14662
2016
eng
49
4
article
1
2017-03-31
--
--
Adenosine-generating ovarian cancer cells attract myeloid cells which differentiate into adenosine-generating tumor associated macrophages - a self-amplifying, CD39- and CD73-dependent mechanism for tumor immune escape
Background
Ovarian cancer (OvCA) tissues show abundant expression of the ectonucleotidases CD39 and CD73 which generate immunomodulatory adenosine, thereby inhibiting cytotoxic lymphocytes. Little, however, is known about the effect of adenosine on myeloid cells. Considering that tumor associated macrophages (TAM) and myeloid-derived suppressor cells (MDSC) constitute up to 20 % of OvCA tissue, we investigated the effect of adenosine on myeloid cells and explored a possible contribution of myeloid cells to adenosine generation in vitro and ex vivo.
Methods
Monocytes were used as human blood-derived myeloid cells. After co-incubation with SK-OV-3 or OAW-42 OvCA cells, monocyte migration was determined in transwell assays. For conversion into M2-polarized “TAM-like” macrophages, monocytes were co-incubated with OAW-42 cells. Ex vivo TAMs were obtained from OvCA ascites. Macrophage phenotypes were investigated by intracellular staining for IL-10 and IL-12. CD39 and CD73 expression were assessed by FACS analysis both on in vitro-induced TAM-like macrophages and on ascites-derived ex situ-TAMs. Myeloid cells in solid tumor tissue were analyzed by immunohistochemistry. Generation of biologically active adenosine by TAM-like macrophages was measured in luciferase-based reporter assays. Functional effects of adenosine were investigated in proliferation-experiments with CD4+ T cells and specific inhibitors.
Results
When CD39 or CD73 activity on OvCA cells were blocked, the migration of monocytes towards OvCA cells was significantly decreased. In vivo, myeloid cells in solid ovarian cancer tissue were found to express CD39 whereas CD73 was mainly detected on stromal fibroblasts. Ex situ-TAMs and in vitro differentiated TAM-like cells, however, upregulated the expression of CD39 and CD73 compared to monocytes or M1 macrophages. Expression of ectonucleotidases also translated into increased levels of biologically active adenosine. Accordingly, co-incubation with these TAMs suppressed CD4+ T cell proliferation which could be rescued via blockade of CD39 or CD73.
Conclusion
Adenosine generated by OvCA cells likely contributes to the recruitment of TAMs which further amplify adenosine-dependent immunosuppression via additional ectonucleotidase activity. In solid ovarian cancer tissue, TAMs express CD39 while CD73 is found on stromal fibroblasts. Accordingly, small molecule inhibitors of CD39 or CD73 could improve immune responses in ovarian cancer.
Journal for ImmunoTherapy of Cancer
10.1186/s40425-016-0154-9
urn:nbn:de:bvb:20-opus-146624
Journal for ImmunoTherapy of Cancer (2016) 4:49 DOI 10.1186/s40425-016-0154-9
Itsaso Montalbán del Barrio
Cornelia Penski
Laura Schlahsa
Roland G. Stein
Joachim Diessner
Achim Wöckel
Johannes Dietl
Manfred B. Lutz
Michel Mittelbronn
Jörg Wischhusen
Sebastian F. M. Häusler
eng
uncontrolled
ovarian cancer
eng
uncontrolled
adenosine
eng
uncontrolled
CD39
eng
uncontrolled
tumor associated macrophages
eng
uncontrolled
immune escape
eng
uncontrolled
CD73
Gynäkologie, Geburtsmedizin, Pädiatrie, Geriatrie
open_access
Frauenklinik und Poliklinik
Institut für Virologie und Immunbiologie
Förderzeitraum 2016
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/14662/Barrio_10.1186_s40425-016-0154-9.pdf
14837
2015
eng
12319
5
article
1
2017-05-11
--
--
PRC2 inhibition counteracts the culture-associated loss of engraftment potential of human cord blood-derived hematopoietic stem and progenitor cells
Cord blood hematopoietic stem cells (CB-HSCs) are an outstanding source for transplantation approaches. However, the amount of cells per donor is limited and culture expansion of CB-HSCs is accompanied by a loss of engraftment potential. In order to analyze the molecular mechanisms leading to this impaired potential we profiled global and local epigenotypes during the expansion of human CB hematopoietic stem and progenitor cells (HPSCs). Human CB-derived CD34+ cells were cultured in serum-free medium together with SCF, TPO, FGF, with or without Igfbp2 and Angptl5 (STF/STFIA cocktails). As compared to the STF cocktail, the STFIA cocktail maintains in vivo repopulation capacity of cultured CD34+ cells. Upon expansion, CD34+ cells genome-wide remodel their epigenotype and depending on the cytokine cocktail, cells show different HK4me3 and H3K27me3 levels. Expanding cells without Igfbp2 and Angptl5 leads to higher global H3K27me3 levels. ChIPseq analyses reveal a cytokine cocktail-dependent redistribution of H3K27me3 profiles. Inhibition of the PRC2 component EZH2 counteracts the culture-associated loss of NOD scid gamma (NSG) engraftment potential. Collectively, our data reveal chromatin dynamics that underlie the culture-associated loss of engraftment potential. We identify PRC2 component EZH2 as being involved in the loss of engraftment potential during the in vitro expansion of HPSCs.
Scientific Reports
10.1038/srep12319
urn:nbn:de:bvb:20-opus-148374
Scientific Reports 5:12319 (2015). DOI: 10.1038/srep12319
CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International
Linda Varagnolo
Quiong Lin
Nadine Obier
Christoph Plass
Johannes Dietl
Martin Zenke
Rainer Claus
Albrecht M. Müller
eng
uncontrolled
ex vivo expansion
eng
uncontrolled
epigenomic landscapes
eng
uncontrolled
in vivo polycomb
eng
uncontrolled
transplantation states genes
eng
uncontrolled
EZH2 differentiation trichostatin
Medizin und Gesundheit
open_access
Institut für Medizinische Strahlenkunde und Zellforschung
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/14837/016_Varagnolo_Scientific_Reports.pdf
5612
2010
eng
article
1
2012-02-24
--
--
Expression pattern of matrix metalloproteinases in human gynecological cancer cell lines
Background: Matrix metalloproteinases (MMPs) are involved in the degradation of protein components of the extracellular matrix and thus play an important role in tumor invasion and metastasis. Their expression is related to the progression of gynecological cancers (e.g. endometrial, cervical or ovarian carcinoma). In this study we investigated the expression pattern of the 23 MMPs, currently known in humans, in different gynecological cancer cell lines. Methods: In total, cell lines from three endometrium carcinomas (Ishikawa, HEC-1-A, AN3 CA), three cervical carcinomas (HeLa, Caski, SiHa), three chorioncarcinomas (JEG, JAR, BeWo), two ovarian cancers (BG-1, OAW-42) and one teratocarcinoma (PA-1) were examined. The expression of MMPs was analyzed by RT-PCR, Western blot and gelatin zymography. Results: We demonstrated that the cell lines examined can constitutively express a wide variety of MMPs on mRNA and protein level. While MMP-2, -11, -14 and -24 were widely expressed, no expression was seen for MMP-12, -16, -20, -25, -26, -27 in any of the cell lines. A broad range of 16 MMPs could be found in the PA1 cells and thus this cell line could be used as a positive control for general MMP experiments. While the three cervical cancer cell lines expressed 10-14 different MMPs, the median expression in endometrial and choriocarcinoma cells was 7 different enzymes. The two investigated ovarian cancer cell lines showed a distinctive difference in the number of expressed MMPs (2 vs. 10). Conclusions: Ishikawa, Caski, OAW-42 and BeWo cell lines could be the best choice for all future experiments on MMP regulation and their role in endometrial, cervical, ovarian or choriocarcinoma development, whereas the teratocarcinoma cell line PA1 could be used as a positive control for general MMP experiments.
urn:nbn:de:bvb:20-opus-67880
6788
BMC CANCER (2010) 10, DOI: 10.1186/1471-2407-10-553
Andrea Schroepfer
Ulrike Kämmerer
Michaela Kapp
Johannes Dietl
Sonja Feix
Jelena Anacker
deu
swd
Krebs <Medizin>
eng
uncontrolled
Matrix metalloproteinases
eng
uncontrolled
MMP
Medizin und Gesundheit
open_access
Frauenklinik und Poliklinik
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/5612/Anacker_1471_2407_10_553.pdf
12001
2014
eng
129–139
2
6
article
1
2015-10-08
--
--
Anti-CD39 and anti-CD73 antibodies A1 and 7G2 improve targeted therapy in ovarian cancer by blocking adenosine-dependent immune evasion
The ectonucleotidases CD39 and CD73 degrade ATP to adenosine which inhibits immune responses via the \(A_{2A}\) adenosine receptor (ADORA2A) on T and NK cells. The current study investigates the potential therapeutic use of the specific anti CD39- and anti CD73-antibodies A1 (CD39) and 7G2 (CD73) as these two ectonucleotidases are overexpressed in ovarian cancer (OvCA). As expected, NK cell cytotoxicity against the human ovarian cancer cell lines OAW-42 or SK-OV-3 was significantly increased in the presence of A1 or 7G2 antibody. While this might partly be due to antibody-dependent cell-mediated cytotoxicity, a luciferase-dependent assay for quantifying biologically active adenosine further showed that A1 and 7G2 can inhibit CD39 and CD73-dependent adenosine-generation. In turn, the reduction in adenosine levels achieved by addition of A1 and 7G2 to OAW-42 or SK-OV-3 cells was found to de-inhibit the proliferation of \(CD4^+\) T cells in coculture with OvCA cells. Likewise, blocking of CD39 and CD73 on OvCA cells via A1 and 7G2 led to an increased cytotoxicity of alloreactive primed T cells. Thus, antibodies like A1 and 7G2 could improve targeted therapy in ovarian cancer not only by specifically labeling overexpressed antigens but also by blocking adenosine-dependent immune evasion in this immunogenic malignancy.
American Journal of Translational Research
1943-8141
PMC3902223
urn:nbn:de:bvb:20-opus-120016
American Journal of Translational Research 2014;6(2):129-139
Sebastian F. M. Häusler
Itsaso Montalbán del Barrio
Joachim Diessner
Roland G. Stein
Jenny Strohschein
Arnd Hönig
Johannes Dietl
Jörg Wischhusen
eng
uncontrolled
ovarian cancer
eng
uncontrolled
immune escape
eng
uncontrolled
adenosine
eng
uncontrolled
CD39
eng
uncontrolled
CD73
Gynäkologie, Geburtsmedizin, Pädiatrie, Geriatrie
open_access
Frauenklinik und Poliklinik
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/12001/070_Haeusler_AMERICAN_JOURNAL_OF_TRANSLATIONAL_RESEARCH.pdf
12863
2013
eng
221-220
2
3
article
1
2016-03-04
--
--
Targeting breast cancer stem cells with HER2-specific antibodies and natural killer cells
Breast cancer is the most common cancer among women worldwide. Every year, nearly 1.4 million new cases of breast cancer are diagnosed, and about 450.000 women die of the disease. Approximately 15-25% of breast cancer cases exhibit increased quantities of the trans-membrane receptor tyrosine kinase human epidermal growth factor receptor 2 (HER2) on the tumor cell surface. Previous studies showed that blockade of this HER2 proto-oncogene with the antibody trastuzumab substantially improved the overall survival of patients with this aggressive type of breast cancer. Recruitment of natural killer (NK) cells and subsequent induction of antibody-dependent cell-mediated cytotoxicity (ADCC) contributed to this beneficial effect. We hypothesized that antibody binding to HER2-positive breast cancer cells and thus ADCC might be further improved by synergistically applying two different HER2-specific antibodies, trastuzumab and pertuzumab. We found that tumor cell killing via ADCC was increased when the combination of trastuzumab, pertuzumab, and NK cells was applied to HER2-positive breast cancer cells, as compared to the extent of ADCC induced by a single antibody. Furthermore, a subset of \(CD44^{high}CD24^{low}HER2^{low}\) cells, which possessed characteristics of cancer stem cells, could be targeted more efficiently by the combination of two HER2-specific antibodies compared to the efficiency of one antibody. These in vitro results demonstrated the immunotherapeutic benefit achieved by the combined application of trastuzumab and pertuzumab. These findings are consistent with the positive results of the clinical studies, CLEOPATRA and NEOSPHERE, conducted with patients that had HER2-positive breast cancer. Compared to a single antibody treatment, the combined application of trastuzumab and pertuzumab showed a stronger ADCC effect and improved the targeting of breast cancer stem cells.
American Journal of Cancer Research
urn:nbn:de:bvb:20-opus-128633
American Journal of Cancer Research 2013;3(2):211-220
Joachim Diessner
Valentin Bruttel
Kathrin Becker
Miriam Pawlik
Roland Stein
Sebastian Häusler
Johannes Dietl
Jörg Wischhusen
Arnd Hönig
eng
uncontrolled
trastuzumab
eng
uncontrolled
breast cancer
eng
uncontrolled
tumor stem cells
eng
uncontrolled
ADCC
eng
uncontrolled
pertuzumab
Medizin und Gesundheit
open_access
Frauenklinik und Poliklinik
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/12863/007_Diessner_Targeting_breast_cancer.pdf
6931
2010
eng
article
1
2012-02-03
--
--
Perinatal Problems in Multiple Births Inconsistent Terminology
6802
urn:nbn:de:bvb:20-opus-85850
In: DEUTSCHES ARZTEBLATT INTERNATIONAL (2010) 107, 50, 902-902, DOI: 10.3238/arztebl.2010.0902a
Deutsches Urheberrecht
Ursula Zollner
Monika Rehn
Johannes Dietl
Medizin und Gesundheit
open_access
Frauenklinik und Poliklinik
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/6931/Zollner_arztbl-2010-0902a.pdf
6932
2010
eng
article
1
2012-02-03
--
--
The Marketing Effect
6803
urn:nbn:de:bvb:20-opus-85868
In: DEUTSCHES ARZTEBLATT INTERNATIONAL (2010) 107, 45, 797-797, DOI: 10.3238/arztebl.2010.0796b
Deutsches Urheberrecht
Johannes Dietl
Medizin und Gesundheit
open_access
Frauenklinik und Poliklinik
Universität Würzburg
29753
2022
eng
11
10
article
1
--
2022-11-16
--
Ovarian cancer-cell pericellular hyaluronan deposition negatively impacts prognosis of ovarian cancer patients
Background: Hyaluronan (HA), a component of the extracellular matrix, is frequently increased under pathological conditions including cancer. Not only stroma cells but also cancer cells themselves synthesize HA, and the interaction of HA with its cognate receptors promotes malignant progression and metastasis. Methods: In the present study, HA deposition in tissue sections was analyzed by hyaluronan-binding protein (HABP) ligand histochemistry in 17 borderline tumors and 102 primary and 20 recurrent ovarian cancer samples. The intensity and, particularly, localization of the HA deposition were recorded: for the localization, the pericellular deposition around the ovarian cancer cells was distinguished from the deposition within the stromal compartment. These histochemical data were correlated with clinical and pathological parameters. Additionally, within a reduced subgroup of ovarian cancer samples (n = 70), the RNA levels of several HA-associated genes were correlated with the HA localization and intensity. Results: Both stroma-localized and pericellular tumor-cell-associated HA deposition were observed. Cancer-cell pericellular HA deposition, irrespective of its staining intensity, was significantly associated with malignancy, and in the primary ovarian cancer cohort, it represents an independent unfavorable prognostic marker for overall survival. Furthermore, a significant association between high CD44, HAS2 and HAS3 mRNA levels and a cancer-cell pericellular HA-deposition pattern was noted. In contrast, stromal hyaluronan deposition had no impact on ovarian cancer prognosis. Conclusions: In conclusion, the site of HA deposition is of prognostic value, but the amount deposited is not. The significant association of only peritumoral cancer-cell HA deposition with high CD44 mRNA expression levels suggests a pivotal role of the CD44–HA signaling axis for malignant progression in ovarian cancer.
Biomedicines
2227-9059
10.3390/biomedicines10112944
urn:nbn:de:bvb:20-opus-297539
2022-12-15T14:05:56+00:00
sword
swordwue
attachment; filename=deposit.zip
5bac1ee9266e534c518e6581d502834d
Biomedicines (2022) 10:11, 2944. https://doi.org/10.3390/biomedicines10112944
false
true
CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International
Leticia Oliveira-Ferrer
Barbara Schmalfeldt
Johannes Dietl
Catharina Bartmann
Udo Schumacher
Christine Stürken
eng
uncontrolled
ovarian cancer
eng
uncontrolled
stromal hyaluronan
eng
uncontrolled
tumor-associated hyaluronan staining pattern
eng
uncontrolled
hyaluronan-related enzymes
Medizin und Gesundheit
open_access
Frauenklinik und Poliklinik
Import
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/29753/biomedicines-10-02944.pdf