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Article ; Online: Progesterone-induced progesterone receptor membrane component 1 rise-to-decline changes are essential for decidualization.

Liu, Hailun / Franken, André / Bielfeld, Alexandra P / Fehm, Tanja / Niederacher, Dieter / Cheng, Zhongping / Neubauer, Hans / Stamm, Nadia

Reproductive biology and endocrinology : RB&E

2024 Volume 22, Issue 1, Page(s) 20

Abstract: Background: Decidualization of endometrial cells is the prerequisite for embryo implantation and subsequent placenta formation and is induced by rising progesterone levels following ovulation. One of the hormone receptors contributing to endometrial ... ...

Abstract	Background: Decidualization of endometrial cells is the prerequisite for embryo implantation and subsequent placenta formation and is induced by rising progesterone levels following ovulation. One of the hormone receptors contributing to endometrial homeostasis is Progesterone Receptor Membrane Component 1 (PGRMC1), a non-classical membrane-bound progesterone receptor with yet unclear function. In this study, we aimed to investigate how PGRMC1 contributes to human decidualization. Methods: We first analyzed PGRMC1 expression profile during a regular menstrual cycle in RNA-sequencing datasets. To further explore the function of PGRMC1 in human decidualization, we implemented an inducible decidualization system, which is achieved by culturing two human endometrial stromal cell lines in decidualization-inducing medium containing medroxyprogesterone acetate and 8-Br-cAMP. In our system, we measured PGRMC1 expression during hormone induction as well as decidualization status upon PGRMC1 knockdown at different time points. We further conferred proximity ligation assay to identify PGRMC1 interaction partners. Results: In a regular menstrual cycle, PGRMC1 mRNA expression is gradually decreased from the proliferative phase to the secretory phase. In in vitro experiments, we observed that PGRMC1 expression follows a rise-to-decline pattern, in which its expression level initially increased during the first 6 days after induction (PGRMC1 increasing phase) and decreased in the following days (PGRMC1 decreasing phase). Knockdown of PGRMC1 expression before the induction led to a failed decidualization, while its knockdown after induction did not inhibit decidualization, suggesting that the progestin-induced 'PGRMC1 increasing phase' is essential for normal decidualization. Furthermore, we found that the interactions of prohibitin 1 and prohibitin 2 with PGRMC1 were induced upon progestin treatment. Knocking down each of the prohibitins slowed down the decidualization process compared to the control, suggesting that PGRMC1 cooperates with prohibitins to regulate decidualization. Conclusions: According to our findings, PGRMC1 expression followed a progestin-induced rise-to-decline expression pattern during human endometrial decidualization process; and the correct execution of this expression program was crucial for successful decidualization. Thereby, the results of our in vitro model explained how PGRMC1 dysregulation during decidualization may present a new perspective on infertility-related diseases.
MeSH term(s)	Pregnancy ; Female ; Humans ; Prohibitins ; Progesterone/pharmacology ; Progesterone/metabolism ; Decidua/metabolism ; Receptors, Progesterone/genetics ; Progestins/metabolism ; Endometrium/metabolism ; Stromal Cells/metabolism ; Membrane Proteins/genetics ; Membrane Proteins/metabolism
Chemical Substances	Prohibitins ; Progesterone (4G7DS2Q64Y) ; Receptors, Progesterone ; Progestins ; PGRMC1 protein, human ; Membrane Proteins
Language	English
Publishing date	2024-02-03
Publishing country	England
Document type	Journal Article
ZDB-ID	2119215-7
ISSN	1477-7827 ; 1477-7827
ISSN (online)	1477-7827
ISSN	1477-7827
DOI	10.1186/s12958-024-01188-9
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Article: DanioCTC: Analysis of Circulating Tumor Cells from Metastatic Breast Cancer Patients in Zebrafish Xenografts.

Reinhardt, Florian / Coen, Luisa / Rivandi, Mahdi / Franken, André / Setyono, Eunike Sawitning Ayu / Lindenberg, Tobias / Eberhardt, Jens / Fehm, Tanja / Niederacher, Dieter / Knopf, Franziska / Neubauer, Hans

Cancers

2023 Volume 15, Issue 22

Abstract: Circulating tumor cells (CTCs) serve as crucial metastatic precursor cells, but their study in animal models has been hindered by their low numbers. To address this challenge, we present DanioCTC, an innovative xenograft workflow that overcomes the ... ...

Abstract	Circulating tumor cells (CTCs) serve as crucial metastatic precursor cells, but their study in animal models has been hindered by their low numbers. To address this challenge, we present DanioCTC, an innovative xenograft workflow that overcomes the scarcity of patient-derived CTCs in animal models. By combining diagnostic leukapheresis (DLA), the Parsortix microfluidic system, flow cytometry, and the CellCelector setup, DanioCTC effectively enriches and isolates CTCs from metastatic breast cancer (MBC) patients for injection into zebrafish embryos. Validation experiments confirmed that MDA-MB-231 cells, transplanted following the standard protocol, localized frequently in the head and blood-forming regions of the zebrafish host. Notably, when MDA-MB-231 cells spiked (i.e., supplemented) into DLA aliquots were processed using DanioCTC, the cell dissemination patterns remained consistent. Successful xenografting of CTCs from a MBC patient revealed their primary localization in the head and trunk regions of zebrafish embryos. DanioCTC represents a major step forward in the endeavors to study the dissemination of individual and rare patient-derived CTCs, thereby enhancing our understanding of metastatic breast cancer biology and facilitating the development of targeted interventions in MBC. Summary statement: DanioCTC is a novel workflow to inject patient-derived CTCs into zebrafish, enabling studies of the capacity of these rare tumor cells to induce metastases.
Language	English
Publishing date	2023-11-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers15225411
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Article ; Online: A novel cancer risk prediction score for the natural course of FA patients with biallelic BRCA2/FANCD1 mutations.

Radulovic, Ivana / Schündeln, Michael M / Müller, Lisa / Ptok, Johannes / Honisch, Ellen / Niederacher, Dieter / Wiek, Constanze / Scheckenbach, Kathrin / Leblanc, Thierry / Larcher, Lise / Soulier, Jean / Reinhardt, Dirk / Schaal, Heiner / Andreassen, Paul R / Hanenberg, Helmut

Human molecular genetics

2023 Volume 32, Issue 11, Page(s) 1836–1849

Abstract: Biallelic germline mutations in BRCA2 occur in the Fanconi anemia (FA)-D1 subtype of the rare pediatric disorder, FA, characterized clinically by severe congenital abnormalities and a very high propensity to develop malignancies early in life. Clinical ... ...

Abstract	Biallelic germline mutations in BRCA2 occur in the Fanconi anemia (FA)-D1 subtype of the rare pediatric disorder, FA, characterized clinically by severe congenital abnormalities and a very high propensity to develop malignancies early in life. Clinical and genetic data from 96 FA-D1 patients with biallelic BRCA2 mutations were collected and used to develop a new cancer risk prediction score system based on the specific mutations in BRCA2. This score takes into account the location of frameshift/stop and missense mutations relative to exon 11 of BRCA2, which encodes the major sites for interaction with the RAD51 recombinase, and uses the MaxEnt and HBond splicing scores to analyze potential splice site perturbations. Among 75 FA-D1 patients with ascertained BRCA2 mutations, 66 patients developed 102 malignancies, ranging from one to three independent tumors per individual. The median age at the clinical presentation of peripheral embryonal tumors was 1.0, at the onset of hematologic malignancies 1.8 and at the manifestation of CNS tumors 2.7 years, respectively. Patients who received treatment lived longer than those without. Using our novel scoring system, we could distinguish three distinct cancer risk groups among FA-D1 patients: in the first, patients developed their initial malignancy at a median age of 1.3 years (n = 36, 95% CI = 0.9-1.8), in the second group at 2.3 years (n = 17, 95% CI = 1.4-4.4) and in the third group at 23.0 years (n = 22, 95% CI = 4.3-n/a). Therefore, this scoring system allows, for the first time, to predict the cancer manifestation of FA-D1 patients simply based on the type and position of the mutations in BRCA2.
MeSH term(s)	Humans ; Child ; Infant ; Fanconi Anemia/genetics ; BRCA2 Protein/genetics ; Neoplasms/genetics ; Mutation ; Rad51 Recombinase/genetics
Chemical Substances	BRCA2 Protein ; Rad51 Recombinase (EC 2.7.7.-) ; BRCA2 protein, human
Language	English
Publishing date	2023-05-23
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't
ZDB-ID	1108742-0
ISSN	1460-2083 ; 0964-6906
ISSN (online)	1460-2083
ISSN	0964-6906
DOI	10.1093/hmg/ddad017
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Article: Whole Exome Analysis to Select Targeted Therapies for Patients with Metastatic Breast Cancer - A Feasibility Study.

Jaeger, Bernadette Anna Sophia / Krawczyk, Natalia / Japp, Anna Sophia / Honisch, Ellen / Köhrer, Karl / Scheuring, Sibylle / Petzsch, Patrick / Neubauer, Hans / Volkmer, Anne Kathrin / Esposito, Irene / Ruckhäberle, Eugen / Niederacher, Dieter / Fehm, Tanja

Geburtshilfe und Frauenheilkunde

2023 Volume 83, Issue 9, Page(s) 1138–1147

Abstract: Introduction: The purpose of this feasibility study was to select targeted therapies according to "ESMO Scale for Clinical Actionability of molecular Targets (ESCAT)". Data interpretation was further supported by a browser-based Treatment Decision ... ...

Abstract	Introduction: The purpose of this feasibility study was to select targeted therapies according to "ESMO Scale for Clinical Actionability of molecular Targets (ESCAT)". Data interpretation was further supported by a browser-based Treatment Decision Support platform (MH Guide, Molecular Health, Heidelberg, Germany). Patients: We applied next generation sequencing based whole exome sequencing of tumor tissue and peripheral blood of patients with metastatic breast cancer (n = 44) to detect somatic as well as germline mutations. Results: In 32 metastatic breast cancer patients, data interpretation was feasible. We identified 25 genomic alterations with ESCAT Level of Evidence I or II in 18/32 metastatic breast cancer patients, which were available for evaluation: three copy number gains in Conclusions: Resulting treatment options were discussed in a tumor board and could be recommended in a small but relevant proportion of patients with metastatic breast cancer (7/18). Thus, this study is a valuable preliminary work for the establishment of a molecular tumor board within the German initiative "Center for Personalized Medicine" which aims to shorten time for analyses and optimize selection of targeted therapies.
Language	English
Publishing date	2023-09-12
Publishing country	Germany
Document type	Journal Article
ZDB-ID	80111-2
ISSN	1438-8804 ; 0016-5751 ; 1615-3359
ISSN (online)	1438-8804
ISSN	0016-5751 ; 1615-3359
DOI	10.1055/a-2150-9440
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Article ; Online: A workflow for the enrichment, the identification, and the isolation of non-apoptotic single circulating tumor cells for RNA sequencing analysis.

Abramova, Anna / Rivandi, Mahdi / Yang, Liwen / Stamm, Nadia / Cieslik, Jan-Philipp / Honisch, Ellen / Niederacher, Dieter / Fehm, Tanja / Neubauer, Hans / Franken, André

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2023 Volume 105, Issue 4, Page(s) 242–251

Abstract: Circulating tumor cells (CTCs) are constantly shed by tumor tissue and can serve as a valuable analyte for a gene expression analysis from a liquid biopsy. However, a high proportion of CTCs can be apoptotic leading to rapid mRNA decay and challenging ... ...

Abstract	Circulating tumor cells (CTCs) are constantly shed by tumor tissue and can serve as a valuable analyte for a gene expression analysis from a liquid biopsy. However, a high proportion of CTCs can be apoptotic leading to rapid mRNA decay and challenging the analysis of their transcriptome. We established a workflow to enrich, to identify, and to isolate single CTCs including the discrimination of apoptotic and non-apoptotic CTCs for further single CTC transcriptome analysis. Viable tumor cells-we first used cells from breast cancer cell lines followed by CTCs from metastatic breast cancer patients-were enriched with the CellSearch system from diagnostic leukapheresis products, identified by immunofluorescence analysis for neoplastic markers, and isolated by micromanipulation. Then, their cDNA was generated, amplified, and sequenced. In order to exclude early apoptotic tumor cells, staining with Annexin V coupled to a fluorescent dye was used. Annexin V staining intensity was associated with decreased RNA integrity as well as lower numbers of total reads, exon reads, and detected genes in cell line cells and CTCs. A comparative RNA analysis of single cells from MDA-MB-231 and MCF7 cell lines revealed the expected differential transcriptome profiles. Enrichment and staining procedures of cell line cells that were spiked into blood had only little effect on the obtained RNA sequencing data compared to processing of naïve cells. Further, the detection of transcripts of housekeeping genes such as GAPDH was associated with a significantly higher quality of expression data from CTCs. This workflow enables the enrichment, detection, and isolation of single CTCs for individual transcriptome analyses. The discrimination of apoptotic and non-apoptotic cells allows to focus on CTCs with a high RNA integrity to ensure a successful transcriptome analysis.
MeSH term(s)	Humans ; Female ; Neoplastic Cells, Circulating/pathology ; Workflow ; Annexin A5 ; Breast Neoplasms/pathology ; Sequence Analysis, RNA ; RNA ; Biomarkers, Tumor
Chemical Substances	Annexin A5 ; RNA (63231-63-0) ; Biomarkers, Tumor
Language	English
Publishing date	2023-12-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.24816
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Zs.A 1688: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article: Ganz-Exom-Sequenzierung zur Bestimmung zielgerichteter Therapien für Patientinnen mit metastasiertem Mammakarzinom – eine Machbarkeitsstudie

Senologie - Zeitschrift für Mammadiagnostik und -therapie

2024 Volume 21, Issue 01, Page(s) 40–50

Abstract: Einleitung: Ziel dieser Machbarkeitsstudie war es, zielgerichtete Therapien entsprechend der ESCAT-Skala (ESMO Scale for Clinical Actionability of molecular Targets) zu bestimmen. Für die Interpretation der Daten wurde eine browserbasierte Plattform zur ...

Abstract	Einleitung: Ziel dieser Machbarkeitsstudie war es, zielgerichtete Therapien entsprechend der ESCAT-Skala (ESMO Scale for Clinical Actionability of molecular Targets) zu bestimmen. Für die Interpretation der Daten wurde eine browserbasierte Plattform zur Entscheidungsfindung (MH Guide, Molecular Health, Heidelberg, Germany) eingesetzt. Patientinnen: Es wurde eine Exomsequenzierung von Tumorgewebe und peripherem Blut von Patientinnen mit metastasiertem Mammakarzinom (n = 44) durchgeführt, um somatische sowie Keimbahnmutationen zu identifizieren. Ergebnisse: Bei 32 Patientinnen mit metastasiertem Mammakarzinom konnte eine Dateninterpretation durchgeführt werden. Es wurden 25 genomische Veränderungen (ESCAT-Evidenzstufe I oder II) bei 18/32 Patientinnen mit metastasiertem Mammakarzinom identifiziert und abschließend ausgewertet: Darunter fanden sich 3 Fälle mit erhöhter Kopienzahl bei HER2, 2 g BRCA1 -, 2 g BRCA2 -, 6 PIK3CA -, 1 ESR1 -, 3 PTEN -, 1 AKT1 - und 2 HER2 -Mutationen. Dazu kamen noch 5 Proben, die eine hochgradige Mikrosatelliten-Instabilität aufwiesen. Schlussfolgerung: Die daraus abzuleitenden Behandlungsoptionen wurden in einer Tumorkonferenz diskutiert und dann einer kleinen, aber relevanten Anzahl von Patientinnen mit metastasiertem Mammakarzinom (7/18) empfohlen. Die hier vorgestellte Arbeit stellt eine wertvolle Machbarkeitsstudie dar, die dazu beitragen kann, molekulare Tumorboards innerhalb des Deutschen Netzwerks für Personalisierte Medizin zu etablieren. Ziel ist, die für Analysen benötigte Zeit zu verkürzen und die Wahl zielgerichteter Therapien zu optimieren.
Keywords	Mammakarzinom ; zielgerichtete Therapie ; Ganz-Exom-Sequenzierung ; Entscheidungsplattform ; breast cancer ; targeted therapy ; whole exome sequencing ; decision making platform
Language	German
Publishing date	2024-03-01
Publisher	Georg Thieme Verlag KG
Publishing place	Stuttgart ; New York
Document type	Article
ZDB-ID	2174147-5
ISSN	1611-647X ; 1611-6453
ISSN (online)	1611-647X
ISSN	1611-6453
DOI	10.1055/a-2238-9615
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Zs.A 6562: Show issues

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Article ; Online: Implementing microwell slides for detection and isolation of single circulating tumor cells from complex cell suspensions.

Yang, Liwen / Rivandi, Mahdi / Franken, André / Hieltjes, Maarten / van der Zaag, Pieter Jan / Nelep, Constantin / Eberhardt, Jens / Peter, Stefan / Niederacher, Dieter / Fehm, Tanja / Neubauer, Hans

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2022 Volume 101, Issue 12, Page(s) 1057–1067

Abstract: Cell loss during detection and isolation of circulating tumor cells (CTCs) is a challenge especially when label-free pre-enrichment technologies are used without the aid of magnetic particles. Although microfluidic systems can remove the majority of " ... ...

Abstract	Cell loss during detection and isolation of circulating tumor cells (CTCs) is a challenge especially when label-free pre-enrichment technologies are used without the aid of magnetic particles. Although microfluidic systems can remove the majority of "contaminating" white blood cells (WBCs), their remaining numbers are still impeding single CTC isolation, thus making additional separation steps needed. This study aimed to develop a workflow from blood-to-single CTC for complex cell suspensions by testing two microwell formats. In the first step, different cell lines were used to compare the performances of Sievewell™ 370 K (TOK, Japan) and CellCelector™ Nanowell U25 (ALS Automated Lab Solutions, Germany) slides for cell labelling and single-cell micromanipulation. Confounding levels of auto-fluorescence inherent to different plastic materials used to cast the microwells, staining recovery rates, and cell isolation rates were determined. In the second step, three different blood preservation tubes were tested for RNA analysis. Lastly, the established workflow was applied to isolate CTCs from peripheral blood samples obtained from metastasized breast cancer (mBC) patients for single-cell DNA and RNA analysis. The detection of CTCs in Sievewell slides profit from better signal-to-noise ratios in the fluorescence channels mainly used for CTC detection. In addition, due to its design, Sievewell supports direct in situ CTC labelling, which minimizes cell loss and leads to single-cell recovery rates after staining of approx. 94%. Detection of PIK3CA mutations in single CTCs verified the applicability of the workflow for the analysis of genomic DNA of CTCs. Furthermore, combined with blood preservation up to 48 h at room temperature in LBguard tubes, panel RT-PCR transcript analysis was successful for single cell line cells and CTCs, respectively. The combined use of Sievewell microwell slides and CellCelector™ automated micromanipulation system improves single CTC detection, labelling and isolation from complex cell suspensions. This approach is especially valuable when samples of high cellular content are processed.
MeSH term(s)	Humans ; Female ; Neoplastic Cells, Circulating/pathology ; Cell Separation ; Breast Neoplasms/diagnosis ; Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Microfluidics ; RNA ; Cell Line, Tumor
Chemical Substances	RNA (63231-63-0)
Language	English
Publishing date	2022-06-14
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.24660
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Book: Genetische Basis gynäkologischer Karzinome

Niederacher, Dieter

Abschlußbericht des Teilprojekts 02 im BMBF-geförderten Forschungsverbund ; Vorhabensbezeichnung: ... Molekulare Analyse und Validierung von Kandidatengenen, die mit sporadischen Mamma-, Ovarial- und Endometriumkarzinomen assoziiert sind ; Berichtszeitraum: 01.05.2000 - 30.06.2004

2005

Title variant	Genetic basis of sporadic gynecological carcinomas
Author's details	[Autor: Niederacher, Dieter]
Language	German
Size	6 Bl.
Publisher	Frauenklinik des Universitätsklinikums, Heinrich-Heine-Univ
Publishing place	Düsseldorf
Document type	Book
Note	Förderkennzeichen BMBF 01KW9942. - Verbund-Nr. 01018066. - Literaturverz. - Engl. Zsfassung u.d.T.: Genetic basis of sporadic gynecological carcinomas
Database	Library catalogue of the German National Library of Science and Technology (TIB), Hannover

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Article: Whole Exome Analysis to Select Targeted Therapies for Patients with Metastatic Breast Cancer – A Feasibility Study

Geburtshilfe und Frauenheilkunde

2023 Volume 83, Issue 09, Page(s) 1138–1147

Abstract: The purpose of this feasibility study was to select targeted therapies according to “ESMO Scale for Clinical Actionability of molecular Targets (ESCAT)”. Data interpretation was further supported by a browser-based Treatment Decision Support ... ...

Abstract	The purpose of this feasibility study was to select targeted therapies according to “ESMO Scale for Clinical Actionability of molecular Targets (ESCAT)”. Data interpretation was further supported by a browser-based Treatment Decision Support platform (MH Guide, Molecular Health, Heidelberg, Germany). We applied next generation sequencing based whole exome sequencing of tumor tissue and peripheral blood of patients with metastatic breast cancer (n = 44) to detect somatic as well as germline mutations. In 32 metastatic breast cancer patients, data interpretation was feasible. We identified 25 genomic alterations with ESCAT Level of Evidence I or II in 18/32 metastatic breast cancer patients, which were available for evaluation: three copy number gains in HER2, two g BRCA1, two g BRCA2, six PIK3CA, one ESR1, three PTEN, one AKT1 and two HER2 mutations. In addition, five samples displayed Microsatellite instability high-H. Resulting treatment options were discussed in a tumor board and could be recommended in a small but relevant proportion of patients with metastatic breast cancer (7/18). Thus, this study is a valuable preliminary work for the establishment of a molecular tumor board within the German initiative “Center for Personalized Medicine” which aims to shorten time for analyses and optimize selection of targeted therapies.
Keywords	Mammakarzinom ; zielgerichtete Therapie ; Ganz-Exom-Sequenzierung ; Entscheidungsplattform ; breast cancer ; targeted therapy ; whole exome sequencing ; decision making platform
Language	English
Publishing date	2023-09-01
Publisher	Georg Thieme Verlag KG
Publishing place	Stuttgart ; New York
Document type	Article
ZDB-ID	80111-2
ISSN	1438-8804 ; 0016-5751 ; 1615-3359
ISSN (online)	1438-8804
ISSN	0016-5751 ; 1615-3359
DOI	10.1055/a-2150-9440
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Article: 7. Wissenschaftliches Symposium der AGO-TraFo

Niederacher, Dieter / Fasching, Peter A. / Fehm, Tanja N.

Geburtshilfe und Frauenheilkunde

2016 Volume 76, Issue 02, Page(s) 119–121

Language	German
Publishing date	2016-02-01
Publisher	Georg Thieme Verlag KG
Publishing place	Stuttgart ; New York
Document type	Article
ZDB-ID	80111-2
ISSN	1438-8804 ; 0016-5751 ; 1615-3359
ISSN (online)	1438-8804
ISSN	0016-5751 ; 1615-3359
DOI	10.1055/s-0042-101116
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