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Polymorphisms in VEGFA and KDR encoding proteins have been associated with anterior cruciate ligament (ACL) injury risk. We leveraged a collective sample from Sweden, Poland, and Australia to investigate the association of functional polymorphisms in VEGFA and KDR with susceptibility to ACL injury risk. Using a case–control genetic association approach, polymorphisms in VEGFA and KDR were genotyped and haplotypes inferred from 765 controls, and 912 cases clinically diagnosed with ACL rupture. For VEGFA, there was a significant overrepresentation of the rs2010963 CC genotype (p = 0.0001, false discovery rate [FDR]: p = 0.001, odds ratio [OR]: 2.16, 95% confidence interval [CI]: 1.47–3.19) in the combined ACL group (18%) compared to the combined control group (11%). The VEGFA (rs699947 C/A, rs1570360 G/A, rs2010963 G/C) A-A-G haplotype was significantly (p = 0.010, OR: 0.85, 95% CI: 0.69–1.05) underrepresented in the combined ACL group (23%) compared to the combined control group (28%). In addition, the A-G-G construct was significantly (p = 0.036, OR: 0.81, 95% CI: 0.64–1.02) underrepresented in the combined ACL group (12%) compared to the combined CON group (16%). Our findings support the association of the VEGFA rs2010963 CC genotype with increased risk and (ii) the VEGFA A-A-G haplotype with a reduced risk, and are in alignment with the a priori hypothesis. Collectively identifying a genetic interval within VEGFA to be implicated in ACL risk modulation and highlight further the importance of vascular regulation in ligament biology.

The aim of this study was to explore the interactions between the interleukins and the angiogenesis signalling pathway, following a pathway-based approach. Statistical modelling tools were used to develop a preliminary polygenic risk assessment model for anterior cruciate ligament (ACL) ruptures, incorporating the angiogenesis signalling genes (

Objectives Anterior cruciate ligament rupture (ACLR) is a common and severe knee injury which typically occurs as a result of sports participation, primarily via a non-contact mechanism. A number of extrinsic and intrinsic risk factors, including genetics, have been identified thus far. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteases (TIMPs) play a crucial role in extracellular matrix remodeling of ligaments and therefore the genes encoding MMPs and TIMPs are plausible candidates for investigation with ACL rupture risk. Design A case-control genetic association study was conducted on 229 (158 male) individuals with surgically diagnosed primary ACLR, ruptured through non-contact mechanisms and 192 (107 male) apparently healthy participants (CON) without any history of ACLR. All participants were physically active, unrelated, self-reported Caucasians. Methods All participants were genotyped for four single nucleotide polymorphisms (SNP): MMP3 (rs591058C/T, rs679620 G/A), MMP8 (rs11225395C/T), and TIMP2 (rs4789932 G/A) using standard PCR assays. Gene-gene interactions were inferred. Single-locus association analysis was conducted using the Chi-square test. SNP-SNP interaction effects were analysed using multifactor dimensionality reduction (MDR) method. Results Genotype frequencies did not significantly differ between cases and controls, however, the MMP3 rs679620 G and rs591058C alleles were significantly overrepresented in cases compared to controls (p = 0.021, OR = 1.38, 95% CI: 1.05–1.81). Conclusions These results support the hypothesis that genetic variation within MMP3 contributes to inter-individual susceptibility to non-contact ACLR. However, these results need to be explored further in larger, independent sample sets.

Collagen alpha-1(V) chain, encoded by the COL5A1 gene, plays a crucial role in abundant fibrillar collagens supporting many tissues in the body containing type I collagen and appears to regulate the association between heterotypic fibers composed of both type I and type V collagen occurring among others in muscles, tendons and ligaments. Taking this fact into consideration we decided to examine the association between COL5A1 rs12722 and rs13946 polymorphisms, individually and as inferred haplotypes, with anterior cruciate ligament rupture risk (ACLR) in professional soccer players. A total of 134 male professional soccer players with surgically diagnosed primary anterior cruciate ligament ruptures and 211 apparently healthy male professional soccer players, who were without any self-reported history of ligament or tendon injury, were included in the study. Both the cases and the healthy controls were recruited from the same soccer teams, of a similar age category, and had a comparable level of exposure to anterior cruciate ligament injury. Genomic DNA was extracted from oral epithelial cells using GenElute Mammalian Genomic DNA MiniprepKit. All samples were genotyped for the rs12722 and rs13946 polymorphisms using a Rotor-Gene realtime polymerase chain reaction. Statistically significant differences in the genotype frequencies for the COL5A1 rs13946 polymorphisms in dominant modes of inheritance occurred (p = 0.039). Statistically significant differences were documented only in the dominant model under the representation tendency of the C-C haplotype in the ACLR group compared to controls (p = 0.038). Our results suggest that variation in the COL5A1 gene may be one of the non-modifiable factors associated with the ACL injury in professional soccer players. The C-C rs12722-rs13946 haplotype provides a protective effect against the ACL tear.

The evidence implicating genetics in the susceptibility to musculoskeletal soft tissue injuries is growing, with over 80 risk-conferring loci discovered to date. Primarily through the application of candidate gene studies, researchers have highlighted some of the critical extracellular matrix proteins and biological pathways underpinning matrix remodeling proposed to influence injury susceptibility. However, many of these loci have yet to be repeated in independent cohorts. Moreover, limited functional data is available at present to support the implicated genomic regions. Recent technologies have enabled researchers to explore the genome more broadly, though these studies are limited by the unavailability of large groups of well-defined cases and controls. It is essential that researchers collaborate to unravel the complexities of matrix remodeling, tissue capacity, and thereby improve our understanding of injury susceptibility. Through the establishment of international consortia, large cohorts can be attained to aid the identification of clinically relevant loci and molecules that may be targeted for therapeutic applications.

To investigate the functional effect of genetic polymorphisms of the inflammatory pathway on structural extracellular matrix components (ECM) and the susceptibility to an anterior cruciate ligament (ACL) injury.Laboratory study, case-control study.Eight healthy participants were genotyped for interleukin (IL)1B rs16944 CT and IL6 rs1800795 GC and classified into genetic risk profile groups. Differences in type I collagen (COL1A1), type V collagen (COL5A1), biglycan (BGN) and decorin (DCN) gene expression were measured in fibroblasts either unstimulated or following IL-1β, IL-6 or tumor necrosis factor (TNF)-α treatment. Moreover, a genetic association study was conducted in: (i) a Swedish cohort comprised of 116 asymptomatic controls (CON) and 79 ACL ruptures and (ii) a South African cohort of 100 CONs and 98 ACLs. Participants were genotyped for COL5A1 rs12722 CT, IL1B rs16944 CT, IL6 rs1800795 GC and IL6R rs2228145 GC.IL1B high-risk fibroblasts had decreased BGN (p=0.020) and COL5A1 (p=0.012) levels after IL-1β stimulation and expressed less COL5A1 (p=0.042) following TNF-α treatment. Similarly, unstimulated IL6 high-risk fibroblasts had lower COL5A1 (p=0.012) levels than IL6 low-risk fibroblasts. In the genetic association study, the COL5A1-IL1B-IL6 T-C-G (p=0.034, Haplo-score 2.1) and the COL5A1-IL1B-IL6R T-C-A (p=0.044, Haplo-score: 2.0) combinations were associated with an increased susceptibility to ACL injury in the Swedish cohort when only male participants were evaluated.This study shows that polymorphisms within genes of the inflammatory pathway modulate the expression of structural and fibril-associated ECM components in a genetic risk depended manner, contributing to an increased susceptibility to ACL injuries.

Collagen alpha-1(V) chain, encoded by the COL5A1 gene, plays a crucial role in abundant fibrillar collagens supporting many tissues in the body containing type I collagen and appears to regulate the association between heterotypic fibers composed of both type I and type V collagen occurring among others in muscles, tendons and ligaments. Taking this fact into consideration we decided to examine the association between COL5A1 rs12722 and rs13946 polymorphisms, individually and as inferred haplotypes, with anterior cruciate ligament rupture risk (ACLR) in professional soccer players. A total of 134 male professional soccer players with surgically diagnosed primary anterior cruciate ligament ruptures and 211 apparently healthy male professional soccer players, who were without any self-reported history of ligament or tendon injury, were included in the study. Both the cases and the healthy controls were recruited from the same soccer teams, of a similar age category, and had a comparable level of exposure to anterior cruciate ligament injury. Genomic DNA was extracted from oral epithelial cells using GenElute Mammalian Genomic DNA MiniprepKit. All samples were genotyped for the rs12722 and rs13946 polymorphisms using a Rotor-Gene realtime polymerase chain reaction. Statistically significant differences in the genotype frequencies for the COL5A1 rs13946 polymorphisms in dominant modes of inheritance occurred (p = 0.039). Statistically significant differences were documented only in the dominant model under the representation tendency of the C-C haplotype in the ACLR group compared to controls (p = 0.038). Our results suggest that variation in the COL5A1 gene may be one of the non-modifiable factors associated with the ACL injury in professional soccer players. The C-C rs12722-rs13946 haplotype provides a protective effect against the ACL tear.

Angiogenesis-associated signaling is a fundamental component in the remodeling of the extracellular matrix in response to loading. Genes encoding protein components within this signaling cascade are therefore suitable candidates for investigation into ACL injury susceptibility: namely, vascular endothelial growth factor A isoform (VEGFA), kinase insert-domain receptor (KDR), nerve growth factor (NGF), and hypoxia inducible factor-1α (HIF1A). A case-control genetic association study was conducted on 227 asymptomatic control participants and 227 participants with surgically diagnosed ACL ruptures of which 126 participants reported a non-contact mechanism of rupture. All participants were genotyped for seven polymorphisms within the four genes. The VEGFA rs699947 CC genotype (p=0.010, OR: 1.92, 95% CI: 1.17-3.17) was significantly over-represented within participants with non-contact ACL ruptures. The VEGFA rs1570360 GA genotype was significantly over-represented in the CON group (p=0.007, OR: 1.70, 95% CI: 1.16-2.50). Furthermore, the KDR rs2071559 GA genotype was significantly over-represented in the female controls (p=0.023, OR: 2.16, 95% CI: 1.11-4.22). Inferred haplotype analyses also implicated genomic regions spanning the VEGFA and KDR genes. These novel findings suggest that regions within VEGFA and KDR may be implicated in the pathophysiology of ACL ruptures; highlighting the potential biological significance of angiogenesis-associated signaling in the aetiology of ACL ruptures.

Sport and Exercise Medicine is one of the important subspecialties of 21st century healthcare contributing to improving the physical function, health, and vitality of populations while reducing the prevalence of lifestyle-related diseases. Moreover, sport and exercise are associated with injuries such as Achilles tendinopathy, which is a common tendon injury. The angiogenesis-associated signaling pathway plays a key role in extracellular matrix remodeling, with increased levels of angiogenic cytokines reported after cyclic stretching of tendon fibroblasts. We investigated the variants in angiogenesis genes in relation to the risk of Achilles tendinopathy in two population samples drawn independently from South Africa (SA) and the United Kingdom (UK). The study sample comprised 120 SA and 130 UK healthy controls, and 108 SA and 87 UK participants with Achilles tendinopathy. All participants were genotyped for five functional polymorphisms in the vascular endothelial growth factor, A isoform (VEGFA) (rs699947, rs1570360, rs2010963) and kinase insert-domain receptor (KDR) genes (rs1870377, rs2071559). The VEGFA A-G-G inferred haplotype was associated with an increased risk of Achilles tendinopathy in the SA group (15% in controls vs. 20% in cases, p = 0.048) and the combined SA+UK group (14% in controls vs. 20% in cases, p = 0.009). These new findings implicate the VEGFA gene with Achilles tendinopathy risk, while highlighting the potential biological significance of the angiogenesis signaling pathway in the etiology of Achilles tendinopathy. The evidence suggesting a genetic contribution to the susceptibility of sustaining a tendon injury is growing. We anticipate that high-throughput and multi-omics approaches, building on genomics, proteomics, and metabolomics, may soon uncover the pathophysiology of many diseases in the field of Sports and Exercise Medicine, as a new frontier of global precision medicine.

To date, more than 18 genomic intervals, which underpin the complex myriad of extracellular matrix interactions of tendons, have been implicated in risk models for tendinopathy. It is these relationships that most likely regulate the tissue's response to loading and unloading, thereby dictating the overall capacity of tendons and influencing injury susceptibility. The evidence suggesting a genetic contribution to the susceptibility of sustaining a tendon injury is growing. However, only a few of the loci have been repeated in independent studies, of which some have included a range of musculoskeletal soft tissues injuries. Case-control study designs can be effective in capturing risk, provided that the cases and controls are equally well-defined and carefully considered. The genome consists of 3.6 × 10(9) sequences and therefore we realise that we are far from decoding all the genomic signatures. We are indeed fortunate to be living in such exciting times where high-throughput technologies are at our disposal. Through collaboration, our chances of harnessing these "omics" technologies to further our clinical understanding of tendinopathy will increase.

There is mounting evidence suggesting a genetic contribution to the susceptibility of sustaining a musculoskeletal soft-tissue injury. To date, more than 70 loci have been implicated in several injury profiles. The genes implicated through these loci encode a broad spectrum of matrix proteins including collagens and non-collagens. The large majority of these studies have followed a candidate gene case-control study design. A small proportion of these loci have been repeated in independent studies, of which some have included different musculoskeletal injuries. However, the large majority of these studies are underpowered to detect contributions of small effect sizes (odds ratio2.0). It is therefore critical that large data sets are collected and that consortia are established to effectively pool resources to understand the biological significance of these genetic loci and risk susceptibility. We are in the era of omics and high-throughput technologies but it is only through collaborations that we will realize the clinical significance of the genomic revolution and its application to musculoskeletal soft-tissue injury susceptibility.

To date, more than 18 genomic intervals, which underpin the complex myriad of extracellular matrix interactions of tendons, have been implicated in risk models for tendinopathy. It is these relationships that most likely regulate the tissue’s response to loading and unloading, thereby dictating the overall capacity of tendons and influencing injury susceptibility. The evidence suggesting a genetic contribution to the susceptibility of sustaining a tendon injury is growing. However, only a few of the loci have been repeated in independent studies, of which some have included a range of musculoskeletal soft tissues injuries. Case-control study designs can be effective in capturing risk, provided that the cases and controls are equally well-defined and carefully considered. The genome consists of 3.6 × 109 sequences and therefore we realise that we are far from decoding all the genomic signatures. We are indeed fortunate to be living in such exciting times where high-throughput technologies are at our disposal. Through collaboration, our chances of harnessing these “omics” technologies to further our clinical understanding of tendinopathy will increase.

Angiogenesis-associated signaling is a fundamental component in the remodeling of the extracellular matrix in response to loading. Genes encoding protein components within this signaling cascade are therefore suitable candidates for investigation into ACL injury susceptibility: namely, vascular endothelial growth factor A isoform (VEGFA), kinase insert-domain receptor (KDR), nerve growth factor (NGF), and hypoxia inducible factor-1α (HIF1A). A case-control genetic association study was conducted on 227 asymptomatic control participants and 227 participants with surgically diagnosed ACL ruptures of which 126 participants reported a non-contact mechanism of rupture. All participants were genotyped for seven polymorphisms within the four genes. The VEGFA rs699947 CC genotype (p=0.010, OR: 1.92, 95% CI: 1.17-3.17) was significantly over-represented within participants with non-contact ACL ruptures. The VEGFA rs1570360 GA genotype was significantly over-represented in the CON group (p=0.007, OR: 1.70, 95% CI: 1.16-2.50). Furthermore, the KDR rs2071559 GA genotype was significantly over-represented in the female controls (p=0.023, OR: 2.16, 95% CI: 1.11-4.22). Inferred haplotype analyses also implicated genomic regions spanning the VEGFA and KDR genes. These novel findings suggest that regions within VEGFA and KDR may be implicated in the pathophysiology of ACL ruptures; highlighting the potential biological significance of angiogenesis-associated signaling in the aetiology of ACL ruptures.

Introduction Genetic factors have been implicated with risk of Achilles tendinopathy. Angiogenesis plays an important role in extracellular matrix (ECM) remodelling following mechanical loading. It is proposed that dysregulation of this process may alter the mechanical properties of the tissue thereby increasing the risk of injury. Furthermore, increased levels of angiogenic cytokines and growth factors have been reported in injured Achilles tendons [Pufe, 2001] and after cyclic stretching of tendon fibroblasts [Petersen, 2004]. Vascular endothelial growth factor (VEGF) is a potent pro-angiogenic factor and the biological effects of VEGF are mediated through its receptor kinase insert-domain receptor (KDR). Recently, polymorphisms within the VEGFA and KDR genes were found to be associated with risk of anterior cruciate ligament (ACL) ruptures [Rahim, 2014]. Thus, the aim of this study was to investigate if genes encoding proteins involved in angiogenesis-associated signalling are associated with risk of Achilles tendon injuries. Methods A genetic-association study was conducted on 120 control participants (CON), 91 participants with Achilles tendinopathy (TEN) and 44 participants with partial or complete ruptures of the Achilles tendon (RUP). All participants were genotyped for the functional VEGFA rs699947, VEGFA rs1570360, VEGFA rs2010963, KDR rs2071559 and KDR rs1870377 polymorphisms. Haplotypes were also inferred for VEGFA and KDR . Statistical analyses were conducted to determine any significant differences (p Results No independent significant differences were observed in the genotype frequency distributions of any of the polymorphisms between the CON vs. TEN groups. However, there was a trend towards significance (p = 0.053) in the genotype frequencies for the VEGFA rs699947 polymorphism when comparing the CON group vs. the RUP group. The CC genotype was significantly over-represented (p = 0.019) in the CON group (24%) compared to the RUP group (3%). Inferred haplotype analyses showed no significant difference in the frequency distributions of the VEGFA or KDR haplotype combinations between the CON vs. TEN groups or the CON vs. RUP groups. Discussion This pilot study did not find a statistical association with any of the five tested polymorphisms within the VEGFA and KDR genes using the Achilles tendinopathy risk model. However, the preliminary results are suggesting that the VEGFA rs699947 polymorphism may be implicated in acute injuries as noted in the Achilles tendon rupture cases. This hypothesis would be in alignment with the previous ACL risk model observations [Rahim, 2014]. These loci therefore need further interrogation in larger sample sizes. Studies are underway to explore the five polymorphisms within two independent case-control samples sets, in order to elucidate the potential biological significance of the angiogenesis-associated cell signalling pathway in the aetiology of Achilles tendon injuries (overuse and acute). References Petersen et al . J Orthop Res. 2004;22:847–853 Pufe et al . Virchows Arch. 2001;439(4):579–585 Rahim et al . J Orthop Res. 2014 (in press)