The Experts below are selected from a list of 1620 Experts worldwide ranked by ideXlab platform
Marcel Trautmann - One of the best experts on this subject based on the ideXlab platform.
-
phosphatidylinositol 3 kinase pi3k akt signaling is functionally essential in Myxoid Liposarcoma
Molecular Cancer Therapeutics, 2019Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Ilka Isfort, Bianca Altvater, Claudia Rossig, Birte Jeiler, Arne Kruger, Konrad SteinestelAbstract:Myxoid Liposarcoma (MLS) is an aggressive soft-tissue tumor characterized by a specific reciprocal t(12;16) translocation resulting in expression of the chimeric FUS-DDIT3 fusion protein, an oncogenic transcription factor. Similar to other translocation-associated sarcomas, MLS is characterized by a low frequency of somatic mutations, albeit a subset of MLS has previously been shown to be associated with activating PIK3CA mutations. This study was performed to assess the prevalence of PI3K/Akt signaling alterations in MLS and the potential of PI3K-directed therapeutic concepts. In a large cohort of MLS, key components of the PI3K/Akt signaling cascade were evaluated by next generation seqeuncing (NGS), fluorescence in situ hybridization (FISH), and immunohistochemistry (IHC). In three MLS cell lines, PI3K activity was inhibited by RNAi and the small-molecule PI3K inhibitor BKM120 (buparlisib) in vitro An MLS cell line-based avian chorioallantoic membrane model was applied for in vivo confirmation. In total, 26.8% of MLS cases displayed activating alterations in PI3K/Akt signaling components, with PIK3CA gain-of-function mutations representing the most prevalent finding (14.2%). IHC suggested PI3K/Akt activation in a far larger subgroup of MLS, implying alternative mechanisms of pathway activation. PI3K-directed therapeutic interference showed that MLS cell proliferation and viability significantly depended on PI3K-mediated signals in vitro and in vivo Our preclinical study underlines the elementary role of PI3K/Akt signals in MLS tumorigenesis and provides a molecularly based rationale for a PI3K-targeted therapeutic approach which may be particularly effective in the subgroup of tumors carrying activating genetic alterations in PI3K/Akt signaling components.
-
abstract 3939 activation of phosphatidylinositol 3 kinase akt signaling in Myxoid Liposarcoma
Cancer Research, 2018Co-Authors: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga GrunewaldAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and characterized by a high tendency to develop metastases. The molecular hallmark of MLS (≈90%) is a pathognomonic reciprocal t(12;16) (q13;p11) translocation, leading to the specific gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis, although its specific biological function and mechanism of action remain to be substantiated. While radiotherapy and chemotherapy with high-dose ifosfamide and doxorubicin represent established therapeutic options, prognosis in the metastasized situation is poor. Molecularly targeted therapeutic approaches are currently not available. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of phosphatidylinositol-39-kinase (PI3-kinase)/Akt signaling in MLS. Experimental procedures: Immunohistochemical and FISH analyses of PI3-kinase/Akt signaling effectors were performed in a large cohort of clinical MLS tumor specimens. Mutational burden was studied by targeted next-generation sequencing (NGS; Illumina MiSeq). PI3-kinase/Akt-mediated signaling transduction was modulated by specific RNAi knockdown and a pharmacological approach applying the small molecule inhibitor BKM120 (Buparlisib; NVP-BKM120). Cell proliferation and FACS assays were performed in different MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS tumor specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, indicating that activation of PI3-kinase/Akt signaling is a frequent feature in MLS. Activating PIK3CA mutations and loss of PTEN as mechanism for PI3-kinase/Akt activation were detected in ≈15%. PI3-kinase inhibition significantly suppressed the signaling cascade, associated with reduction of MLS cell viability and induction of apoptosis in vitro and in vivo. Conclusions: Our preclinical study emphasizes the pivotal role of the PI3-kinase/Akt signaling cascade in MLS pathogenesis and indicates the occurrence of specific mutational aberrations apart from the pathognomonic FUS-DDIT3 gene fusion. Our in vitro and in vivo results suggest that targeting the PI3-kinase/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga Grunewald, Pierre Aman, Reinhard Buttner, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Activation of phosphatidylinositol-3′-kinase/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3939.
-
abstract b04 functional characterization of igf ir pi3k akt signaling in Myxoid Liposarcoma
Clinical Cancer Research, 2018Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Konrad Steinestel, Jasmin Menzel, Pierre AmanAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and an aggressive disease with particular propensity to develop hematogenic metastases. Ninety percent of MLS are characterized by a reciprocal translocation t(12;16) (q13;p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific mechanism of action remains to be substantiated. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of FUS-DDIT3 in IGF-IR/PI3K/Akt signal transduction. Experimental Procedures: Immunohistochemical analyses of IGF-IR/PI3K/Akt signaling effectors and modulators were performed in a comprehensive cohort of clinical MLS specimens. FUS-DDIT3-dependent activation of the IGF-IR/PI3K/Akt signaling cascade was analyzed by siRNA and immunoblotting in vitro. Cell proliferation and FACS assays were performed in multiple tumor-derived MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, representing a strong indication of activated IGF-IR/PI3K/Akt signaling to be a frequent feature in MLS. IGF-IR inhibition significantly suppressed the IGF-IR/PI3K/Akt signaling cascade, associated with impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 led to dephosphorylation of IGF-IR/PI3K/Akt signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusions: Our preclinical study emphasizes the pivotal role of the IGF-IR/PI3K/Akt signaling pathway in MLS pathogenesis and indicates its functional dependence on the MLS-specific FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR/PI3K/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Alice Cyra, Christian Bertling, Ilka Isfort, Jasmin Menzel, Konrad Steinestel, Inga Grunewald, Bianca Altvater, Claudia Rossig, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Functional characterization of IGF-IR/PI3K/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR Conference on Advances in Sarcomas: From Basic Science to Clinical Translation; May 16-19, 2017; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(2_Suppl):Abstract nr B04.
-
abstract b04 oncogenic relevance of igf ir and pi3k akt gsk3 beta signaling in Myxoid Liposarcoma
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Eva Wardelmann, Pierre Aman, Konrad Steinestel, Jasmin Menzel, Sebastian Huss, Wolfgang HartmannAbstract:Introduction: Myxoid Liposarcoma (MLS) accounts for 30-35% of all LS cases and is the second most common type of Liposarcoma. One third of MLS lesions will become metastatic with tumors spreading to unusual bone and soft tissue locations. Ninety percent of MLS are characterized by a specific reciprocal translocation t (12; 16) (q13; p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific biological function and the mechanism of action remain to be defined. Aiming at the preclinical identification of novel therapeutic options in vitro and in vivo, we investigate the functional relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in primary MLS and tumor-derived cell lines. Methods: Immunohistochemical analyses of IGF-IR and PI3K/AKT/GSK3-beta signaling effectors and modulators were performed in a comprehensive cohort of primary MLS specimens. FUS-DDIT3-dependent activation of the PI3K/AKT/GSK3-beta signaling cascade was analyzed by siRNA knockdown experiments and protein immunoblotting in vitro. Cell proliferation and FACS assays were performed in two different MLS cell lines. An in vivo tumor model was successfully established performing the chicken chorioallantoic membrane (CAM) assay. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of the respective signaling components, indicating that activated IGF-IR and PI3K/AKT/GSK3-beta signaling is a frequent feature in MLS. IGF-IR inhibition significantly suppressed the PI3K/AKT/GSK3-beta downstream cascade, associated with reduced phosphorylation levels of several signaling components, impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 lead to dephosphorylation of PI3K/AKT/GSK3-beta signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusion: Our study emphasizes the pivotal role of IGF-IR and PI3K/AKT/GSK3-beta signaling in MLS pathogenesis and indicates its functional dependence on the characteristic FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR and PI3K/AKT/GSK3-beta signaling pathway might provide a specific, molecular founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Oncogenic relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B04.
-
abstract b05 Myxoid Liposarcoma a molecular and clinicopathological analysis by targeted next generation sequencing and fluorescence in situ hybridization
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Pierre Aman, Birte Jeiler, Arne Kruger, Konrad Steinestel, Jasmin Menzel, Eva WardelmannAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma, accounting for 30-35% of all LS cases. Over 90% of tumors are characterized by a reciprocal translocation t (12; 16) (q13; p11), resulting in a pathogenic gene fusion. The chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis and progression, although the specific biological function and the mechanism of action remain to be defined. We compiled a comprehensive cohort of 105 well-characterized MLS tissue specimens to identify actionable genetic aberrations. Methods: Targeted next-generation sequencing (NGS) using the Illumina MiSeq platform was performed to examine the mutational status of 23 cancer-related genes (covering all exons) known to be frequently mutated across various neoplasms. Furthermore, we examined the amplification/deletion status and characterized the specific chromosomal FUS-DDIT3 rearrangements by FISH and RT-PCR. A multivariate analysis was conducted to investigate the prognostic significance of mutation/amplification/deletion status. Results: Targeted next-generation sequencing drives the potential to generate comprehensive genetic information including less frequent mutated genes relevant for actionable treatments and prognostic assessment. Besides PIK3CA, six additional genes showed at least five mutations, including AKT1, CTNNB1, EGFR, ERBB2, MET and PTEN. Several oncogenic mutations were detected which have not been reported in MLS previously. We demonstrated several gene amplification/deletion events in MLS. Conclusion: Our results indicate the occurrence of mutational aberrations besides the chromosomal FUS-DDIT3 hallmark. These appear not to be related to specific subtypes of FUS-DDIT3 fusion transcripts in terms of a molecular pattern. Molecular screening for actionable mutations might represent a rational tool for the implementation of innovative targeted therapeutic approaches in MLS. To our best knowledge, this study is the most extensive one to yield a detailed map of actionable genetic aberrations across a comprehensive cohort of >100 well-characterized MLS tissue specimens. Moreover, it reveals several molecular alteration-specific targets for innovative therapy strategies. Citation Format: Marcel Trautmann, Arne Kruger, Birte Jeiler, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Myxoid Liposarcoma: A molecular and clinicopathological analysis by targeted next-generation sequencing and fluorescence in situ hybridization [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B05.
Claudia Rossig - One of the best experts on this subject based on the ideXlab platform.
-
phosphatidylinositol 3 kinase pi3k akt signaling is functionally essential in Myxoid Liposarcoma
Molecular Cancer Therapeutics, 2019Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Ilka Isfort, Bianca Altvater, Claudia Rossig, Birte Jeiler, Arne Kruger, Konrad SteinestelAbstract:Myxoid Liposarcoma (MLS) is an aggressive soft-tissue tumor characterized by a specific reciprocal t(12;16) translocation resulting in expression of the chimeric FUS-DDIT3 fusion protein, an oncogenic transcription factor. Similar to other translocation-associated sarcomas, MLS is characterized by a low frequency of somatic mutations, albeit a subset of MLS has previously been shown to be associated with activating PIK3CA mutations. This study was performed to assess the prevalence of PI3K/Akt signaling alterations in MLS and the potential of PI3K-directed therapeutic concepts. In a large cohort of MLS, key components of the PI3K/Akt signaling cascade were evaluated by next generation seqeuncing (NGS), fluorescence in situ hybridization (FISH), and immunohistochemistry (IHC). In three MLS cell lines, PI3K activity was inhibited by RNAi and the small-molecule PI3K inhibitor BKM120 (buparlisib) in vitro An MLS cell line-based avian chorioallantoic membrane model was applied for in vivo confirmation. In total, 26.8% of MLS cases displayed activating alterations in PI3K/Akt signaling components, with PIK3CA gain-of-function mutations representing the most prevalent finding (14.2%). IHC suggested PI3K/Akt activation in a far larger subgroup of MLS, implying alternative mechanisms of pathway activation. PI3K-directed therapeutic interference showed that MLS cell proliferation and viability significantly depended on PI3K-mediated signals in vitro and in vivo Our preclinical study underlines the elementary role of PI3K/Akt signals in MLS tumorigenesis and provides a molecularly based rationale for a PI3K-targeted therapeutic approach which may be particularly effective in the subgroup of tumors carrying activating genetic alterations in PI3K/Akt signaling components.
-
abstract 3939 activation of phosphatidylinositol 3 kinase akt signaling in Myxoid Liposarcoma
Cancer Research, 2018Co-Authors: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga GrunewaldAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and characterized by a high tendency to develop metastases. The molecular hallmark of MLS (≈90%) is a pathognomonic reciprocal t(12;16) (q13;p11) translocation, leading to the specific gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis, although its specific biological function and mechanism of action remain to be substantiated. While radiotherapy and chemotherapy with high-dose ifosfamide and doxorubicin represent established therapeutic options, prognosis in the metastasized situation is poor. Molecularly targeted therapeutic approaches are currently not available. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of phosphatidylinositol-39-kinase (PI3-kinase)/Akt signaling in MLS. Experimental procedures: Immunohistochemical and FISH analyses of PI3-kinase/Akt signaling effectors were performed in a large cohort of clinical MLS tumor specimens. Mutational burden was studied by targeted next-generation sequencing (NGS; Illumina MiSeq). PI3-kinase/Akt-mediated signaling transduction was modulated by specific RNAi knockdown and a pharmacological approach applying the small molecule inhibitor BKM120 (Buparlisib; NVP-BKM120). Cell proliferation and FACS assays were performed in different MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS tumor specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, indicating that activation of PI3-kinase/Akt signaling is a frequent feature in MLS. Activating PIK3CA mutations and loss of PTEN as mechanism for PI3-kinase/Akt activation were detected in ≈15%. PI3-kinase inhibition significantly suppressed the signaling cascade, associated with reduction of MLS cell viability and induction of apoptosis in vitro and in vivo. Conclusions: Our preclinical study emphasizes the pivotal role of the PI3-kinase/Akt signaling cascade in MLS pathogenesis and indicates the occurrence of specific mutational aberrations apart from the pathognomonic FUS-DDIT3 gene fusion. Our in vitro and in vivo results suggest that targeting the PI3-kinase/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga Grunewald, Pierre Aman, Reinhard Buttner, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Activation of phosphatidylinositol-3′-kinase/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3939.
-
abstract b04 functional characterization of igf ir pi3k akt signaling in Myxoid Liposarcoma
Clinical Cancer Research, 2018Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Konrad Steinestel, Jasmin Menzel, Pierre AmanAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and an aggressive disease with particular propensity to develop hematogenic metastases. Ninety percent of MLS are characterized by a reciprocal translocation t(12;16) (q13;p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific mechanism of action remains to be substantiated. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of FUS-DDIT3 in IGF-IR/PI3K/Akt signal transduction. Experimental Procedures: Immunohistochemical analyses of IGF-IR/PI3K/Akt signaling effectors and modulators were performed in a comprehensive cohort of clinical MLS specimens. FUS-DDIT3-dependent activation of the IGF-IR/PI3K/Akt signaling cascade was analyzed by siRNA and immunoblotting in vitro. Cell proliferation and FACS assays were performed in multiple tumor-derived MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, representing a strong indication of activated IGF-IR/PI3K/Akt signaling to be a frequent feature in MLS. IGF-IR inhibition significantly suppressed the IGF-IR/PI3K/Akt signaling cascade, associated with impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 led to dephosphorylation of IGF-IR/PI3K/Akt signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusions: Our preclinical study emphasizes the pivotal role of the IGF-IR/PI3K/Akt signaling pathway in MLS pathogenesis and indicates its functional dependence on the MLS-specific FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR/PI3K/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Alice Cyra, Christian Bertling, Ilka Isfort, Jasmin Menzel, Konrad Steinestel, Inga Grunewald, Bianca Altvater, Claudia Rossig, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Functional characterization of IGF-IR/PI3K/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR Conference on Advances in Sarcomas: From Basic Science to Clinical Translation; May 16-19, 2017; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(2_Suppl):Abstract nr B04.
Pierre Aman - One of the best experts on this subject based on the ideXlab platform.
-
abstract b04 functional characterization of igf ir pi3k akt signaling in Myxoid Liposarcoma
Clinical Cancer Research, 2018Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Konrad Steinestel, Jasmin Menzel, Pierre AmanAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and an aggressive disease with particular propensity to develop hematogenic metastases. Ninety percent of MLS are characterized by a reciprocal translocation t(12;16) (q13;p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific mechanism of action remains to be substantiated. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of FUS-DDIT3 in IGF-IR/PI3K/Akt signal transduction. Experimental Procedures: Immunohistochemical analyses of IGF-IR/PI3K/Akt signaling effectors and modulators were performed in a comprehensive cohort of clinical MLS specimens. FUS-DDIT3-dependent activation of the IGF-IR/PI3K/Akt signaling cascade was analyzed by siRNA and immunoblotting in vitro. Cell proliferation and FACS assays were performed in multiple tumor-derived MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, representing a strong indication of activated IGF-IR/PI3K/Akt signaling to be a frequent feature in MLS. IGF-IR inhibition significantly suppressed the IGF-IR/PI3K/Akt signaling cascade, associated with impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 led to dephosphorylation of IGF-IR/PI3K/Akt signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusions: Our preclinical study emphasizes the pivotal role of the IGF-IR/PI3K/Akt signaling pathway in MLS pathogenesis and indicates its functional dependence on the MLS-specific FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR/PI3K/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Alice Cyra, Christian Bertling, Ilka Isfort, Jasmin Menzel, Konrad Steinestel, Inga Grunewald, Bianca Altvater, Claudia Rossig, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Functional characterization of IGF-IR/PI3K/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR Conference on Advances in Sarcomas: From Basic Science to Clinical Translation; May 16-19, 2017; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(2_Suppl):Abstract nr B04.
-
abstract b04 oncogenic relevance of igf ir and pi3k akt gsk3 beta signaling in Myxoid Liposarcoma
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Eva Wardelmann, Pierre Aman, Konrad Steinestel, Jasmin Menzel, Sebastian Huss, Wolfgang HartmannAbstract:Introduction: Myxoid Liposarcoma (MLS) accounts for 30-35% of all LS cases and is the second most common type of Liposarcoma. One third of MLS lesions will become metastatic with tumors spreading to unusual bone and soft tissue locations. Ninety percent of MLS are characterized by a specific reciprocal translocation t (12; 16) (q13; p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific biological function and the mechanism of action remain to be defined. Aiming at the preclinical identification of novel therapeutic options in vitro and in vivo, we investigate the functional relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in primary MLS and tumor-derived cell lines. Methods: Immunohistochemical analyses of IGF-IR and PI3K/AKT/GSK3-beta signaling effectors and modulators were performed in a comprehensive cohort of primary MLS specimens. FUS-DDIT3-dependent activation of the PI3K/AKT/GSK3-beta signaling cascade was analyzed by siRNA knockdown experiments and protein immunoblotting in vitro. Cell proliferation and FACS assays were performed in two different MLS cell lines. An in vivo tumor model was successfully established performing the chicken chorioallantoic membrane (CAM) assay. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of the respective signaling components, indicating that activated IGF-IR and PI3K/AKT/GSK3-beta signaling is a frequent feature in MLS. IGF-IR inhibition significantly suppressed the PI3K/AKT/GSK3-beta downstream cascade, associated with reduced phosphorylation levels of several signaling components, impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 lead to dephosphorylation of PI3K/AKT/GSK3-beta signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusion: Our study emphasizes the pivotal role of IGF-IR and PI3K/AKT/GSK3-beta signaling in MLS pathogenesis and indicates its functional dependence on the characteristic FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR and PI3K/AKT/GSK3-beta signaling pathway might provide a specific, molecular founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Oncogenic relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B04.
-
abstract b05 Myxoid Liposarcoma a molecular and clinicopathological analysis by targeted next generation sequencing and fluorescence in situ hybridization
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Pierre Aman, Birte Jeiler, Arne Kruger, Konrad Steinestel, Jasmin Menzel, Eva WardelmannAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma, accounting for 30-35% of all LS cases. Over 90% of tumors are characterized by a reciprocal translocation t (12; 16) (q13; p11), resulting in a pathogenic gene fusion. The chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis and progression, although the specific biological function and the mechanism of action remain to be defined. We compiled a comprehensive cohort of 105 well-characterized MLS tissue specimens to identify actionable genetic aberrations. Methods: Targeted next-generation sequencing (NGS) using the Illumina MiSeq platform was performed to examine the mutational status of 23 cancer-related genes (covering all exons) known to be frequently mutated across various neoplasms. Furthermore, we examined the amplification/deletion status and characterized the specific chromosomal FUS-DDIT3 rearrangements by FISH and RT-PCR. A multivariate analysis was conducted to investigate the prognostic significance of mutation/amplification/deletion status. Results: Targeted next-generation sequencing drives the potential to generate comprehensive genetic information including less frequent mutated genes relevant for actionable treatments and prognostic assessment. Besides PIK3CA, six additional genes showed at least five mutations, including AKT1, CTNNB1, EGFR, ERBB2, MET and PTEN. Several oncogenic mutations were detected which have not been reported in MLS previously. We demonstrated several gene amplification/deletion events in MLS. Conclusion: Our results indicate the occurrence of mutational aberrations besides the chromosomal FUS-DDIT3 hallmark. These appear not to be related to specific subtypes of FUS-DDIT3 fusion transcripts in terms of a molecular pattern. Molecular screening for actionable mutations might represent a rational tool for the implementation of innovative targeted therapeutic approaches in MLS. To our best knowledge, this study is the most extensive one to yield a detailed map of actionable genetic aberrations across a comprehensive cohort of >100 well-characterized MLS tissue specimens. Moreover, it reveals several molecular alteration-specific targets for innovative therapy strategies. Citation Format: Marcel Trautmann, Arne Kruger, Birte Jeiler, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Myxoid Liposarcoma: A molecular and clinicopathological analysis by targeted next-generation sequencing and fluorescence in situ hybridization [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B05.
-
trabectedin et 743 promotes differentiation in Myxoid Liposarcoma tumors
Molecular Cancer Therapeutics, 2009Co-Authors: Claudia Forni, Alessandro Gronchi, Mario Minuzzo, Emanuela Virdis, Elena Tamborini, Matteo Simone, Michele Tavecchio, Eugenio Erba, Federica Grosso, Pierre AmanAbstract:Differentiation is a complex set of events that can be blocked by rearrangements of regulatory genes producing fusion proteins with altered properties. In the case of Myxoid Liposarcoma (MLS) tumors, the causative abnormality is a fusion between the CHOP transcription factor and the FUS or EWS genes. CHOP belongs to and is a negative regulator of the large CAAT/enhancer binding protein family whose α, β,and δ members are master genes of adipogenesis. Recent clinical data indicate a peculiar sensitivity of these tumors to the natural marine compound trabectedin. One hypothesis is that the activity of trabectedin is related to the inactivation of the FUS-CHOP oncogene. We find that trabectedin causes detachment of the FUS-CHOP chimera from targeted promoters. Reverse transcription-PCR and chromatin immunoprecipitation analysis in a MLS line and surgical specimens of MLS patients in vivo show activation of the CAAT/enhancer binding protein–mediated transcriptional program that leads to morphologic changes of terminal adipogenesis. The activity is observed in cells with type 1 but not type 8 fusions. Hence, the drug induces maturation of MLS lipoblasts in vivo by targeting the FUS-CHOP–mediated transcriptional block. These data provide a rationale for the specific activity of trabectedin and open the perspective of combinatorial treatments with drugs acting on lipogenic pathways. [Mol Cancer Ther 2009;8(2):449–57]
-
Trabectedin (ET-743) promotes differentiation in Myxoid Liposarcoma tumors
'American Association for Cancer Research (AACR)', 2009Co-Authors: Claudia Forni, Alessandro Gronchi, Mario Minuzzo, Emanuela Virdis, Elena Tamborini, Matteo Simone, Michele Tavecchio, Eugenio Erba, Federica Grosso, Pierre AmanAbstract:Differentiation is a complex set of events that can be blocked by rearrangements of regulatory genes producing fusion proteins with altered properties. In the case of Myxoid Liposarcoma (MLS) tumors, the causative abnormality is a fusion between the CHOP transcription factor and the FUS or EWS genes. CHOP belongs to and is a negative regulator of the large CAAT/enhancer binding protein family whose alpha, beta,and delta members are master genes of adipogenesis. Recent clinical data indicate a peculiar sensitivity of these tumors to the natural marine compound trabectedin. One hypothesis is that the activity of trabectedin is related to the inactivation of the FUS-CHOP oncogene. We find that trabectedin causes detachment of the FUS-CHOP chimera from targeted promoters. Reverse transcription-PCR and chromatin immunoprecipitation analysis in a MLS line and surgical specimens of MLS patients in vivo show activation of the CAAT/enhancer binding protein-mediated transcriptional program that leads to morphologic changes of terminal adipogenesis. The activity is observed in cells with type 1 but not type 8 fusions. Hence, the drug induces maturation of MLS lipoblasts in vivo by targeting the FUS-CHOP-mediated transcriptional block. These data provide a rationale for the specific activity of trabectedin and open the perspective of combinatorial treatments with drugs acting on lipogenic pathways. [Mol Cancer Ther 2009;8(2):449-57]
Inga Grunewald - One of the best experts on this subject based on the ideXlab platform.
-
phosphatidylinositol 3 kinase pi3k akt signaling is functionally essential in Myxoid Liposarcoma
Molecular Cancer Therapeutics, 2019Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Ilka Isfort, Bianca Altvater, Claudia Rossig, Birte Jeiler, Arne Kruger, Konrad SteinestelAbstract:Myxoid Liposarcoma (MLS) is an aggressive soft-tissue tumor characterized by a specific reciprocal t(12;16) translocation resulting in expression of the chimeric FUS-DDIT3 fusion protein, an oncogenic transcription factor. Similar to other translocation-associated sarcomas, MLS is characterized by a low frequency of somatic mutations, albeit a subset of MLS has previously been shown to be associated with activating PIK3CA mutations. This study was performed to assess the prevalence of PI3K/Akt signaling alterations in MLS and the potential of PI3K-directed therapeutic concepts. In a large cohort of MLS, key components of the PI3K/Akt signaling cascade were evaluated by next generation seqeuncing (NGS), fluorescence in situ hybridization (FISH), and immunohistochemistry (IHC). In three MLS cell lines, PI3K activity was inhibited by RNAi and the small-molecule PI3K inhibitor BKM120 (buparlisib) in vitro An MLS cell line-based avian chorioallantoic membrane model was applied for in vivo confirmation. In total, 26.8% of MLS cases displayed activating alterations in PI3K/Akt signaling components, with PIK3CA gain-of-function mutations representing the most prevalent finding (14.2%). IHC suggested PI3K/Akt activation in a far larger subgroup of MLS, implying alternative mechanisms of pathway activation. PI3K-directed therapeutic interference showed that MLS cell proliferation and viability significantly depended on PI3K-mediated signals in vitro and in vivo Our preclinical study underlines the elementary role of PI3K/Akt signals in MLS tumorigenesis and provides a molecularly based rationale for a PI3K-targeted therapeutic approach which may be particularly effective in the subgroup of tumors carrying activating genetic alterations in PI3K/Akt signaling components.
-
abstract 3939 activation of phosphatidylinositol 3 kinase akt signaling in Myxoid Liposarcoma
Cancer Research, 2018Co-Authors: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga GrunewaldAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and characterized by a high tendency to develop metastases. The molecular hallmark of MLS (≈90%) is a pathognomonic reciprocal t(12;16) (q13;p11) translocation, leading to the specific gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis, although its specific biological function and mechanism of action remain to be substantiated. While radiotherapy and chemotherapy with high-dose ifosfamide and doxorubicin represent established therapeutic options, prognosis in the metastasized situation is poor. Molecularly targeted therapeutic approaches are currently not available. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of phosphatidylinositol-39-kinase (PI3-kinase)/Akt signaling in MLS. Experimental procedures: Immunohistochemical and FISH analyses of PI3-kinase/Akt signaling effectors were performed in a large cohort of clinical MLS tumor specimens. Mutational burden was studied by targeted next-generation sequencing (NGS; Illumina MiSeq). PI3-kinase/Akt-mediated signaling transduction was modulated by specific RNAi knockdown and a pharmacological approach applying the small molecule inhibitor BKM120 (Buparlisib; NVP-BKM120). Cell proliferation and FACS assays were performed in different MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS tumor specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, indicating that activation of PI3-kinase/Akt signaling is a frequent feature in MLS. Activating PIK3CA mutations and loss of PTEN as mechanism for PI3-kinase/Akt activation were detected in ≈15%. PI3-kinase inhibition significantly suppressed the signaling cascade, associated with reduction of MLS cell viability and induction of apoptosis in vitro and in vivo. Conclusions: Our preclinical study emphasizes the pivotal role of the PI3-kinase/Akt signaling cascade in MLS pathogenesis and indicates the occurrence of specific mutational aberrations apart from the pathognomonic FUS-DDIT3 gene fusion. Our in vitro and in vivo results suggest that targeting the PI3-kinase/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga Grunewald, Pierre Aman, Reinhard Buttner, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Activation of phosphatidylinositol-3′-kinase/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3939.
-
abstract b04 functional characterization of igf ir pi3k akt signaling in Myxoid Liposarcoma
Clinical Cancer Research, 2018Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Konrad Steinestel, Jasmin Menzel, Pierre AmanAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and an aggressive disease with particular propensity to develop hematogenic metastases. Ninety percent of MLS are characterized by a reciprocal translocation t(12;16) (q13;p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific mechanism of action remains to be substantiated. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of FUS-DDIT3 in IGF-IR/PI3K/Akt signal transduction. Experimental Procedures: Immunohistochemical analyses of IGF-IR/PI3K/Akt signaling effectors and modulators were performed in a comprehensive cohort of clinical MLS specimens. FUS-DDIT3-dependent activation of the IGF-IR/PI3K/Akt signaling cascade was analyzed by siRNA and immunoblotting in vitro. Cell proliferation and FACS assays were performed in multiple tumor-derived MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, representing a strong indication of activated IGF-IR/PI3K/Akt signaling to be a frequent feature in MLS. IGF-IR inhibition significantly suppressed the IGF-IR/PI3K/Akt signaling cascade, associated with impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 led to dephosphorylation of IGF-IR/PI3K/Akt signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusions: Our preclinical study emphasizes the pivotal role of the IGF-IR/PI3K/Akt signaling pathway in MLS pathogenesis and indicates its functional dependence on the MLS-specific FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR/PI3K/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Alice Cyra, Christian Bertling, Ilka Isfort, Jasmin Menzel, Konrad Steinestel, Inga Grunewald, Bianca Altvater, Claudia Rossig, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Functional characterization of IGF-IR/PI3K/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR Conference on Advances in Sarcomas: From Basic Science to Clinical Translation; May 16-19, 2017; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(2_Suppl):Abstract nr B04.
-
abstract b04 oncogenic relevance of igf ir and pi3k akt gsk3 beta signaling in Myxoid Liposarcoma
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Eva Wardelmann, Pierre Aman, Konrad Steinestel, Jasmin Menzel, Sebastian Huss, Wolfgang HartmannAbstract:Introduction: Myxoid Liposarcoma (MLS) accounts for 30-35% of all LS cases and is the second most common type of Liposarcoma. One third of MLS lesions will become metastatic with tumors spreading to unusual bone and soft tissue locations. Ninety percent of MLS are characterized by a specific reciprocal translocation t (12; 16) (q13; p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific biological function and the mechanism of action remain to be defined. Aiming at the preclinical identification of novel therapeutic options in vitro and in vivo, we investigate the functional relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in primary MLS and tumor-derived cell lines. Methods: Immunohistochemical analyses of IGF-IR and PI3K/AKT/GSK3-beta signaling effectors and modulators were performed in a comprehensive cohort of primary MLS specimens. FUS-DDIT3-dependent activation of the PI3K/AKT/GSK3-beta signaling cascade was analyzed by siRNA knockdown experiments and protein immunoblotting in vitro. Cell proliferation and FACS assays were performed in two different MLS cell lines. An in vivo tumor model was successfully established performing the chicken chorioallantoic membrane (CAM) assay. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of the respective signaling components, indicating that activated IGF-IR and PI3K/AKT/GSK3-beta signaling is a frequent feature in MLS. IGF-IR inhibition significantly suppressed the PI3K/AKT/GSK3-beta downstream cascade, associated with reduced phosphorylation levels of several signaling components, impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 lead to dephosphorylation of PI3K/AKT/GSK3-beta signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusion: Our study emphasizes the pivotal role of IGF-IR and PI3K/AKT/GSK3-beta signaling in MLS pathogenesis and indicates its functional dependence on the characteristic FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR and PI3K/AKT/GSK3-beta signaling pathway might provide a specific, molecular founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Oncogenic relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B04.
-
abstract b05 Myxoid Liposarcoma a molecular and clinicopathological analysis by targeted next generation sequencing and fluorescence in situ hybridization
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Pierre Aman, Birte Jeiler, Arne Kruger, Konrad Steinestel, Jasmin Menzel, Eva WardelmannAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma, accounting for 30-35% of all LS cases. Over 90% of tumors are characterized by a reciprocal translocation t (12; 16) (q13; p11), resulting in a pathogenic gene fusion. The chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis and progression, although the specific biological function and the mechanism of action remain to be defined. We compiled a comprehensive cohort of 105 well-characterized MLS tissue specimens to identify actionable genetic aberrations. Methods: Targeted next-generation sequencing (NGS) using the Illumina MiSeq platform was performed to examine the mutational status of 23 cancer-related genes (covering all exons) known to be frequently mutated across various neoplasms. Furthermore, we examined the amplification/deletion status and characterized the specific chromosomal FUS-DDIT3 rearrangements by FISH and RT-PCR. A multivariate analysis was conducted to investigate the prognostic significance of mutation/amplification/deletion status. Results: Targeted next-generation sequencing drives the potential to generate comprehensive genetic information including less frequent mutated genes relevant for actionable treatments and prognostic assessment. Besides PIK3CA, six additional genes showed at least five mutations, including AKT1, CTNNB1, EGFR, ERBB2, MET and PTEN. Several oncogenic mutations were detected which have not been reported in MLS previously. We demonstrated several gene amplification/deletion events in MLS. Conclusion: Our results indicate the occurrence of mutational aberrations besides the chromosomal FUS-DDIT3 hallmark. These appear not to be related to specific subtypes of FUS-DDIT3 fusion transcripts in terms of a molecular pattern. Molecular screening for actionable mutations might represent a rational tool for the implementation of innovative targeted therapeutic approaches in MLS. To our best knowledge, this study is the most extensive one to yield a detailed map of actionable genetic aberrations across a comprehensive cohort of >100 well-characterized MLS tissue specimens. Moreover, it reveals several molecular alteration-specific targets for innovative therapy strategies. Citation Format: Marcel Trautmann, Arne Kruger, Birte Jeiler, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Myxoid Liposarcoma: A molecular and clinicopathological analysis by targeted next-generation sequencing and fluorescence in situ hybridization [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B05.
Magdalene Cyra - One of the best experts on this subject based on the ideXlab platform.
-
phosphatidylinositol 3 kinase pi3k akt signaling is functionally essential in Myxoid Liposarcoma
Molecular Cancer Therapeutics, 2019Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Ilka Isfort, Bianca Altvater, Claudia Rossig, Birte Jeiler, Arne Kruger, Konrad SteinestelAbstract:Myxoid Liposarcoma (MLS) is an aggressive soft-tissue tumor characterized by a specific reciprocal t(12;16) translocation resulting in expression of the chimeric FUS-DDIT3 fusion protein, an oncogenic transcription factor. Similar to other translocation-associated sarcomas, MLS is characterized by a low frequency of somatic mutations, albeit a subset of MLS has previously been shown to be associated with activating PIK3CA mutations. This study was performed to assess the prevalence of PI3K/Akt signaling alterations in MLS and the potential of PI3K-directed therapeutic concepts. In a large cohort of MLS, key components of the PI3K/Akt signaling cascade were evaluated by next generation seqeuncing (NGS), fluorescence in situ hybridization (FISH), and immunohistochemistry (IHC). In three MLS cell lines, PI3K activity was inhibited by RNAi and the small-molecule PI3K inhibitor BKM120 (buparlisib) in vitro An MLS cell line-based avian chorioallantoic membrane model was applied for in vivo confirmation. In total, 26.8% of MLS cases displayed activating alterations in PI3K/Akt signaling components, with PIK3CA gain-of-function mutations representing the most prevalent finding (14.2%). IHC suggested PI3K/Akt activation in a far larger subgroup of MLS, implying alternative mechanisms of pathway activation. PI3K-directed therapeutic interference showed that MLS cell proliferation and viability significantly depended on PI3K-mediated signals in vitro and in vivo Our preclinical study underlines the elementary role of PI3K/Akt signals in MLS tumorigenesis and provides a molecularly based rationale for a PI3K-targeted therapeutic approach which may be particularly effective in the subgroup of tumors carrying activating genetic alterations in PI3K/Akt signaling components.
-
abstract 3939 activation of phosphatidylinositol 3 kinase akt signaling in Myxoid Liposarcoma
Cancer Research, 2018Co-Authors: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga GrunewaldAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and characterized by a high tendency to develop metastases. The molecular hallmark of MLS (≈90%) is a pathognomonic reciprocal t(12;16) (q13;p11) translocation, leading to the specific gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis, although its specific biological function and mechanism of action remain to be substantiated. While radiotherapy and chemotherapy with high-dose ifosfamide and doxorubicin represent established therapeutic options, prognosis in the metastasized situation is poor. Molecularly targeted therapeutic approaches are currently not available. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of phosphatidylinositol-39-kinase (PI3-kinase)/Akt signaling in MLS. Experimental procedures: Immunohistochemical and FISH analyses of PI3-kinase/Akt signaling effectors were performed in a large cohort of clinical MLS tumor specimens. Mutational burden was studied by targeted next-generation sequencing (NGS; Illumina MiSeq). PI3-kinase/Akt-mediated signaling transduction was modulated by specific RNAi knockdown and a pharmacological approach applying the small molecule inhibitor BKM120 (Buparlisib; NVP-BKM120). Cell proliferation and FACS assays were performed in different MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS tumor specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, indicating that activation of PI3-kinase/Akt signaling is a frequent feature in MLS. Activating PIK3CA mutations and loss of PTEN as mechanism for PI3-kinase/Akt activation were detected in ≈15%. PI3-kinase inhibition significantly suppressed the signaling cascade, associated with reduction of MLS cell viability and induction of apoptosis in vitro and in vivo. Conclusions: Our preclinical study emphasizes the pivotal role of the PI3-kinase/Akt signaling cascade in MLS pathogenesis and indicates the occurrence of specific mutational aberrations apart from the pathognomonic FUS-DDIT3 gene fusion. Our in vitro and in vivo results suggest that targeting the PI3-kinase/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Susanne Hafner, Thomas Simmet, Jessica Becker, Inga Grunewald, Pierre Aman, Reinhard Buttner, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Activation of phosphatidylinositol-3′-kinase/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3939.
-
abstract b04 functional characterization of igf ir pi3k akt signaling in Myxoid Liposarcoma
Clinical Cancer Research, 2018Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Ilka Isfort, Bianca Altvater, Claudia Rossig, Konrad Steinestel, Jasmin Menzel, Pierre AmanAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma and an aggressive disease with particular propensity to develop hematogenic metastases. Ninety percent of MLS are characterized by a reciprocal translocation t(12;16) (q13;p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific mechanism of action remains to be substantiated. Aiming at the preclinical identification of novel therapeutic options, we here investigate the functional relevance of FUS-DDIT3 in IGF-IR/PI3K/Akt signal transduction. Experimental Procedures: Immunohistochemical analyses of IGF-IR/PI3K/Akt signaling effectors and modulators were performed in a comprehensive cohort of clinical MLS specimens. FUS-DDIT3-dependent activation of the IGF-IR/PI3K/Akt signaling cascade was analyzed by siRNA and immunoblotting in vitro. Cell proliferation and FACS assays were performed in multiple tumor-derived MLS cell lines. An established MLS chorioallantoic membrane model (CAM) was employed for in vivo confirmation of the preclinical in vitro data. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of various signaling components, representing a strong indication of activated IGF-IR/PI3K/Akt signaling to be a frequent feature in MLS. IGF-IR inhibition significantly suppressed the IGF-IR/PI3K/Akt signaling cascade, associated with impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 led to dephosphorylation of IGF-IR/PI3K/Akt signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusions: Our preclinical study emphasizes the pivotal role of the IGF-IR/PI3K/Akt signaling pathway in MLS pathogenesis and indicates its functional dependence on the MLS-specific FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR/PI3K/Akt signaling pathway provides a rational, molecularly founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Magdalene Alice Cyra, Christian Bertling, Ilka Isfort, Jasmin Menzel, Konrad Steinestel, Inga Grunewald, Bianca Altvater, Claudia Rossig, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Functional characterization of IGF-IR/PI3K/Akt signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR Conference on Advances in Sarcomas: From Basic Science to Clinical Translation; May 16-19, 2017; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(2_Suppl):Abstract nr B04.
-
abstract b04 oncogenic relevance of igf ir and pi3k akt gsk3 beta signaling in Myxoid Liposarcoma
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Eva Wardelmann, Pierre Aman, Konrad Steinestel, Jasmin Menzel, Sebastian Huss, Wolfgang HartmannAbstract:Introduction: Myxoid Liposarcoma (MLS) accounts for 30-35% of all LS cases and is the second most common type of Liposarcoma. One third of MLS lesions will become metastatic with tumors spreading to unusual bone and soft tissue locations. Ninety percent of MLS are characterized by a specific reciprocal translocation t (12; 16) (q13; p11), leading to the pathogenic gene fusion of FUS and DDIT3. The resulting chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS pathogenesis, although the specific biological function and the mechanism of action remain to be defined. Aiming at the preclinical identification of novel therapeutic options in vitro and in vivo, we investigate the functional relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in primary MLS and tumor-derived cell lines. Methods: Immunohistochemical analyses of IGF-IR and PI3K/AKT/GSK3-beta signaling effectors and modulators were performed in a comprehensive cohort of primary MLS specimens. FUS-DDIT3-dependent activation of the PI3K/AKT/GSK3-beta signaling cascade was analyzed by siRNA knockdown experiments and protein immunoblotting in vitro. Cell proliferation and FACS assays were performed in two different MLS cell lines. An in vivo tumor model was successfully established performing the chicken chorioallantoic membrane (CAM) assay. Results: In a significant subset of MLS specimens, immunohistochemical staining revealed elevated phosphorylation levels of the respective signaling components, indicating that activated IGF-IR and PI3K/AKT/GSK3-beta signaling is a frequent feature in MLS. IGF-IR inhibition significantly suppressed the PI3K/AKT/GSK3-beta downstream cascade, associated with reduced phosphorylation levels of several signaling components, impairment of MLS cell viability and induction of apoptosis in vitro and in vivo. Furthermore, siRNA-mediated knockdown of FUS-DDIT3 lead to dephosphorylation of PI3K/AKT/GSK3-beta signaling components, implying that the FUS-DDIT3 fusion protein is involved in the IGF-IR regulated signaling cascade. Conclusion: Our study emphasizes the pivotal role of IGF-IR and PI3K/AKT/GSK3-beta signaling in MLS pathogenesis and indicates its functional dependence on the characteristic FUS-DDIT3 fusion protein. Furthermore, our in vitro and in vivo results demonstrate that targeting the IGF-IR and PI3K/AKT/GSK3-beta signaling pathway might provide a specific, molecular founded therapeutic strategy in the treatment of MLS. Citation Format: Marcel Trautmann, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Oncogenic relevance of IGF-IR and PI3K/AKT/GSK3-beta signaling in Myxoid Liposarcoma [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B04.
-
abstract b05 Myxoid Liposarcoma a molecular and clinicopathological analysis by targeted next generation sequencing and fluorescence in situ hybridization
Cancer Research, 2017Co-Authors: Marcel Trautmann, Inga Grunewald, Magdalene Cyra, Christian Bertling, Pierre Aman, Birte Jeiler, Arne Kruger, Konrad Steinestel, Jasmin Menzel, Eva WardelmannAbstract:Introduction: Myxoid Liposarcoma (MLS) is the second most common type of Liposarcoma, accounting for 30-35% of all LS cases. Over 90% of tumors are characterized by a reciprocal translocation t (12; 16) (q13; p11), resulting in a pathogenic gene fusion. The chimeric FUS-DDIT3 fusion protein is suggested to play a crucial role in MLS tumorigenesis and progression, although the specific biological function and the mechanism of action remain to be defined. We compiled a comprehensive cohort of 105 well-characterized MLS tissue specimens to identify actionable genetic aberrations. Methods: Targeted next-generation sequencing (NGS) using the Illumina MiSeq platform was performed to examine the mutational status of 23 cancer-related genes (covering all exons) known to be frequently mutated across various neoplasms. Furthermore, we examined the amplification/deletion status and characterized the specific chromosomal FUS-DDIT3 rearrangements by FISH and RT-PCR. A multivariate analysis was conducted to investigate the prognostic significance of mutation/amplification/deletion status. Results: Targeted next-generation sequencing drives the potential to generate comprehensive genetic information including less frequent mutated genes relevant for actionable treatments and prognostic assessment. Besides PIK3CA, six additional genes showed at least five mutations, including AKT1, CTNNB1, EGFR, ERBB2, MET and PTEN. Several oncogenic mutations were detected which have not been reported in MLS previously. We demonstrated several gene amplification/deletion events in MLS. Conclusion: Our results indicate the occurrence of mutational aberrations besides the chromosomal FUS-DDIT3 hallmark. These appear not to be related to specific subtypes of FUS-DDIT3 fusion transcripts in terms of a molecular pattern. Molecular screening for actionable mutations might represent a rational tool for the implementation of innovative targeted therapeutic approaches in MLS. To our best knowledge, this study is the most extensive one to yield a detailed map of actionable genetic aberrations across a comprehensive cohort of >100 well-characterized MLS tissue specimens. Moreover, it reveals several molecular alteration-specific targets for innovative therapy strategies. Citation Format: Marcel Trautmann, Arne Kruger, Birte Jeiler, Christian Bertling, Jasmin Menzel, Magdalene Cyra, Konrad Steinestel, Inga Grunewald, Pierre Aman, Eva Wardelmann, Sebastian Huss, Wolfgang Hartmann. Myxoid Liposarcoma: A molecular and clinicopathological analysis by targeted next-generation sequencing and fluorescence in situ hybridization [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B05.
