Association analysis of gene polymorphisms COL1A, MCT1, COL12A1 with sports hernia in football players

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O.O. Bogomoletz National Medical University

О.O. Bogomoletz Institute of Physiology National Academy of Sciences of Ukraine

Private higher educational establishment «Kyiv Medical University»

National University of Physical Education and Sports of Ukraine

Association analysis of gene polymorphisms COL1A, MCT1, COL12A1 with sports hernia in football players

O.Y. Ioffe, A.V. Omelchenko, S.V. Goncharov, D.O. Stroy, S.B. Drozdovska, O.P. Stetsenko, M.S. Kryvopustov, Y.A. Dibrova, T.M. Galyga, A.V. Vasilyev, V.M. Perepadya,T.V. Tarasiuk, V.E. Dosenko

Kyiv

Sports hernias are diagnosed in over 80% of athletes presenting with chronic groin pain. The genotype distributions of the examined polymorphisms were studied in 95 football players, all males, using the Copenhagen Hip and Groin Outcome Score (HAGOS) questionnaire. DNA extracted from buccal epithelium. Statistical calculations were performed in the R statistical environment (r-project.org) and Odds ratios (OR) were used to estimate genotype associations. Multifactor dimensionality reduction was used to identify multiple sports hernia susceptibility loci.Two single nucleotide polymorphisms MCT1 rs1049434 and COL1A1 rs1800012 are associated with sports hernias (63% testing accuracy shown by random forest). According to the HAGOS questionnaire outcomes, exercise-related pain, reduced mobility, and range of motion in the groin and thigh are typically associated with sports hernia diagnosis (75% testing accuracy shown by random forest).The combination of genetic research and HAGOS questionnaire helped obtain a sensitivity of 93% for the detection of sports hernia. Active engagement in sport is the most important risk factor for sports hernia, so the identification genes SNP in footballers may contribute to taking timely preventive actions. This study is the first demonstration of an association between sports hernia and COL1A1 rs1800012 genotype. The study discovered a synergistic interaction between the indicated polymorphism and the gene polymorphism MCT1 rs1049434. Further investigations are required to study the association between COL12A1 rs240736 gene SNP and sports hernia.

Key words: single nucleotide polymorphism; genetics;sports hernia; football player; COL1A; MCT1; COL12A1.

О.Ю. Іоффе, А.В. Омельченко, С.В. Гончаров, Д.О. Строй, С.Б. Дроздовська, О.П. Стеценко, М.С.Кривопустов, Ю.А. Діброва, Т.М. Галига, А.В. Васильєв, В.М. Перепадя, Т.В. Тарасюк, В.Є. Досенко

Національний медичний університет ім. О.О. Богомольця, Київ;

Інститут фізіології ім. О.О. Богомольця НАН України, Київ; Приватний вищий навчальний заклад "Київський медичний університет";

Національний університет фізичного виховання і спорту

ВИЯВЛЕННЯ ПОЛІМОРФІЗМІВ ГЕНІВ COL1A1, MCT1, COL12A ПРИ СПОРТИВНІЙ ГРИЖІ У ФУТБОЛІСТІВ

Спортивна грижа діагностується у 80% спортсменів з хронічними болями в пахвинній ділянці. Нами вивчено зв'язок між виникненням спортивної грижі та поліморфізмами, які було досліджено з букального епітелію 95 футболістів чоловічої статі, а також проведено оцінку якості життя за опитувальником HAGOS. Статистичні розрахунки проводили у пакеті R (r-project.org), для аналізу генотипів оцінювали відношення шансів. Визначено два однонуклеотидні поліморфізми MCT1 rs1049434 та COL1A1 rs1800012, які пов'язані зі спортивною грижею (63% за методом random forest). Біль під час фізичних вправ та обмеження рухів у пахвинній ділянці та стегні мають найбільший вплив на якість життя футболіста відповідно до результатів опитування HAGOS (75% за методом random forest). При поєднанні генетичного дослідження та анкетування отримано до 93% за методом random forest для виявлення спортивної грижі. Активні заняття спортом є фактором ризику її виникнення, а виявлення поліморфізмів генів SNP MCT1 rs1049434, COL1A1 rs1800012, COL12A1 rs240736 може сприяти наданню спортсменам своєчасних профілактичних заходів. У дослідженні вперше визначено зв'язок між спортивною грижею та COL1A1 га!800012. Дослідження виявило синергічну взаємодію поліморфізмами генів COL1A1 rs1800012 та MCT1 rs1049434.

Ключові слова: однонуклеотидний поліморфізм; генетика; спортивна грижа; футболіст; COL1A; MCT1; COL12A1.

А.Ю. Иоффе, А.В. Омельченко, С.В. Гончаров, Д.О. Строй, С.Б. Дроздовская, А.П. Стеценко, Н.С. Кривопустов, Ю.А. Диброва, Т.Н. Галыга, В. Васильєв, В.Н. Перепадя, Т.В. Тарасюк, Е. Досенко

ВИЯВЛЕНИЕ ПОЛИМОРФИЗМОВ ГЕНОВ COL1A1, MCT1, COL12A1 ПРИ СПОРТИВНОЙ ГРЫЖИ У ФУТБОЛИСТОВ

Спортивная грыжа диагностируется у 80% спортсменов с хроническими болями в паховой области. Нами изучена связь между возникновением спортивной грыжи и полиморфизмами из букального эпителия 95 футболистов мужского пола, также проведена оценка качества жизни по анкетированию HAGOS. Статистические расчеты проводили в среде R (r-project.org), для анализа ассоциаций генотипов оценивали отношения шансов.Определены два однонуклеотидные полиморфизмы MCT1 rs1049434 и COL1A1 rs1800012, связанные со спортивной грыжей (63% по методу random forest). Боль во время физических упражнений и ограничение объема движений в паховой области и бедре имеют наибольшее влияние на качество жизни футболиста согласно результатам опроса HAGOS (75% по методу random forest). При сочетании генетического исследования и анкетирования получено до 93% по методу random forest для выявления спортивной грыжи. Активные занятия спортом являются весомым фактором риска ее возникновения, а выявление полиморфизмов генов MCT1 rs1049434, COL1A1 rs1800012, COL12A1 rs240736 может способствовать своевременным профилактическим мероприятиям. В исследовании впервые определено значение между спортивной грыжей и COL1A1 rs1800012. Также выявлено синергическое взаимодействие между полиморфизмов генов COL1A1 rs1800012 и MCT1 rs1049434.

Ключевые слова: однонуклеотидный полиморфизм; генетика; спортивная грыжа; футболист; COL1A; MCT1; COL12A1.

Introduction

The study of polymorphism of various genes has recently significantly improved the prediction and treatment of various diseases. However, the study of gene polymorphism, including the detection of genes associated with sports injuries, remains poorly understood, and the polymorphisms of genes associated with sports hernias have not been identified yet. Therefore, early diagnosis and appropriate treatment can be challenging in the management of sports hernia.

Sports hernia is not a common groin injury, accounting for just up to 4% of all injuries to the hip and groin in high-profile male football players. Almost 50% of sports hernias in highprofile male football players result in more than a 4-week cessation of training as well as participation in football matches, and the injury time is almost double that of the injuries to the adductors [1].

However, a considerable number of studies suggest the association between specific genetic polymorphisms and elite athletic performance [2-5]. The COL1A1 (G^-T) gene determines collagen formation in cartilage, bone, skin and connective tissue. Moreover, collagen is the main structural component of tendons and ligaments. In the current study, it has been found that genetic variants in the COL1A1 gene, which encodes various collagen types, are associated with reduced risk of sports-related injuries. The MCT1 gene influences the lactate transport capacity and the intensity of sports performance, particularly rs1049434 polymorphism that has been related to athletic performance [6-10], injuries and body composition in football players [11, 12]. The association of the COL12 gene with the risk of sports injury and the presence of sports hernia has not been described in the literature, so it is necessary to establish whether the polymorphism of the COL12 gene can be used to predict sports hernia in football players. Carriers of the minor MCT1T allele have lactate transport rates reduced by 60-65% [13] and higher blood lactate accumulation during high-intensity circuit weight training, compared with carriers of the MCT1A allele [14]. Based on the known relationship between blood lactate accumulation and the MCT1 T allele and the assumption that football players, who had greater lactate transport rates, demonstrated a higher capacity to maintain their performance at intense effort levels, it was hypothesized that the MCT1 A1470T polymorphism could be associated with sports hernia. Therefore, the purpose of the study is to determine MCT1 rs1049434, COL1A1 rs1800012, COL12A1 rs240736 SNPs association with a risk of sports hernias in football players.

Methods

Participants: The buccal epithelium collected from 95 football players, all males and highprofile football players between ages 17 and 33, who were examined on the basis of the Department of General Surgery No. 2 at O.O. Bo- gomoletz National Medical University. The athletes included 41 football players with pain in the groin at the time of inclusion (experimental group) and 54 healthy youth football players (control group). Additional research methods were also used: assessment of anamnesis data; the HAGOS questionnaire; general clinical examination, inguinal examination; instrumental methods - ultrasound and MRI of the groin (Fig. 1).

Fig. 1. MRI groin scan in a patient with sports hernia. The white arrow indicate increased MR signal intensity observed from the structures of the inguinal canal, yellow arrow indicates increased lymph nodes on the side of groin pain

The Copenhagen Hip and Groin Outcome Score (HAGOS) questionnaire was used in this study. The HAGOS is a measure of pain and function in athletes, and reference values have already been obtained in football players. It consists of six subscales that are scored separately: «Pain» (P) - 10 items; «Symptoms» (S) - 7 items; «Physical function in daily living» (A) - 5 items; «Physical function in sport and recreation» (SP) - 8 items; «Participation in physical activities» (SP) - 2 items and «hip and/or groin-related quality of life» (Q) - 5 items. Each question gets a score from 0 to 4, where 0 indicates no problem. Raw scores are then transformed to a 0-100 scale, with zero representing extreme hip and/or groin issues and 100 representing no hip and/or groin pain; scores between 0 and 100 represent the percentage of the total possible score achieved [15]. The duration of the study was 1 year.

DNA collection and isolation. Genotyping: The buccal epithelium was collected from all football players and treated with means of a set of reagents NeoPrep DNA (“NeoPrep DNA”, Ukraine). All DNA samples were genotyped using an allelic determination assay on a 7500 Fast Real-Time PCR System (“Applied Biosystems”, “Foster City”, USA) with TaqMan probes. For the discrimination of theMCT1 rs1049434, CO- L1A1 rs1800012, COL12A1 rs240736 TaqMan® Pre-designed SNP Genotyping Assays were used 7500 Fast Real-Time PCR System (“Applied Biosystems”, USA; assay ID: C_2017662_30, C_7477170_30,C_3278190_10). According to the literature data, we found a significant role of these genes COL1A1, MTC1and COL12A1 in the occurrence of sports hernia (Table).

Genetic variants associated with groin disruption injury [24]

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