditional biomarkers (e.g., troponins) increase after the onset of myocardial necrosis, whereas new molecules (TRAIL, TGFβ1) can serve as earlier risk predictors, allowing for the initiation of preventive measures before the development of irreversible changes

[14] .

[0025] The aforementioned deficiencies of the prototype method can be eliminated in the claimed method. The technical result achieved by implementing the proposed invention is an increase in the accuracy of predicting acute myocardial infarction in young patients with COVID-19.

[0026] The claimed technical result is achieved by determining the percentage of TRAIL in peripheral blood monocytes of young patients with COVID-19 using flow cytometry using APC (Allophycocyanin)-labeled monoclonal antibodies to TRAIL and TGFβ1 using PE (R-phycoerythrin)-labeled monoclonal antibodies to TGFβ1, in accordance with the recommendations of the kit developer (Biolegend, USA). The results are recorded using a BD Facs Canto II flow cytometer (USA).

[0027] The obtained values ​​are used to calculate the prognostic index (PI) using a formula developed using the discriminant analysis method: PI = -13.197 + 0.355 × TRAIL + 0.276 × TGFβ1, where PI is the discriminant function with a cutoff value of 12.90. When the PI is equal to or greater than the cutoff value, the risk of developing acute myocardial infarction in young patients with COVID-19 is predicted. When the PI is less than the cutoff value, no risk of developing acute myocardial infarction is predicted in young patients with COVID-19.

[0028] The novelty of the proposed method lies in the determination of the prognostic index (PI), taking into account TRAIL and TGFβ1 values ​​in peripheral blood monocytes in young patients with COVID-19. This index, relative to the cutoff value of the discriminant function, is used to predict the risk of developing acute myocardial infarction in young patients with COVID-19. Previously, the combination of these indicators has not been used to predict the risk of developing acute myocardial infarction in young patients with COVID-19.

[0029] The method contains the following techniques:

[0030] 1) In young patients with COVID-19, the percentage of TRAIL in peripheral blood taken by venipuncture is determined using flow cytometry in peripheral blood monocytes using APC (Allophycocyanin)-labeled monoclonal antibodies to TRAIL and TGFβ1 using PE (R-phycoerythrin)-labeled monoclonal antibodies to TGFβ1;

[0031] using the discriminant equation, the prognostic index PI = -13.197 + 0.355 × TRAIL + 0.276 × TGFβ1 is calculated, where PI is the discriminant function, the boundary value of which is 12.90.

[0032] 2) determines the risk of developing acute myocardial infarction in young patients with COVID-19: if the PI is equal to or greater than the cutoff value, the risk of developing acute myocardial infarction in young patients with COVID-19 is predicted. If the PI is less than the cutoff value, the risk of developing acute myocardial infarction in young patients with COVID-19 is predicted to be absent. The probability of a correct prediction is 89.5%.

[0033] The use of this method is confirmed by specific examples:

[0034] Example 1. Patient Ch., 44, was hospitalized in a COVID-19 hospital with a diagnosis of novel coronavirus infection (COVID-19) (confirmed). Upon admission, he complained of a fever of 38.2°C, cough, runny nose, and shortness of breath when walking.

[0035] Objectively: body temperature upon admission is 37.7°C, respiratory rate is 24 / min, shortness of breath when walking, heart rate is 87 beats per minute, SpO2 is 94%.

[0036] Patient Ch. received therapy according to the temporary guidelines “Prevention, diagnosis, and treatment of new coronavirus infection (COVID-19)”.

[0037] A nasopharyngeal (both nasal passages) and oropharyngeal swab tested positive for SARS-CoV-2 RNA using the polymerase chain reaction method. The patient had not previously been seen by a cardiologist and denies a history of cardiovascular disease.

[0038] Considering the high risk of developing acute myocardial infarction, the patient's peripheral blood was collected and the percentage of TRAIL and TGFβ1 was determined on monocytes. The following indicators were obtained using the claimed method: TRAIL - 60.10% and TGFβ1 - 18.62%. The prognostic index, calculated using the formula PI = -13.197 + 0.355 × TRAIL + 0.276 × TGFβ1, was 13.28.

[0039] The development of acute myocardial infarction is predicted, therefore the patient was assigned to a high-risk group for the development of this disease.

[0040] On the 14th day of hospitalization, the patient developed a primary posterolateral left ventricular myocardial infarction with ST-segment elevation, without a Q wave. The prognosis according to the stated method was confirmed.

[0041] Example 2. Patient M., 38, was hospitalized in a COVID-19 hospital with a diagnosis of novel coronavirus infection (COVID-19) (confirmed). Upon admission, he complained of a fever of 38.5°C, weakness, and shortness of breath during physical exertion.

[0042] Objectively: body temperature upon admission is 38.0°C, respiratory rate is 23 / min, shortness of breath when walking, heart rate is 89 beats per minute, SpO2 is 93%.

[0043] Patient M. received therapy according to the interim guidelines "Prevention, diagnosis, and treatment of novel coronavirus infection (COVID-19)"

[0044] A nasopharyngeal (both nasal passages) and oropharyngeal swab tested positive for SARS-CoV-2 RNA using the polymerase chain reaction method. The patient had not previously been seen by a cardiologist and denies a history of cardiovascular disease.

[0045] Peripheral blood was collected from the patient and the percentage of TRAIL and TGFβ1 was determined on monocytes. The following indicators were obtained using the stated method: TRAIL - 13.49%, TGFβ1 - 4.0%. The prognostic index, calculated using the formula PI = -13.197 + 0.355 × TRAIL + 0.276 × TGFβ1 was (-7.303).

[0046] Conclusion: The development of acute myocardial infarction is not predicted.

[0047] These examples clearly demonstrate the accuracy of predicting the development of acute myocardial infarction in young patients with COVID-19.

[0048] The method has undergone clinical testing. It was used to predict acute myocardial infarction in 52 young patients with COVID-19. The correct prognosis was determined in 89.5% of cases, confirming the high efficacy of the proposed method.

[0049] List of references

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Claims

A method for predicting acute myocardial infarction in young patients with COVID-19, which consists in determining the percentage of TRAIL in peripheral blood monocytes of patients with COVID-19 using flow cytofluorimetry, characterized in that the percentage of TGFβ1 in peripheral blood monocytes is additionally determined, after which the prognostic index (PI) is calculated using the formula: PI = -13.197 + 0.355 × TRAIL + 0.276 × TGFβ1, with a PI value equal to or greater than 12.90, a high risk of developing acute myocardial infarction is predicted, and with a PI value less than 12.90, no risk of developing acute myocardial infarction is predicted.