Our high-content screening offering involves exposing cells to chemical (e.g. compound libraries) or biological molecules (e.g. RNAi or antibody) and looking for changes in cell morphology, protein levels and/or localization by image capture and analysis.

A key aspect of compound profiling is the ability to study a target gene in its endogenous context. With our rAAV GENESIS™ gene-editing program, we have created an extensive menu of genetically-defined X-MAN™ cell lines including many that contain endogenous pathway reporters that allow researchers to follow changes in protein levels/localization. We can also support customer research efforts in this area through specialist 2D and 3D cell-based assay development and compound profiling services. 

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

If your cell line of interest is not available consider the generation
of a custom cell line  

Publications

• Kohli, M. et al, SMAC/Diablo-dependent apoptosis induced by nonsteroidal antiinflammatory drugs (NSAIDs) in colon cancer cells

Posters

• Astley H. et al, X MAN™ reporter cell lines: Enabling the study of endogenous promoter activity and protein dynamics
• Morrill, P. et al, New Cellular Reporter Technologies Using X-MAN™ Cell Models

Synthetic lethality is a process that takes advantage of cellular mechanisms whereby the loss of any one of two genes can be compensated by the normal function of the other. By having one gene non-functional a range of other genes can be explored by rendering them non-functional through drug inhibition. When cell death occurs, it indicates that the siRNA or shRNA was interfering with a critical pathway partner of the non-functional gene, suggesting these are potential gene targets for future therapeutics.

We have developed a large panel of genetically-defined X-MAN™ cell lines that can be deployed in cell-based phenotypic screens with siRNA and shRNA libraries designed to represent the drugable genome. The SyntheTx platform can be accessed by partners in a collaborative research or fee for service basis.

Learn more about our target identification platform  

Application Notes

• Compound screening in BRCA2 null XMAN cell lines: A model for synthetic lethality (PDF)
• Identifying new synthetically lethal therapeutic targets (PDF)

Publications

• Foster, R. et al, Multiple Metabolic Alterations Exist in Mutant PI3K Cancers, but Only Glucose Is Essential as a Nutrient Source
• Oh, S. et al, Human LIGIV is synthetically lethal with the loss of Rad54B-dependent recombination and is required for certain chromosome fusion events induced by telomere dysfunction
• Brough, R. et al, Functional Viability Profiles of Breast Cancer

• Li, H.F. et al, A high-throughput screen with isogenic PTEN+/+ and PTEN-/- cells identifies CID1340132 as a novel compound that induces apoptosis in PTEN and PIK3CA mutant human cancer cells
• Mendes-Pereira, A.M. et al , Synthetic lethal targeting of PTEN mutant cells with PARP inhibitors
• Sur, S. et al, A panel of isogenic human cancer cells suggests a therapeutic approach for cancers with inactivated p53
• Torrance, C.J. et al, Use of isogenic human cancer cells for high-throughput screening and drug discovery

  See more...

Posters

• Tsuji, T. et al, Development of a 3-dimensional synthetic lethality screening approach targeting KRas-mut cells
• Howes, R., Simple and precise targeted editing of the human genome using rAAV-mediated homologous recombination

Comparative profiling compares multiple samples, often differentiated by disease state, to identify the molecular factors (genetic or epigenetic) which differ between the samples. These genes or proteins can be candidates for therapeutic development.

Multiple samples will come from different genetic backgrounds and it can be difficult to determine precisely which changes are associated with the disease state.

Introducing the genetic changes in a clean genetic background,  as can be achieved using rAAV GENESIS™ in creating genetically-defined X-MAN™ cell lines, allows the investigation of the specific changes associated with a precise genetic change using genomic, proteomic, metabolomic and other approaches.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Publications

• Foster, R. et al, Multiple Metabolic Alterations Exist in Mutant PI3K Cancers, but Only Glucose Is Essential as a Nutrient Source
• Di Nicolantonio, F. et al, Isogenic mutant human cells: a new tool for personalized cancer medicine
• Fattah, F. et al, Ku regulates the non-homologous end joining pathway choice of DNA double-strand break repair in human somatic cells

• Fattah, K.R. et al, Mutations to Ku reveal differences in human somatic cell lines
• Grim, J.E. et al, Isoform- and cell cycle-dependent substrate degradation by the Fbw7 ubiquitin ligase
• Arena, S. et al, Knock-in of oncogenic Kras does not transform mouse somatic cells but triggers a transcriptional response that classifies human cancers
• Arena, S. et al, Genetic targeting of the kinase activity of the Met receptor in cancer cells

  See more...

Posters

• Griffin, S. et al, Horizon Discovery X-MAN™ cell lines reveal how PI3K mutation can result in EMT switch and increased invasiveness
• Wiggins, C. et al, Using Rac1 isogenics to model tumour metastasis
• Christopherson, C. et al, Differential impact of gefitinib and PLX4720 on proliferation of MCF10A and isogenic lines as measured with a metastasis expression score

• Goodall, J.E. et al, Isogenic K-Ras mutant cancer cells: A novel platform for drug profiling
• Rigby, S. et al, The use of PI3K isogenic cell lines in conditions that model the tumour microenvironment provides a relevant system in which to test PI3K inhibitor profiles

  See more...

Biological processes are orchestrated by many specific cellular pathways which consist of interacting genes and proteins. In any given disease state, many of these pathways are altered and the ability to understand these pathways and how the various proteins within it interact can lead to the identification of novel ways to intervene in the pathway signalling.

A crucial element for pathway analysis is that interactions between members of the pathway are at the correct levels. Genetically-defined X-MAN™ cell lines can be created with rAAV GENESIS™ that have target genes tagged with pathway and transcriptional reports at their endogenous loci resulting in relevant expression levels of the protein and interactions with other members of the pathway to be measured. These tagged-proteins can be used to monitor activity of the pathway as well as identifying directly interacting proteins.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Profiling ALK inhibition in Cell Lines Harbouring Genomic ALK Alterations (PDF)
• Characterization of IDH mutations in X-MAN™ isogenic cell lines (PDF)
• Pathway activation analysis in X-MAN™ SW48 K-Ras cell lines: A tool for predicting patient response (PDF)

• CEM2n APOBEC3 X-MAN™ Cell Lines: New model systems for HIV & host factor biology (PDF)

  See more...

Publications

• Wang, G. et al, Single copies of mutant KRAS and mutant PIK3CA cooperate in immortalized human epithelial cells to induce tumor formation
• Grassian, A. et al, Isocitrate Dehydrogenase (IDH) Mutations Promote a Reversible ZEB1/mir-200-Dependent Epithelial-Mesenchymal (EMT) Transition
• Wallin, J.J. et al, Active PI3K pathway causes an invasive phenotype which can be reversed or promoted by blocking the pathway at divergent nodes

• Ericson, K. et al, Genetic inactivation of AKT1, AKT2, and PDPK1 in human colorectal cancer cells clarifies their roles in tumor growth regulation
• Fattah, F. et al, Ku regulates the non-homologous end joining pathway choice of DNA double-strand break repair in human somatic cells
• Lauring, J. et al, Knock in of the AKT1 E17K mutation in human breast epithelial cells does not recapitulate oncogenic PIK3CA mutations
• Vitolo, M.I. et al, Deletion of PTEN promotes tumorigenic signaling, resistance to anoikis, and altered response to chemotherapeutic agents in human mammary epithelial cells
• Grim, J.E. et al, Isoform- and cell cycle-dependent substrate degradation by the Fbw7 ubiquitin ligase
• Benvenuti, S. et al, Identification of cancer genes by mutational profiling of tumor genomes
• Samuels, Y. et al, Mutant PIK3CA promotes cell growth and invasion of human cancer cells
• Kohli, M. et al, SMAC/Diablo-dependent apoptosis induced by nonsteroidal antiinflammatory drugs (NSAIDs) in colon cancer cells

  See more...

Posters

• Astley H. et al, X MAN™ reporter cell lines: Enabling the study of endogenous promoter activity and protein dynamics
• Griffin, S. et al, The use of novel endogenous reporter technology in large scale siRNA library screens provides a simple system for rapid identification of potential drug targets
• Mudd C. et al, 3 Dimensional growth reveals KRAS dependency

• Grooby, S. et al, X-MAN™ NanoLuc™ Reporter Cell Lines: Applications to drug-discovery
• Kopish, K. et al, NanoLuc™: A Smaller, Brighter, and More Versatile Luciferase Reporter
• Morrill, P. et al, New Cellular Reporter Technologies Using X-MAN™ Cell Models
• Wiggins, C. et al, Using Rac1 isogenics to model tumour metastasis
• Foster, R.E. et al, Profiling the metabolic dependencies of PI3Kα mutant cancers
• Goodall, J.E. et al, The use of X-MAN™ isogenic cell lines to define PI3-kinase inhibitor activity profiles
• Guan, J. et al, Endogenous expression of an oncogenic PI3K mutation leads to activated PI3K pathway signalling and an invasive phenotype

  See more...

Many novel targets for drug discovery are identified by knocking down their expression with siRNA or shRNA’s.  A standard approach to show that the effect is specific to the gene in which expression is reduced is to rescue the effect by exogenous expression of the cDNA of the target gene. This is not always successful as it is difficult to achieve the correct levels of expression of the cDNA.

The binding site of the siRNA/shRNA can easily be identified and we can use rAAV GENESIS™ to create X-MAN™ cell lines that insert multiple point mutations at this binding site to prevent binding of the siRNA/shRNA but not alter the open reading frame of the gene. This will create an allele which is resistant to the effects of degradation. As this allele is expressed at endogenous levels it provides an ideal method to rescue the effects of a siRNA/shRNA and hence validate the target gene. 

Alternatively, rAAV GENESIS™ can be used to knock-in activating mutations or knock-out of target to look for phenotype/biomarkers, and to engineer kinase dead cell lines to specifically address whether kinase activity of a putative target is critical.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our target identification platform  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Characterization of IDH mutations in X-MAN™ isogenic cell lines (PDF)

Publications

• Wang, G. et al, Single copies of mutant KRAS and mutant PIK3CA cooperate in immortalized human epithelial cells to induce tumor formation
• Grassian, A. et al, Isocitrate Dehydrogenase (IDH) Mutations Promote a Reversible ZEB1/mir-200-Dependent Epithelial-Mesenchymal (EMT) Transition
• Prahallad, A. et al, Unresponsiveness of colon cancer to BRAF(V600E) inhibition through feedback activation of EGFR

• Brough, R. et al, Functional Viability Profiles of Breast Cancer
• Konishi, H. et al, Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells
• Weiss, M.B. et al, Deletion of p53 in human mammary epithelial cells causes chromosomal instability and altered therapeutic response
• Wang, Y. et al, Ku86 represses lethal telomere deletion events in human somatic cells
• Arena, S. and Bardelli, A., Understanding how kinase-targeted therapies work
• Fattah, F. et al, Ku70, an essential gene, modulates the frequency of rAAV-mediated gene targeting in human somatic cells
• Cummins, J.M. et al, The colorectal microRNAome
• Ghosh, G. et al, The lethality of Ku86 (XRCC5) loss-of-function mutations in human cells is independent of p53 (TP53)
• Samuels, Y. et al, Mutant PIK3CA promotes cell growth and invasion of human cancer cells
• Bachman, K.E. et al, The PIK3CA gene is mutated with high frequency in human breast cancers
• Kohli, M. et al, SMAC/Diablo-dependent apoptosis induced by nonsteroidal antiinflammatory drugs (NSAIDs) in colon cancer cells

  See more...

Posters

• Sun, Y. et al, Utilization of in vivo human isogenic cancer models in the new era of targeted therapies
• Foster, R.E. et al, Investigation into the influence of the PI3Kα (H1047R) mutation on the bioenergetic dependency of a cell
• Foster, R.E. et al, Profiling the metabolic dependencies of PI3Kα mutant cancers

• Guan, J. et al, Endogenous expression of an oncogenic PI3K mutation leads to activated PI3K pathway signalling and an invasive phenotype

  See more...

The complex microenvironment that surrounds a solid tumor can impact not only gene and protein expression but also affect the way drugs will behave. By manipulating in vitro assay conditions, different aspects of the tumor microenvironment can be examined.

X-MAN™ cell lines contain disease-relevant mutations at their endogenous loci. These endogenous mutations can be placed under selective pressure to illicit true disease phenotypes & drug responses. We have developed a large panel of 2D and 3D phenotypic assays and have the specialist equipment and knowledge that allow evaluation of aspects of the tumor microenvironment.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• The use of PI3K XMAN cell lines in conditions that model the tumour microenvironment (PDF)
• A multicellular tumour model: Applications for evaluating drug signalling pathways in a 3D system (PDF)

Publications

• Bottos, A. et al, Targeting oncogenic serine/threonine-protein kinase BRAF in cancer cells inhibits angiogenesis and abrogates hypoxia
• Deyle, D.R. et al, Normal Collagen and Bone Production by Gene-targeted Human Osteogenesis Imperfecta iPSCs
• Foster, R. et al, Multiple Metabolic Alterations Exist in Mutant PI3K Cancers, but Only Glucose Is Essential as a Nutrient Source

• Wallin, J.J. et al, Active PI3K pathway causes an invasive phenotype which can be reversed or promoted by blocking the pathway at divergent nodes
• Ericson, K. et al, Genetic inactivation of AKT1, AKT2, and PDPK1 in human colorectal cancer cells clarifies their roles in tumor growth regulation
• Arena, S. et al, Genetic targeting of the kinase activity of the Met receptor in cancer cells

  See more...

Posters

• Bartholomeusz, G. et al, A 3D high throughput RNAi screen identifies the TLR4 kinase which alters spheroid architecture: A promising therapeutic stratergy for pancreatic cancer
• Mudd C. et al, 3 Dimensional growth reveals KRAS dependency
• Tsuji, T. et al, Development of a 3-dimensional synthetic lethality screening approach targeting KRas-mut cells

• Griffin, S. et al, Horizon Discovery X-MAN™ cell lines reveal how PI3K mutation can result in EMT switch and increased invasiveness
• Foster, R.E. et al, Investigation into the influence of the PI3Kα (H1047R) mutation on the bioenergetic dependency of a cell
• Foster, R.E. et al, Development of a multicellular tumour spheroid model to study the hypoxic tumour microenvironment
• Goodall, J.E. et al, The use of X-MAN™ isogenic cell lines to define PI3-kinase inhibitor activity profiles
• Rigby, S. et al, The use of PI3K isogenic cell lines in conditions that model the tumour microenvironment provides a relevant system in which to test PI3K inhibitor profiles
• Foster, R.E. et al, The Effects of Hypoxia on the Sensitivity of Glioma Cells to Gemcitabine Treatment

  See more...

When trying to address specific research questions it is critical to use not only patient relevant cell line models but also to apply the most appropriate assay format. 

Genetically-defined human disease models, such as X-MAN™ cell lines can be used across many in vitro assay formats, and in some instances particular assay formats may be required in order to reveal the specific phenotypic consequence of a genetic alteration.  Extensive in vitro assay development expertise can be accessed via Horizon’s contract research group on a fee for service basis.  As well an existing panel of optimized assays, we also have experience with bespoke assay development.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Immunofluorescent microscopy: Applications for secondary profiling of compounds that target the cytoskeleton (PDF)
• MCF10A XMAN cell lines endogenously expressing mutant or wild type PI3K; A model system for screening mTOR targeted agents (PDF)
• Compound Screening in DNA Repair X-MAN Cell Lines (PDF)

Publications

• Deyle, D.R. et al, Normal Collagen and Bone Production by Gene-targeted Human Osteogenesis Imperfecta iPSCs
• Foster, R. et al, Multiple Metabolic Alterations Exist in Mutant PI3K Cancers, but Only Glucose Is Essential as a Nutrient Source
• Ericson, K. et al, Genetic inactivation of AKT1, AKT2, and PDPK1 in human colorectal cancer cells clarifies their roles in tumor growth regulation

• Vitolo, M.I. et al, Deletion of PTEN promotes tumorigenic signaling, resistance to anoikis, and altered response to chemotherapeutic agents in human mammary epithelial cells
• Arena, S. et al, Genetic targeting of the kinase activity of the Met receptor in cancer cells

  See more...

Posters

• Astley H. et al, X MAN™ reporter cell lines: Enabling the study of endogenous promoter activity and protein dynamics
• Bartholomeusz, G. et al, A 3D high throughput RNAi screen identifies the TLR4 kinase which alters spheroid architecture: A promising therapeutic stratergy for pancreatic cancer
• Mudd C. et al, 3 Dimensional growth reveals KRAS dependency

• Tsuji, T. et al, Development of a 3-dimensional synthetic lethality screening approach targeting KRas-mut cells
• Griffin, S. et al, Horizon Discovery X-MAN™ cell lines reveal how PI3K mutation can result in EMT switch and increased invasiveness
• Howes, R., Simple and precise targeted editing of the human genome using rAAV-mediated homologous recombination
• Foster, R.E. et al, Investigation into the influence of the PI3Kα (H1047R) mutation on the bioenergetic dependency of a cell
• Foster, R.E. et al, Profiling the metabolic dependencies of PI3Kα mutant cancers
• Foster, R.E. et al, Development of a multicellular tumour spheroid model to study the hypoxic tumour microenvironment
• Goodall, J.E. et al, The use of X-MAN™ isogenic cell lines to define PI3-kinase inhibitor activity profiles
• Guan, J. et al, Endogenous expression of an oncogenic PI3K mutation leads to activated PI3K pathway signalling and an invasive phenotype
• Rigby, S. et al, The use of PI3K isogenic cell lines in conditions that model the tumour microenvironment provides a relevant system in which to test PI3K inhibitor profiles
• Foster, R.E. et al, Investigation into the mechanism of action of the ruthenium(II) organometallic complex, ONCO 4417

  See more...

A number of cases, particularly in oncology, have now been identified where a patient’s genetics can impact the efficacy and safety of therapeutics. A key example of this is the variable response of patients with K-Ras mutations to Erbitux and Vectibix, two drugs that target EGFR mutant patient populations in colorectal cancer.  Particularly given the cost of such treatments, it is increasingly important to ensure that drugs are safe and effective and so genetic resistance mechanisms are increasingly being looked for.

Patient-relevant X-MAN™ isogenic cell models have been successful at predicting such genetic-based clinical outcomes for single and combinatorial treatment regimens, guiding focused project development, clinical trial design and the recruitment and marketing/co-marketing of more effective therapeutic regimens.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Characterizing cellular response to BRAF inhibitors in Isogenic Cell Lines (PDF)
• Investigating compound selectivity in EGFR mutant XMAN cell lines (PDF)
• Profiling patient responses: Not all K-Ras mutations are equal (PDF)

Publications

• Misale, S. et al, Emergence of KRAS mutations and acquired resistance to anti-EGFR therapy in colorectal cancer
• Prahallad, A. et al, Unresponsiveness of colon cancer to BRAF(V600E) inhibition through feedback activation of EGFR
• Higgins, M.J. et al, PIK3CA mutations and EGFR overexpression predict for lithium sensitivity in human breast epithelial cells

• Martini, M. et al, Targeted therapies: how personal should we go?
• Rosa, R. et al, Toll-like Receptor 9 Agonist IMO Cooperates with Cetuximab in K-Ras Mutant Colorectal and Pancreatic Cancers
• Bardelli, A. and Siena, S., Molecular mechanisms of resistance to cetuximab and panitumumab in colorectal cancer
• Di Nicolantonio, F. et al, Deregulation of the PI3K and KRAS signaling pathways in human cancer cells determines their response to everolimus
• Mohseni, M. and Park, B.H., PIK3CA and KRAS mutations predict for response to everolimus therapy: now that's RAD001
• Vitolo, M.I. et al, Deletion of PTEN promotes tumorigenic signaling, resistance to anoikis, and altered response to chemotherapeutic agents in human mammary epithelial cells
• Di Nicolantonio, F. et al, Replacement of normal with mutant alleles in the genome of normal human cells unveils mutation-specific drug responses
• Di Nicolantonio, F. et al, Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer
• Benvenuti, S. et al, Oncogenic activation of the RAS/RAF signaling pathway impairs the response of metastatic colorectal cancers to anti-epidermal growth factor receptor antibody therapies
• Bachman, K.E. et al, p21(WAF1/CIP1) mediates the growth response to TGF-beta in human epithelial cells
• Bunz, F. et al, Disruption of p53 in human cancer cells alters the responses to therapeutic agents

  See more...

Posters

• Bartholomeusz, G. et al, A 3D high throughput RNAi screen identifies the TLR4 kinase which alters spheroid architecture: A promising therapeutic stratergy for pancreatic cancer
• Howes, R., Simple and precise targeted editing of the human genome using rAAV-mediated homologous recombination
• Goodall, J.E. et al, Isogenic K-Ras mutant cancer cells: A novel platform for drug profiling

• Rigby, S. et al, The use of PI3K isogenic cell lines in conditions that model the tumour microenvironment provides a relevant system in which to test PI3K inhibitor profiles

  See more...

Careful study of genomic markers is causing clinical data to be re-examined to understand whether there are subtleties in patient responses to drugs. This is allowing drugs to be re-profiled for new patient populations or indications.  An example is Lithium Chloride, for use in breast cancer types carrying the mutant PI3K gene. 

X-MAN™ cell lines are being used as a powerful and definitive system to explore and predict patient drug-sensitivity and resistance profiles based on their genetics.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Drug Repurposing: LiCl as PI3K mutant selective inhibitor (PDF)

Publications

• Prahallad, A. et al, Unresponsiveness of colon cancer to BRAF(V600E) inhibition through feedback activation of EGFR
• De Roock, W. et al, Association of KRAS p.G13D mutation with outcome in patients with chemotherapy-refractory metastatic colorectal cancer treated with cetuximab
• Gustin, J.P. et al, Knockin of mutant PIK3CA activates multiple oncogenic pathways

• Mendes-Pereira, A.M. et al , Synthetic lethal targeting of PTEN mutant cells with PARP inhibitors
• Di Nicolantonio, F. et al, Replacement of normal with mutant alleles in the genome of normal human cells unveils mutation-specific drug responses

  See more...

Compound screening is a widely-used approach to drug discovery involving the performance of chemical, genetic or pharmacological tests in parallel on compound libraries. Through this process researchers can quickly identify active compounds, antibodies or genes which modulate a particular biomolecular pathway, providing a starting point for the identification of promising lead compounds. Compound screening of large numbers of compounds is known as High Throughput Screening.

Genetically-defined X-MAN™ cell lines can be deployed in these assays to identify novel drug-candidates that selectively impact a chosen genotype. Moreover, by using isogenic cell lines researchers will inherently understand if altering that gene has selective effects on a disease vs. normal cell type – thus feeding the drug discovery process with candidates that have higher chance of success. At Horizon, our screening lab can handle small to medium sized compound (small molecule or RNAi) screening projects on a fee for service basis.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• The Evaluation of Drug Combinations (PDF)
• Profiling ALK inhibition in Cell Lines Harbouring Genomic ALK Alterations (PDF)
• Characterizing cellular response to BRAF inhibitors in Isogenic Cell Lines (PDF)

• Investigating compound selectivity in EGFR mutant XMAN cell lines (PDF)
• DLD-1 p53 XMAN cell lines as a model system for screening checkpoint targeted agents (Chk1) (PDF)
• MCF10A XMAN cell lines endogenously expressing mutant or wild type PI3K; A model system for screening mTOR targeted agents (PDF)
• The use of PI3K XMAN cell lines in conditions that model the tumour microenvironment (PDF)
• A multicellular tumour model: Applications for evaluating drug signalling pathways in a 3D system (PDF)
• Compound screening in BRCA2 null XMAN cell lines: A model for synthetic lethality (PDF)
• Pathway activation analysis in X-MAN™ SW48 K-Ras cell lines: A tool for predicting patient response (PDF)
• Profiling patient responses: Not all K-Ras mutations are equal (PDF)
• Compound Screening in DNA Repair X-MAN Cell Lines (PDF)

  See more...

Publications

• Li, H.F. et al, A high-throughput screen with isogenic PTEN+/+ and PTEN-/- cells identifies CID1340132 as a novel compound that induces apoptosis in PTEN and PIK3CA mutant human cancer cells
• De Roock, W. et al, Effects of KRAS, BRAF, NRAS, and PIK3CA mutations on the efficacy of cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal cancer: a retrospective consortium analysis
• Sur, S. et al, A panel of isogenic human cancer cells suggests a therapeutic approach for cancers with inactivated p53

• Torrance, C.J. et al, Use of isogenic human cancer cells for high-throughput screening and drug discovery

  See more...

Posters

• Lowe, C. et al, X-MAN™ NanoLuc™ Reporter Cell Lines: Tools for High Throughput Screening
• Grooby, S. et al, X-MAN™ NanoLuc™ Reporter Cell Lines: Applications to drug-discovery
• Kopish, K. et al, NanoLuc™: A Smaller, Brighter, and More Versatile Luciferase Reporter

• Goodall, J.E. et al, Isogenic K-Ras mutant cancer cells: A novel platform for drug profiling

  See more...

Patient stratification involves identifying the subsets within a patient population that respond differently to a given drug. Stratification can be a critical component to transforming a clinical trial from a negative or neutral outcome to one with a positive outcome by identifying the subset of the population most likely to respond to a novel therapy.

A panel of genetically defined X-MAN™ cell lines covering a wide range of human disease genotypes enables drug discovery researchers to determine the optimal patient populations to target with their drug candidates in the expensive clinical trial stage of the drug development process.

The availability of such data enables researchers to determine the full range of sensitive and resistant patient populations to the disease target in clinical trials. This will enable the design of smaller more efficient clinical trials that read-out quicker and be more likely to succeed and reduce the high-rate of attrition currently observed at this critical phase of drug development.

Biomarker discovery and drug profiling can be performed on a fee for service basis.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Characterizing cellular response to BRAF inhibitors in Isogenic Cell Lines (PDF)
• Investigating compound selectivity in EGFR mutant XMAN cell lines (PDF)
• Pathway activation analysis in X-MAN™ SW48 K-Ras cell lines: A tool for predicting patient response (PDF)

• Profiling patient responses: Not all K-Ras mutations are equal (PDF)
• CEM2n APOBEC3 X-MAN™ Cell Lines: New model systems for HIV & host factor biology (PDF)

  See more...

Publications

• Tejpar, S. et al, Association of KRAS G13D Tumor Mutations With Outcome in Patients With Metastatic Colorectal Cancer Treated With First-Line Chemotherapy With or Without Cetuximab
• Brough, R. et al, Functional Viability Profiles of Breast Cancer
• Higgins, M.J. et al, PIK3CA mutations and EGFR overexpression predict for lithium sensitivity in human breast epithelial cells

• Martini, M. et al, Targeted therapies: how personal should we go?
• De Roock, W. et al, Effects of KRAS, BRAF, NRAS, and PIK3CA mutations on the efficacy of cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal cancer: a retrospective consortium analysis
• De Roock, W. et al, Association of KRAS p.G13D mutation with outcome in patients with chemotherapy-refractory metastatic colorectal cancer treated with cetuximab
• Di Nicolantonio, F. et al, Isogenic mutant human cells: a new tool for personalized cancer medicine
• Di Nicolantonio, F. et al, Deregulation of the PI3K and KRAS signaling pathways in human cancer cells determines their response to everolimus
• Lauring, J. et al, Knock in of the AKT1 E17K mutation in human breast epithelial cells does not recapitulate oncogenic PIK3CA mutations
• Mohseni, M. and Park, B.H., PIK3CA and KRAS mutations predict for response to everolimus therapy: now that's RAD001
• Mendes-Pereira, A.M. et al , Synthetic lethal targeting of PTEN mutant cells with PARP inhibitors
• Vitolo, M.I. et al, Deletion of PTEN promotes tumorigenic signaling, resistance to anoikis, and altered response to chemotherapeutic agents in human mammary epithelial cells
• Di Nicolantonio, F. et al, Replacement of normal with mutant alleles in the genome of normal human cells unveils mutation-specific drug responses
• Di Nicolantonio, F. et al, Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer
• Benvenuti, S. et al, Oncogenic activation of the RAS/RAF signaling pathway impairs the response of metastatic colorectal cancers to anti-epidermal growth factor receptor antibody therapies
• Bleeker, F.E. and Bardelli, A., Genomic landscapes of cancers: prospects for targeted therapies
• Disley, D., Impact of the new biology: From genomics to personalized medicine
• Benvenuti, S. et al, Identification of cancer genes by mutational profiling of tumor genomes
• Samuels, Y. et al, Mutant PIK3CA promotes cell growth and invasion of human cancer cells
• Bunz, F. et al, Disruption of p53 in human cancer cells alters the responses to therapeutic agents

  See more...

Posters

• Howes, R., Simple and precise targeted editing of the human genome using rAAV-mediated homologous recombination
• Rigby, S. et al, The use of PI3K isogenic cell lines in conditions that model the tumour microenvironment provides a relevant system in which to test PI3K inhibitor profiles

During drug development it is crucial to understand the mode of action of a novel drug. Secondary profiling seeks to understand the mode of action from activity of the drug on its primary target through to a response in a disease model.

By using a panel of genetically-defined X-MAN™ cell lines that contain disease-causing mutations at endogenous loci, the activity of a drug can be profiled as it is developed. This allows the determination of on vs. off-target activity of the drug and so desired activities can be incorporated into the final drug design. 

We can support these efforts on a fee for service basis via performance of detailed mechanism of action assays (e.g. pathway signaling, apoptosis, FACS, IF etc.) to evaluate the mode of action of a drug and we can study on/off target effects. 

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• Profiling ALK inhibition in Cell Lines Harbouring Genomic ALK Alterations (PDF)
• Immunofluorescent microscopy: Applications for secondary profiling of compounds that target the cytoskeleton (PDF)

Publications

• Bottos, A. et al, Targeting oncogenic serine/threonine-protein kinase BRAF in cancer cells inhibits angiogenesis and abrogates hypoxia
• Wallin, J.J. et al, Active PI3K pathway causes an invasive phenotype which can be reversed or promoted by blocking the pathway at divergent nodes
• Higgins, M.J. et al, PIK3CA mutations and EGFR overexpression predict for lithium sensitivity in human breast epithelial cells

• Bardelli, A. and Siena, S., Molecular mechanisms of resistance to cetuximab and panitumumab in colorectal cancer
• De Roock, W. et al, Effects of KRAS, BRAF, NRAS, and PIK3CA mutations on the efficacy of cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal cancer: a retrospective consortium analysis
• Arena, S. et al, Genetic targeting of the kinase activity of the Met receptor in cancer cells

  See more...

Posters

• Griffin, S. et al, The use of novel endogenous reporter technology in large scale siRNA library screens provides a simple system for rapid identification of potential drug targets
• Griffin, S. et al, Horizon Discovery X-MAN™ cell lines reveal how PI3K mutation can result in EMT switch and increased invasiveness
• Grooby, S. et al, X-MAN™ NanoLuc™ Reporter Cell Lines: Applications to drug-discovery

• Kopish, K. et al, NanoLuc™: A Smaller, Brighter, and More Versatile Luciferase Reporter
• Christopherson, C. et al, Differential impact of gefitinib and PLX4720 on proliferation of MCF10A and isogenic lines as measured with a metastasis expression score
• Buron, N. et al, Endogenous expression of an oncogenic PI3K mutation leads to accumulation of anti-apoptotic proteins at mitochondria
• Foster, R.E. et al, Investigation into the mechanism of action of the ruthenium(II) organometallic complex, ONCO 4417

  See more...

Many drugs entering the market have limited efficacy with resistance occurring after a few months of treatment. Combinations of drugs with well-understood modes of action will probably be the future of treatment and we will need to understand which combinations are most suited for specific disease genotypes especially for the treatment of resistant genotypes.

Creation of genetically-defined X-MAN™ cell lines, which contain the disease causing mutations at the endogenous locus, provide a clean system to investigate drug sensitivity, resistance and combinations of drugs which provide efficacy.   We can perform fee for service work looking at different combination assay formats:

• Simple potentiation assays where one compound in not effective alone
• Multiplexed formats to look at combinations of  two or more compounds that are individually effective, to see if synergistic when combined.

Learn more about our menu of X-MAN™ cell lines and X-MAN™ Reporter Kits  

Learn more about our contract research services  

If your cell line of interest is not available consider the generation
of a custom cell line  

Application Notes

• The Evaluation of Drug Combinations (PDF)
• DLD-1 p53 XMAN cell lines as a model system for screening checkpoint targeted agents (Chk1) (PDF)

Posters

• Goodall, J.E. et al, Isogenic K-Ras mutant cancer cells: A novel platform for drug profiling

Bioproduction is the production of biologics-based reagents (typically proteins) in a cell-based system at large volume. A small number of cell lines have been refined for this application, most notably CHO cells, yielding large quantities of high-quality protein.

Efforts are continually underway to further improve yield, quality and purity, and the current focus is on targeted gene editing of CHO cells.  Our rAAV GENESIS™ platform is ideal for development of custom CHO cell lines that modify productivity factors.  For example, there are many factors that can limit the ability to produce high levels of biologics including confounding enzymatic activities. We can remove the genes that encode for these limiting factors creating a cell line that can increase the yield/purity of the biologic produced.

Explore our custom Bioproduction cell line production service  

Application Notes

• CHO Cell Engineering for Bioproduction using rAAV (PDF)

Posters

• Kapp, J. et al, Use of rAAV-mediated homologous recombination to engineer CHO cells

Increasingly, drugs are being mandated to confirm through theranostics that patients must conform to specific genotypes before they are able to receive the drug in order to ensure safety and efficacy.

This has led to a range of molecular diagnostic tests being developed across a broad number of platforms all with the goal of screening patients.  In order to set up an assay in a lab and to make sure that the sensitivity and specificity of these tests is maintained, regular quality control is required.

We have generated a menu of off-the-shelf reference standards capable of being used as controls that are available in defined allelic ratios between mutant and wild-type forms and in gDNA and FFPE formats. 

Learn about Quantitative Molecular Reference Standards  

Application Notes

• Next Generation Sequencing: A highly Sensitive Test for Mutation Detection (PDF)
• DNA Extraction from Low Cell Number K-Ras G12D FFPE Samples (PDF)
• Genetically Defined B-Raf, K-Ras and EGFR Reference Standards used to validate the sensitivity of Droplet Digital PCR™ (PDF)

Posters

• Frampton, J. et al, Development of Quantitative B-Raf, EGFR, K-Ras and PI3Kα FFPE Molecular Reference Standards
• Howes, R., Simple and precise targeted editing of the human genome using rAAV-mediated homologous recombination