Theresa Vilsmaier

Background: One in fifty infants is conceived through assisted reproductive technologies (ART). To date, data on the cardiovascular morbidity of ART individuals is ambiguous. This study investigated the vascular function of young ART subjects using 24 hour ambulatory blood pressure monitoring (24 h ABPM). Methods: ART subjects and spontaneously conceived controls matched in age as well as sex were enrolled. A 24 h blood pressure profile including pulse wave analysis was executed in all study participants. Blood pressure readings were assessed every 15 min during daytime and every 30 min during nighttime. The 24 h systolic/diastolic blood pressure (SBP/DBP) as well as central blood pressure, nocturnal blood pressure decrease, and 24 h pulse wave velocity (PWV) were analyzed. Results: A total of 41 ART individuals and 46 spontaneously conceived peers were included in the final analysis (mean age: 15.37 ± 5.46 years vs. 16.48 ± 5.23 years, p = 0.338). The 24 h SBP (112.74 ± 9.24 mmHg vs. 112.73 ± 6.70 mmHg, p = 0.997), 24 h DBP (65.61 ± 7.98 mmHg vs. 66.57 ± 7.03 mmHg, p = 0.550), 24 h central blood pressure, nocturnal blood pressure decrease, and 24 h PWV did not demonstrate significant differences between the ART and control group. Conclusions: In contrast to previous studies, no significant differences in 24 h blood pressure were demonstrated between ART subjects and spontaneously conceived peers. Hence, the results of this study do not indicate an unfavorable blood pressure profile in ART offspring. Larger multi-centric studies are needed to validate these results in the future.

The use of assisted reproductive technologies (ART) for the treatment of infertility is gaining popularity. Limited data on the overall vascular health of females with history of ART are available. This pilot study aimed to investigate the overall vascular health of females with history of ART compared to individuals who conceived spontaneously. The assessment of overall vascular health included the measurement of brachial blood pressure, central blood pressure, and pulse wave velocity, as well as the evaluation of the arterial stiffness and carotid intima-media thickness (cIMT) of the common carotid arteries. Conventional blood lipids including lipoprotein a (Lp(a)) were also determined. In total, 45 females with history of ART and 52 females who conceived spontaneously were included (mean age: 47.72 ± 5.96 years vs. 46.84 ± 7.43 years, p = 0.525). An initial comparison revealed a significantly higher prevalence of elevated Lp(a) in ART females (p = 0.011). However, after multiple comparison correction, the significant result disappeared (p = 0.132). Within the cohort of ART females, no significantly higher cardiovascular risk was detected regarding vascular function. The potentially higher prevalence of elevated Lp(a) in ART females must be further investigated in future studies, as it might contribute to the impaired reproductive process in this cohort.

The aim of this retrospective study was to assess the respective prognostic values of cytoplasmic and nuclear TRα, TRα1, and TRα2 expression in breast cancer (BC) tissue samples and correlate the results with clinico-pathological parameters. In 249 BC patients, the expression patterns of general TRα and the α1 and α2 isoforms were evaluated via immuno-histochemistry. Prognosis-determining aspects were calculated via univariate, as well as multivariate, analysis. Univariate Cox-regression analysis revealed no association between nuclear TRα expression and overall survival (OS) (p = 0.126), whereas cytoplasmic TRα expression was significantly correlated with a poor outcome for both OS (p = 0.034) and ten-year survival (p = 0.009). Strengthening these results, cytoplasmic TRα was found to be an independent marker of OS (p = 0.010) when adjusted to fit clinico-pathological parameters. Analyses of the TRα-subgroups revealed that TRα1 had no prognostic relevance, whereas nuclear TRα2 expression was positively associated with OS (p = 0.014), ten-year survival (p = 0.029), and DFS (p = 0.043). Additionally, nuclear TRα2 expression was found to be an independent positive prognosticator (p = 0.030) when adjusted to fit clinico-pathological parameters. Overall, our results support the hypothesis that subcellular localization of TRα and its isoforms plays an important role in the carcinogenesis and prognosis of breast cancer. Cytoplasmic TRα expression correlates with more aggressive disease progression, whereas nuclear TRα2 expression appears to be a protective factor. These data may help us to prioritize high-risk BC subgroups for possible targeted tumor therapy.

Subjects conceived through assisted reproductive technologies (ART) potentially suffer from impaired left ventricular (LV) function due to premature vascular aging. This study aimed to evaluate whether subtle differences in LV diastolic function can be observed echocardiographically between young ART subjects and their spontaneously conceived peers. The echocardiographic assessment included the measurement of LV dimensions, mitral inflow velocities, and myocardial velocity at early diastole (E’, cm/s) at the LV wall and the interventricular septum (IVS). An average from E/E’LV and E/E’IVS (E/E’AVG) was derived. In total, 66 ART subjects and 83 controls (12.85 ± 5.80 years vs. 13.25 ± 5.89 years, p = 0.677) were included. The ART subjects demonstrated a significantly lower E’LV (19.29 ± 3.29 cm/s vs. 20.67 ± 3.78 cm/s, p = 0.020) compared to their spontaneously conceived peers. Study participants of ≥ 10 years of age displayed a significantly higher E/E’AVG (6.50 ± 0.97 vs. 6.05 ± 0.99, p = 0.035) within the ART cohort. The results of this study demonstrate a significantly lower LV diastolic function in the ART subjects. However, no significant changes in LV diastolic function were observed between the two groups when the results were adjusted for age, birth weight percentile, and gestational age. Those ART subjects born preterm might have an elevated risk of developing LV diastolic alterations and could therefore profit from close echocardiographic monitoring.

Multiple studies reported signs of vascular dysfunction in subjects conceived through assisted reproductive technologies (ART). The assessment of arterial stiffness in this cohort seems beneficial for risk stratification. Regional arterial stiffness of the abdominal aorta (AAO) and the common carotid arteries (CCA) was evaluated sonographically using two-dimensional speckle tracking in subjects conceived through ART and spontaneously conceived peers. Global arterial stiffness was assessed utilizing an oscillometric blood pressure device. The cohorts of 67 ART subjects and 86 spontaneously conceived peers (11.31 (8.10–18.20) years vs. 11.85 (8.72–18.27) years, p = 0.43) did not differ significantly in parameters of regional and global arterial stiffness. In the sub-analysis of study participants ≥10 years of age, markers of arterial stiffness did not display significant differences between both groups. However, a higher tendency of brachial systolic blood pressure was demonstrated in the ART cohort compared to the control group (120.18 ± 9.57 mmHg vs. 116.55 ± 8.05 mmHg, p = 0.050). The present study displayed no significant differences in arterial stiffness between ART subjects and spontaneously conceived peers. Moreover, this study suggests that arterial stiffness does not elevate more profoundly in ART subjects with increasing age. Further studies are required for a more detailed cardiovascular risk stratification of the ART cohort.

The prognostic impact of tumor-infiltrating lymphocytes (TILs) is intensively investigated in breast cancer (BC). It is already known that triple-negative breast cancer (TNBC), the most aggressive type of BC, has the highest percentage of TILs. In addition, there is an influence of steroid hormone receptor expression (type I nuclear receptors) on TIL subpopulations in breast cancer tissue. The link between type II nuclear receptors and the level of TILs is unclear. Therefore, the aim of this study was to quantify TILs in a panel of 264 sporadic breast cancers and investigate the correlation of TIL levels with type I and II nuclear receptors expression. TIL levels were significantly increased in the subgroup of TNBC. By contrast, they decreased in estrogen (ER)- or progesterone receptor (PR)-positive cases. Moreover, TIL levels were correlated with type II nuclear receptors, including PPARγ, with a significant inverse correlation of the nuclear form (r = −0.727, p < 0.001) and a weak positive correlation of the cytoplasmic form (r = 0.202, p < 0.002). Surprisingly, BC cases with a TIL Salgado score of >15% showed a significantly decreased overall survival. In addition, peritumoral inflammation was also quantified in BC tissue samples. In our cohort, although the level of peritumoral inflammation was not correlated with OS, it determined the prognostic value of ER, PR, and PPARγ in BC. Altogether, the present study provides a differentiated overview of the relations between nuclear receptor expression, TIL levels, peritumoral inflammation, and prognosis in BC.

Macrophages are dynamic cells susceptible to the local microenvironment which includes tumor-associated macrophages (TAMs) in cancers. TAMs are a collection of heterogeneous macrophages, including M1 and M2 subtypes, shaped by various activation modes and labeled with various markers in different tumors. CCL22+-infiltrating cells are thought to be significantly associated with the prognosis of cervical cancer patients. Moreover, CCL22 is an established marker of M2a macrophages. Although the phenotypic identification of M1 and M2 macrophages is well established in mice and human macrophages cultured in a medium with fetal calf serum (FCS), fewer studies have focused on M2 subtypes. In addition, the question of whether CCL22 affects polarization of M2a macrophages remains unanswered. This study constructed a co-culture system to shape TAMs in vitro. We found that CCL22 was mainly secreted by TAMs but not cervical cancer cell lines. Human peripheral blood monocytes were differentiated into uncommitted macrophages (M0) and then polarized to M1, M2a, M2b, and M2c macrophages using LPS plus IFNr, IL-4, LPS plus IL1β, and IL-10, respectively. Using flowcytometry, we found CD80++ was the marker of M1 and M2b, CD206++ was the marker of M2a, and CD163++ was the marker of M2c, compared with M0 macrophages. By regulating CCL22, we found that the mean fluorescence intensity (MFI) of CD206 in TAMs was significantly affected compared to the control group. Therefore, CCL22 could polarize TAMs of cervical cancer toward M2a macrophages. In conclusion, our study revealed that CCL22 could be a therapeutic target for cervical cancer, which might be because of its role in regulating macrophage polarization.

Einleitung Kernrezeptoren wie Östrogen- (ÖR) oder Progesteron-Rezeptor (PR) sind etablierte prognostische Marker bei Brustkrebs und dienen als Ansatzpunkt für zielgerichtete Tumortherapie. Darüber hinaus rücken weitere Kernrezeptoren wie zum Beispiel der Vitamin-D-Rezeptor (VDR) in den Fokus der Forschung [1], ebenso neue prognostische Marker wie beispielsweise das Vorhandensein von Tumor-Infiltrierenden Lymphozyten (TILs). Insbesondere bei triple-negativen und HER-2 überexprimierenden Brustkrebssubtypen konnte ein Überlebensvorteil durch eine hohe Rate an TILs gezeigt werden [2]. Stimuli für das Einströmen der TILs in das Tumorareal sind Gegenstand laufender Untersuchungen. Material und Methoden In dieser Studie wurden die Zusammenhänge zwischen nukleärer Expression der genannten Kernrezeptoren (ÖR und VDR), Quantifizierung von TILs in Brustkrebsgewebe und prognostischer Bedeutung untersucht. Grundlage bildete ein bekanntes Brustkrebs-Kollektiv von 320 Fällen, die zwischen 2000 und 2002 an der Klinik für Frauenheilkunde und Geburtshilfe der Ludwigs-Maximilians-Universität München behandelt wurden [3]. Die Quantifizierung der TILs erfolgte am Institut national de la santé et de la recherche médicale (INSERM) in Montpellier nach den Klintrup-Kriterien [4]. Hiernach erfolgten Spearman’s-Rho-Korrelationsanalysen zur Korrelation mit der Kernrezeptor-Expression und Überlebenszeitanalysen mittels Kaplan-Meier-Analysen. Alle Berechnungen wurden mit IBM SPSS Statistics (Version 24) durchgeführt. Ergebnisse In der vorliegenden Untersuchung korreliert der Einstrom von TILs in das Tumorgewebe negativ mit der Expression von ÖR und VDR. Dies führt jedoch zu unterschiedlichen Effekten in Bezug auf das Überleben. Es zeigt sich, dass die Expression von ÖR ein positiver prognostischer Marker nur bei geringer und mittlerer TIL-Infiltration ist, so liegt das 5-Jahres-Überleben bei 20 % für ÖR-negative Fälle und 90 % für ÖR-positive Fälle (p < 0,005) bei geringer TIL-Infiltration (s. [Abb. 1a]). Bei hoher TIL-Infiltration hingegen spielt die Expression von ÖR keine Rolle für das Überleben (5-Jahres-Überleben von 80% bei ÖR-positiven vs. 85% bei ÖR-negativen Fällen, p = 0,64) (s. [Abb. 1b]). Gegensätzliche Ergebnisse zeigt die VDR-Überlebensanalyse. Bei geringer TIL-Infiltration ist die VDR-Expression mit einem signifikant schlechteren krankheitsfreien Überleben vergesellschaftet (5-Jahres-krankheitsfreies-Überleben bei 65 % vs. 88 % bei fehlender VDR-Expression, p = 0,037) (s. [Abb. 2a]). Bei hoher TIL-Infiltration zeigt sich auch hier kein signifikanter Unterschied im krankheitsfreien Überleben in Abhängigkeit von der VDR-Expression (5-Jahres-krankheitsfreies-Überleben bei 65 % bei VDR-positiven vs. 70 % bei VDR-negativen Fällen, p = 0,708) (s. [Abb. 2b]). Zusammenfassung Die vorliegende Untersuchung zeigt ein differenziertes Bild individueller Zusammenhänge zwischen TILs, jeweiligem Kernrezeptor und Einfluss auf das Überleben. Insbesondere der Zusammenhang zwischen fehlender TIL-Infiltration und signifikant erniedrigtem rezidivfreien Überleben bei hoher VDR-Expression war bisher nicht bekannt. Diese Daten unterstützen die These, dass die Kernexpression von ÖR und VDR über bisher ungeklärte Mechanismen ein Einströmen von TILs in das Tumorareal beeinflusst. Da sich hieraus ein signifikanter Effekt auf das Überleben von Mammakarzinompatientinnen zu ergeben scheint, können mit diesem Wissen perspektivisch neue Therapieansätze entwickelt werden. Dies ist von besonderem Interesse in der Behandlung von Frauen mit triple-negativem Brustkrebs, bei denen derzeit nur begrenzte systemtherapeutische Optionen bestehen. Article published online: 19 May 2022 © 2022. Thieme. All rights reserved. Georg Thieme Verlag Rüdigerstraße 14, 70469 Stuttgart, Germany

Retinoid X receptor α (RXRα) is a nuclear receptor (NR) which functions as the primary heterodimeric partner of other NRs including the peroxisome proliferator-activated receptor γ (PPARγ). We previously reported that, in breast cancers (BC), the subcellular localization of these two receptors was strongly associated with patient prognosis. In the present work, we investigated the prognosis value of the combined cytoplasmic expression of RXRα and PPARγ using a retrospective cohort of 250 BC samples. Patients with tumors expressing both NRs in tumor cell cytoplasm exhibited a significant shorter overall (OS) and disease-free survival (DFS). This was also observed for patients with stage 1 tumors. Cox univariate analysis indicated that patients with tumors coexpressing RXRα and PPARγ in the cytoplasm of tumor cells have a decreased 5 y OS rate. Cytoplasmic co-expression of the two NRs significantly correlated with HER2 positivity and with NCAD and CD133, two markers of tumor aggressiveness. Finally, in Cox multivariate analysis, the co-expression of RXRα and PPARγ in the cytoplasm appeared as an independent OS prognosticator. Altogether, this study demonstrates that the cytoplasmic co-expression of RXRα and PPARγ could be of relevance for clinicians by identifying high-risk BC patients, especially amongst those with early and node-negative disease.

The aim of this retrospective study was to assess the prognostic value of cytoplasmic versus nuclear RXRα expression in breast cancer (BC) tissue samples and to correlate the results with clinicopathological parameters. In 319 BC patients, the expression of RXRα was evaluated via immunohistochemistry. Prognosis-determining aspects were calculated through uni- and multivariate analyses. Correlation analysis revealed a trend association with nuclear RXRα expression regarding an improved overall survival (OS) (p = 0.078), whereas cytoplasmic RXRα expression was significantly correlated with a poor outcomes in terms of both OS (p = 0.038) and disease-free survival (DFS) (p = 0.037). Strengthening these results, cytoplasmic RXRα was found to be an independent marker for DFS (p = 0.023), when adjusted to clinicopathological parameters, whereas nuclear RXRα expression was positively associated with lower TNM-staging, i.e., pT (p = 0.01), pN (p = 0.029) and pM (p = 0.001). Additionally, cytoplasmic RXRα expression was positively associated with a higher histopathological tumor grading (p = 0.02). Cytoplasmic RXRα was also found to be a negative prognosticator for Her-2neu-negative and triple-negative patients. Altogether, these findings support the hypothesis that the subcellular localization of RXRα plays an important role in carcinogenesis and the prognosis of BC. The expression of cytoplasmic RXRα is correlated with a more aggressive course of the disease, whereas nuclear RXRα expression appears to be a protective factor. These data may help to identify high-risk BC subgroups in order to find possible specific options in targeted tumor therapy.