Male breast cancer (MBC), a rare disease accounting for less than 1% of all cancers in men and approximately 1% of all breast cancers globally, exhibits unique biological and clinical characteristics that distinguish it from female breast cancer (FBC). To identify the latest developments and trends in MBC research, we performed a comprehensive analysis of relevant literature and generated visual representations of the data. Building on these outcomes, we delve into the insights gained, aiming to enhance the ongoing comprehension of MBC. We conducted a search of the Web of Science Core Collection (WoSCC) for English-language articles and reviews on MBC published from 1982 to 2024. Following a rigorous dual-researcher screening process, we refined the dataset to 6802 relevant records. Subsequently, we employed Microsoft Excel for initial data organization and processing. For in-depth analysis and visualization of journal, country/region, author/institution trends, as well as keyword patterns, we utilized a combination of R Package, VOSviewer, and CiteSpace. This multi-tool approach enabled us to extract and present meaningful insights from the data. MBC research has risen since 1996, with a 2024 peak at 503 publications, reflecting advances like BRCA1/2 gene discoveries. The USA leads in publications and citations, with North America and Europe being the most collaborative regions. Key journals include Breast Cancer Research and Treatment, and leading contributors are Ottini, Laura and Van Diest, Paul J. Research focuses on MBC's genetic links (e.g. BRCA mutations) and clinical features, with emerging topics like precision medicine and immunotherapy. MBC research has grown but remains concentrated in high-income countries. Future efforts should explore MBC's unique biology, improve diagnostic/therapeutic strategies, and involve global, interdisciplinary collaboration to enhance patient outcomes.

ObjectivesSociocultural standards that emphasize idealized appearance and promote the objectification of women’s bodies have been consistently associated with negative body image outcomes and increased interest in cosmetic surgery (Fredrickson & Roberts; Calogero et al.). Constructs such as body image flexibility and intuitive eating have been discussed in the literature as psychological resources that promote well-being in body image–related contexts (Sandoz et al.; Tylka & Kroon Van Diest). However, their specific role in the relationship between objectification and cosmetic surgery interest remains underexplored. Recent evidence suggests that body image flexibility may function as a protective factor in this relationship (Huang et al.), while intuitive eating has been associated with greater psychological well-being and reduced body-related distress (Tylka and Wilcox), suggesting its potential relevance. Building on this background, the present study investigates the mediating roles of body image flexibility and intuitive eating in the relationship between body objectification and the desire to undergo cosmetic surgery for intrapersonal and social reasons, as well as the likelihood of pursuing such procedures in the future.MethodsThe sample consisted of 555 Romanian women (M = 29.61 years, SD = 13.396), who completed validated scales measuring body objectification, body image flexibility, intuitive eating, and attitudes toward cosmetic surgery. Data were analyzed using parallel mediation models, controlling for age, educational status, ethnicity, relationship status, and body mass index.FindingsThe findings indicated that body objectification was negatively associated with body image flexibility and intuitive eating. While intuitive eating did not mediate the relationship between body objectification and the desire for cosmetic surgery in any of the tested models, body image flexibility emerged as a partial mediator in the model related to social motivations and a full mediator in the model predicting future consideration of cosmetic procedures. These results are consistent with theoretical frameworks emphasizing self-perception and sociocultural context—such as self-verification and self-affirmation theories—that help explain how women’s behaviors are shaped by societal expectations and patriarchal cultural norms.Conclusion and recommendationsThe study highlights the relevance of cultural context in understanding adaptive factors that may buffer the psychological impact of objectification. The results suggest that body image flexibility may function as a protective factor in reducing the desire for cosmetic surgery. While these findings may suggest potential directions for intervention, such as promoting positive body image and intuitive eating, we emphasize that further longitudinal research is needed before such psychoeducational programs can be designed or implemented. This study contributes to the growing body of literature by shedding light on culturally specific dynamics influencing cosmetic surgery motivations.

BackgroundSerotonin has been attributed both an anxiety-reducing and an anxiety-enhancing influence. For example, acute administration of selective serotonin reuptake inhibitors (SSRIs) as well as of a serotonin releaser, fenfluramine, may provoke anxiety in susceptible individuals (Targum and Marshall, 1989). In contrast, there are reports on acute administration of 5-HTP alleviating anxiety (Maron et al., 2004; Schruers et al., 2002). We have previously shown that acute administration of an SSRI to rat exacerbates unconditioned freezing behaviour without influencing context-conditioned freezing (Hagsater et al., 2019).Aims & ObjectivesTo explore the possible impact of compounds (5-HTP and fenfluramine) exerting a more robust (and more non-physiological) increase in extracellular serotonin concentrations than that obtained with an SSRI (escitalopram) on acquisition and expression of context-conditioned fear.MethodMale Sprague Dawley rats were tested in a paradigm of conditioned fear. Electric foot shocks (amperage: 0.6 mA, duration: 1 s, inter-shock intervals: 30 s) were used as unconditioned stimuli while the context constituted the conditioned stimulus. The effects of 5-HTP, fenfluramine and escitalopram administered i) before acquisition of contextual-conditioned fear, ii) immediately after acquisition of contextual- conditioned fear and iii) before expression of context-conditioned fear, were explored.ResultsBoth 5-HTP (> 25 mg/kg) and fenfluramine (> 0.5 mg/kg) decreased expression of conditioned fear in a dose-dependent manner. At higher dosage, both 5-HTP (> 250 mg/kg) and fenfluramine (5 mg/kg) administered prior to acquisition of context-conditioned fear eliminated freezing in subsequent testing. Neither 5-HTP (500 mg/kg) nor fenfluramine (5 mg/kg) administered immediately after fear conditioning impaired freezing in subsequent testing. No effect on acquisition or expression of context-conditioned fear was observed for escitalopram (30 mg/kg).Discussion & ConclusionThe results suggest 5-HTP and fenfluramine at an intermediate dose to disrupt fear conditioning when administered prior to acquisition or prior to expression of conditioned fear. Since there, in contrast, was no effect of 5-HTP or fenfluramine when administered immediately after acquisition, the effects on conditioned freezing are not likely due to drug-induced memory impairment. The effects of the drugs on acquisition and expression of conditioned freezing suggest that the augmentation of the serotonergic output and/or the disruption of normal serotonergic transmission induced by 5-HTP and fenfluramine interfere with the processes underlying fear conditioning.ReferencesHagsater SM, Thoren J, Pettersson R, Eriksson E. Selective serotonin reuptake inhibition increases noise burst-induced unconditioned and context-conditioned freezing. Acta neuropsychiatrica. 2019 Feb;31(1):46-51.Maron E, Toru I, Vasar V, Shlik J (2004) The effect of 5-hydroxytryptophan on cholecystokinin-4-induced panic attacks in healthy volunteers. Journal of psychopharmacology 18:194-199.Schruers K, van Diest R, Overbeek T, Griez E (2002) Acute L-5-hydroxytryptophan administration inhibits carbon dioxide-induced panic in panic disorder patients. Psychiatry research 113:237-243. Targum SD, Marshall LE (1989) Fenfluramine provocation of anxiety in patients with panic disorder. Psychiatry research 28:295-306.

Abstract Rationale: Pregnancy-Associated Breast Cancer (PABC), often defined as breast cancer during pregnancy (PrBC) or in the first year postpartum (PPBC), is known for its aggressive histopathology and higher stage at diagnosis. In a previous study of the nationwide Dutch PABC Cohort, we observed an impaired survival for patients diagnosed in the second and third trimester of pregnancy. However, the etiology of these differences, and how this relates to the patients diagnosed with breast cancer in the postpartum period (PPBC) or after an abortion (AABC), remains to be elucidated. While we are exploring the etiologic hypotheses, we updated and expanded our database to compare these specific PABC subgroups. Methods: All pathology reports of invasive breast carcinoma between January 1st 1988 and July 1st 2022 were screened for pregnancy-related keywords to find all patients diagnosed with PrBC, PPBC or AABC within one year after childbirth or an interrupted pregnancy (induced abortion or miscarriage before a gestational age of 24 weeks). A local patient series provided 22 additional cases. Pregnant patients were subdivided by gestational trimester at time of diagnosis. The different PrBC, PPBC and AABC subgroups were compared by histopathology, clinical characteristics and outcome. Results: 787 patients were included, of whom 60% were pregnant during their diagnosis. Median age at diagnosis was 34 years. In line with our previous findings, a large majority (68%) of patients had a Bloom & Richardson (B&R) grade III tumor and 37% of tumors were triple negative. Moreover, over a third of patients presented with lymph node metastases (38%). We observed two distinct groups based on histopathology and prognosis: group A, consisting of three patient groups: (I) patients diagnosed in the first trimester of pregnancy, (II) patients diagnosed between 6-12 months postpartum, and (III) patients diagnosed after an abortion, and group B, consisting of two patient groups, (I) patients diagnosed in the second or third trimester of pregnancy or (II) within 6 months after childbirth. Compared to group A, patients in group B more often had B&R grade III tumors: (73% vs. 60%, p=0.002) and more frequently had triple negative tumors (43% vs. 28% in group A, p< 0,001). Survival was significantly worse for patients in group B (5-year OS 67,3% versus 88.2%, p< 0.001). In a multivariable analysis, corrected for grade, type of surgery and overall stage, these differences upheld with an HR of 1.515 (95% CI 1,012 – 2,268). Conclusions: With this extensive update of our nationwide Dutch PABC cohort, we show that especially PrBC patients diagnosed in the second and third trimesters and PPBC patients within six months after childbirth exhibit unfavorable tumor characteristics, with an associated worse prognosis. This highlights the need for in-depth analyses in these specific groups of PABC patients to elucidate the etiologic mechanisms involved. Citation Format: Carsten Bakhuis, Carmen Van Dooijeweert, Britt Suelmann, Janneke Verloop, Pieter Westenend, Sabine Linn, Paul Van Diest, Elsken Van der Wall. Worse Prognosis for Breast Cancer in the Second and Third Trimesters and Shortly Postpartum:An Update of the Dutch Pregnancy-Associated Breast Cancer Cohort [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PS13-01.

Abstract Background Metastases in sentinel lymph nodes (SN) of breast cancer (BC) patients are strongly associated with a worse survival and consequently guide treatment. If metastases are absent upon pathologist’ assessment of the regular hematoxylin and eosin (HE)-slide, additional immunohistochemistry (IHC)-stains are performed to ensure that no metastases are missed. However, these stains come at high additional costs, which may exceed specimen reimbursement. Fortunately, digital pathology is becoming more common, thereby creating an avenue of opportunities for artificial intelligence (AI) assistance. Although the number of studies on (promising) AI-algorithms increases exponentially, studies on actual clinical implementation are lacking. In this single-center prospective trial, we investigated to which extent an artificial intelligence (AI)-assisted clinical workflow for the detection of SN-metastases reduces IHC-use, while maintaining current diagnostic safety standards. Methods We enrolled 190 SN-specimens of 182 patients with invasive or in situ BC from September 2022 to May 2023. SN-specimens were allocated bi-weekly to either the control-arm (n=90) or the intervention-arm (n=100). In the control-arm, SN-specimens were digitally assessed according to the current clinical workflow, while pathologists in the intervention-arm assessed the SN-specimen with the output the ‘Metastasis-Detection-App’ (Visiopharm©) available. In both groups, IHC was performed in all morphologically negative cases. Main outcome was the relative risk (RR) of IHC-use per detected case of SN-metastases. Case-mix adjustment was performed by log-binomial regression. Results Overall, 59 SN-specimens contained metastases (31.1%). AI-assistance resulted in a significantly lower risk of IHC-use per detected case of SN-metastases (adjusted RR: 0.680, 95% CI: 0.347-0.878). Besides preventing IHC-use, thereby reducing costs, AI-assisted pathologists also spent significantly less time on their assessment of the SN-specimen (3m:41s vs. 6m:04s, p = 0.028). Furthermore, the sensitivity of AI-assisted pathologists was up to 30% higher. The AI-assisted pathologists missed two cases of micro-metastases in the intervention arm, one of which was in retrospect highlighted by the algorithm, while in the other case the tumor cells were located in a heavily cauterized area of the HE-slide and therefore only visible on the (serial) IHC-slides. In the control arm, the algorithm in retrospect picked up all micro- and macro-metastases and nearly half of the isolated tumor cells (ITC). In addition, all participating pathologists stated that AI was easy to use, that they felt confident using AI, and that besides saving them time, AI made their work more enjoyable. Cost reductions on IHC by AI-assistance depend on laboratory policy (i.e. when and on how many levels IHC is performed), but, at a cost of €25,- per IHC-stain, range from €1.500 to €3.500 per 100 SN’s in a scenario where IHC is performed in all morphologically negative cases. In a scenario where IHC is only performed in patients in whom finding ITC has clinical consequences (i.e. patients who received neoadjuvant treatment), cost savings on IHC range from €7.500-€12.500 per 100 SNs, depending on laboratory policy. Conclusion AI-implementation for the detection of SN-metastases in BC-patients leads to a significant reduction of IHC-use and subsequent costs, while saving pathologists time and making their work more enjoyable. Importantly, AI-implementation during this trial was safe and patients were not at risk of an inferior diagnosis. By doing this trial alone, an estimated €3,000 on IHC-use was saved. Such tangible cost savings are crucial to build a viable business case for AI implementation in diagnostic pathology. Citation Format: Carmen van Dooijeweert, Rachel Flach, Natalie ter Hoeve, Celien Vreuls, Roel Goldschmeding, Jan-Erik Freund, Paul Pham, Tri Nguyen, Elsken Van der Wall, Geert Frederix, Nikolas Stathonikos, Paul Van Diest. Clinical implementation of artificial-intelligence-assisted detection of breast cancer metastases in sentinel lymph nodes: saving costs and time (the CONFIDENT-B trial) [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PS03-06.

Abstract Rationale: Breast cancer is the most common malignancy in pregnant women, occurring in approximately 1:3000 pregnancies. Breast cancer diagnosed during pregnancy or after childbirth is also known as Pregnancy-Associated Breast Cancer (PABC). PABC is known for exhibiting several unfavorable prognostic tumor characteristics, such as an advanced tumor stage at diagnosis, high histologic grade and frequent hormone receptor negativity. Most probably, several interplaying mechanisms may promote the tumor progression and adverse clinical outcome of PABC. One of these mechanisms may be the presence of hypoxia, a relative state of low intra-tumoral oxygen levels which is a known marker of adverse outcome in breast cancer. However, the occurrence and prognostic consequences of the presence of intra-tumoral hypoxia in PABC have not been studied yet. Methods: We constructed a cohort with histopathological, clinical, and outcome data of patients diagnosed with breast cancer during pregnancy (PrBC) or within one year after childbirth (PPBC) in the Netherlands between 1988 and 2022. Next, tissue blocks of all patients were collected from the participating Dutch pathology laboratories. Using H&E stained slides of the tumor tissue blocks, we selected suitable donor sites for a tissue micro array (TMA). Slides from these TMAs were stained for three important hypoxia-associated proteins: glucose transporter-1 (Glut-1), carbonic anhydrase IX (CAIX) and hypoxia-inducible factor-1α (HIF-1α). Expression was scored by an expert breast pathologist blinded by clinicopathologic data. Results: For a total of 195 PrBC and PPBC patients, we were able to assess the expression of Glut-1, CAIX and HIF-1α on tumor cells. Patients had a median age of 33 years at diagnosis, whilst a large majority had a Bloom & Richardson grade III tumor (76%) with frequent hormone receptor negativity (50%). Expression of hypoxia-associated proteins was frequent, with 61% of the tumors expressing Glut-1, 30% expressing CAIX and 56% expressing HIF-1α. In total, 153 (78%) of the tumors expressed at least one of the hypoxia-associated proteins. We observed a significantly worse 5-year overall survival for patients with intra-tumoral hypoxia in comparison to patients without intra-tumoral hypoxia (70% vs. 90%, p=0.046). Conclusions: We show that the presence of intra-tumoral hypoxia in PABC is common, with 78% of tumors expressing at least one of the studied hypoxia-associated proteins. Importantly, patients with tumors overexpressing hypoxia-markers have a significantly worse survival. This shows that intra-tumoral hypoxia may be an important underlying carcinogenic mechanism in PABC, and might be a promising novel therapeutic target in this patient group. Citation Format: Carsten Bakhuis, Carmen Van Dooijeweert, Britt Suelmann, Pieter Westenend, Sabine Linn, Elsken Van der Wall, Paul Van Diest. Frequent Intra-Tumoral Hypoxia in Pregnancy-Associated Breast Cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PS13-04.

Abstract SY45-02: Engineering strategies based on receptors derived from gdT cells

Slow-paced breathing at an individual’s resonance frequency (RF) is a common element of heart rate variability (HRV) biofeedback training (Laborde et al. in Psychophysiology 59:e13952, 2022). Although there is strong empirical support for teaching clients to slow their respiration rate (RR) to the adult RF range between 4.5 and 6.5 bpm (Lehrer & Gevirtz, 2014), there have been no definitive findings regarding the best inhalation-to-exhalation (IE) ratio to increase HRV when breathing within this range. Three methodological challenges have frustrated previous studies: ensuring participants breathed at the target RR, IE ratio, and the same RR during each IE ratio. The reviewed studies disagreed regarding the effect of IE ratios. Three studies found no IE ratio effect (Cappo & Holmes in J Psychosom Res 28:265-273, 1984; Edmonds et al. in Biofeedback 37:141-146, 2009; Klintworth et al. in Physiol Meas 33:1717-1731, 2012). One reported an advantage for equal inhalations and exhalations (Lin et al. in Int J Psychophysiol 91:206?211, 2014). Four studies observed an advantage for longer exhalations than inhalations (Bae et al. in Psychophysiology 58:e13905, 2021; Laborde et al. in Sustainability 13:7775, 2021; Strauss-Blasche et al. in Clin Exp Pharmacol Physiol 27:601?60, 2000; Van Diest et al. in Appl Psychophysiol Biofeedback 39:171?180, 2014). One study reported an advantage for longer inhalations than exhalations (Paprika et al. in Acta Physiol Hung 101:273?281, 2014). We conducted original (N = 26) and replication (N = 16) studies to determine whether a 1:2 IE ratio produces different HRV time-domain, frequency-domain, or nonlinear metrics than a 1:1 ratio when breathing at 6 bpm. Our original study found that IE ratio did not affect HRV time- and frequency-domain metrics. The replication study confirmed these results and found no effect on HRV nonlinear measurements.

PurposeIntuitive Eating (IE) is an approach to eating characterised by attunement to intrinsic cues, and using those cues to guide behaviours related to food and eating. Tylka and Kroon Van Diest's (2008) Intuitive Eating Scale (IES-2) is well-validated in adults, but not yet among early adolescents. The current study was designed to adapt and validate a version suited for use in early adolescence (IES-2-EA). MethodsData collected from two independent samples of adolescents aged 11 to 13 (N = 471) were structurally examined using exploratory factor analysis (EFA), with a subsequent confirmatory factor analysis (CFA) to confirm hypothesised model fit. Relationships between scores on the IES-2-EA and validated measures of actual-ideal body size discrepancy, body appreciation, interoceptive awareness, and psychological distress were also examined. ResultsThe adapted 17-item IES-2-EA had a three-factor structure with several key differences from the original version developed for adults. Moderate-to-strong correlations were found between scores on the IES-2-EA, body appreciation, interoceptive awareness, actual-ideal body size discrepancy, and psychological distress in the first sample of adolescents (n = 245). Secondary CFA showed good model fit in the second sample of adolescents (n = 226). ConclusionThe IES-2-EA is well-suited to measure intuitive eating behaviour among early adolescents. The 17 items reflect a three-component structure similar to that seen in adults completing the IES-2. These early data suggest the adapted IES-2-EA has evidence of reliability and validity; it may be an effective measure for research and clinical use.

Abstract This study describes the expression of an intracellular variant of the Zona Pellucida glycoprotein 3 (ZP3) that is predominantly expressed in cancer. ZP3 is an important constituent of the Zona Pellucida (ZP), the extracellular layer surrounding oocytes. It is highly expressed in maturing oocytes, but not in other adult human tissues. As ZP3 protein has also been detected in cancer, it may serve as an interesting diagnostic and therapeutic target. Here, we extend the cancer-related ZP3 expression data using immunohistochemistry (IHC) of tumor biopsies, interrogating publicly available RNA-sequencing (RNA-seq) data of cancer cell lines (CCLs) and tumor and normal tissues, as well as computational analysis and real-time quantitative PCR (qPCR) of CCLs. IHC data for several cancer types shows abundant ZP3 protein expression, which is confined to the cytoplasm, contradicting the extracellular ZP3 localization in oocytes. An alternative ZP3 mRNA variant, which we term ‘ZP3-Cancer’, is annotated in the NCBI and Ensembl databases, and lacks the genetic information encoding the N-terminal signal peptide that governs extracellular secretion. Analysis of publicly available RNA-seq data of 1339 CCLs indicates that ZP3-Cancer is the dominant variant as compared to ZP3-Oocyte, which was validated by independent computational analysis. Expression of ZP3-Cancer in CCLs was confirmed by qPCR. Publicly available RNAseq data of tumor and normal tissues confirms strongly enhanced expression of ZP3-Cancer in cancer cells. In addition, ZP3-Cancer expression is upregulated in advanced stages and shows a relationship with patient survival in a number of cancer types. The folliculogenesis specific transcription factor FIGLA, which activates transcription of ZP3-Oocyte in maturing oocytes, appears absent in cancer as inferred from publicly available transcriptome data, indicating alternative ZP3-Cancer transcriptional activation mechanisms in this disease. The cancer restricted expression of this ZP3 transcript variant renders it an attractive tumor specific antigen for the development of a therapeutic cancer vaccine, particularly using mRNA technology. Citation Format: Iman J. Schultz, Yvette Zimmerman, Cathy B. Moelans, Marcin Chrusciel, Jan Krijgh, Paul J. van Diest, Ilpo T. Huhtaniemi, Herjan J. Coelingh Bennink. An intracellular variant of zona pellucida glycoprotein 3 is expressed in cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3937.

Abstract Introduction: Our current understanding of the molecular changes predisposing Ductal carcinoma in situ (DCIS) to progress to invasive breast cancer (IBC) is limited. The aim of this study is to characterize the molecular changes of DCIS to improve DCIS risk assessment. Methods: We obtained paraffin-embedded tissue of 210 DCIS samples with no concurrent or antecedent IBC from 5 cancer institutions, and extracted DNA and RNA. Transcriptome analysis: Illumina’s TruSeq RNA Exome kit was used for library construction, followed by sequencing. Methylome analysis: Bisulfite treated genomic DNA was restored and arrayed using the Illumina 450K methylation chips. DNA Copy number (CNV) analysis: CNV was estimated from the methylation data set using the EpiCopy R package. Results: Samples passing Q/C metrics: Transcriptome: 59 cases, 63 controls. Methylome & CNV: 93 cases, 98 controls. We classified samples into their intrinsic PAM50 subsets. Cases showed an increase in the Her2 subtype, and the controls were enriched in LuminalA (LumA) samples, while Basal and LuminalB (LumB) subtypes were evenly distributed. Compared to the TCGA IBC dataset, proportions of Her2 and LumB subtypes in DCIS were increased while the LumA cohort was decreased. Unsupervised clustering of the transcriptome data resulted in 3 clusters with key differences between PAM50 subtypes: one cluster predominated in Basal and Her2 subtypes, and a second was enriched in hormone-positive samples (LumA and LumB). For the methylome data, the optimal number of clusters was 6. Again, several clusters showed correlation with PAM50 subtype, e.g., one was enriched for hormone-negative subtypes (Basal and Her2), while another was enriched for hormone-positive subtypes (LumA and LumB). For the CNV data, the optimal number of clusters was 4. Here, one CNV cluster appeared predominantly in the Basal subtype, and there were multiple regions showing significant subtype-specific differences with the TCGA IBC data set. While the well-known heterogeneity of DCIS prevents the above broad molecular categories from strongly stratifying DCIS by outcome, we are continuing to analyze our data sets for predictive molecular signatures. Since the long-term outcome of all DCIS in this cohort is known, we are in the unique position to pursue additional questions such as PAM50 subtype differences in times-to-events or lineage fidelity (i.e., whether the subtype of the subsequent IBC matches the preceding DCIS), and it appears that the time to IBC diagnosis is shortest in Basal DCIS, while at the same time, lineage fidelity is lowest in Basal DCIS, as has been previously reported. Conclusions: Our subtype-stratified analyses identified multiple molecular differences both between intrinsic subtypes as well as between DCIS and IBC that suggest subtype-specific characteristics that may be exploitable for risk stratification of DCIS. Citation Format: Marija Debeljak, Soonweng Cho, Bradley Downs, Michael Considine, Brittany Avin-McKelvey, Yongchun Wang, William Grizzle, Katherine Hoadley, Charles Lynch, Brenda Hernandez, Paul van Diest, Wendy Cozen, Ann Hamilton, Debra Hawes, Edward Gabrielson, Ashley Cimino-Mathews, Liliana Florea, Leslie Cope, Christopher B. Umbricht. Multimodal genome-wide survey of progressing and non-progressing DCIS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3126.

Psychometric properties of a Greek translation of the Intuitive Eating Scale-2 (IES-2) in adults from Cyprus

MP43-13 INTERLABORATORY GLEASON GRADING VARIATION AFFECTS TREATMENT: A DUTCH HISTORIC COHORT STUDY IN 30,509 PROSTATE CANCER PATIENTS

Abstract BACKGROUND Pregnancy-associated breast cancer (PABC), although most commonly defined as breast cancer diagnosed during pregnancy or within one year following delivery, knows a variety of definitions, likely related to the diversity of reported clinicopathological features and prognosis. More insight into the different breast cancer subgroups during pregnancy, the time after delivery and the postpartum period is therefore warranted. METHODS Patients with breast cancer diagnosed during pregnancy or within six months after delivery between January 1, 1988 and July 1, 2019, were included. Pregnant patients were subdivided according to gestational trimester, and postpartum patients according to lactational status. To investigate the influence of pregnancy and lactation on the histopathologic profile, these subgroups were compared to non-PABC patients matched for age at diagnosis, year at diagnosis, grade and ER status. Pearson Chi-square tests were used to compare clinicopathologic characteristics, while Kaplan-Meier/logrank/Cox regression methods were used to perform overall survival (OS) analysis. RESULTS Overall, 662 PABC patients were included, of which 73.6% were diagnosed during pregnancy. Median age at diagnosis was 34 years, with a median follow-up of 6.5 years. Overall, PABC patients as a group showed an advanced stage at diagnosis and an inferior OS at 5-years (75.4% vs. 83.2%, p = 0.000) compared to 1,392 non-PABC patients. In subgroup analysis, PABC patients within their first trimester showed a significantly lower tumor size and stage as compared to other trimesters. Patients diagnosed during the first trimester and postpartum non-lactating patients had a relatively good OS (81.3% and 77.9%, respectively) versus patients diagnosed during the second and third trimesters and during lactation (OS 60.0%, 64.9% and 65.6%, respectively, p = 0.003). In multivariate Cox regression, trimester of diagnosis, year of diagnosis, PR status, stage at diagnosis and surgery-or-not were significant contributors to OS. CONCLUSION In this large PABC cohort, uniquely specified by gestational trimester, an inferior outcome was found for patients diagnosed within the second and third gestational trimesters and during postpartum lactation, compared to first trimester and non-lactating postpartum patients. These findings indicate that PABC is clinically a heterogeneous group of breast cancer patients that should be redefined based on trimester of diagnosis and postpartum lactational status. Citation Format: Britt BM Suelmann, Carsten FJ Bakhuis, Carmen van Dooijeweert, Janneke Verloop, Ronald P Zweemer, Sabine C Linn, Elsken van der Wall, Paul J van Diest. Prognosis of pregnancy-associated breast cancer: Inferior outcome in patients diagnosed during second and third gestational trimesters and lactation [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-12-06.

Abstract Background. ILC represents the second most common histological type of breast cancer (BC), accounting for approximately 15% of all invasive BCs. Loss of cell-cell adhesion due to genomic alterations of CDH1,. the gene coding for E-cadherin, is the hallmark of ILC. So far, in the WHO guidelines, it is essential to recognize the dispersed or linear discohesive cells but it is not mandatory to demonstrate E-cadherin loss by immunohistochemistry (IHC) for diagnosing ILC. Recent central pathology revisions of clinical trials have demonstrated overdiagnosis of ILC in local pathological diagnosis, as only ~60% of the locally diagnosed ILCs were confirmed by central pathology. To understand the possible underlying reasons, we undertook a worldwide survey on the currently used histopathological diagnostic criteria for ILC. Materials and Methods. A survey was drafted using the online tool SurveyMonkey by a panel of pathologists and researchers from the European Lobular Breast Cancer Consortium (ELBCC). This survey was circulated to pathologists from December 14, 2020 until July, 1 2021. The main goals were to register the use of E-cadherin as a diagnostic marker for ILC and the systematic reporting of the ILC subtypes. Results. A total of 149 entries were recorded from 34 different countries from 6 continents. Pathologists declared working in a large tertiary (30%, 44/149) or university hospital (56%, 84/149), with an average yearly volume of BC samples >300 in 111/149 (74%) and >500 in 80/149 (54%) respondents. 117/149 (79%) are specialized in breast pathology. About half of the pathologists systematically perform IHC for ILC diagnosis (52%, 77/149), whilst others only perform staining in case of doubt (43%, 64/149) or for differentiating DCIS from LCIS (3%, 4/149). There was no association between the systematic use of IHC, the volume of BC samples, the type of institution (academic, large tertiary, private), and the number of pathologists in the institution. Concerning the use of IHC, 141/145(97%) participants use E-cadherin, 35/145 (24%) use β-catenin and 49/145 (34%) use p120-catenin. The majority (50%, 73/145) uses only E-cadherin, 13% (19/145) use E-cadherin in combination with β-catenin or 23% (33/145) use E-cadherin with p120-catenin, while 11% (16/145) use all 3 antibodies. For E-cadherin, 11 different clones were reported, of which the NCH-38 is the most frequently used (45%, 39/86), followed by Clone 36 (17%, 15/86) and EP700Y (16%, 14/86). Heterogeneity is reported regarding the used concentration per clone. The most frequently used modality of antigen retrieval is the heat induced one. Similar findings were observed for β-catenin and p120-catenin with each 4 different clones reported, again with variable concentrations. Only 4/104 (4%) respondents reported to perform DNA sequencing for CDH1 for diagnosing ILC. Most special lobular types are systematically reported by the vast majority of the pathologists: classic (149/149, 100%), pleomorphic (140/149, 94%), solid (108/149, 72%), histiocytoid/apocrine (90/149, 60%), alveolar (90/149, 60%), trabecular (54/149, 36%), mixed non-classic (54/149, 36%) and mucinous (51/149, 34%). Conclusions. We report the results of the first worldwide survey concerning diagnosis of ILC in pathological practice. The results demonstrate that ~half of the institutions systematically perform E-cadherin IHC to support the diagnosis of ILC. There is a great variability in E-cadherin antibody clones used as well as their concentrations, which might result in differences in staining results and their interpretation. As ILC-specific therapeutic avenues are currently being explored, some of which already in the context of clinical trials, it is of utmost importance to further improve the standardization of ILC diagnosis at the pathology level. Citation Format: Maxim De Schepper, Anne Vincent-Salomon, Matthias Christgen, Karen Van Baelen, Hitoshi Tsuda, Sasagu Kurozumi, Maria Jose Brito, Gabor Cserni, Stuart Schnitt, Denis Larsimont, Janina Kulka, Pedro Luis Fernandez, Paula Rodriguez, Ana Aula, Cristina Mendelez, Mieke Van Bockstal, Aniko Kovacs, Zsuzsanna Varga, Jelle Wesseling, Rohit Bhargava, Pia Boström, Camille Franchet, Blessing Zambuko, Gustavo Matute, Anca Berghian, Paul van Diest, Steffi Oesterreich, Patrick WB Derksen, Giuseppe Floris, Christine Desmedt. Results of a worldwide survey on the currently used histopathological diagnostic criteria for invasive lobular breast cancer (ILC) [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-02-09.

Abstract Invasive lobular breast carcinoma (ILC) is a low grade and mostly chemo-refractory luminal-type breast cancer that has been linked to sustained proliferative quiescence and long-term latency relapses (15-20 years). Loss of E-cadherin causes metastatic lobular breast cancer, partly through acquisition of anchorage independence. It is however still unknown how ILC cells control the balance between proliferative indolence and cell cycle re-entry at the metastatic site. We show here that E-cadherin loss leads to upregulation of Id2 through p120-catenin/Kaiso-dependent transcriptional derepression. Anchorage independent conditions further exacerbate p120-driven Id2 expression, leading to a sustained G0/G1 cell cycle arrest through binding of cytosolic Id2 to hypo-phosphorylated Rb. Intriguingly, we find that E-cadherin inactivation causes increased sensitivity to CDK4/6 inhibition in mouse and human breast cancer cell lines and primary tumor organoids. Finally, we find that Id2 expression is elevated in human ILC when compared to ductal breast cancers. Based on these data, we propose that combined E-cadherin loss and cytosolic Id2 expression can be used for the differential diagnosis of ILC and represent a candidate predictive biomarker pair for cell cycle targeting drug efficacy. Citation Format: Max Antonius Klaus Rätze, Thijs Koorman, Thijmen Sijnesael, Blessing Bassey-Archibong, Robert van de Ven, Lotte Enserink, Daan Visser, Sridevi Jaksani, Elvira Bakker, François Richard, Andrew Tutt, Rebecca Steele, Stephen Pettitt, Christopher J. Lord, Amanda Fitzpatrick, Clare Isacke, Paul J. van Diest, Christine Desmedt, Juliet M. Daniel, Patrick W.B. Derksen. E-cadherin loss drives Id2-dependent dampening of cell cycle progression and predicts increased susceptibility to CDK4/6 inhibition in lobular breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB246.

Abstract Background: Pregnancy associated breast cancer (PABC) comprises nearly 4% of all breast cancers. Compared to age-matched non-pregnant breast cancer patients, PABC is generally characterized by a particularly aggressive histopathologic profile. Whether these tumors arise before or during pregnancy, and whether they are stimulated by pregnancy-related hormones and/or epigenetic changes that by itself may affect subsequent hormone concentrations, remains to be elucidated. It is currently even unknown whether the histopathologic profile within PABC patients is affected by gestational age at diagnosis. In addition, some small studies observed a poorer outcome in lactating patients for patients with a postpartum PABC diagnosis. To fill this gap in knowledge, the present study assesses the influence of gestational age and lactation status at diagnosis on the histopathologic profile of PABC in a large population-based cohort.Methods:We identified 741 patients with PABC, between 1988 and 2019, in the nationwide Dutch Pathology Registry (PALGA). Histopathologic features (grade, ER-, PR-, and HER2-receptor status) were compared between pregnant- and postpartum PABC patients. Within pregnant PABC patients, histopathologic features were compared between the three gestational trimesters. For PABC patients with a postpartum diagnosis we compared histopathologic features of lactating versus non-lactating women.Results:Mean age at diagnosis was 34.2 years and the majority of breast cancers were diagnosed during pregnancy (74.2%); of which nearly half during the third trimester (47.3%). Histopathologic features did not differ between pregnant (n=550) and post-partum (n=191) PABC patients. Within pregnant patients, a significantly higher percentage of tumors were ER-receptor negative in the second (56.8%) and third trimester (57.3%), as compared to the first trimester (41.9%) (p=0.0015). A similar pattern was observed for PR; PR-negative receptor-status within first, second and third trimester: 48.0%, 57.7%, 56.5%, though these differences were not statistically significant (p=0.233). For histologic grade, we observed a higher proportion of grade III tumors with the second (87.4%) and third trimester (79.2%), as compared to the first trimester (73.7%) (p=0.055). For postpartum PABC patients no differences were observed for histologic grade between lactating (n=83) and non-lactating patients (n=94). However, tumors of lactating PABC patients were more often ER-negative (62.7% vs 48.9%, p=0.179), PR-negative (66.3% vs. 57.4%, p=0.258) and HER2-negative (74.7% vs. 61.7%, p=0.100). Conclusion:This unique, large population-based study of 741 PABC patients, demonstrates surprising histopathologic differences by gestational age at diagnosis for pregnant PABC-patients, and by lactation status for post-partum PABC-patients, which has not been reported before. Pregnant patients diagnosed at an advanced gestational age and postpartum PABC-patients who are lactating, seem to have a more aggressive histopathologic profile. This renders interesting clues for further studies, that will be conducted to unravel the molecular and genetic background and to investigate whether this has consequences on outcome. Citation Format: Britt BerendineMaria Suelmann, Carmen van Dooijeweert, Carsten Bakhuis, Elsken van der Wall, Sabine Linn, Paul van Diest. The histopathologic profile of pregnancy associated breast cancer by gestational age and lactation: Analysis of the nationwide Dutch Pathology Registry [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS7-14.

Response to “The Use of Non-Invasive Vagus Nerve Stimulation to Treat Respiratory Symptoms Associated with COVID-19: A Theoretical Hypothesis and Early Clinical Experience”

Response to Letter to the Editor Regarding: “The Use of Non-Invasive Vagus Nerve Stimulation to Treat Respiratory Symptoms Associated With COVID-19: A Theoretical Hypothesis and Early Clinical Experience”

Abstract Background: Early stage breast cancer patients are often confronted with uncertainty regarding the decision to undergo adjuvant chemotherapy (CT). Gene-expression profiles (GEP) are used to gain additional prognostic information and can influence adjuvant systemic treatment (AST) decisions. Little is known about patients’ experience with systemic treatment decisions and the impact of GEP use. Patients and Methods: In a prospective, observational, multicenter study in estrogen receptor (ER) positive (+) patients in whom a 70-gene signature (70-GS) was used to support the decision to administer adjuvant chemotherapy, patients were asked to fill out an online questionnaire including their preference for systemic treatment regimens. The patient reported uncertainty regarding the choice to undergo adjuvant systemic treatment was measured prior and after the 70-GS test result with the 16-item decisional conflict scale (DCS) scale. Results: Between 1 January 2017 and 31 December 2018, 106 patients were enrolled. Fifty-four percent of patients had N0, grade I/II, HER2-negative breast cancers, while 40% had N+ cancers. Before the 70-GS results were available, 58% of patients formulated a clear treatment preference, whereas 42% of patients felt unsure regarding their systemic treatment decisions (Fig 1). After disclosure of the 70-GS test result the percentage of patients who felt unsure about their treatment preference decreased considerably (from 42% to 5%, Fig 2). In addition, the patients’ final treatment decision was changed to the opposite in 34% of patients (CT to no CT or vice versa). Prior to the 70-GS test result, the mean total DCS-score at baseline was 34 (out of 100) and decreased to 23 after release of the 70-GS test result (P<0.001, table 1). Conclusion: In this prospective, multicenter study in ER+/HER2- breast cancer patients, the use of the 70-GS strongly decreased the percentage of patients who felt unsure about their treatment preference, and changed patients’ CT treatment decisions for 34% of patients. Furthermore, use of the 70-GS resulted in a significant decrease in decisional conflict regarding their final treatment plan. Table 1 CT preference and DCS-score prior and after release of the 70-GS test resultTreatment preference at baselineNo. of patientsTreatment preference after the 70-GS No. (%)DCS-score prior to 70-GSDCS-score after 70-GSP- value*No CT CT UnsureTotal1063423<0.001No CT5339(74)13(25)1(1)CT98(89)1(11)-Unsure4431(71)9(20)4(9)Abbreviations: CT, chemotherapy, 70-GS, 70-gene signature, DCS, Decisional Conflict Scale. There was a change in 34% of patients who had a clear pretest CT preference (ie, yes or no CT). The mean total DCS-score decreased from 34 of out 100 to 23 after release of the 70-GS test result. *P-value represents a paired T-test. Citation Format: Julia E.C. van Steenhoven, Bianca den Dekker, Anne Kuijer, Sjoerd G. Elias, Paul J. van Diest, Sabine Siesling, Thijs van Dalen. Patients’ experience with 70-gene signature testing on adjuvant systemic treatment decisions: Results of a prospective cohort study [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P2-14-19.