ºÝºÝߣshows by User: JanDetmers / http://www.slideshare.net/images/logo.gif ºÝºÝߣshows by User: JanDetmers / Mon, 11 Jan 2021 16:55:16 GMT ºÝºÝߣShare feed for ºÝºÝߣshows by User: JanDetmers Precision Oncology /slideshow/precision-oncology-241198730/241198730 precisiononcology-210111165516
Precision medicine’s goal of achieving a better response rate by avoiding ineffective therapies has sparked new approaches, including the testing of patient-derived 3d (PD3Ds) tumor cultures for modeling individual patient response, and the use of various molecular pathology techniques for advanced tumor profiling. Well-established genomics methodologies cannot directly assess cell-signaling activity within the tumor cells defined by the phosphorylation status of cellular signaling pathway networks. Here we present the development of a robust protein profiling strategy utilizing the DigiWest immuno-assay platform, to obtain data on the activation status of key cellular signaling networks implicated in cancer, and on proteins targeted by FDA-approved drugs including a number of targeted cancer therapies for e.g. EGFR, HER2, PI3K, mTOR, ALK and AKT. We compiled a list of relevant pathway nodes and their phosphorylation sites that yield activity information on RAS/RAF/ERK, PI3K/AKT and mTOR signaling pathways. Based on this, we validated 242 total and phospho-antibodies in a pre-set oncoproteomic DigiWest panel that yields information on the activity of these signaling networks from the receptor level down to transcription factors, apoptosis and proliferation. This oncoproteomic panel can be utilized for elucidating drug response in pre-clinical cell models, in PD3D organoid models or in clinical tumor samples. Exemplary, we show differential effects of PI3K kinase inhibitor copanlisib vs MEK inhibitor trametinib at various levels within their signaling networks. Also, we validated this panel in PD3D organoids that were subjected to screening against common targeted therapies.]]>
Precision medicine’s goal of achieving a better response rate by avoiding ineffective therapies has sparked new approaches, including the testing of patient-derived 3d (PD3Ds) tumor cultures for modeling individual patient response, and the use of various molecular pathology techniques for advanced tumor profiling. Well-established genomics methodologies cannot directly assess cell-signaling activity within the tumor cells defined by the phosphorylation status of cellular signaling pathway networks. Here we present the development of a robust protein profiling strategy utilizing the DigiWest immuno-assay platform, to obtain data on the activation status of key cellular signaling networks implicated in cancer, and on proteins targeted by FDA-approved drugs including a number of targeted cancer therapies for e.g. EGFR, HER2, PI3K, mTOR, ALK and AKT. We compiled a list of relevant pathway nodes and their phosphorylation sites that yield activity information on RAS/RAF/ERK, PI3K/AKT and mTOR signaling pathways. Based on this, we validated 242 total and phospho-antibodies in a pre-set oncoproteomic DigiWest panel that yields information on the activity of these signaling networks from the receptor level down to transcription factors, apoptosis and proliferation. This oncoproteomic panel can be utilized for elucidating drug response in pre-clinical cell models, in PD3D organoid models or in clinical tumor samples. Exemplary, we show differential effects of PI3K kinase inhibitor copanlisib vs MEK inhibitor trametinib at various levels within their signaling networks. Also, we validated this panel in PD3D organoids that were subjected to screening against common targeted therapies.]]> Mon, 11 Jan 2021 16:55:16 GMT /slideshow/precision-oncology-241198730/241198730 JanDetmers@slideshare.net(JanDetmers) Precision Oncology JanDetmers Precision medicine’s goal of achieving a better response rate by avoiding ineffective therapies has sparked new approaches, including the testing of patient-derived 3d (PD3Ds) tumor cultures for modeling individual patient response, and the use of various molecular pathology techniques for advanced tumor profiling. Well-established genomics methodologies cannot directly assess cell-signaling activity within the tumor cells defined by the phosphorylation status of cellular signaling pathway networks. Here we present the development of a robust protein profiling strategy utilizing the DigiWest immuno-assay platform, to obtain data on the activation status of key cellular signaling networks implicated in cancer, and on proteins targeted by FDA-approved drugs including a number of targeted cancer therapies for e.g. EGFR, HER2, PI3K, mTOR, ALK and AKT. We compiled a list of relevant pathway nodes and their phosphorylation sites that yield activity information on RAS/RAF/ERK, PI3K/AKT and mTOR signaling pathways. Based on this, we validated 242 total and phospho-antibodies in a pre-set oncoproteomic DigiWest panel that yields information on the activity of these signaling networks from the receptor level down to transcription factors, apoptosis and proliferation. This oncoproteomic panel can be utilized for elucidating drug response in pre-clinical cell models, in PD3D organoid models or in clinical tumor samples. Exemplary, we show differential effects of PI3K kinase inhibitor copanlisib vs MEK inhibitor trametinib at various levels within their signaling networks. Also, we validated this panel in PD3D organoids that were subjected to screening against common targeted therapies. <img style="border:1px solid #C3E6D8;float:right;" alt="" src="https://cdn.slidesharecdn.com/ss_thumbnails/precisiononcology-210111165516-thumbnail.jpg?width=120&amp;height=120&amp;fit=bounds" /><br> Precision medicine’s goal of achieving a better response rate by avoiding ineffective therapies has sparked new approaches, including the testing of patient-derived 3d (PD3Ds) tumor cultures for modeling individual patient response, and the use of various molecular pathology techniques for advanced tumor profiling. Well-established genomics methodologies cannot directly assess cell-signaling activity within the tumor cells defined by the phosphorylation status of cellular signaling pathway networks. Here we present the development of a robust protein profiling strategy utilizing the DigiWest immuno-assay platform, to obtain data on the activation status of key cellular signaling networks implicated in cancer, and on proteins targeted by FDA-approved drugs including a number of targeted cancer therapies for e.g. EGFR, HER2, PI3K, mTOR, ALK and AKT. We compiled a list of relevant pathway nodes and their phosphorylation sites that yield activity information on RAS/RAF/ERK, PI3K/AKT and mTOR signaling pathways. Based on this, we validated 242 total and phospho-antibodies in a pre-set oncoproteomic DigiWest panel that yields information on the activity of these signaling networks from the receptor level down to transcription factors, apoptosis and proliferation. This oncoproteomic panel can be utilized for elucidating drug response in pre-clinical cell models, in PD3D organoid models or in clinical tumor samples. Exemplary, we show differential effects of PI3K kinase inhibitor copanlisib vs MEK inhibitor trametinib at various levels within their signaling networks. Also, we validated this panel in PD3D organoids that were subjected to screening against common targeted therapies.
Precision Oncology from Assay Engineers
]]> 160 0 https://cdn.slidesharecdn.com/ss_thumbnails/precisiononcology-210111165516-thumbnail.jpg?width=120&height=120&fit=bounds document Black http://activitystrea.ms/schema/1.0/post http://activitystrea.ms/schema/1.0/posted 0 Colorectal Cancer / Nature Communications /slideshow/colorectal-cancer-nature-communications/241198606 colorectalcancerinnaturecommunications-210111164156
Wnt signaling is one of the key cascades regulating development and stemness and has also been tightly associated with cancer. The role of Wnt signaling in carcinogenesis has most prominently been described for colorectal cancer, but aberrant Wnt signaling is observed in many more cancer entities. In colorectal cancer (CRC), aberrant Wnt signaling is essential for tumorigenesis and maintenance of cancer stem cells. However, how other oncogenic pathways converge on Wnt signaling to modulate stem cell homeostasis in CRC currently remains poorly understood. Using large-scale compound screens in CRC, we identify MEK1/2 inhibitors as potent activators of Wnt/β-catenin signaling. Targeting MEK increases Wnt activity in different CRC cell lines and murine intestine in vivo. Truncating mutations of APC generated by CRISPR/Cas9 strongly synergize with MEK inhibitors in enhancing Wnt responses in isogenic CRC models. Mechanistically, we demonstrate that MEK inhibition induces a rapid downregulation of AXIN1. Using patient-derived CRC organoids, we show that MEK inhibition leads to increased Wnt activity and elevated LGR5 levels using DigiWest, a novel high parameter immunoblot platform. Our study demonstrates that clinically used MEK inhibitors inadvertently induce stem cell plasticity, revealing an unknown side effect of RAS pathway inhibition.]]>
Wnt signaling is one of the key cascades regulating development and stemness and has also been tightly associated with cancer. The role of Wnt signaling in carcinogenesis has most prominently been described for colorectal cancer, but aberrant Wnt signaling is observed in many more cancer entities. In colorectal cancer (CRC), aberrant Wnt signaling is essential for tumorigenesis and maintenance of cancer stem cells. However, how other oncogenic pathways converge on Wnt signaling to modulate stem cell homeostasis in CRC currently remains poorly understood. Using large-scale compound screens in CRC, we identify MEK1/2 inhibitors as potent activators of Wnt/β-catenin signaling. Targeting MEK increases Wnt activity in different CRC cell lines and murine intestine in vivo. Truncating mutations of APC generated by CRISPR/Cas9 strongly synergize with MEK inhibitors in enhancing Wnt responses in isogenic CRC models. Mechanistically, we demonstrate that MEK inhibition induces a rapid downregulation of AXIN1. Using patient-derived CRC organoids, we show that MEK inhibition leads to increased Wnt activity and elevated LGR5 levels using DigiWest, a novel high parameter immunoblot platform. Our study demonstrates that clinically used MEK inhibitors inadvertently induce stem cell plasticity, revealing an unknown side effect of RAS pathway inhibition.]]> Mon, 11 Jan 2021 16:41:56 GMT /slideshow/colorectal-cancer-nature-communications/241198606 JanDetmers@slideshare.net(JanDetmers) Colorectal Cancer / Nature Communications JanDetmers Wnt signaling is one of the key cascades regulating development and stemness and has also been tightly associated with cancer. The role of Wnt signaling in carcinogenesis has most prominently been described for colorectal cancer, but aberrant Wnt signaling is observed in many more cancer entities. In colorectal cancer (CRC), aberrant Wnt signaling is essential for tumorigenesis and maintenance of cancer stem cells. However, how other oncogenic pathways converge on Wnt signaling to modulate stem cell homeostasis in CRC currently remains poorly understood. Using large-scale compound screens in CRC, we identify MEK1/2 inhibitors as potent activators of Wnt/β-catenin signaling. Targeting MEK increases Wnt activity in different CRC cell lines and murine intestine in vivo. Truncating mutations of APC generated by CRISPR/Cas9 strongly synergize with MEK inhibitors in enhancing Wnt responses in isogenic CRC models. Mechanistically, we demonstrate that MEK inhibition induces a rapid downregulation of AXIN1. Using patient-derived CRC organoids, we show that MEK inhibition leads to increased Wnt activity and elevated LGR5 levels using DigiWest, a novel high parameter immunoblot platform. Our study demonstrates that clinically used MEK inhibitors inadvertently induce stem cell plasticity, revealing an unknown side effect of RAS pathway inhibition. <img style="border:1px solid #C3E6D8;float:right;" alt="" src="https://cdn.slidesharecdn.com/ss_thumbnails/colorectalcancerinnaturecommunications-210111164156-thumbnail.jpg?width=120&amp;height=120&amp;fit=bounds" /><br> Wnt signaling is one of the key cascades regulating development and stemness and has also been tightly associated with cancer. The role of Wnt signaling in carcinogenesis has most prominently been described for colorectal cancer, but aberrant Wnt signaling is observed in many more cancer entities. In colorectal cancer (CRC), aberrant Wnt signaling is essential for tumorigenesis and maintenance of cancer stem cells. However, how other oncogenic pathways converge on Wnt signaling to modulate stem cell homeostasis in CRC currently remains poorly understood. Using large-scale compound screens in CRC, we identify MEK1/2 inhibitors as potent activators of Wnt/β-catenin signaling. Targeting MEK increases Wnt activity in different CRC cell lines and murine intestine in vivo. Truncating mutations of APC generated by CRISPR/Cas9 strongly synergize with MEK inhibitors in enhancing Wnt responses in isogenic CRC models. Mechanistically, we demonstrate that MEK inhibition induces a rapid downregulation of AXIN1. Using patient-derived CRC organoids, we show that MEK inhibition leads to increased Wnt activity and elevated LGR5 levels using DigiWest, a novel high parameter immunoblot platform. Our study demonstrates that clinically used MEK inhibitors inadvertently induce stem cell plasticity, revealing an unknown side effect of RAS pathway inhibition.
Colorectal Cancer / Nature Communications from Assay Engineers
]]> 179 0 https://cdn.slidesharecdn.com/ss_thumbnails/colorectalcancerinnaturecommunications-210111164156-thumbnail.jpg?width=120&height=120&fit=bounds document Black http://activitystrea.ms/schema/1.0/post http://activitystrea.ms/schema/1.0/posted 0 https://cdn.slidesharecdn.com/profile-photo-JanDetmers-48x48.jpg?cb=1610790242 Assay Engineers is a platform agnostic CRO company specialized in hyperplex bioanalysis. We support precision medicine through translational biomarker services ranging from biomarker identification through validation and patient stratification. Based on a concise description of the biomarker's specified use (COU) we help selecting the appropriate bioanalysis platform using a fit-for-purpose approach. www.assay-engineers.com https://cdn.slidesharecdn.com/ss_thumbnails/precisiononcology-210111165516-thumbnail.jpg?width=320&height=320&fit=bounds slideshow/precision-oncology-241198730/241198730 Precision Oncology https://cdn.slidesharecdn.com/ss_thumbnails/colorectalcancerinnaturecommunications-210111164156-thumbnail.jpg?width=320&height=320&fit=bounds slideshow/colorectal-cancer-nature-communications/241198606 Colorectal Cancer / Na...