Final Report Summary - BIOMARKERIGF (Identification of Clinically Useful Biomarkers for IGF-I Receptor Signalling n Cancer)
Biomarker IGF Summary Report October 2014
Background and approaches taken:
The Biomarker-IGF project addressed the problem of developing clinically relevant biomarkers for the Insulin-like Growth Factor (IGF) signalling pathway in cancer. Activity of this growth and survival signaling pathway in cells is strongly associated with cancer development and progression. In the last 10-15 years many researchers and pharmaceutical companies have tested two kinds of agents that inhibit the signalling pathway by targeting the IGF-I Receptor on cancer cells. These agents displayed great promise in pre-clinical testing in cell models and xenograft tumour models. However, with the exception of some notable sporadic responses the results of clinical trials in a range of cancers have overall been disappointing, and further testing was stopped in many cases. These clinical results highlighted the urgent need for biomarkers that measure IGF-1R activity in cancer, principally to select patients that might benefit most from IGF-1R inhibition. Moreover, it became apparent that the IGF-1R participates in adaptive resistance to inhibitors of its own signalling pathway as well as to other drugs. A number of key challenges have become apparent in developing ways to effectively target the IGF-1R in cancer. In general, there may be no relationship between levels of IGF-1R expression, its activity level, activation of the signaling pathway, or the response to IGF-1R inhibitors. Thus, there is no reliable method to predict which patients or kinds of cancer will respond to IGF-1R inhibitors.
The Biomarker-IGF project took two approaches to addressing this problem: A) to develop, models and reagents for testing candidate biomarkers of IGF activity in cancer and B) to carry out an unbiased genomic screen followed by validation in clinical samples for identification of IGF signatures. To do this we used proprietary disease-specific arrays and genomic data sets available in Almac as part of their proven capability in validating clinically important signatures in cancer. We also developed an isogenic ovarian cancer cell model for IGF-1 signaling and used cell models, gene lists and candidate IGF-1R-associated proteins that were available at UCC. The project was carried out by two experienced researchers recruited to UCC. They interacted closely with four early stage researchers and four experienced researchers who were seconded to Almac. They were supervised by the experimental medicine research team at Almac..
Results of the project:
In the first approach to the project (section A) candidate biomarkers from a panel of IGF-1R regulated genes that had been identified in UCC was carried out in the Almac ovarian cancer-associated-dataset. From this analysis, the PDLIM2 gene appeared to be the only IGF-1 regulated gene to be associated with poor survival, although no significant association between PDLIM2 with clinical parameters was found. PDLIM2 is a PDZ-LIM domain protein that shuttles between the cytoskeleton and the nucleus to regulate transcription factors stability, and thereby cell phenotype. High PDLIM2 levels are associated with poor survival in Triple Negative breast cancers. A panel of anti-PDLIM2 specific antibodies for detecting its expression and location in cells and tissues were generated (see Figure 1), and PDLIM2 activity was further explored in breast cancer in a series of cell and murine models. These studies demonstrated that suppression of PDLIM2 was found to disrupt normal breast epithelium polarity and also to prevent polarized cell migration and an invasive phenotype in breast cancer cell lines. An orthotopic model of breast cancer metastasis indicated that primary and secondary tumour formation was reduced in the absence of PDLIM2 expression.
In the second approach (section B) of the project we generated two pairs of isogenic ovarian cell lines with high or low IGF-1R expression. RNA microarray profiling was carried out on these, and differentially expressed genes were identified and used to generate a gene list that might represent a true IGF signalling pathway in ovarian cancer. These genes were further explored by extensive bioinformatic analysis of the Almac ovarian clinical sample dataset , but we could not establish the existence of significant clusters of patients that can be grouped using this list mostly due to technical factors associated with the tissues. However, this gene list remains available for analysis in other clinical data sets. The IGF isogenic cell models were also used to identify mediators of resistance to IGF-1R inhibition using phosphoprotein arrays. This identified a subset of receptor and cellular kinases that mediate resistance to and cooperate with IGF signalling. Candidates were identified, and will be further tested for contribution to drug resistance.
Impact of the project:
The BiomarkerIGF project has enabled a valuable intersectoral collaboration in an area of cancer research that has the potential to impact many people. The IGF system is active in a large number of cancers. Moreover IGF signaling and its regulation by genetic, environmental and cellular factors is critical to maintaining good health and longevity. Thus, identifying biomarkers and modulators of this pathway has potential to impact on healthy ageing in a large segment of the population.
The project has produced a valuable source of reagents and data on biomarkers of IGF signaling and IGF-1R activity in cancer cells. Further clinical utilities of data and reagents will be exploited in further collaborative research programs. Importantly, the project has contributed significantly to the scientific knowledge and discussion in IGF biology and has enabled a valuable collaboration between industry, clinical and basic researchers in this field that will allow future interaction.
The Intersect oral training for ESRs and ERs has had a very positive impact on all of the researchers associated with the project with all of them continuing in research (three in Industry, three in academia in Ireland, one in US and two in Europe). The results of the project have already been and will be presented at meetings and conferences in Europe, USA and Asia, highlighting the research and Marie Curie actions. A snapshot of the Biomarker-IGF was featured on the Horizon 2020 website https://ec.europa.eu/programmes/horizon2020/en/news/laying-down-markers-future-cancer-treatmentsAll of the secondees gained valuable training in bioinformatics, genomic approaches to biomarker development, functional screens, clinical validation of biomarkers and of generally working in Industry. Six of the secondees returned to UCC for further training and two were recruited directly to employment in Industry.
Contact: Prof. Rosemary O’Connor, University College Cork. r.oconnor@ucc.ie
Background and approaches taken:
The Biomarker-IGF project addressed the problem of developing clinically relevant biomarkers for the Insulin-like Growth Factor (IGF) signalling pathway in cancer. Activity of this growth and survival signaling pathway in cells is strongly associated with cancer development and progression. In the last 10-15 years many researchers and pharmaceutical companies have tested two kinds of agents that inhibit the signalling pathway by targeting the IGF-I Receptor on cancer cells. These agents displayed great promise in pre-clinical testing in cell models and xenograft tumour models. However, with the exception of some notable sporadic responses the results of clinical trials in a range of cancers have overall been disappointing, and further testing was stopped in many cases. These clinical results highlighted the urgent need for biomarkers that measure IGF-1R activity in cancer, principally to select patients that might benefit most from IGF-1R inhibition. Moreover, it became apparent that the IGF-1R participates in adaptive resistance to inhibitors of its own signalling pathway as well as to other drugs. A number of key challenges have become apparent in developing ways to effectively target the IGF-1R in cancer. In general, there may be no relationship between levels of IGF-1R expression, its activity level, activation of the signaling pathway, or the response to IGF-1R inhibitors. Thus, there is no reliable method to predict which patients or kinds of cancer will respond to IGF-1R inhibitors.
The Biomarker-IGF project took two approaches to addressing this problem: A) to develop, models and reagents for testing candidate biomarkers of IGF activity in cancer and B) to carry out an unbiased genomic screen followed by validation in clinical samples for identification of IGF signatures. To do this we used proprietary disease-specific arrays and genomic data sets available in Almac as part of their proven capability in validating clinically important signatures in cancer. We also developed an isogenic ovarian cancer cell model for IGF-1 signaling and used cell models, gene lists and candidate IGF-1R-associated proteins that were available at UCC. The project was carried out by two experienced researchers recruited to UCC. They interacted closely with four early stage researchers and four experienced researchers who were seconded to Almac. They were supervised by the experimental medicine research team at Almac..
Results of the project:
In the first approach to the project (section A) candidate biomarkers from a panel of IGF-1R regulated genes that had been identified in UCC was carried out in the Almac ovarian cancer-associated-dataset. From this analysis, the PDLIM2 gene appeared to be the only IGF-1 regulated gene to be associated with poor survival, although no significant association between PDLIM2 with clinical parameters was found. PDLIM2 is a PDZ-LIM domain protein that shuttles between the cytoskeleton and the nucleus to regulate transcription factors stability, and thereby cell phenotype. High PDLIM2 levels are associated with poor survival in Triple Negative breast cancers. A panel of anti-PDLIM2 specific antibodies for detecting its expression and location in cells and tissues were generated (see Figure 1), and PDLIM2 activity was further explored in breast cancer in a series of cell and murine models. These studies demonstrated that suppression of PDLIM2 was found to disrupt normal breast epithelium polarity and also to prevent polarized cell migration and an invasive phenotype in breast cancer cell lines. An orthotopic model of breast cancer metastasis indicated that primary and secondary tumour formation was reduced in the absence of PDLIM2 expression.
In the second approach (section B) of the project we generated two pairs of isogenic ovarian cell lines with high or low IGF-1R expression. RNA microarray profiling was carried out on these, and differentially expressed genes were identified and used to generate a gene list that might represent a true IGF signalling pathway in ovarian cancer. These genes were further explored by extensive bioinformatic analysis of the Almac ovarian clinical sample dataset , but we could not establish the existence of significant clusters of patients that can be grouped using this list mostly due to technical factors associated with the tissues. However, this gene list remains available for analysis in other clinical data sets. The IGF isogenic cell models were also used to identify mediators of resistance to IGF-1R inhibition using phosphoprotein arrays. This identified a subset of receptor and cellular kinases that mediate resistance to and cooperate with IGF signalling. Candidates were identified, and will be further tested for contribution to drug resistance.
Impact of the project:
The BiomarkerIGF project has enabled a valuable intersectoral collaboration in an area of cancer research that has the potential to impact many people. The IGF system is active in a large number of cancers. Moreover IGF signaling and its regulation by genetic, environmental and cellular factors is critical to maintaining good health and longevity. Thus, identifying biomarkers and modulators of this pathway has potential to impact on healthy ageing in a large segment of the population.
The project has produced a valuable source of reagents and data on biomarkers of IGF signaling and IGF-1R activity in cancer cells. Further clinical utilities of data and reagents will be exploited in further collaborative research programs. Importantly, the project has contributed significantly to the scientific knowledge and discussion in IGF biology and has enabled a valuable collaboration between industry, clinical and basic researchers in this field that will allow future interaction.
The Intersect oral training for ESRs and ERs has had a very positive impact on all of the researchers associated with the project with all of them continuing in research (three in Industry, three in academia in Ireland, one in US and two in Europe). The results of the project have already been and will be presented at meetings and conferences in Europe, USA and Asia, highlighting the research and Marie Curie actions. A snapshot of the Biomarker-IGF was featured on the Horizon 2020 website https://ec.europa.eu/programmes/horizon2020/en/news/laying-down-markers-future-cancer-treatmentsAll of the secondees gained valuable training in bioinformatics, genomic approaches to biomarker development, functional screens, clinical validation of biomarkers and of generally working in Industry. Six of the secondees returned to UCC for further training and two were recruited directly to employment in Industry.
Contact: Prof. Rosemary O’Connor, University College Cork. r.oconnor@ucc.ie