- Email: [email protected]
Novel associations in mitochondria-related genes and mitochondria-nucleus interactions. Role in osteoarthritis
S210
Abstracts / Osteoarthritis and Cartilage 25 (2017) S76eS444
annotated as predicted secreted proteins and have potential value as biomarkers of OA. We performed an in silico DrugBank search and found ten compounds that target genes highlighted in the pilot study at stringent confidence levels, and which have FDA marketing authorisation for an existing clinical indication. Three genes are consistently differentially regulated across all 3 omics levels: AQP1, COL1A1 and CLEC3B. For example, AQP1 encodes aquaporin-1, which facilitates water transport across biological membranes. Chondrocyte swelling and increased cartilage hydration has been suggested as an important mechanism in OA. The carbonic anhydrase inhibitor Acetazolamide is a known inhibitor of aquaporin-1, indicating the potential of functional genomics to identify opportunities for drug repurposing, shortening the investigative pipeline to use in OA. We used independent datasets for knee OA and hip OA to show that the direction of change replicates for over 90% of differentially expressed genes and differentially methylated probes, and to demonstrate good agreement in the global estimates of the fold-changes. Integrated pathway analysis implicates the involvement of extracellular matrix degradation, collagen catabolism and angiogenesis in disease progression. Conclusions: This work provides a comprehensive view of the molecular landscape of human primary chondrocytes in OA progression. All data from these experiments are freely available as a resource for the scientific community. 335 NOVEL ASSOCIATIONS IN MITOCHONDRIA-RELATED GENES AND MITOCHONDRIA-NUCLEUS INTERACTIONS. ROLE IN OSTEOARTHRITIS s-Pereira y, M. Ferna ndez-Moreno y, I. Rego-Perez y, E. Corte zquez-Mosquera y, S. Relan ~ o y, S. Pe rtega z, A. Go nza lez x, M. Va ndez-Lo pez y, F. Blanco y. y Servicio L. Vidal-Bralo x, N. Oreiro y, C. Ferna n Biom ~ a (INIBIC). de Reumatología. Inst. de Investigacio edica de A Corun ~ a (CHUAC), Sergas. Univ.e da Complexo Hosp.ario Univ.rio de A Corun ~ a (UDC). As Xubias, 15006, A Corun ~ a, Spain; z Unidad de Corun n Biom Epidemiología Clínica y Bioestadística. Inst. de Investigacio edica ~ a (INIBIC). Complexo Hosp.ario Univ.rio de A Corun ~a de A Corun ~ a (UDC). As Xubias, 15006, A Corun ~ a, (CHUAC), Sergas. Univ.e da Corun n 10 y Unidad de Reumatología. Inst. Spain; x Laboratorio de Investigacio n Sanitaria de Santiago (IDIS), Hosp. Clínico Univ.rio de de Investigacio Santiago, Travesía Choupana s/n, 15706, Santiago de Compostela, Spain Purpose: To analyze the potential influence of specific gene polymorphisms of mitochondria-related genes involved in cellular detoxification and to explore the mitochondria-nucleus interactions in the development of hip OA Methods: For this work we used two well-characterized cohorts of ~ a and patients from Galicia (Spain) including patients from A Corun Santiago reaching a total of 944 subjects, of which 601 were hip OA with KL grade IV and 343 age-matched radiological healthy controls (KL grade 0-I). The mtDNA haplogroups were assigned to all the subjects and grouped into mtDNA clusters. In a first subset of subjects we analyzed a set of mitochondria-related gene polymorphisms involved in the cellular detoxification: CAT (rs659366), GPX1 (rs2234693), UCP2 (rs659366), SOD2 (rs4880) and NOS2A (microsatellite (CCTTT)n in the promoter region of the gene), as well as their potential interaction with mtDNA clusters. In a second set of subjects we analyzed the potential interactions between the mtDNA clusters and a set of previously proposed gene polymorphisms related to OA: GDF-5 (rs143383), CILP (rs2073711), PAI (rs2227631), MMP-1 (rs514921) and MMP-2 (rs243866). The obtained data were appropriately analyzed using PowerMarker and SPSS (v19) Results: We found that the TT genotype of rs659366 in CAT gene was significantly underrepresented in hip OA patients (OR ¼ 0.512;95% CI ¼ 0.321-0.817;p ¼ 0.004). In relation to NOS2A microsatellite, the 13repetition allele was significantly underrepresented in hip OA patients too (OR ¼ 0.675;95%CI ¼ 0.509-0.895;p ¼ 0.006) meanwhile the 11 repetition-allele was significantly associated with an increased risk of hip OA (OR ¼ 1.524;95%CI ¼ 1.133-2.049;p ¼ 0.005). The subsequent regression model adjusting for the confounder variables of gender and age confirmed the above described associations as well as the mtDNA cluster TJ as a protective factor against hip OA (OR ¼ 0.672; 95% CI ¼ 0.455-0.993;p ¼ 0.046) (Table 1). In terms of mitochondria-nucleus interactions, a significant interaction between the SNP rs2234693 in GPX1 and the mtDNA cluster TJ (p ¼ 0.003) and HV (p ¼ 0.05) was discovered; the combination of cluster TJ and CC genotype emerged as a
protective factor (OR ¼ 0.427;95%CI ¼ 0.249-0.742;p ¼ 0.002), and the combination of cluster HV and TT genotype resulted in a protective factor too (OR ¼ 0.535;95%CI ¼ 0.307-0.934;p ¼ 0.028). In addition, we found significant interactions between the mtDNA clusters HV and/or TJ and other specific nuclear polymorphisms in PAI (p < 0.001 for TJ and p ¼ 0.049 for HV) and MMP1 (p ¼ 0.016 for HV) genes. The combination of cluster TJ with TT genotype of PAI significantly associates with a decreased risk of hip OA (OR ¼ 0.183;95%CI ¼ 0.068-0.490;p ¼ 0.001), and the combination of cluster HV with no-TT genotype associated with decreased risk too (OR ¼ 0.496;95%CI ¼ 0.291-0.848;p ¼ 0.010). In relation to MMP1, the combination of cluster HV with AA genotype associates with an increased risk of hip OA (OR ¼ 1.770;95%CI ¼ 1.0243.058;p ¼ 0.041) (Table 2). Finally, the interactions involving both CILP (p ¼ 0.058 for HV) and MMP-2.1575 (p ¼ 0.062 for TJ) border-lined the statistical significance Conclusions: The associations described in this work indicate that mitochondrial genetic background determines the behavior of specific nuclear variants in the development of OA
Table 1 Variables
p-value
OR
95%CI
Gender (male) Age NOS2A-11 NOS2A-13 CAT-TT mtDNA clusters (n ¼ 944) HV (n ¼ 496) KU (n ¼ 222) TJ (n ¼ 147) Others (n ¼ 79)
0.098 0.034 0.041 0.016 0.009
1.275 0.987 1.387 0.694 0.523
0.956-1.700 0.974-0.999 1.014-1.897 0.515-0.935 0.321-0.853
0.693 0.046 0.385
1 1.073 0.672 1.277
0.756-1.522 0.455-0.993 0.735-2.216
Table 2 Cluster TJ
Cluster HV
Genotypes (TJ/HV)
p-value
OR
95% CI
p-value
OR
95% CI
GPX1. CC/TT PAI. TT/no-TT MMP1. -/AA
0.002 0.001 0.041
0.427 0.183 1.770
0.249-0.742 0.068-0.490 1.024-3.058
0.028 0.010
0.535 0.496
0.307-0.934 0.291-0.848
336 FUNCTIONAL GENETIC POLYMORPHISM OF IL-1RN ENCODING THE IL-1 RECEPTOR ANTAGONIST PREDICTS RADIOGRAPHIC SEVERITY OF SYMPTOMATIC KNEE OA M. Attur y, S. Ma y, J. Samuels y, S. Krasnokutsky Samuels y, H. Zhou y, J.T. Bencardino y, M.C. Hochberg z, B.D. Mitchell z, V.B. Kraus x, J.M. Jordan k, S.B. Abramson y. y New York Univ., New York, NY, USA; z Univ. of Maryland Sch. of Med., Baltimore, MD, USA; x Duke Univ. Med. Ctr., Durham, NC, USA; k Univ. of North Carolina, Chapel Hill, NC, USA Purpose: Growing numbers of studies show increased expression in Osteoarthritis (OA) of inflammatory cytokines, such as IL-1b and TNFa, in joint tissues and peripheral blood mononuclear (PBM) cells. The IL1 receptor antagonist (IL1RN) gene cluster region has been associated with susceptibility to knee OA, thereby further implicating inflammation in OA pathogenesis. In these studies we examined the association of IL-1RN haplotype with the radiographic severity of symptomatic knee OA (SKOA). Methods: Genomic DNA from SKOA patients from three cohorts (NYU I, NYU-II and OAI) in this prospective analysis were used to genotype three single nucleotide polymorphisms (SNPs) [rs419598, rs315952 & rs9005) based on our prior publications. Genotyping was accomplished by PCR using validated SNP primers and probes (Applied Biosystems) along with detection using allelic discrimination computation. Genotypes were scored blinded to all patients
Abstracts / Osteoarthritis and Cartilage 25 (2017) S76eS444
annotated as predicted secreted proteins and have potential value as biomarkers of OA. We performed an in silico DrugBank search and found ten compounds that target genes highlighted in the pilot study at stringent confidence levels, and which have FDA marketing authorisation for an existing clinical indication. Three genes are consistently differentially regulated across all 3 omics levels: AQP1, COL1A1 and CLEC3B. For example, AQP1 encodes aquaporin-1, which facilitates water transport across biological membranes. Chondrocyte swelling and increased cartilage hydration has been suggested as an important mechanism in OA. The carbonic anhydrase inhibitor Acetazolamide is a known inhibitor of aquaporin-1, indicating the potential of functional genomics to identify opportunities for drug repurposing, shortening the investigative pipeline to use in OA. We used independent datasets for knee OA and hip OA to show that the direction of change replicates for over 90% of differentially expressed genes and differentially methylated probes, and to demonstrate good agreement in the global estimates of the fold-changes. Integrated pathway analysis implicates the involvement of extracellular matrix degradation, collagen catabolism and angiogenesis in disease progression. Conclusions: This work provides a comprehensive view of the molecular landscape of human primary chondrocytes in OA progression. All data from these experiments are freely available as a resource for the scientific community. 335 NOVEL ASSOCIATIONS IN MITOCHONDRIA-RELATED GENES AND MITOCHONDRIA-NUCLEUS INTERACTIONS. ROLE IN OSTEOARTHRITIS s-Pereira y, M. Ferna ndez-Moreno y, I. Rego-Perez y, E. Corte zquez-Mosquera y, S. Relan ~ o y, S. Pe rtega z, A. Go nza lez x, M. Va ndez-Lo pez y, F. Blanco y. y Servicio L. Vidal-Bralo x, N. Oreiro y, C. Ferna n Biom ~ a (INIBIC). de Reumatología. Inst. de Investigacio edica de A Corun ~ a (CHUAC), Sergas. Univ.e da Complexo Hosp.ario Univ.rio de A Corun ~ a (UDC). As Xubias, 15006, A Corun ~ a, Spain; z Unidad de Corun n Biom Epidemiología Clínica y Bioestadística. Inst. de Investigacio edica ~ a (INIBIC). Complexo Hosp.ario Univ.rio de A Corun ~a de A Corun ~ a (UDC). As Xubias, 15006, A Corun ~ a, (CHUAC), Sergas. Univ.e da Corun n 10 y Unidad de Reumatología. Inst. Spain; x Laboratorio de Investigacio n Sanitaria de Santiago (IDIS), Hosp. Clínico Univ.rio de de Investigacio Santiago, Travesía Choupana s/n, 15706, Santiago de Compostela, Spain Purpose: To analyze the potential influence of specific gene polymorphisms of mitochondria-related genes involved in cellular detoxification and to explore the mitochondria-nucleus interactions in the development of hip OA Methods: For this work we used two well-characterized cohorts of ~ a and patients from Galicia (Spain) including patients from A Corun Santiago reaching a total of 944 subjects, of which 601 were hip OA with KL grade IV and 343 age-matched radiological healthy controls (KL grade 0-I). The mtDNA haplogroups were assigned to all the subjects and grouped into mtDNA clusters. In a first subset of subjects we analyzed a set of mitochondria-related gene polymorphisms involved in the cellular detoxification: CAT (rs659366), GPX1 (rs2234693), UCP2 (rs659366), SOD2 (rs4880) and NOS2A (microsatellite (CCTTT)n in the promoter region of the gene), as well as their potential interaction with mtDNA clusters. In a second set of subjects we analyzed the potential interactions between the mtDNA clusters and a set of previously proposed gene polymorphisms related to OA: GDF-5 (rs143383), CILP (rs2073711), PAI (rs2227631), MMP-1 (rs514921) and MMP-2 (rs243866). The obtained data were appropriately analyzed using PowerMarker and SPSS (v19) Results: We found that the TT genotype of rs659366 in CAT gene was significantly underrepresented in hip OA patients (OR ¼ 0.512;95% CI ¼ 0.321-0.817;p ¼ 0.004). In relation to NOS2A microsatellite, the 13repetition allele was significantly underrepresented in hip OA patients too (OR ¼ 0.675;95%CI ¼ 0.509-0.895;p ¼ 0.006) meanwhile the 11 repetition-allele was significantly associated with an increased risk of hip OA (OR ¼ 1.524;95%CI ¼ 1.133-2.049;p ¼ 0.005). The subsequent regression model adjusting for the confounder variables of gender and age confirmed the above described associations as well as the mtDNA cluster TJ as a protective factor against hip OA (OR ¼ 0.672; 95% CI ¼ 0.455-0.993;p ¼ 0.046) (Table 1). In terms of mitochondria-nucleus interactions, a significant interaction between the SNP rs2234693 in GPX1 and the mtDNA cluster TJ (p ¼ 0.003) and HV (p ¼ 0.05) was discovered; the combination of cluster TJ and CC genotype emerged as a
protective factor (OR ¼ 0.427;95%CI ¼ 0.249-0.742;p ¼ 0.002), and the combination of cluster HV and TT genotype resulted in a protective factor too (OR ¼ 0.535;95%CI ¼ 0.307-0.934;p ¼ 0.028). In addition, we found significant interactions between the mtDNA clusters HV and/or TJ and other specific nuclear polymorphisms in PAI (p < 0.001 for TJ and p ¼ 0.049 for HV) and MMP1 (p ¼ 0.016 for HV) genes. The combination of cluster TJ with TT genotype of PAI significantly associates with a decreased risk of hip OA (OR ¼ 0.183;95%CI ¼ 0.068-0.490;p ¼ 0.001), and the combination of cluster HV with no-TT genotype associated with decreased risk too (OR ¼ 0.496;95%CI ¼ 0.291-0.848;p ¼ 0.010). In relation to MMP1, the combination of cluster HV with AA genotype associates with an increased risk of hip OA (OR ¼ 1.770;95%CI ¼ 1.0243.058;p ¼ 0.041) (Table 2). Finally, the interactions involving both CILP (p ¼ 0.058 for HV) and MMP-2.1575 (p ¼ 0.062 for TJ) border-lined the statistical significance Conclusions: The associations described in this work indicate that mitochondrial genetic background determines the behavior of specific nuclear variants in the development of OA
Table 1 Variables
p-value
OR
95%CI
Gender (male) Age NOS2A-11 NOS2A-13 CAT-TT mtDNA clusters (n ¼ 944) HV (n ¼ 496) KU (n ¼ 222) TJ (n ¼ 147) Others (n ¼ 79)
0.098 0.034 0.041 0.016 0.009
1.275 0.987 1.387 0.694 0.523
0.956-1.700 0.974-0.999 1.014-1.897 0.515-0.935 0.321-0.853
0.693 0.046 0.385
1 1.073 0.672 1.277
0.756-1.522 0.455-0.993 0.735-2.216
Table 2 Cluster TJ
Cluster HV
Genotypes (TJ/HV)
p-value
OR
95% CI
p-value
OR
95% CI
GPX1. CC/TT PAI. TT/no-TT MMP1. -/AA
0.002 0.001 0.041
0.427 0.183 1.770
0.249-0.742 0.068-0.490 1.024-3.058
0.028 0.010
0.535 0.496
0.307-0.934 0.291-0.848
336 FUNCTIONAL GENETIC POLYMORPHISM OF IL-1RN ENCODING THE IL-1 RECEPTOR ANTAGONIST PREDICTS RADIOGRAPHIC SEVERITY OF SYMPTOMATIC KNEE OA M. Attur y, S. Ma y, J. Samuels y, S. Krasnokutsky Samuels y, H. Zhou y, J.T. Bencardino y, M.C. Hochberg z, B.D. Mitchell z, V.B. Kraus x, J.M. Jordan k, S.B. Abramson y. y New York Univ., New York, NY, USA; z Univ. of Maryland Sch. of Med., Baltimore, MD, USA; x Duke Univ. Med. Ctr., Durham, NC, USA; k Univ. of North Carolina, Chapel Hill, NC, USA Purpose: Growing numbers of studies show increased expression in Osteoarthritis (OA) of inflammatory cytokines, such as IL-1b and TNFa, in joint tissues and peripheral blood mononuclear (PBM) cells. The IL1 receptor antagonist (IL1RN) gene cluster region has been associated with susceptibility to knee OA, thereby further implicating inflammation in OA pathogenesis. In these studies we examined the association of IL-1RN haplotype with the radiographic severity of symptomatic knee OA (SKOA). Methods: Genomic DNA from SKOA patients from three cohorts (NYU I, NYU-II and OAI) in this prospective analysis were used to genotype three single nucleotide polymorphisms (SNPs) [rs419598, rs315952 & rs9005) based on our prior publications. Genotyping was accomplished by PCR using validated SNP primers and probes (Applied Biosystems) along with detection using allelic discrimination computation. Genotypes were scored blinded to all patients