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Aquila BioMedical

Edinburgh, Midlothian, GB

Aquila BioMedical is an innovative pre-clinical contract research organisation that offers clients world-leading research expertise in Immuno-Oncology, Immunology and Multiplex Histology.

Bespoke services combine advanced models with protocolled techniques, to provide high-value data, defining both efficacy and the mechanism of action of drug candidate compounds. A key feature of the Aquila offering is the partnering of services with world-leading academics to provide expert advice and interpretation of the data.

Aquila has developed novel technologies to help you better understand the cellular and molecular events that occur with compound administration. Our assays allow both phenotypic screening and target based research methods to enable hit identification and optimisation of compound and target selection, directly increasing the compound success rate and reducing the overall cost associated with drug development.

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ADCC and CDC Assay Services
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Tumour Killing Assays

Understanding whether a test biologic or small molecule can accelerate tumour killing is a key component of drug development. Tumour cell killing by immune cell populations can be mediated either directly or as antibody-dependent cytotoxicity (ADCC).

Aquila uses the IncuCyte ZOOM® imaging system to... Show more »

Tumour Killing Assays

Understanding whether a test biologic or small molecule can accelerate tumour killing is a key component of drug development. Tumour cell killing by immune cell populations can be mediated either directly or as antibody-dependent cytotoxicity (ADCC).

Aquila uses the IncuCyte ZOOM® imaging system to provide an automated analysis. Tumour cells are seeded in a 96- well plate format incubated with immune cell populations. Incorporation of a caspase3/7-sensitive dye allows real-time visualization of tumour cell apoptosis, to complement measurements of their proliferative rate. Supernatants can also be collected to assess for levels of cytokines e.g. IFN-γ and cells can be analysed by flow cytometry.

The data shown below are derived from assays with an ovarian cancer cell line exposed to PBMC stimulated with anti-CD3. The system is adaptable to a range of tumour types and immune cell populations (T cells, NK cell, macrophages, neutrophils) and the inclusion of anti-TAA antibodies. More traditional label-release cytotoxicity assays are also available.

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Macrophage Suppression Assay
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M2-mediated Suppression of T Cell Responses:
Human M2 macrophages, differentiated in the presence of M-CSF and activated for a short period of time with LPS, are very potent suppressors of autologous T cell (PBMC) responses. In contrast, M1 macrophages boost activation of T cells. This assay tests the effects of macrophages on... Show more »

M2-mediated Suppression of T Cell Responses:
Human M2 macrophages, differentiated in the presence of M-CSF and activated for a short period of time with LPS, are very potent suppressors of autologous T cell (PBMC) responses. In contrast, M1 macrophages boost activation of T cells. This assay tests the effects of macrophages on autologous PBMC stimulated with anti-CD3 (+/- anti-CD28), using IFN-γ production as the routine readout. M2s suppress T cells via multiple mechanisms such as
PD-1 ligation and increased IL-10 expression. Addition of small molecule drugs capable of repolarizing M2 macrophages can significantly reverse M2-mediated suppression of T cells. This effect can work synergistically when combined with anti-PD-1.

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Immuno-oncology Assays
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Immuno-Oncology is a relatively new area of medicine that focuses on the manipulation of the immune response to effectively target tumours.

Until recently, immunotherapeutic agents had provided very limited evidence of clinical success; however, over the last decade, we have witnessed a paradigm shift in the treatment of... Show more »

Immuno-Oncology is a relatively new area of medicine that focuses on the manipulation of the immune response to effectively target tumours.

Until recently, immunotherapeutic agents had provided very limited evidence of clinical success; however, over the last decade, we have witnessed a paradigm shift in the treatment of cancer. Now, many think immunotherapy should be considered alongside surgery, chemotherapy and radiotherapy as the fourth cornerstone of anticancer treatment.

Although still in its infancy, immunotherapy has been yielding some promising clinical data from checkpoint modulators. In 2011, Ipilimumab, a fully human monoclonal antibody against cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4), became the first agent approved in the EU for the treatment of adult patients with un-resectable or metastatic melanoma. More recently, Nivolumab, an immunomodulatory compound that blocks the activation of program cell death 1 (PD-1), was approved for metastatic melanoma.

Understanding the immunomodulatory effects of your treatment(s) within the tumour microenvironment is of critical importance for progress on the path to the clinic, and questions on how these might play out in combination with other already marketed checkpoint modulators is exactly where Aquila can help.

Aquila has extensive in vitro and in vivo immunology, immuno-oncology and multiplex histology experience, which can be specifically tailored for any project. The suite of available immuno-oncology services and assays include but are not limited to:

• An Exhausted CD4+ T Cell Assay
• Human PBMC & T Cell Activation Assays,
• Human Mixed Lymphocyte Reactions,
• Human DC, Macrophage and Co-culture Assays,
• T Cell Killing Assay

All relevant to investigating compound mechanism of action for cancer immunotherapy.

Further human ex vivo patient assays are also in development and should be available later Q3 2017.

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T Cell Activation and Proliferation Assays
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Aquila can provide a range of in vitro human cell assays, tailored to address specific questions on novel compound effects and mechanism of action in the human immune system. The use of flow cytometry enables multiple physiologically relevant readouts from a given population of primary cells allowing phenotypic screening of... Show more »

Aquila can provide a range of in vitro human cell assays, tailored to address specific questions on novel compound effects and mechanism of action in the human immune system. The use of flow cytometry enables multiple physiologically relevant readouts from a given population of primary cells allowing phenotypic screening of compounds. Screening candidate compounds using short, predictive in vitro assays identifies those that show efficacy on target pathways, as well as demonstrating absence of unwanted off-target effects, consequently compounds have the potential to reach the clinic faster and with a greater chance of success.

The development of assays utilising PBMCs from healthy donors, as well as from patients with various diseases, allows for therapeutic investigation of normal and abnormal functions in human immune cells.

PBMCs or isolated T cells can be activated or polarised, with responses profiled following the application of a test drug in comparison to relevant reference controls.

Our assays include:

  • LPS-induced cytokine production
  • PBMC stimulation with anti CD3/28
  • T cells assays to evaluate effects on occurrence and/or activity of pathogenic T cell subsets (Th1, Th2 and Th17)

Read-outs include:

  • Cytokine production by ELISA/multiplex array (eg. IFN-γ, IL 17, IL-10 and more)
  • Proliferation (CFSE, Ki67 or thymidine incorporation)
  • Multi parameter flow cytometry (cytokines, transcription factors, activation markers, proliferation, viability)
  • Cytotoxicity assays (LDH)
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T-Cell Exhaustion Assay
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In cancer, the tumour microenvironment evolves over time, driven in part by an exhaustion of T cells, with a reduced capacity to mount an effective immune response to the proliferating tumour. Modulation of checkpoint targets (e.g. PD-1, PD-L1, CTLA-4, PX-40, LAG-3, Tim-3 and others) on immune cells can help to restore effector... Show more »

In cancer, the tumour microenvironment evolves over time, driven in part by an exhaustion of T cells, with a reduced capacity to mount an effective immune response to the proliferating tumour. Modulation of checkpoint targets (e.g. PD-1, PD-L1, CTLA-4, PX-40, LAG-3, Tim-3 and others) on immune cells can help to restore effector function.

Aquila has established a murine in vitro CD4+ T cell culture system with an exhausted phenotype, to model T cell exhaustion noted in patients with cancer, in which hyporesponsive T cells have a reduced capacity to target cancer cells. The exhausted CD4+ T cell assay can be used for high-throughput screening (HTS) in 96-well plate format. Compound effects are determined by assessment of cytokine production by ELISA as well as capacity to include additional readouts such as proliferation using flow cytometry. An analogous human system is in development as is a CD8+ version of this assay.

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Human
Mouse
Cancer
in vitro
Macrophage Activation Assay
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Human Macrophages Assays

The tumour microenvironment (TME) prompts macrophages to acquire pro-tumoral phenotype. Tumour Associated Macrophages (TAMs) are immunosuppressive, resembling type 2 macrophages (M2), rather than the more proinflammatory M1 phenotype. In the TME, TAMs can induce proliferation and survival of tumour... Show more »

Human Macrophages Assays

The tumour microenvironment (TME) prompts macrophages to acquire pro-tumoral phenotype. Tumour Associated Macrophages (TAMs) are immunosuppressive, resembling type 2 macrophages (M2), rather than the more proinflammatory M1 phenotype. In the TME, TAMs can induce proliferation and survival of tumour cells, facilitate angiogenesis, and supress immune responses via expression of inhibitory molecules (e.g. PD-L1, B7-H4) and cytokines (e.g. IL-10, TGF-β). TAM polarisation status is a consequence of M2-promoting signals dominating over M1-promoting signals in the TME. Human monocytes differentiated with M-CSF are skewed towards an M2 phenotype, whereas GM-CSF leads to M1 polarization. M2 macrophages are characterized by expression of CD163 (M1s lack this receptor) and the production of high IL-10, and low IL-12 in response to LPS. M1 macrophages show the opposite cytokine balance, with IL-12 more dominant. Therapeutic “re-direction” of M2 macrophages towards an anti-tumour/immunostimulatory M1 function is desirable. Aquila has a suite of protocols that can be used to screen libraries of small molecules or biologics for their ability to induce an M2—M1 switch.

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Histological Staining
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Specialist Multiplex Histology

A key asset to an immuno-oncology program is the preclinical and clinical assessment of immune cell biomarkers in the tumour microenvironment, enabling patient stratification and ultimately personalised medicine.

Aquila Histoplex deliver specialised histological and multiplex tissue probing... Show more »

Specialist Multiplex Histology

A key asset to an immuno-oncology program is the preclinical and clinical assessment of immune cell biomarkers in the tumour microenvironment, enabling patient stratification and ultimately personalised medicine.

Aquila Histoplex deliver specialised histological and multiplex tissue probing and staining technologies, using a range of specialised histological techniques including automated multiplex immunofluorescence (IF), automated colourmetric immunohistochemistry, in-situ hybridisation for mRNA using ACD Bio RNAscope technology (as Europe’s first ACD Bio accredited service provider). Other specialised histological techniques can also be delivered including Tissue Micro Array production, sterile sectioning of tissue matrices for culture. High precision image analysis and quantification is managed through collaboration with OracleBio Ltd.

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Mixed Lymphocyte Reactions
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Mixed leukocyte reaction (MLR) assays are commonly used to understand the effects of biologics or small molecules upon of T cell activation, indicating the potential ability of the drug to boost anti-tumour T cell immunity. Allogeneic MLRs can be established using monocytederived dendritic cells (Mo-DC), which can be either... Show more »

Mixed leukocyte reaction (MLR) assays are commonly used to understand the effects of biologics or small molecules upon of T cell activation, indicating the potential ability of the drug to boost anti-tumour T cell immunity. Allogeneic MLRs can be established using monocytederived dendritic cells (Mo-DC), which can be either immature, or matured under varying conditions, depending on target of interest. Responder T cells are isolated from HLA-mismatched donors, providing a source of alloreactive T cells.

Assay readouts include:
Proliferation by CFSE, Ki-67 (flow cytometry)
Cytokine expression e.g. IL-2, IFN-γ etc. (ELISA, multiplex arrays, or flow cytometry)
Activation markers e.g. CD80, CD83, CD86 (flow cytometry).

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Cell-Based Assays
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Pharmacology & Toxicology
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Immune Cell Assays
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Cytotoxicity Assays
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Biology
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Clinical and Anatomic Pathology
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Clinical Laboratory Services
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Bioanalysis
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Functional & Cell Type Specific Assays
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Toxicology
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Immunoassays
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Clinical Research
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