Search

PDF file - 49K, Clinicopathological features and molecular findings of the index patients with a mutation in EPCAM or MSH2 gene.

PDF file - 198K

Perspective on This Article from Characterization of New Founder Alu-Mediated Rearrangements in MSH2 Gene Associated with a Lynch Syndrome Phenotype

It has been reported that large genomic deletions in the MLH1 and MSH2 genes are a frequent cause of Lynch syndrome in certain populations. Here, a cohort has been screened and two new founder rearrangements have been found in the MSH2 gene. These mutations have been characterized by break point determination, haplotype analysis, and genotype–phenotype correlation. Mutations have been identified in the MLH1, MSH2, and MSH6 genes in 303 subjects from 160 suspected Lynch syndrome unrelated families. All subjects were tested using heteroduplex analysis by capillary array electrophoresis. Multiplex ligation-dependent probe amplification was used to detect rearrangements in mutation-negative index patients and confirmed by reverse transcriptase PCR. The break point of the deletions was further characterized by the array comparative genomic hybridization method. Immunohistochemical staining and microsatellite instability were studied in tumor samples. Hereditary nonpolyposis colorectal cancer–related phenotypes were evaluated. More than 16% (24 of 160) of the families had pathogenic mutations (8 MLH1, 15 MSH2, and 1 MSH6). Twelve of these families (50%) are carriers of a novel mutation. Seven of the 15 positive MSH2 families (47%) are carriers of a rearrangement. The exon 7 deletion and exon 4 to 8 deletion of MSH2 are new founder mutations. The segregation of a common haplotype, a similar phenotype, and anticipation effects were observed in these families. These findings will greatly simplify the diagnosis, counseling, and clinical care in suspected Lynch syndrome families and not just in specific geographic areas, so wide distribution may be explained by migration patterns. Cancer Prev Res; 4(10); 1546–55. ©2011 AACR.

Lynch syndrome is caused by germline mutations in MSH2, MLH1, MSH6, and PMS2 mismatch repair genes and leads to a high risk of colorectal and endometrial cancer. It was recently shown that constitutional 3′ end deletions of EPCAM could cause Lynch syndrome in tissues with MSH2 deficiency. We aim to establish the spectrum of mutations in MSH2-associated Lynch syndrome cases and their clinical implications. Probands from 159 families suspected of having Lynch syndrome were enrolled in the study. Immunohistochemistry and microsatellite instability (MSI) analyses were used on the probands of all families. Eighteen cases with MSH2 loss were identified: eight had point mutations in MSH2. In 10 Lynch syndrome families without MSH2 mutations, EPCAM-MSH2genomic rearrangement screening was carried out with the use of multiplex ligation–dependent probe amplification and reverse transcriptase PCR. We report that large germline deletions, encompassing one or more exons of the MSH2 gene, cosegregate with the Lynch syndrome phenotype in 23% (8 of 35) of MSI families tested. A new combined deletion EPCAM-MSH2 was identified and characterized by break point analysis, encompassing from the 3′ end region of EPCAM to the 5′ initial sequences of the MSH2 (c.859-1860_MSH2:646-254del). EPCAM-MSH2 fusion transcript was isolated. The tumors of the carriers show high-level MSI and MSH2 protein loss. The clinical correlation provided evidence that the type of mutation and the extension of the deletions involving the MSH2 gene could have different implications in cancer predisposition. Thus, the identification of EPCAM-MSH2 rearrangements and their comprehensive characterization should be included in the routine mutation screening protocols for Lynch syndrome. Cancer Prev Res; 4(10); 1556–62. ©2011 AACR.

Perspective on This Article from Frequency of Rearrangements in Lynch Syndrome Cases Associated with MSH2: Characterization of a New Deletion Involving both EPCAM and the 5′ Part of MSH2

PDF file - 98K, Experimental conditions of EPCAM-MSH2 deletion by a routine PCR.

PDF file - 90K

Purpose: Most BRCA1/2 mutations are of unknown clinical relevance. An increasing amount of evidence indicates that there can be deleterious effects through the disruption of the splicing process. We have investigated the effect of aberrant splicing of BRCA1/2 on hereditary breast/ovarian cancer (HBOC).Experimental Design: DNA variants were analyzed with splicing prediction programs to select putative splicing mutations. Splicing assays of 57 genetic variants were done by lymphocyte reverse transcription-PCR and/or hybrid minigenes in HeLa and nontumor breast epithelial cells.Results: Twenty-four BRCA1/2 variants of Spanish HBOC patients were bioinformatically preselected. Functional assays showed that 12 variants induced anomalous splicing patterns, 6 of which accounted for 58.5% of BRCA1 families. To further evaluate the defective splicing of BRCA1/2, we analyzed 31 Breast Cancer Information Core Database (BIC) and two artificial variants that were generated by mutagenesis. Sixteen variants induced different degrees of aberrant splicing. Altogether, anomalous splicing was caused by 28 BRCA1/2 variants of all types, indicating that any DNA change can disrupt pre-mRNA processing. We show that a wide range of regulatory elements can be involved, including the canonical and cryptic splice sites, the polypyrimidine tract, and splicing enhancers/silencers. Twenty mutations were predicted to truncate the BRCA proteins and/or to delete essential domains, thus supporting a role in HBOC.Conclusions: An important fraction of DNA variants of BRCA1/2 presents splicing aberrations that may represent a relevant disease-causing mechanism in HBOC. The identification of splicing disruptions by functional assays is a valuable tool to discriminate between benign polymorphisms and pathogenic mutations. Clin Cancer Res; 16(6); 1957–67

Background: Genome-wide association studies (GWAS) identified variants at 19p13.1 and ZNF365 (10q21.2) as risk factors for breast cancer among BRCA1 and BRCA2 mutation carriers, respectively. We explored associations with ovarian cancer and with breast cancer by tumor histopathology for these variants in mutation carriers from the Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA).Methods: Genotyping data for 12,599 BRCA1 and 7,132 BRCA2 mutation carriers from 40 studies were combined.Results: We confirmed associations between rs8170 at 19p13.1 and breast cancer risk for BRCA1 mutation carriers [HR, 1.17; 95% confidence interval (CI), 1.07–1.27; P = 7.42 × 10−4] and between rs16917302 at ZNF365 (HR, 0.84; 95% CI, 0.73–0.97; P = 0.017) but not rs311499 at 20q13.3 (HR, 1.11; 95% CI, 0.94–1.31; P = 0.22) and breast cancer risk for BRCA2 mutation carriers. Analyses based on tumor histopathology showed that 19p13 variants were predominantly associated with estrogen receptor (ER)-negative breast cancer for both BRCA1 and BRCA2 mutation carriers, whereas rs16917302 at ZNF365 was mainly associated with ER-positive breast cancer for both BRCA1 and BRCA2 mutation carriers. We also found for the first time that rs67397200 at 19p13.1 was associated with an increased risk of ovarian cancer for BRCA1 (HR, 1.16; 95% CI, 1.05–1.29; P = 3.8 × 10−4) and BRCA2 mutation carriers (HR, 1.30; 95% CI, 1.10–1.52; P = 1.8 × 10−3).Conclusions: 19p13.1 and ZNF365 are susceptibility loci for ovarian cancer and ER subtypes of breast cancer among BRCA1 and BRCA2 mutation carriers.Impact: These findings can lead to an improved understanding of tumor development and may prove useful for breast and ovarian cancer risk prediction for BRCA1 and BRCA2 mutation carriers. Cancer Epidemiol Biomarkers Prev; 21(4); 645–57. ©2012 AACR.

Supplementary Data from A High Proportion of DNA Variants of BRCA1 and BRCA2 Is Associated with Aberrant Splicing in Breast/Ovarian Cancer Patients

Supplementary Methods, Tables 1-3, Figure 1 from Common Breast Cancer Susceptibility Alleles and the Risk of Breast Cancer for BRCA1 and BRCA2 Mutation Carriers: Implications for Risk Prediction

Objective The virulence of COVID-19 has been particularly problematic in countries such as Spain. This led the government to decide that the population should be locked down at home to reduce the spread of the disease and avoid the collapse of the health system. Considering this, this study analyzed the changes in intimate relationships that occurred during lockdown in terms of dyadic adjustment, conflict, and quality of the relationship, as well as their relationship with anxiety symptoms. Method Cross-sectional questionnaire-based study with adults (N = 342) aged 20-67 years who lived in Spain. Each participant completed self-report measures of anxiety (State-Trait Anxiety Inventory state and trait subscales), dyadic adjustment (Dyadic Adjustment Scale), relationship conflict and quality, and sociodemographic variables. Results The results showed significant levels of state anxiety, which was associated with poorer dyadic adjustment and a decrease in the perceived quality of relationships since the start of lockdown. Increased partner conflict seems to be an important predictor of dyadic adjustment and relationship quality during social isolation. Conclusions This study suggests that the COVID-19 pandemic has negatively affected the mental health of the population, especially women. This finding is closely associated with difficulties with one's cohabiting partner (e.g., worse dyadic adjustment), but the most determining factor seems to be the previous state of the relationship. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Around 50% of the familial breast cancer (BC) cases are estimated to be caused by germline variants in known low-, moderate-, and high-risk susceptibility genes, while the other half is of unknown genetic origin. In the present study, we wanted to evaluate the role of the RECQ helicases, some of which have been studied in the past as candidates, with unclear results about their role in the disease. Using next-generation sequencing (NGS) technology, we analyzed the whole coding sequence of BLM, RECQL1, RECQL4, RECQL5, and WRN in almost 2000 index cases from BC Spanish families that had previously tested negative for the known BC susceptibility genes (BRCAX) and compared the results with the controls extracted from gnomAD. Our results suggest that BLM, RECQL1, RECQL4, and WRN do not play a major role in BC susceptibility. However, in the combined analysis, joining the present results with those previously reported in a series of 1334 BC Spanish patients and controls, we found a statistically significant association between Loss of Function (LoF) variants in RECQL5 and BC risk, with an OR of 2.56 (p = 0.009; 95% CI, 1.18–4.98). Our findings support our previous work and places the RECQL5 gene as a new moderate-risk BC gene., A.O. is partially funded by FIS PI19/00640 supported by FEDER funds and the Spanish Network on Rare Diseases (CIBERER). M.d.l.H. is partially funded by FIS PI20/00110 supported by FEDER funds.

The sending of sexually explicit images by men to women without prior request, a practice commonly referred to as sending or receiving a “dick pic,” is a fairly common manifestation of sexual cyber-violence that has grown in recent times. As research on this type of sexual cyber-violence is limited, the current study analyzed the prevalence of this phenomenon in a sample of 347 Spanish women between 18 and 30 years of age, studying the factors that influence the emotional impact reported by women if they received an unsolicited dick pic (using a hypothetical scenario) and exploring the various coping strategies that women would use in that situation. Results showed a significant prevalence of this type of cyber-violence in the sample, as 48.1% of the participants had received an unsolicited dick pic from an unknown man at some point. Women with lower levels of hostile sexism—but not of benevolent sexism—reported a higher depressed and angry/annoyed emotional impact of the sexual cyber-violence scenario. This was also the case for women with a less conservative political ideology, with less religious beliefs, as well as those women who perceived that their female friends receive this type of images frequently (descriptive norm) and who perceived that their female friends are less accepting of these situations (injunctive norm). In addition, from the strategies presented to the participants to cope with this situation of sexual cyber-violence, it was observed that a significant percentage of women would choose strategies, such as talking about the incident with other people and blocking the sender’s access. Yet, fewer women would employ effective strategies, such as reporting the perpetrator’s profile to the managers or administrators of the social network or reporting the incident to the police. This study is one of the first studies in Spain that addresses this new form of sexual cyber-violence against women by unknown men and suggests that, in online social networks, women experience the same situations of abuse, harassment, and sexual objectification that they have faced offline in everyday life. Therefore, more work needs to be done to raise awareness and try to prevent these situations, while also providing more support to these women so that they can adopt effective coping strategies.

The development of nursing knowledge requires a close relationship between theory, research, and practice. The purpose of the analysis of the concept of "parental decision-making in pediatric critical care" is to facilitate nurses' therapeutic care of critically ill children and their families. To construct, structure, and give meaning to the concept, we use our experience in the field, critical reading of the literature, and careful analysis of data that have emerged about parental decision-making in pediatric intensive care. Several factors affect parent's ability to act as decision-makers: the psychosocial and physical disorders they develop, the subordination of their parental roles by the health care team, and the child's critical state of health. While different disciplines, including nursing, have well described the decision-making concept, parental decision-making in the context of pediatric intensive care has not been as well delineated. Nursing science recognizes the importance of decision-making and has incorporated the concept as an essential domain of its philosophical and disciplinary interests. Following the method proposed by Walker and Avant, the concept was analyzed, attributes, background, and consequences described. A model case was presented and discussed. An operational definition emerges, providing knowledge for professional nursing practice and will be the basis for an essential theoretical development around this phenomenon. Parents' recognition, the promotion of family-centered care, and shared decisions are ideal for encouraging parental participation.

The ataxia telangiectasia-mutated (ATM) protein is a major coordinator of the DNA damage response pathway. ATM loss-of-function variants are associated with 2-fold increased breast cancer risk. We aimed at identifying and classifying spliceogenic ATM variants detected in subjects of the large-scale sequencing project BRIDGES. A total of 381 variants at the intron-exon boundaries were identified, 128 of which were predicted to be spliceogenic. After further filtering, we ended up selecting 56 variants for splicing analysis. Four functional minigenes (mgATM) spanning exons 4-9, 11-17, 25-29, and 49-52 were constructed in the splicing plasmid pSAD. Selected variants were genetically engineered into the four constructs and assayed in MCF-7/HeLa cells. Forty-eight variants (85.7%) impaired splicing, 32 of which did not show any trace of the full-length (FL) transcript. A total of 43 transcripts were identified where the most prevalent event was exon/multi-exon skipping. Twenty-seven transcripts were predicted to truncate the ATM protein. A tentative ACMG/AMP (American College of Medical Genetics and Genomics/Association for Molecular Pathology)-based classification scheme that integrates mgATM data allowed us to classify 29 ATM variants as pathogenic/likely pathogenic and seven variants as likely benign. Interestingly, the likely pathogenic variant c.1898+2T>G generated 13% of the minigene FL-transcript due to the use of a noncanonical GG-5'-splice-site (0.014% of human donor sites). Circumstantial evidence in three ATM variants (leakiness uncovered by our mgATM analysis together with clinical data) provides some support for a dosage-sensitive expression model in which variants producing ¿30% of FL-transcripts would be predicted benign, while variants producing ¿13% of FL-transcripts might be pathogenic., PD, MPGV, DFE, MdlH and EAV received funding from the European Union's Horizon 2020 research and innovation program under grant agreement no. 634935. The EAV lab is supported by grants from the Spanish Ministry of Science and Innovation, Plan Nacional de I+D+I 2013-2016, ISCIII (PI17/00227 and PI20/00225) co-funded by the FEDER from Regional Development European Funds (European Union) and from the Consejería de Educación, Junta de Castilla y León, ref. CSI242P18 (actuación cofinanciada P.O. FEDER 2014-2020 de Castilla y León). The MdlH lab is supported by a grant from the Spanish Ministry of Science and Innovation, Plan Nacional de I+D+I 2013-2016, ISCIII (PI20/00110) co-funded by FEDER from Regional Development European Funds (European Union). Programa Estratégico Instituto de Biología y Genética Molecular (IBGM), Escalera de Excelencia, Junta de Castilla y León (Ref. CLU-2019-2002). EB-M is a postdoctoral researcher funded by the University of Valladolid (POSTDOC-UVA05, 2022-2025). AV-P and IL-B are supported by predoctoral fellowships from the Consejería de Educación, Junta de Castilla y León. LS-M is supported by a predoctoral fellowship from the AECC-Scientific Foundation, Sede Provincial de Valladolid (2019–2023). AE-S is supported through the Operational Program for Youth Employment and Youth Employment Initiative (YEI), called by the Community of Madrid in 2020, and co-financed by the European Social Fund.

Background: In patients with advanced high-grade serous ovarian cancer (HGSOC) and prostate adenocarcinoma, the identification of somatic/germline BRCA1/2 mutations allows new therapeutic opportunities. To estimate the prevalence of somatic and germline BRCA1/2 mutations in non-mucinous high grade ovarian/fallopian tube/peritoneal extraovarian cancer (NMHGOC) and prostate adenocarcinoma. Methods: Prevalence was established by analyzing patients with NMHGOC or prostate adenocarcinoma, with a BRCA1/2 study in the tumor between 2017 and 2018. Whether a germline study had been carried out was subsequently reviewed. Results: 10 patients out of 43 (23.3%) with NMHGOC had a BRCA1/2 mutation in the tumor. 9 patients (20.9%) presented a BRCA1/2 mutation in the germline setting (2 without tumor result due to limited tissue sample). 3 patients (6.9%) had only somatic mutations. 30% of the mutations in the tumor were, therefore, somatic mutations. Of the 9 patients with prostate adenocarcinoma, 2 (22.2%) had a BRCA2 mutation in the tumor. While 1 (11.1%) had the mutation in the germline setting, 1 patient (11.1%) had only somatic mutations. Conclusion: In our series, the prevalence of somatic and germline BRCA1/2 mutations in NMHGOC is similar to that reported in the literature. Whereas somatic mutations are only present at the neoplastic tissue, the rate of mutations in the tumor is higher than in the germline setting. A more effective diagnostic and predictive strategy could be achieved with tumor BRCA analysis as the first attempt. Initial results in prostate adenocarcinoma point to the same conclusion for this tumor.

En los últimos años ha aumentado el interés por el estudio de la ciberviolencia en la pareja, sin embargo es necesario examinar la metodología de investigación que sustenta las evidencias obtenidas hasta el momento. Este artículo presenta una revisión sistemática de la literatura científica que analiza 30 instrumentos de medida de ciberviolencia en la pareja de adolescentes y jóvenes adultos utilizados a lo largo de estos años. Los principales resultados muestran una elevada pluralidad metodológica, conceptual y terminológica, observándose un reducido número de instrumentos con suficientes garantías psicométricas. Predominan los trabajos de procedencia estadounidense y, entre los instrumentos aplicados en muestras españolas, destaca una infrarrepresentación de indicadores de ciberviolencia de tipo sexual, lo que limita la comprensión de esta problemática. Esta investigación aporta información sobre el estado actual en el estudio de la ciberviolencia en la pareja y pone de relieve deficiencias metodológicas en la construcción del conocimiento en este campo de estudio. Este trabajo permite una mejor comprensión de la disparidad de resultados señalada por investigaciones previas, especialmente referidas a prevalencia, frecuencia y diferencias de género en este tipo de comportamientos violentos, además de sentar las bases para abordar el fenómeno desde el rigor científico. The interest for the study of the cyber dating abuse has increased in the last years; however, it is necessary to examine the research methodology that supports the evidence obtained so far. This paper presents a systematic review of the scientific literature that analyzes 30 measuring instruments of cyber dating abuse in adolescent and young adults used along these years. The main results show a methodological, conceptual and terminological plurality, observing a small number of instruments with sufficient psychometric guarantees. The instruments come mostly from the United States, and of the instruments applied to Spanish samples, few include indicators of sexual cyber dating abuse, which limits the understanding of this problem. This research provides information on the current status of the study of cyber dating abuse and highlights methodological shortcomings in the construction of knowledge in this field of study. This study allows a better understanding of the disparity of results indicated by previous research, especially referring to prevalence, frequency and gender differences in this type of violent behavior, as well as laying the foundations for addressing the phenomenon from scientific rigor.

Sexist attitudes have been identified as an important variable which fosters a climate of social permissiveness towards sexual violence towards women. However, its impact on social reactions to sce...

© The Author(s) 2021., Weight gain is a frequent and severe adverse reaction in patients taking antipsychotics. The objective was to further investigate in a natural setting influential risk factors associated with clinically significant weight gain. An observational follow-up study was conducted. Patients when initiating treatment with whatever antipsychotic were included; a structured questionnaire was applied at baseline, 3 and 6 months later; a blood sample was obtained. In a nested case–control approach, patients with an increase ≥ 7% of their initial weight were considered as cases, the remaining, as controls. The results showed that, out of 185 patients, 137 completed the 6-month follow-up (cases, 38; controls, 99). Weight gain gradually and significantly increased in cases (baseline, 65.0 kg; 6 months, 74.0 kg) but not in controls (65.6 kg and 65.8 kg, respectively). Age (adjusted OR = 0.97, 95% CI = 0.96–0.99, p = 0.004), olanzapine (adjusted OR = 2.98, 95% CI = 1.13–7.80, p = 0.027) and quetiapine (adjusted OR = 0.25, 95% = 0.07–0.92, p = 0.037) significantly associated with weight gain. An association was also found for the CNR1 (rs1049353) and INSIG2 (rs7566605) polymorphisms. In conclusion, an increased risk of antipsychotics-induced weight gain was observed for younger age and olanzapine, and a relative lower risk for quetiapine. A potential role of CNR1 rs1049353 and INSIG2 rs7566605 polymorphisms is suggested., Funding for this study was provided by the Ministry of Education of the Regional Government of Castile and Leon [GR129 and VA315U14]; Ministry of Health of the Regional Government of Castile and Leon [BIO/VA20/13] and Spanish Ministry of Health [EC11-395].

El modo en que los medios de comunicación abordan determinados problemas sociales, como es el caso del ciberacoso en la adolescencia, condiciona en gran medida el interés mediático y su percepción social, lo que en último término afecta a su correcto abordaje y erradicación. Por esta razón, el presente estudio analiza la imagen que dos importantes medios de comunicación españoles, el periódico El País y el periódico El Mundo, trasmiten del ciberacoso adolescente. Se analizaron mediante la metodología análisis de contenido 26 piezas informativas publicadas en un período de dos años. Entre los resultados destacados se observó una mayor tendencia al análisis del ciberacoso frente a la noticiabilidad o sensacionalismo. A pesar de valorarse de manera suficiente el tratamiento que realizan del ciberacoso adolescente, se recomienda a ambos periódicos mayores esfuerzos en la lucha contra este problema, así como proporcionar información fidedigna, recursos de asistencia y fomentar su denuncia. The way in which the mass media tackle certain social problems, such as cyberbullying in adolescence, greatly conditions media interest and social perception and ultimately affects its right approach and eradication. For this reason, the present study analyzes the image of cyberbullying which transmit two important Spanish media, the newspaper El País and the newspaper El Mundo. Twenty-six journalistic texts published over a period of two years were analyzed by the content analysis methodology. Among the outstanding results was observed a greater tendency to the analysis of cyberbullying against sensationalism. In spite of valuing the treatment that they make of the adolescent cyberbullying how sufficient, it is recommended to both newspapers greater efforts in the struggle against this problem, as well as providing reliable information, resources of assistance and encouraging its denunciation.

In this study, we aim to gain insight in the germline mutation spectrum of ATM, BARD1, BRIP1, ERCC4, PALB2, RAD51C and RAD51D in breast and ovarian cancer families from Spain. We have selected 180 index cases in whom a germline mutation in BRCA1 and BRCA2 was previously ruled out. The importance of disease-causing variants in these genes lies in the fact that they may have possible therapeutic implications according to clinical guidelines. All variants were assessed by combined annotation dependent depletion (CADD) for scoring their deleteriousness. In addition, we used the cancer genome interpreter to explore the implications of some variants in drug response. Finally, we compiled and evaluated the family history to assess whether carrying a pathogenic mutation was associated with age at diagnosis, tumour diversity of the pedigree and total number of cancer cases in the family. Eight unequivocal pathogenic mutations were found and another fourteen were prioritized as possible causal variants. Some of these molecular results could contribute to cancer diagnosis, treatment selection and prevention. We found a statistically significant association between tumour diversity in the family and carrying a variant with a high score predicting pathogenicity (p = 0.0003)., This research was funded by the Regional Government of Castilla Y León through the University of Valladolid, Valladolid (Spain) and BOF15/GOA/011/Universiteit Gent. C.V. was supported by a predoctoral fellowship from the Regional Government of Castilla y León (Orden EDU/1083/2013) co-financed by the European Social Fund (ESF). Special Research Fund (BOF) scholarship of Ghent University to F.A.C. (BOF.DOC.2017.0026.01).

Simple Summary Hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome is a very rare hereditary disorder characterized by cutaneous leiomyomas (CLMs), uterine leiomyomas (ULMs), renal cysts (RCys) and renal cell cancer (RCC), with no data on its prevalence worldwide. No genotype-phenotype associations have been described. The aim of our study was to describe the genotypic and phenotypic features of the largest series of patients with HLRCC from Spain reported to date. Of 27 FH germline pathogenic variants, 12 were not previously reported in databases. Patients with missense pathogenic variants showed higher frequencies of CLMs, ULMs and RCys, than those with loss-of-function variants. The frequency of RCCs (10.9%) was lower than those reported in the previously published series. Hereditary leiomyomatosis and renal cell cancer syndrome (HLRCC) is a very rare hereditary disorder characterized by cutaneous leiomyomas (CLMs), uterine leiomyomas (ULMs), renal cysts (RCys) and renal cell cancers (RCCs). We aimed to describe the genetics, clinical features and potential genotype-phenotype associations in the largest cohort of fumarate hydratase enzyme mutation carriers known from Spain using a multicentre, retrospective study of individuals with a genetic or clinical diagnosis of HLRCC. We collected clinical information from medical records, analysed genetic variants and looked for genotype-phenotype associations. Analyses were performed using R 3.6.0. software. We included 197 individuals: 74 index cases and 123 relatives. CLMs were diagnosed in 65% of patients, ULMs in 90% of women, RCys in 37% and RCC in 10.9%. Twenty-seven different pathogenic variants were detected, 12 (44%) of them not reported previously. Patients with missense pathogenic variants showed higher frequencies of CLMs, ULMs and RCys, than those with loss-of-function variants (p = 0.0380, p = 0.0015 and p = 0.024, respectively). This is the first report of patients with HLRCC from Spain. The frequency of RCCs was lower than those reported in the previously published series. Individuals with missense pathogenic variants had higher frequencies of CLMs, ULMs and RCys.

BRIP1 is a component of the Fanconi Anemia/BRCA pathway responsible for DNA reparation via helicase activity. Some heterozygous variants in BRIP1 could contribute to Hereditary Breast Cancer through a defective DNA repair. The clinical utility of BRIP1 mutations in a familial cancer context is compromised by the conflicting interpretation of “variants of uncertain significance” (VUS). Defining the clinical significance of variants identified in genetic tests is a major challenge; therefore, studies that evaluate the biological effect of these variants are definitely necessary. To contribute to this purpose, we have characterized the variant c.550G>T of BRIP1, a missense mutation with little evidence about its pathogenicity. Since Human Splicing FinderTM predicts the creation of a new exonic splicing enhancer site we decided to perform cDNA analysis revealing that the c.550G>T mutation located in exon 6 led to an aberrant transcript causing exon 5 skipping. Our results demonstrate that the c.550G>T BRIP1 variant disrupts normal splicing, causing exon 5 skipping. Considering that the exon 5 encodes the helicase domain of BRIP1, it is expected an alteration of the function. This finding enhances the interpretation of this VUS, suggesting a potential pathogenic effect., This work was supported by the University of Valladolid, Valladolid (Spain) and The Regional Government of Castilla y León.

[Purpose]: Promoter mutations may affect transcription and can be associated with human diseases. However, the promoters of the breast cancer (BC) genes are not regularly screened. Our goal was to investigate the BRCA2 promoter in order to study a possible correlation between impaired transcription and disease. [Methods]: The proximal and core promoter of the BRCA2 gene was sequenced in 95 high-risk BC patients. A BRCA2-promoter insert [- 938 to + 312 from the transcription start site (TSS)] was generated and cloned into the firefly luciferase vector pGL4.10. Promoter variants and deletions were introduced by site-directed mutagenesis and quantified by Dual-Luciferase assays and semi-quantitative RT-PCR. [Results]: Three different variants were detected in high-risk BC patients: rs3092989, rs206118, and rs563971900. Functional mapping of 13 overlapping deletions revealed four down-regulating segments (TSS positions): -59_-10del/µdel3 (16% of activity of the wild-type construct), -104_-55del/µdel4 (62%), -239_-190del/µdel7 (39%), -464_-415/µdel12 (78%), suggesting the presence therein of putative transcriptional activator motifs. Additionally, six microdeletions rendered luciferase overexpression: +32_+81del/µdel1 (356%), -14_+36del/µdel2 (180%), -194_-145del/µdel6 (154%), -284_-235del/µdel8 (168%), -329_-280del/µdel9 (111%), and -509_-460del/µdel13 (139%), which is indicative of repressor elements. Functional assays of 15 promoter variants (including those detected in patients) showed that ten of them significantly altered expression with seven up-regulating (113-163%) and three down-regulating (rs551887850_G, rs570548398_T, rs55880202_T; 72-83%) SNPs. Eight of them were located in an ENCODE-DNase Hypersensitive Cluster (TSS - 185 to + 105) where most active transcriptional motifs are known to be placed. [Conclusions]: BRCA2 expression is highly sensitive to promoter variations as most of them induced relevant changes. Moreover, we mapped critical regions of the BRCA2 promoter that may constitute potential targets for regulatory variants. Three SNPs moderately decreased luciferase activity, but confirmation of its potential pathogenicity requires further analysis. These data reinforce the need to screen the promoter regions of breast cancer genes with a view to discovering novel deleterious mutations., EAV’s lab was supported by grants from the Spanish Ministry of Economy and Competitivity, Plan Nacional de I+ D +I 2013–2016, ISCIII (Fis: PI13/01749) co-funded by FEDER from Regional Development European Funds (European Union), and Grant CSI090U14 from the Consejería de Educación (ORDEN EDU/122/2014), Junta de Castilla y León. EFB was supported by a predoctoral fellowship from the University of Valladolid and Banco de Santander (2015–2019).