المؤلفون: Velina S. Atanasova, Crhistian de Jesus Cardona, Václav Hejret, Andreas Tiefenbacher, Theresia Mair, Loan Tran, Janette Pfneissl, Kristina Draganić, Carina Binder, Julijan Kabiljo, Janik Clement, Katharina Woeran, Barbara Neudert, Sabrina Wohlhaupter, Astrid Haase, Sandra Domazet, Markus Hengstschläger, Markus Mitterhauser, Leonhard Müllauer, Boris Tichý, Michael Bergmann, Gabriele Schweikert, Markus Hartl, Helmut Dolznig, Gerda Egger

المصدر: Cellular and Molecular Gastroenterology and Hepatology, Vol 15, Iss 6, Pp 1391-1419 (2023)

مصطلحات موضوعية: Cancer, Co-cultures, Colorectal Cancer, Fibroblasts, Organoids, Diseases of the digestive system. Gastroenterology, RC799-869

الوصف: Background & Aims: Patient-derived organoid cancer models are generated from epithelial tumor cells and reflect tumor characteristics. However, they lack the complexity of the tumor microenvironment, which is a key driver of tumorigenesis and therapy response. Here, we developed a colorectal cancer organoid model that incorporates matched epithelial cells and stromal fibroblasts. Methods: Primary fibroblasts and tumor cells were isolated from colorectal cancer specimens. Fibroblasts were characterized for their proteome, secretome, and gene expression signatures. Fibroblast/organoid co-cultures were analyzed by immunohistochemistry and compared with their tissue of origin, as well as on gene expression levels compared with standard organoid models. Bioinformatics deconvolution was used to calculate cellular proportions of cell subsets in organoids based on single-cell RNA sequencing data. Results: Normal primary fibroblasts, isolated from tumor adjacent tissue, and cancer associated fibroblasts retained their molecular characteristics in vitro, including higher motility of cancer associated compared with normal fibroblasts. Importantly, both cancer-associated fibroblasts and normal fibroblasts supported cancer cell proliferation in 3D co-cultures, without the addition of classical niche factors. Organoids grown together with fibroblasts displayed a larger cellular heterogeneity of tumor cells compared with mono-cultures and closely resembled the in vivo tumor morphology. Additionally, we observed a mutual crosstalk between tumor cells and fibroblasts in the co-cultures. This was manifested by considerably deregulated pathways such as cell-cell communication and extracellular matrix remodeling in the organoids. Thrombospondin-1 was identified as a critical factor for fibroblast invasiveness. Conclusion: We developed a physiological tumor/stroma model, which will be vital as a personalized tumor model to study disease mechanisms and therapy response in colorectal cancer.

وصف الملف: electronic resource

Relation: http://www.sciencedirect.com/science/article/pii/S2352345X2300036X; https://doaj.org/toc/2352-345X

URL الوصول: https://doaj.org/article/4731d5078fae4db69082355f0d4fbdc9

المؤلفون: Alexander Ries, Daniela Flehberger, Astrid Slany, Christine Pirker, Johanna C. Mader, Thomas Mohr, Karin Schelch, Katharina Sinn, Berta Mosleh, Mir Alireza Hoda, Balazs Dome, Helmut Dolznig, Georg Krupitza, Leonhard Müllauer, Christopher Gerner, Walter Berger, Michael Grusch

المصدر: Journal of Experimental & Clinical Cancer Research, Vol 42, Iss 1, Pp 1-20 (2023)

مصطلحات موضوعية: Pleural mesothelioma, Cancer-associated fibroblasts, Co-cultures, Tumor microenvironment, WNT, PI3K, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Abstract Background Pleural mesothelioma (PM) is an aggressive malignancy with poor prognosis. Unlike many other cancers, PM is mostly characterized by inactivation of tumor suppressor genes. Its highly malignant nature in absence of tumor driving oncogene mutations indicates an extrinsic supply of stimulating signals by cells of the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) are an abundant cell type of the TME and have been shown to drive the progression of several malignancies. The aim of the current study was to isolate and characterize patient-derived mesothelioma-associated fibroblasts (Meso-CAFs), and evaluate their impact on PM cells. Methods Meso-CAFs were isolated from surgical specimens of PM patients and analyzed by array comparative genomic hybridization, next generation sequencing, transcriptomics and proteomics. Human PM cell lines were retrovirally transduced with GFP. The impact of Meso-CAFs on tumor cell growth, migration, as well as the response to small molecule inhibitors, cisplatin and pemetrexed treatment was investigated in 2D and 3D co-culture models by videomicroscopy and automated image analysis. Results Meso-CAFs show a normal diploid genotype without gene copy number aberrations typical for PM cells. They express CAF markers and lack PM marker expression. Their proteome and secretome profiles clearly differ from normal lung fibroblasts with particularly strong differences in actively secreted proteins. The presence of Meso-CAFs in co-culture resulted in significantly increased proliferation and migration of PM cells. A similar effect on PM cell growth and migration was induced by Meso-CAF-conditioned medium. Inhibition of c-Met with crizotinib, PI3K with LY-2940002 or WNT signaling with WNT-C59 significantly impaired the Meso-CAF-mediated growth stimulation of PM cells in co-culture at concentrations not affecting the PM cells alone. Meso-CAFs did not provide protection of PM cells against cisplatin but showed significant protection against the EGFR inhibitor erlotinib. Conclusions Our study provides the first characterization of human patient-derived Meso-CAFs and demonstrates a strong impact of Meso-CAFs on PM cell growth and migration, two key characteristics of PM aggressiveness, indicating a major role of Meso-CAFs in driving PM progression. Moreover, we identify signaling pathways required for Meso-CAF-mediated growth stimulation. These data could be relevant for novel therapeutic strategies against PM.

وصف الملف: electronic resource

Relation: https://doaj.org/toc/1756-9966

URL الوصول: https://doaj.org/article/0584feb1baa7455e98b30e32f6b10aef

المؤلفون: Natalia Stepien, Daniel Senfter, Julia Furtner, Christine Haberler, Christian Dorfer, Thomas Czech, Daniela Lötsch-Gojo, Lisa Mayr, Cora Hedrich, Alicia Baumgartner, Maria Aliotti-Lippolis, Hannah Schned, Johannes Holler, Katharina Bruckner, Irene Slavc, Amedeo A. Azizi, Andreas Peyrl, Leonhard Müllauer, Sibylle Madlener, Johannes Gojo

المصدر: Cancers, Vol 15, Iss 9, p 2525 (2023)

مصطلحات موضوعية: liquid biopsy, droplet digital PCR, medulloblastoma, group 3, MYC amplification, minimal residual disease, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Background: Liquid biopsy diagnostic methods are an emerging complementary tool to imaging and pathology techniques across various cancer types. However, there is still no established method for the detection of molecular alterations and disease monitoring in MB, the most common malignant CNS tumor in the pediatric population. In the presented study, we investigated droplet digital polymerase chain reaction (ddPCR) as a highly sensitive method for the detection of MYC amplification in bodily fluids of group 3 MB patients. Methods: We identified a cohort of five MYC-amplified MBs by methylation array and FISH. Predesigned and wet-lab validated probes for ddPCR were used to establish the detection method and were validated in two MYC-amplified MB cell lines as well as tumor tissue of the MYC-amplified cohort. Finally, a total of 49 longitudinal CSF samples were analyzed at multiple timepoints during the course of the disease. Results: Detection of MYC amplification by ddPCR in CSF showed a sensitivity and specificity of 90% and 100%, respectively. We observed a steep increase in amplification rate (AR) at disease progression in 3/5 cases. ddPCR was proven to be more sensitive than cytology for the detection of residual disease. In contrast to CSF, MYC amplification was not detectable by ddPCR in blood samples. Conclusions: ddPCR proves to be a sensitive and specific method for the detection of MYC amplification in the CSF of MB patients. These results warrant implementation of liquid biopsy in future prospective clinical trials to validate the potential for improved diagnosis, disease staging and monitoring.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2072-6694/15/9/2525; https://doaj.org/toc/2072-6694

URL الوصول: https://doaj.org/article/7053f498d8194fd1aefe0d23a5b59375

المؤلفون: Matthias Preusser, Anna Sophie Berghoff, Thorsten Fuereder, Leonhard Müllauer, Gerwin Heller, Angelika Martina Starzer, Katharina Feldmann, Erwin Tomasich, Teresa Hatziioannou, Stefan Traint, Thomas Melchardt, Christoph Minichsdorfer, Ursula Schwarz-Nemec, Maja Nackenhorst

المصدر: Journal for ImmunoTherapy of Cancer, Vol 10, Iss 3 (2022)

مصطلحات موضوعية: Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Background Biomarkers for response prediction to anti-programmed cell death 1 (PD-1) immune checkpoint inhibitors (ICI) in patients with head and neck squamous cell carcinoma (HNSCC) are urgently needed for a personalized therapy approach. We investigated the predictive potential of inflammatory parameters and DNA methylation profiling in patients with HNSCC treated with anti-PD-1 ICI.Methods We identified patients with HNSCC that were treated with anti-PD-1 ICI therapy in the recurrent or metastatic setting after progression to platinum-based chemotherapy in two independent centers. We analyzed DNA methylation profiles of >850.000 CpG sites in tumor specimens of these patients by Infinium MethylationEPIC microarrays, immune cell density in the tumor microenvironment (CD8, CD3, CD45RO, forkhead box P3 (FOXP3), CD68), PD-1 and programmed cell death ligand 1 (PD-L1) expression by immunohistochemistry, and blood inflammation markers (platelet-to-lymphocyte ratio, leucocyte-to-lymphocyte ratio, monocyte-to-lymphocyte ratio, neutrophil-to-lymphocyte ratio). DNA methylation profiles and immunological markers were bioinformatically and statistically correlated with radiological response to anti-PD-1 ICI.Results 37 patients with HNSCC (median age of 62 years; range 49–83; 8 (21.6%) women, 29 (78.4%) men) were included (Center 1 N=26, 70.3%; Center 2 N=11, 29.7%). Median number of prior systemic therapies was 1 (range 1–4). Five out of 37 (13.5%) patients achieved an objective response to ICI. Median progression-free survival and median overall survival times were 3.7 months (range 0–22.9) and 9.0 months (range 0–38.8), respectively. Microarray analyses revealed a methylation signature including both hypomethylation and hypermethylation which was predictive for response to ICI and included several genes involved in cancer-related molecular pathways. Over-represented differentially methylated genes between responders and non-responders were associated with ‘Axon guidance’, ‘Hippo signaling’, ‘Pathways in cancer’ and ‘MAPK signaling’. A statistically significant correlation of PD-L1 expression and response was present (p=0.0498).Conclusions Our findings suggest that tumor DNA methylation profiling may be useful to predict response to ICI in patients with HNSCC.

وصف الملف: electronic resource

Relation: https://jitc.bmj.com/content/10/3/e003420.full; https://doaj.org/toc/2051-1426

URL الوصول: https://doaj.org/article/e6550fd49f974f6795b61c4497460f1c

المؤلفون: Natalia Stepien, Christine Haberler, Sarah Theurer, Maria-Theresa Schmook, Carola Lütgendorf-Caucig, Leonhard Müllauer, Johannes Gojo, Amedeo A. Azizi, Thomas Czech, Irene Slavc, Andreas Peyrl

المصدر: Frontiers in Neuroscience, Vol 16 (2022)

مصطلحات موضوعية: pediatric brain tumor, neuroendocrine carcinoma (NEC), primary CNS tumor, rare entities, neuroendocrine tumors, Neurosciences. Biological psychiatry. Neuropsychiatry, RC321-571

الوصف: Neuroendocrine tumors (NETs) are rare neoplasms predominantly arising in the gastrointestinal-tract or the lungs of adults. To date, only ten cases of primary central nervous system (CNS) NETs have been reported, with just three of them describing a neuroendocrine carcinoma (NECA) and none occurring in a child. We report on a previously healthy 5-year-old boy, who presented with headaches, nausea and vomiting, and was diagnosed with a left cerebellar solid mass with a cystic component. After gross-total resection, histology revealed a neuroendocrine carcinoma. Molecular analysis of the tumor tissue showed a KRAS-splice-site mutation (c451-3C > T). The KRAS-mutation was discovered to be a maternal germline mutation, previously described as likely benign. After extensive search for an extracranial primary tumor, including Ga-68 DOTANOC-PET-CT, the diagnosis of a primary CNS NECA was established, and proton irradiation was performed. Unfortunately, the patient developed an in-field recurrence just 5 weeks after the end of radiotherapy. The tumor was re-resected with vital tumor tissue. Six cycles of chemotherapy were initiated, consisting of cisplatin, carboplatin, etoposide and ifosfamide. The patient remains disease free 22 months after the end of treatment, supporting the beneficial effect of platinum- and etoposide-based chemotherapy for this tumor entity.

وصف الملف: electronic resource

Relation: https://www.frontiersin.org/articles/10.3389/fnins.2022.810645/full; https://doaj.org/toc/1662-453X

URL الوصول: https://doaj.org/article/687fe07356514865bdee0dc6cfc84df4

المؤلفون: Bahil Ghanim, Dina Baier, Christine Pirker, Leonhard Müllauer, Katharina Sinn, Gyoergy Lang, Konrad Hoetzenecker, Walter Berger

المصدر: Cancers, Vol 14, Iss 22, p 5602 (2022)

مصطلحات موضوعية: solitary fibrous tumor, NAB2-STAT6 gene fusion, targeted therapy, trabectedin, ponatinib, dasatinib, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Solitary fibrous tumor of the pleura (SFT) is a rare disease. Besides surgery combined with radiotherapy in nondisseminated stages, curative options are currently absent. Out of fourteen primo-cell cultures, established from surgical SFT specimens, two showed stable in vitro growth. Both cell models harbored the characteristic NAB2-STAT6 fusion and were further investigated by different preclinical methods assessing cell viability, clone formation, and protein regulation upon single-drug treatment or in response to selected treatment combinations. Both fusion-positive cell models showed—in line with the clinical experience and the literature—a low to moderate response to most of the tested cytotoxic and targeted agents. However, the multi-tyrosine kinase inhibitors ponatinib and dasatinib, as well as the anti-sarcoma compound trabectedin, revealed promising activity against SFT growth. Furthermore, both cell models spontaneously presented strong FGFR downstream signaling targetable by ponatinib. Most interestingly, the combination of either ponatinib or dasatinib with trabectedin showed synergistic effects. In conclusion, this study identified novel trabectedin-based treatment combinations with clinically approved tyrosine kinase inhibitors, using two newly established NAB2-STAT6 fusion-positive cell models. These findings can be the basis for anti-SFT drug repurposing approaches in this rare and therapy-refractory disease.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2072-6694/14/22/5602; https://doaj.org/toc/2072-6694

URL الوصول: https://doaj.org/article/4071f3b507dc4cc2b5e2a31b2ca0711c

المؤلفون: Hossein Taghizadeh, Sabine Zöchbauer‐Müller, Robert M. Mader, Leonhard Müllauer, Thomas Klikovits, Thomas Bachleitner‐Hofmann, Mir A. Hoda, Gerald W. Prager

المصدر: Thoracic Cancer, Vol 11, Iss 7, Pp 1979-1988 (2020)

مصطلحات موضوعية: Genomic aberrations, malignant mesothelioma, molecular profiling, mutations, precision medicine, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Abstract Background Malignant mesothelioma is an aggressive cancer and has a poor prognosis. Here, we analyzed the feasibility, molecular and gender aspects of targeted therapy recommendations for malignant mesothelioma based on the individual molecular tumor profile. Methods In this single‐center, real‐world retrospective analysis of our platform for precision medicine, we evaluated the molecular profiling of malignant mesothelioma in 14 patients, including nine men and five women. Tumor samples of the patients were examined with a 50 gene next‐generation sequencing (NGS) panel, immunohistochemistry, and fluorescence in situ hybridization, to detect possible molecular aberrations which may be targeted by off‐label therapy custom‐tailored to the individual patient. Results In total, we identified 11 mutations in six of the 14 patients, including BAP1, FANCA, NF1, NF2, PD‐L1, RAD52D, SETD2, SRC, and TP53. No mutation was detected in eight of the 14 patients. Targeted therapy was recommended for 11 out of the 14 patients. All recommendations were mainly based on the molecular characteristics determined by immunohistochemistry. Targeted therapy recommendations were significantly more often for men than women due to gender‐specific differences in PDGFRα expression. Eventually, four patients received the targeted therapy, of whom one patient subsequently achieved stable disease. Conclusions Our observations suggest that a molecular‐guided treatment approach is feasible for the management of advanced malignant mesothelioma. Our analysis revealed gender specific differences in PDGFRα expression that should be further evaluated in clinical trials.

وصف الملف: electronic resource

Relation: https://doaj.org/toc/1759-7706; https://doaj.org/toc/1759-7714

URL الوصول: https://doaj.org/article/a04bb54214904b2682b9ee2765ed5071

المؤلفون: Christina T. Grech, Dietmar Pils, Stefanie Aust, Christoph Grimm, Stephan Polterauer, Alexander Reinthaller, Leonhard Müllauer, Theresa Reischer, Christine Bekos

المصدر: Journal of Personalized Medicine, Vol 12, Iss 9, p 1467 (2022)

مصطلحات موضوعية: BRCA, PARP-inhibitor, ovarian cancer, allele frequency, Medicine

الوصف: PARP inhibitors (PARPi) have increased treatment options in ovarian cancer, particularly in patients with BRCA1/2 mutations, although there are still marked differences in the duration of patients’ response to this targeted therapy. BRCA testing is routinely performed in tumor tissue of ovarian cancer patients. The resulting molecular pathological findings include the genetic nomenclature of the mutation, the frequency of the mutated allele (variant allele frequency, VAF), and the tumor cell content. VAF measures the percentage of mutated alleles from the total alleles in the cells of the examined tissue. The aim of this study was to investigate the significance of VAF on the therapeutic response to PARPis in ovarian cancer patients. Epithelial ovarian cancer patients harboring BRCA1/2 tumor mutations, who underwent germline testing and received PARPi therapy at the Medical University of Vienna (n = 41) were included in the study. Corrected VAF (cVAF) was calculated based on VAF, tumor cell content, and germline mutation. Patients were divided into two groups based on their cVAF. Median PFS under PARPi in patients with low cVAF was 13.0 months (IQR [10.3-not reached]) and was not reached in the high cVAF group. High cVAF was significantly associated with longer PFS in the multivariate analysis (HR = 0.07; 95% CI [0.01–0.63]; p = 0.017). In conclusion, high cVAF was associated with a significantly better response to PARPi in this study population.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2075-4426/12/9/1467; https://doaj.org/toc/2075-4426

URL الوصول: https://doaj.org/article/26c908172d184f9d8c12e0e01a739942

المؤلفون: Matthias Preusser, Leonhard Müllauer, Anna S Berghoff, Sebastian Bauer, Gerwin Heller, Thomas Brodowicz, Wolfgang Lamm, Katharina Feldmann, Angelika M Starzer, Rainer Hamacher, Erwin Tomasich, Teresa Hatziioannou, Stefan Traint, Iris M Noebauer-Huhmann, Julia Furtner, Gabriele Amann, Hans-Ulrich Schildhaus

المصدر: Journal for ImmunoTherapy of Cancer, Vol 9, Iss 3 (2021)

مصطلحات موضوعية: Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Background Some sarcomas respond to immune checkpoint inhibition, but predictive biomarkers are unknown. We analyzed tumor DNA methylation profiles in relation to immunological parameters and response to anti-programmed cell death 1 (anti-PD-1) immune checkpoint inhibitor (ICI) therapy in patients with sarcoma.Patients and methods We retrospectively identified adult patients who had received anti-PD-1 ICI therapy for recurrent sarcoma in two independent centers. We performed (1) blinded radiological response evaluation according to immune response evaluation criteria in solid tumors (iRECIST) ; (2) tumor DNA methylation profiling of >850,000 probes using Infinium MethylationEPIC microarrays; (3) analysis of tumor-infiltrating immune cell subsets (CD3, CD8, CD45RO, FOXP3) and intratumoral expression of immune checkpoint molecules (PD-L1, PD-1, LAG-3) using immunohistochemistry; and (4) evaluation of blood-based systemic inflammation scores (neutrophil-to-lymphocyte ratio, leucocyte-to-lymphocyte ratio, monocyte-to-lymphocyte ratio, platelet-to-lymphocyte ratio). Response to anti-PD-1 ICI therapy was bioinformatically and statistically correlated with DNA methylation profiles and immunological data.Results 35 patients (median age of 50 (23–81) years; 18 females, 17 males; 27 soft tissue sarcomas; 8 osteosarcomas) were included in this study. The objective response rate to anti-PD-1 ICI therapy was 22.9% with complete responses in 3 out of 35 and partial responses in 5 out of 35 patients. Adjustment of DNA methylation data for tumor-infiltrating immune cells resulted in identification of methylation differences between responders and non-responders to anti-PD-1 ICI. 2453 differentially methylated CpG sites (DMPs; 2043 with decreased and 410 with increased methylation) were identified. Clustering of sarcoma samples based on these DMPs revealed two main clusters: methylation cluster 1 (MC1) consisted of 73% responders and methylation cluster 2 (MC2) contained only non-responders to anti-PD-1 ICI. Median progression-free survival from anti-PD-1 therapy start of MC1 and MC2 patients was 16.5 and 1.9 months, respectively (p=0.001). Median overall survival of these patients was 34.4 and 8.0 months, respectively (p=0.029). The most prominent DNA methylation differences were found in pathways implicated in Rap1 signaling, focal adhesion, adherens junction Phosphoinositide 3-kinase (PI3K)-Akt signaling and extracellular matrix (ECM)–receptor interaction.Conclusions Our data demonstrate that tumor DNA methylation profiles may serve as a predictive marker for response to anti-PD-1 ICI therapy in sarcoma.

وصف الملف: electronic resource

Relation: https://jitc.bmj.com/content/9/3/e001458.full; https://doaj.org/toc/2051-1426

URL الوصول: https://doaj.org/article/a79916ad1b1542e9b504874a3913dd32

المؤلفون: Hossein Taghizadeh, Agnieszka Maj-Hes, Gerald W. Prager, Leonhard Müllauer, Robert M. Mader

المصدر: Cancers, Vol 14, Iss 8, p 1936 (2022)

مصطلحات موضوعية: targeted therapy, molecular oncology, mTOR, tissue-agnostic, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: In this analysis, we examined the efficacy, feasibility, and limitations of the application of mTOR inhibitors based on the individual molecular profiles of pretreated cancer patients after the failure of all standard treatments in the palliative setting. In this single-center, real-world analysis of our platform for precision medicine, we analyzed the molecular characteristics of 71 cancer patients. The tumor samples of the patients were analyzed using next-generation sequencing panels of mutation hotspots, microsatellite stability testing, and immunohistochemistry. All profiles were reviewed by a multidisciplinary team to provide a targeted treatment recommendation after a consensus discussion. Seventy-one cancer patients with activation of the mTOR pathway were offered an mTORC1-inhibitor-based targeted therapy, and twenty-three (32.4%) of them eventually received the targeted therapy. Only three patients (4.2%) achieved stable disease, of whom one experienced progressive disease again after 9.1 months. The median time to treatment failure was 2.8 months. In total, 110 mutations were detected in 60 patients (84.5%). The three most frequent mutations were found in TP53, PTEN, and KRAS, which accounted for over 50% (56.4%) of all mutations. In sum, in selected patients with heavily pretreated solid tumors with activation of the mTOR pathway, the antitumoral activity of mTORC1 inhibition was weak.

وصف الملف: electronic resource

Relation: https://www.mdpi.com/2072-6694/14/8/1936; https://doaj.org/toc/2072-6694

URL الوصول: https://doaj.org/article/c8ce19e1dc34416cb3b60458f2e85e8b