Mademoiselle de Neuville; Ide de Chaudfontaine; Béatrix

266 p

Functional mechanisms underlying pleiotropic risk alleles at the 19p13.1 breast-ovarian cancer susceptibility locus

A locus at 19p13 is associated with breast cancer (BC) and ovarian cancer (OC) risk. Here we analyse 438 SNPs in this region in 46,451 BC and 15,438 OC cases, 15,252 BRCA1 mutation carriers and 73,444 controls and identify 13 candidate causal SNPs associated with serous OC (P=9.2 × 10-20), ER-negative BC (P=1.1 × 10-13), BRCA1-associated BC (P=7.7 × 10-16) and triple negative BC (P-diff=2 × 10-5). Genotype-gene expression associations are identified for candidate target genes ANKLE1 (P=2 × 10-3) and ABHD8 (P<2 × 10-3). Chromosome conformation capture identifies interactions between four candidate SNPs and ABHD8, and luciferase assays indicate six risk alleles increased transactivation of the ADHD8 promoter. Targeted deletion of a region containing risk SNP rs56069439 in a putative enhancer induces ANKLE1 downregulation; and mRNA stability assays indicate functional effects for an ANKLE1 3′-UTR SNP. Altogether, these data suggest that multiple SNPs at 19p13 regulate ABHD8 and perhaps ANKLE1 expression, and indicate common mechanisms underlying breast and ovarian cancer risk

The FANCM:p.Arg658* truncating variant is associated with risk of triple-negative breast cancer

Abstract: Breast cancer is a common disease partially caused by genetic risk factors. Germline pathogenic variants in DNA repair genes BRCA1, BRCA2, PALB2, ATM, and CHEK2 are associated with breast cancer risk. FANCM, which encodes for a DNA translocase, has been proposed as a breast cancer predisposition gene, with greater effects for the ER-negative and triple-negative breast cancer (TNBC) subtypes. We tested the three recurrent protein-truncating variants FANCM:p.Arg658*, p.Gln1701*, and p.Arg1931* for association with breast cancer risk in 67,112 cases, 53,766 controls, and 26,662 carriers of pathogenic variants of BRCA1 or BRCA2. These three variants were also studied functionally by measuring survival and chromosome fragility in FANCM−/− patient-derived immortalized fibroblasts treated with diepoxybutane or olaparib. We observed that FANCM:p.Arg658* was associated with increased risk of ER-negative disease and TNBC (OR = 2.44, P = 0.034 and OR = 3.79; P = 0.009, respectively). In a country-restricted analysis, we confirmed the associations detected for FANCM:p.Arg658* and found that also FANCM:p.Arg1931* was associated with ER-negative breast cancer risk (OR = 1.96; P = 0.006). The functional results indicated that all three variants were deleterious affecting cell survival and chromosome stability with FANCM:p.Arg658* causing more severe phenotypes. In conclusion, we confirmed that the two rare FANCM deleterious variants p.Arg658* and p.Arg1931* are risk factors for ER-negative and TNBC subtypes. Overall our data suggest that the effect of truncating variants on breast cancer risk may depend on their position in the gene. Cell sensitivity to olaparib exposure, identifies a possible therapeutic option to treat FANCM-associated tumors

A case-only study to identify genetic modifiers of breast cancer risk for BRCA1/BRCA2 mutation carriers

Abstract: Breast cancer (BC) risk for BRCA1 and BRCA2 mutation carriers varies by genetic and familial factors. About 50 common variants have been shown to modify BC risk for mutation carriers. All but three, were identified in general population studies. Other mutation carrier-specific susceptibility variants may exist but studies of mutation carriers have so far been underpowered. We conduct a novel case-only genome-wide association study comparing genotype frequencies between 60,212 general population BC cases and 13,007 cases with BRCA1 or BRCA2 mutations. We identify robust novel associations for 2 variants with BC for BRCA1 and 3 for BRCA2 mutation carriers, P < 10−8, at 5 loci, which are not associated with risk in the general population. They include rs60882887 at 11p11.2 where MADD, SP11 and EIF1, genes previously implicated in BC biology, are predicted as potential targets. These findings will contribute towards customising BC polygenic risk scores for BRCA1 and BRCA2 mutation carriers

Functional mechanisms underlying pleiotropic risk alleles at the 19p13.1 breast-ovarian cancer susceptibility locus

A locus at 19p13 is associated with breast cancer (BC) and ovarian cancer (OC) risk. Here we analyse 438 SNPs in this region in 46,451 BC and 15,438 OC cases, 15,252 BRCA1 mutation carriers and 73,444 controls and identify 13 candidate causal SNPs associated with serous OC (P = 9.2 x 10(-20)), ER-negative BC (P = 1.1 x 10(-13)), BRCA1-associated BC (P = 7.7 x 10(-16)) and triple negative BC (P-diff = 2 x 10(-5)). Genotype-gene expression associations are identified for candidate target genes ANKLE1 (P = 2 x 10(-3)) and ABHD8 (PPeer reviewe

A case-only study to identify genetic modifiers of breast cancer risk for BRCA1/BRCA2 mutation carriers

Breast cancer (BC) risk for BRCA1 and BRCA2 mutation carriers varies by genetic and familial factors. About 50 common variants have been shown to modify BC risk for mutation carriers. All but three, were identified in general population studies. Other mutation carrier-specific susceptibility variants may exist but studies of mutation carriers have so far been underpowered. We conduct a novel case-only genome-wide association study comparing genotype frequencies between 60,212 general population BC cases and 13,007 cases with BRCA1 or BRCA2 mutations. We identify robust novel associations for 2 variants with BC for BRCA1 and 3 for BRCA2 mutation carriers, P < 10−8, at 5 loci, which are not associated with risk in the general population. They include rs60882887 at 11p11.2 where MADD, SP11 and EIF1, genes previously implicated in BC biology, are predicted as potential targets. These findings will contribute towards customising BC polygenic risk scores for BRCA1 and BRCA2 mutation carriers

Adenomyosis : immune system involvement

L'adénomyose est une pathologie chronique bénigne de l'utérus caractérisée par la présence de glandes endométriales et de stroma au sein du myomètre. C'est une maladie hétérogène, présentant des formes cliniques distinctes selon la localisation des lésions dans le myomètre. Cette maladie fréquente peut être responsable de symptômes invalidants comme des dysménorrhées, des saignements utérins anormaux et/ou une infertilité. Sa pathogenèse est très peu comprise aujourd'hui et plusieurs théories ont été proposées. Certaines suggèrent que le développement de l'adénomyose est de novo, à partir de reliquats müllériens ou de cellules souches. Néanmoins, selon la théorie la plus communément admise, l'adénomyose trouverait son origine dans la couche basale de l'endomètre avec une invagination de cellules entre les faisceaux musculaires. De multiples facteurs pourraient être impliqués dans l'initiation de cette invasion, et notamment des modifications immunitaires. Afin d'explorer l'implication du système immunitaire dans la pathogenèse de l'adénomyose, nous avons réalisé une revue systématique de la littérature portant sur les modifications immunitaires décrites dans l'adénomyose chez l'Humain. Cette analyse a montré la présence de changements immunitaire tant innés qu'adaptatifs, au niveau local utérin ou systémique chez les femmes atteintes d'adénomyose par rapport aux contrôles. Ensuite, nous avons étudié la concentration sérique systémique de cytokines selon la présence d'une adénomyose. Nous avons montré une diminution des taux sériques de cytokines pro-inflammatoires et immunorégulatrices (IL17F, IL23, IL25, IL31 et IL33) chez les femmes atteintes d'adénomyose par rapport aux contrôles.Nous avons ensuite étudié dans le sérum, le profil métabolique de femmes avec adénomyose. Celles-ci avaient un profil métabolique différent par rapport à celui des contrôles. Les métabolites différentiellement retrouvés semblaient impliqués dans l'activation immunitaire ainsi que dans la prolifération et la migration cellulaire. De plus, nos résultats ont suggéré qu'un certain nombre de voies métaboliques étaient spécifiques selon le phénotype d'adénomyose. Enfin, nous avons reproduit un modèle d'adénomyose murin, précédemment décrite par l'équipe de Guo S-W. Nous avons cherché à étudier dans ce modèle, le rôle des cellules immunitaires et de la voie NOTCH1 dans le développement de l'adénomyose, notamment via l'activation d'une transition épithélio-mésenchymateuse (TEM), phénomène décrit dans l'invasion du myomètre par des cellules endométriales dans l'adénomyose. Nous avons montré une augmentation de certains marqueurs de cellules immunitaires, notamment les macrophages M1 et les cellules Natural Killer à 30 jours et une augmentation des marqueurs des cellules T CD4+ à 60 jours de vie, dans l'utérus de souris adénomyosiques par rapport aux contrôles. Parallèlement, la protéine NOTCH1 activé ainsi que certains marqueurs de la TEM étaient augmentés, suggérant que les modifications immunitaires et l'activation NOTCH1 dans l'utérus pourraient être en lien avec le développement des lésions d'adénomyose. En conclusion, nous avons confirmé la présence de modifications immunitaires dans la pathogenèse de l'adénomyose, tant au niveau systémique que local au niveau de l'utérus. Nous avons mis en évidence dans l'adénomyose, un déséquilibre de la balance des cytokines systémiques anti-inflammatoires/pro-inflammatoires ainsi qu'un profil métabolique différent par rapport aux contrôles. Nous avons également montré une expression aberrante de plusieurs marqueurs des cellules immunitaires au sein de l'utérus de souris adénomyosiques au cours du développement de la maladie ainsi qu'une activation de la voie NOTCH1 et des signaux de TEM. Ces résultats confirment que certains médiateurs immuns sont impliqués dans la pathogenèse de l'adénomyose et probablement dans le processus d'invasion cellulaire des cellules endométriales vers le myomètre.Adenomyosis is a chronic benign uterine disease characterized by the presence of endometrial glands and stroma within the myometrium. It is a heterogeneous disease, presenting various clinical forms, depending on the location of the ectopic lesions within the myometrium. Adenomyosis can be responsible for several symptoms such as dysmenorrhea, abnormal uterine bleeding and/or infertility. Its pathogenesis is a real conundrum and several theories have been proposed: development of adenomyosis lesion could initiate de novo from Mullerian rests or from stem cells. However, according to the most widely accepted theory, adenomyosis originates in the basal layer of the endometrium that invaginate between smooth muscle cells bundles into the myometrium. Multiple factors could be involved in initiating this invasion, including specific immune changes. In order to explore the involvement of the immune system in the pathogenesis of adenomyosis, we first carried out a systematic review on the immune changes described in adenomyosis in Humans. This analysis showed the presence of changes for both innate and adaptive immunity, locally, within the uterus or at a systemic level, in adenomyosis women compared to controls. Next, we studied the systemic serum concentration of cytokines according to the presence of adenomyosis. We have shown a decrease in serum levels of pro-inflammatory and immunoregulatory cytokines (IL17F, IL23, IL25, IL31 and IL33) in women with adenomyosis compared to controls. Then, we have studied in the serum, the metabolic profile of women with adenomyosis. Women with adenomyosis presented a different metabolic profile compared to disease-free counterparts. The metabolites with altered levels were involved in immune activation as well as cells proliferation and migration. In addition, our results suggested that a number of metabolic pathways were specific according to the adenomyosis phenotype. Finally, we have reproduced a murine model of adenomyosis, as previously described by the Guo S-W et al. We have investigated in this model the role of immune cells and NOTCH1 pathway in the development of adenomyosis, namely via the epithelial to mesenchymal transition (EMT), this latter phenomenon being described as involved in the invasion of endometrial cells within the myometrium in adenomyosis. We have shown an increase in the expression of some markers of immune cells, in particular M1 macrophages and Natural Killer cells at 30 days and an increase in CD4 + T cells markers at 60 days of life, in the uterus of adenomyosis affected mice compared to controls. At the same time, the activated NOTCH1 as well as some TEM markers were increased, suggesting that immune changes and NOTCH1 activation could be linked to the development of adenomyosis lesions in the uterus. In conclusion, we confirmed the presence of immune changes in adenomyosis pathogenesis - both at a systemic and at a local uterine level. We have demonstrated that an imbalance of anti-inflammatory / pro-inflammatory systemic cytokines occurred in adenomyosis women, in addition to a different metabolic profile, compared to controls. We have also shown an aberrant expression of several markers of immune cells in the uterus of adenomyosis affected mice during the development of the disease as well as an activation of the NOTCH1 pathway and TEM signals. These results confirm that some immune mediators are involved in the pathogenesis of adenomyosis and probably in the process of cellular invasion of endometrial cells within the myometrium