Antien L. Mooyaart

Genetic associations in diabetic nephropathy: a meta-analysis

Aims/hypothesisThis meta-analysis assessed the pooled effect of each genetic variant reproducibly associated with diabetic nephropathy.MethodsPubMed, EMBASE and Web of Science were searched for articles assessing the association between genes and diabetic nephropathy. All genetic variants statistically associated with diabetic nephropathy in an initial study, then independently reproduced in at least one additional study, were selected. Subsequently, all studies assessing these variants were included. The association between these variants and diabetic nephropathy (defined as macroalbuminuria/proteinuria or end-stage renal disease [ESRD]) was calculated at the allele level and the main measure of effect was a pooled odds ratio. Pre-specified subgroup analyses were performed, stratifying for type 1/type 2 diabetes mellitus, proteinuria/ESRD and ethnic group.ResultsThe literature search yielded 3,455 citations, of which 671 were genetic association studies investigating diabetic nephropathy. We identified 34 replicated genetic variants. Of these, 21 remained significantly associated with diabetic nephropathy in a random-effects meta-analysis. These variants were in or near the following genes: ACE, AKR1B1 (two variants), APOC1, APOE, EPO, NOS3 (two variants), HSPG2, VEGFA, FRMD3 (two variants), CARS (two variants), UNC13B, CPVL and CHN2, and GREM1, plus four variants not near genes. The odds ratios of associated genetic variants ranged from 0.48 to 1.70. Additional variants were detected in subgroup analyses: ELMO1 (Asians), CCR5 (Asians) and CNDP1 (type 2 diabetes).Conclusions/interpretationThis meta-analysis found 24 genetic variants associated with diabetic nephropathy. The relative contribution and relevance of the identified genes in the pathogenesis of diabetic nephropathy should be the focus of future studies.

Genetic variants in pre-eclampsia: a meta-analysis

BACKGROUND Pre-eclampsia has a clear familial component, suggesting that the condition may be partly attributable to genetic susceptibility. The search for susceptibility genes has led to a drastic increase in the number of published studies associating genetic factors with pre-eclampsia. However, attempts to replicate these findings have yielded inconsistent results. This meta-analysis assessed the pooled effect of each genetic variant that is reproducibly associated with pre-eclampsia. METHODS Studies that assessed the association between genes and pre-eclampsia were searched in PubMed, Embase and Web of Science. We selected all genetic variants that were significantly associated with pre-eclampsia in an initial study and were subsequently independently reproduced in at least one additional study. All studies that assessed these reproduced variants were then included. The association between genetic variants and pre-eclampsia was calculated at the allele level, and the main measure of effect was a pooled odds ratio in a random-effects model. RESULTS The literature search yielded 2965 articles, of which 542 investigated genetic associations in pre-eclampsia. We identified 22 replicated genetic variants, of which 7 remained significantly associated with pre-eclampsia following meta-analysis. These variants were in or near the following genes: ACE, CTLA4, F2, FV, LPL and SERPINE1. CONCLUSIONS This meta-analysis identified seven genetic variants associated with pre-eclampsia. Importantly, many of these variants are also risk factors for developing cardiovascular disease, revealing that pre-eclampsia and cardiovascular disease have shared genetic risk factors. The contribution of the identified genetic variants in the pathogenesis of pre-eclampsia should be the focus of future studies.

Vegetarianism, female gender and increasing age, but not CNDP1 genotype, are associated with reduced muscle carnosine levels in humans

Carnosine is found in high concentrations in skeletal muscles, where it is involved in several physiological functions. The muscle carnosine content measured within a population can vary by a factor 4. The aim of this study was to further characterize suggested determinants of the muscle carnosine content (diet, gender and age) and to identify new determinants (plasma carnosinase activity and testosterone). We investigated a group of 149 healthy subjects, which consisted of 94 men (12 vegetarians) and 55 women. Muscle carnosine was quantified in M. soleus, gastrocnemius and tibialis anterior using magnetic resonance proton spectroscopy and blood samples were collected to determine CNDP1 genotype, plasma carnosinase activity and testosterone concentrations. Compared to women, men have 36, 28 and 82% higher carnosine concentrations in M. soleus, gastrocnemius and tibialis anterior muscle, respectively, whereas circulating testosterone concentrations were unrelated to muscle carnosine levels in healthy men. The carnosine content of the M. soleus is negatively related to the subjects’ age. Vegetarians have a lower carnosine content of 26% in gastrocnemius compared to omnivores. In contrast, there is no difference in muscle carnosine content between omnivores with a high or low ingestion of β-alanine. Muscle carnosine levels are not related to the polymorphism of the CNDP1 gene or to the enzymatic activity of the plasma carnosinase. In conclusion, neither CNDP1 genotype nor the normal variation in circulating testosterone levels affects the muscular carnosine content, whereas vegetarianism, female gender and increasing age are the factors associated with reduced muscle carnosine stores.

Lower frequency of the 5/5 homozygous CNDP1 genotype in South Asian Surinamese

We investigated the frequency of the 5/5 homozygous CNDP1 (carnosinase) genotype, which was found to be associated with a reduced risk of developing diabetic nephropathy, in three ethnic groups in The Netherlands. Particularly interesting were the South Asian Surinamese, who have a high prevalence of diabetic nephropathy. Furthermore, we investigated the association between this gene and carnosinase activity in South Asian Surinamese and whether carnosinase was expressed in the kidney. We genotyped 290 South Asian Surinamese, 532 African Surinamese, and 472 White Dutch in a cross-sectional population study. Furthermore, an independent cohort of South Asian Surinamese was genotyped. In this population, carnosinase activity was measured in serum. Immunostaining and in situ hybridization for CNDP1 were performed on kidney tissue. Both South Asian populations had lower frequencies of the 5/5 homozygous genotype than African Surinamese and White Dutch (23.0%, 27.2%, 38.2%, and 41.3%, respectively; chi-square, p<0.001). This genotype showed a lower carnosinase activity in South Asian Surinamese (Wilcoxon rank-sum, p=0.03). CNDP1 was expressed in the kidney. South Asian Surinamese have a lower frequency of the 5/5 homozygous genotype, which was associated with lower carnosinase activity. Our study provides an indication that South Asian Surinamese are genetically at risk for developing diabetic nephropathy.

Genetic associations in diabetic nephropathy

Diabetic nephropathy is a complex disease, caused by both environmental and genetic factors. As in most complex diseases, genetic association studies in diabetic nephropathy showed inconsistent results. In retrospect, studies with small sample sizes, given what are now known to be small odds ratios, were partially responsible for this poor replication record. Furthermore, the low prior probability in complex genetics and multiple testing played a role. Results become more consistent when one only considers those that were replicated. In a large meta-analysis study including only replicated associated genetic variants, 24 genetic variants in 16 genes were found to be associated with diabetic nephropathy. These genetic variants may provide novel biological insight. In particular, rare variants with a large effect found by hypothesis-free approaches (genome-wide association scans, next-generation sequencing) may open new avenues of discovery.

The clinical prognostic value of molecular intrinsic tumor subtypes in older breast cancer patients: A FOCUS study analysis

Introduction: It was recently proposed that the molecular breast tumor subtypes are differently distributed in the elderly breast cancer patients, and also lack prognostic value. Given the limited number of elderly patients in previous studies, the aim of this study was to determine the prognostic effect of the molecular intrinsic subtypes in a large older breast cancer population.

Erratum to: Genetic associations in diabetic nephropathy: a meta-analysis

Erratum to: Diabetologia DOI 10.1007/s00125-010-1996-1 In Table 1, Fig. 2a and Fig. 3a, the minor allele of the single nucleotide polymorphism rs1617640 in the EPO promoter was incorrectly stated as being the T allele, whereas it should have been the G allele. This change does not affect the values reported and the authors apologise for any confusion this may have caused.

Long-term prognosis of young breast cancer patients (≤40 years) who did not receive adjuvant systemic treatment: protocol for the PARADIGM initiative cohort study

Introduction Currently used tools for breast cancer prognostication and prediction may not adequately reflect a young patient’s prognosis or likely treatment benefit because they were not adequately validated in young patients. Since breast cancers diagnosed at a young age are considered prognostically unfavourable, many treatment guidelines recommend adjuvant systemic treatment for all young patients. Patients cured by locoregional treatment alone are, therefore, overtreated. Lack of prognosticators for young breast cancer patients represents an unmet medical need and has led to the initiation of the PAtients with bReAst cancer DIaGnosed preMenopausally (PARADIGM) initiative. Our aim is to reduce overtreatment of women diagnosed with breast cancer aged ≤40 years. Methods and analysis All young, adjuvant systemic treatment naive breast cancer patients, who had no prior malignancy and were diagnosed between 1989 and 2000, were identified using the population based Netherlands Cancer Registry (n=3525). Archival tumour tissues were retrieved through linkage with the Dutch nationwide pathology registry. Tissue slides will be digitalised and placed on an online image database platform for clinicopathological revision by an international team of breast pathologists. Immunohistochemical subtype will be assessed using tissue microarrays. Tumour RNA will be isolated and subjected to next-generation sequencing. Differences in gene expression found between patients with a favourable and those with a less favourable prognosis will be used to establish a prognostic classifier, using the triple negative patients as proof of principle. Ethics and dissemination Observational data from the Netherlands Cancer Registry and left over archival patient material are used. Therefore, the Dutch law on Research Involving Human Subjects Act (WMO) is not applicable. The PARADIGM study received a ‘non-WMO’ declaration from the Medical Ethics Committee of the Netherlands Cancer Institute - Antoni van Leeuwenhoek hospital, waiving individual patient consent. All data and material used are stored in a coded way. Study results will be presented at international (breast cancer) conferences and published in peer-reviewed, open-access journals.

Abstract P3-07-54: Insulin-like growth factor 1 receptor expression and polymorphism are associated with response to neoadjuvant chemotherapy in breast cancer patients: Results from the NEOZOTAC trial (BOOG 2010-01)

Background The insulin-like growth factor 1 (IGF-1) pathway is involved in cell growth, proliferation and cell cycle progression and associated with tumor genesis and therapy resistance. This study aims to elucidate whether variation in the IGF-1 pathway is predictive for pathologic response in early breast cancer (BC) patients taking part in the phase III NEOZOTAC trial, randomizing between 6 cycles of neoadjuvant TAC chemotherapy with or without zoledronic acid. Method Formalin-fixed paraffin-embedded (FFPE) tissue samples of pre-chemotherapy biopsies and operation specimens were collected for analysis of IGF-1 receptor (IGF-1R) expression using IHC (n=216) and for analysis of 8 candidate SNPs in genes of the IGF-1 pathway (n=184) using OpenArray® RealTime PCR. Optionally, blood samples were collected immediately before chemotherapy for determination of glucose, insulin, IGF-1, IGF-2 and IGF-BP3. Associations with patient and tumor characteristics and chemotherapy response according to Miller and Payne (MP) pathologic response were performed using chi square and logistic regression analyses. Results High IGF-1R expression was associated with estrogen receptor expression (P=0.001). During chemotherapy, a significant number of the tumors (47.2%) showed a decrease in IGF-1R expression, while in a small number of the tumors an upregulation was seen (15.1%). IGF-1R expression before treatment was not associated with pathological response, however absence of IGF-1R expression after treatment was associated with a better response in multivariate analyses (P=0.012) and patients with a decrease in expression during treatment showed a better response to chemotherapy as well (P=0.008). Moreover, the variant T allele of 3129G>T in IGF-1R (rs2016347) was associated with a better pathological response in multivariate analyses (P=0.032). In addition, high glucose and insulin levels were associated with positive lymph node status before chemotherapy in multivariate analysis (P=0.019) and (P=0.031), respectively. Conclusion Neoadjuvant chemotherapy induced changes in the IGF-1R expression in most of the tumors. Absence or diminished expression of IGF-1R after treatment was associated with a better pathological response. Additionally, we found a SNP (rs2016347) in IGF-1R as a potential predictive marker for chemotherapy efficacy in BC patients treated with TAC. These findings may help to select patients who might benefit from (co-)treatment with an IGF-1 pathway inhibitor. Citation Format: de Groot S, Charehbili A, van Laarhoven HWM, Mooyaart AL, Dekker-Ensink NG, van de Ven S, Janssen LGM, Swen JJ, Smit VTHBM, Heijns JB, Kessels LW, van der Straaten RJHM, Bhringer S, Gelderblom AJ, van der Hoeven JJM, Guchelaar HJ, Pijl H, Kroep JR. Insulin-like growth factor 1 receptor expression and polymorphism are associated with response to neoadjuvant chemotherapy in breast cancer patients: Results from the NEOZOTAC trial (BOOG 2010-01). [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P3-07-54.

OS050. Genetic variants in pre-eclampsia: a meta-analysis

INTRODUCTION Preeclampsia has a clear familial component, suggesting that the syndrome may be partly attributable to genetic susceptibility. The search for susceptibility genes has lead to a massive increase in the number of published studies involving genetic associations in preeclampsia. However, attempts to replicate these findings have yielded inconsistent results. This meta-analysis aims to assess the pooled effect of each genetic variant that is reproducibly associated with preeclampsia. OBJECTIVES To create an overview of the genetic variants that are reproducibly associated with preeclampsia. METHODS Studies assessing the association between genes and preeclampsia were searched in PubMed, EMBASE and Web of Science. We selected all genetic variants that were significantly associated with preeclampsia in an initial study and then independently reproduced in at least one additional study. Subsequently, all studies assessing these reproduced variants were included. The association between these variants and preeclampsia was calculated at the allele level and the main measure of effect was a pooled odds ratio. RESULTS The literature search resulted in 2965 citations, of which 542 were genetic association studies investigating preeclampsia. We identified 23 replicated genetic variants, of which 8 remained significantly associated with preeclampsia in a random-effects meta-analysis. These variants were in or near the following genes: ACE,AGT,CTLA4,F2,FV (two variants),LPL and SERPINE1. CONCLUSION This meta-analysis found 8 genetic variants associated with preeclampsia. Most of these variants are in the renin-angiotensin and the coagulation system. Importantly, many of the variants that were associated with preeclampsia are known to be risk factors for the development of cardiovascular disease, indicating that preeclampsia and cardiovascular disease have shared genetic risk factors. The relative contribution and relevance of the identified genes in the pathogenesis of preeclampsia should be the focus of future studies.