Cytokine and Cancer Signalling Group
Our research focus
Gastrointestinal Cancer models
With the advancement of immunotherapies, new cancer models are needed that mimic both the tumour cells as well as the normal body cells present in cancer tissues to test the effectiveness of therapies. We develop novel murine cancer models by establishing organoids, 3-D miniature organ-like structures, from gastric and colorectal cancers tissues. These organoids represent new models enabling the testing of genetic variation in the tumour cells or the tumour microenvironment selectively and the testing of new immunotherapies.
Cytokine signalling
Cytokines are signalling molecules that mediate the crosstalk between cancer cells and the normal cells within cancerous tissue. Depending on the source of the cytokine and the responding cell type, the impact can either be tumour promoting or tumour inhibitory. We are studying, two cytokine signalling pathways, IL33/ST2 and IL6/IL11/STAT3. Both are highly active in gastro-intestinal cancers and generate an immunosuppressive microenvironment. We study how they promote primary tumour growth, metastasis formation and responses to (immuno)therapies.
Immunotherapy
Immunotherapies are drugs that manipulate the body’s own immune cells to attack cancers. One class of immunotherapies, the immune checkpoint inhibitors, has strongly improved therapy outcomes for many different cancer types. However, in gastric and colorectal cancer only a minority of patients respond to these therapies. We study the role of cytokine signalling in the anti-tumour responses to immune checkpoint blockade. Additionally, we investigate, if inhibition of these cytokine signalling pathways can improve the efficacy of immunotherapies against gastric and colorectal cancers.
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Recent publications
ScienceDirect
Generation of gene-of-interest knockouts in murine organoids using CRISPR-Cas9
DOI: 10.1016/j.xpro.2023.102076
17 March 2023
Frontiers
IL33 and Mast Cells-The Key Regulators of Immune Responses in Gastrointestinal Cancers?
DOI: 10.3389/fimmu.2020.01389
3 July 2020
Nature
IL-33-mediated mast cell activation promotes gastric cancer through macrophage mobilization
DOI: 10.1038/s41467-019-10676-1
21 June 2019
Our team
Meet our researchers
- Dr Moritz Eissmann - Head, Cytokine and Cancer Signalling Group Publications
- Amr Allam - Postdoctoral Research Fellow
- Anne Huber – Postdoctoral Research Fellow
- Saumya Jacob – Research Assistant
- Josh Konecnik – Research Assistant
Blood Cancer and Immunotherapy Laboratory
Our research focus
Human cancers harbour defects in processes that inhibit or enhance tumour growth, so-called tumour suppressor and oncogenes. Importantly, these abnormalities render the tumour cells resistant to many therapies.
Our research is focused on understanding how cancers develop and finding new therapy approaches, including targeted drugs as well as immune therapies to treat these cancers.
We aim to identify novel genes involved in the development of normal and malignant haematopoiesis. We are using in vivo CRISPR approaches to systematically identify new candidate genes, with the ultimate goal of using these targets as novel anti-cancer therapies.
We have recently also started to understand the mechanisms of therapy resistance in cancers and how we can overcome this by reducing the fitness of the cancer cells or enhancing immune therapies using different CRISPR approaches.
Recent publications
CELL DEATH & DIFFERENTIATION
Deletion of the transcriptional regulator TFAP4 accelerates c-MYC-driven lymphomagenesis
DOI: 10.1038/s41418-023-01145-w
9 March 2023
Frontiers Immunology
An arrayed CRISPR screen of primary B cells reveals the essential elements of the antibody secretion pathway
DOI: 10.3389/fimmu.2023.1089243
13 February 2023
EMBO Journal
Caspase-8-driven apoptotic and pyroptotic crosstalk causes cell death and IL-1β release in X-linked inhibitor of apoptosis (XIAP) deficiency
DOI: 10.15252/embj.2021110468
17 January 2023
Our team
Meet our researchers
- Prof Marco Herold - Head, Blood Cancer and Immunotherapy Laboratory Publications
- Andrew Kueh - Platform Lead
- Emily Lelliott - Postdoctoral Research Fellow
Molecular Immunology Laboratory
Our research focus
Identification of tumour immune evasion mechanisms
Cancer immune evasion is a major hurdle in for the success of current immunotherapies, both in the context of adoptive cellular therapy (ACT) and immune checkpoint blockade. Despite the clinical success of diverse immunotherapies, many patients do not respond or ultimately relapse, likely due to tumours evolving to escape sufficient recognition by the immune system. Although considerable progress has been made in understanding how cancers evade immunity, measures to counteract tumour immune escape are lacking. We employ cutting-edge screening mechanisms to identify tumour immune evasion mechanisms and avenues to sensitise tumour cells to T cell-mediated killing.
Genetic/epigenetic control of effective anti-tumour T cell responses
Despite the success of adoptive cellular therapy (ACT) in the context of haematological malignancies, response rates against solid malignancies are poor, likely due to tumour-associated immunosuppression and subsequent T cell exhaustion. Indeed, it is becoming clear that the failure of T cells to elicit a successful and long-term anti-tumour immune response is controlled by transcriptional, epigenetic and post-translational modifications. However, our current understanding of the molecules involved in these processes is limited. We use cutting-edge technology including in vitro and in vivo CRISPR screens, high throughput drug screens and novel single cell sequencing protocols to identify novel underlying molecular mechanisms leading to T cell dysfunction in cancer and identify mechanisms to improve T cell function in this context. We employ a variety of pre-clinical models of CAR T cell therapy and re-directed TCR T cell therapy to validate novel immunotherapy targets for translation into the clinic.
Identification of novel immunotherapy approaches.
High-throughput screening has been a staple in drug discovery in recent decades. Target-based drug discovery relies heavily on singular readouts such as reporter gene expression or perturbation of enzymatic activity in response to small molecule treatment. However, with a recent renewed focus on phenotypic based drug discovery, there is an increased interest in more comprehensive and less biased screening methods that combine aspects of both target-based and phenotypic screening, such as RNA-seq. To complement our genetic screens, we also perform high-throughput drug screens for agents that promote favourable states of T cell differentiation for anti-tumour immunity and agents that increase the immunogenicity of cancer cells. Importantly, identification of such agents would allow us to enhance current immunotherapy approaches.
Fast facts
Adoptive cellular therapy, also known as cellular immunotherapy, is a form of treatment that uses the cells of our immune system to eliminate cancer. Some of these approaches involve directly isolating our own immune cells and simply expanding their numbers and re-introducing into a patient, whereas others involve genetically engineering our immune cells (via gene therapy) to enhance their anti-cancer functions.
CD8+ T cells (often called cytotoxic T lymphocytes, or CTLs) are critical for immune defence against pathogens including viruses and bacteria, but also for detecting and killing cells that have become cancerous. When a CD8+ T cell recognises its antigen and becomes activated, it can directly kill pathogen infected cells or cancer cells through direct contact, but also release soluble factors called cytokines which alert other cells of the immune system.
CRISPR is a powerful tool for editing genomes, meaning it allows researchers to easily alter DNA sequences and modify gene function. CRISPR technology was adapted from the natural immune defence mechanisms of bacteria and archaea, species of relatively simple single-celled microorganisms.
Recent publications
PNAS
Rationally designed chimeric PI3K-BET bromodomain inhibitors elicit curative responses in MYC-driven lymphoma
DOI: 10.1073/pnas.2306414120
29 August 2023
Cell Reports
CRISPR-Cas9 screening identifies an IRF1-SOCS1-mediated negative feedback loop that limits CXCL9 expression and antitumor immunity
DOI: 10.1016/j.celrep.2023.113014
20 August 2023
Science Advances
Eprenetapopt triggers ferroptosis, inhibits NFS1 cysteine desulfurase, and synergizes with serine and glycine dietary restriction
DOI: 10.1126/sciadv.abm9427
14 September 2022
Our team
Meet our researchers
- Dr Conor J Kearney - Head, Molecular Immunology Laboratory Publications
- Mario Djajawi - Postdoctoral Research Fellow
- Akash Srivaths - PhD Student
Tissue and Tumour Immunity Laboratory
Our research focus
Tumour infiltrating regulatory T cells
Our body’s immune system can recognise and effectively eliminate tumours. To escape immune attack, tumours hijack a specialised immune cell population known as regulatory T cells (Tregs), which potently suppresses anti-tumour immune cells to promote tumour growth. Targeting Tregs, therefore, is an attractive strategy to revert immune suppression in tumours and boost the function of anti-tumour immune cells. Our laboratory aims to discover transcriptional and epigenetic mechanisms employed by Tregs to infiltrate tumours in vital organs such as the gut, lung, liver and brain. These mechanisms will be harnessed to target Tregs for the treatment of solid tumours.
Stromal-immune cell crosstalk
Tumour microenvironment is composed of actively proliferating tumour cells, stromal cells, blood vessels and a plethora of immune cells. Stromal cells are known to produce a variety of growth factors and immunomodulatory molecules that directly shapes the immune landscape of tumours. We use cutting-edge molecular tools and microscopy to understand the molecular makeup of stromal cells in diverse tissues and tumours as well as their crosstalk with immune cells, in particular Tregs.
Metabolic regulation of tumour immunity
Cellular metabolism is critical for the differentiation, growth and function of tumour cells and immune cells. Metabolic intermediates serve as catalysts for several cellular processes including gene transcription. Systemic metabolic changes also influence anti-tumour immune responses and immunotherapy outcomes. Our laboratory aims to understand how cellular metabolism and tumour derived ‘oncometabolites’ shape the transcriptional landscape of tumour infiltrating immune cells. We also investigate how changes in systemic metabolism affects anti-tumour immunity and immunotherapy outcomes.
Fast facts
Cancer cells have hijacked for their own benefit the inflammatory processes that help support wound healing of normal tissues.
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A gene or protein which is identified to cause, or play a major role, in the disease.
A drug which attacks a specific protein of the cancer. Such drugs therefore only work on cancer where such a protein confers a specific benefit for a particular cancer to grow and spread.
Recent publications
Trends in Cancer
Gut microbiota - a double-edged sword in cancer immunotherapy
DOI: 10.1016/j.trecan.2022.08.003
8 September 2022
Clinical & Translational Medicine
Sex-bias in CD8+ T-cell stemness and exhaustion in cancer
DOI: 10.1002/cti2.1414
26 August 2022
Cellular & Molecular Immunology
Resident and migratory adipose immune cells control systemic metabolism and thermogenesis
DOI: 10.1038/s41423-021-00804-7
26 November 2021
Our team
Meet our researchers
- Dr Ajith Vasanthakumar - Head, Tissue and Tumour Immunity Laboratory Publications
- Tabinda Hussain - Postdoctoral Research Fellow
- Pathum Thilakasiri - Research Assistant Publications
- Adelynn Tang - PhD Student
- Jian Wu - PhD Student
Flow Cytometry Core Facility
Flow Cytometry Core Facility
The ONJCRI Flow Cytometry Core Facility is a shared resource laboratory which provides state-of-the-art analytical cytometry and high speed cell sorting services to the research community of our Institute, affiliates, and external users. Our goal is to provide high quality service to researchers and proactively introduce flow cytometric methods into new research areas. An important part of our mission is to teach this technology to students, staff, and investigators.
Cell sorter is operated by facility staff from 10:00am-5:30pm Wednesday to Friday. Extended sorting hours are available upon request. Self-service sorting is also available for trained researchers. Analysers available 24 hours / 7 days a week for self-service use (for suitably trained users only).
Services and Instrumentation
Our Flow Cytometry Core Facility provides:
- Operator assisted Cell Sorting
- Independent acquisition and analysis (users must go through the training process first)
- Operator assisted acquisition and analysis
- Consultation and assistance with experimental design and data analysis
Currently we house four instruments:
- BD FACSARIA III is a 4 laser, 12 detector cell sorter capable of measuring 10 colours simultaneously plus cell size and complexity parameter. The lasers in this instrument are 488nm (Blue), 633nm (Red), 561nm (Yellow-Green) and 405nm (Violet). It can sort up to 4 populations simultaneously into different size tubes, tissue culture plates, and onto individual slides. Sorting on the Aria can be performed using high-pressure (70 µm), intermediate-pressure (85 µm), or low-pressure (100 µm) nozzle. Instrument operates on a PC-based platform using FACSDiva (version 8) software.
- 2x BD FACSCanto II is three lasers, 10 detector analysis flow cytometer, capable of measuring 8 colors simultaneously plus cell size and complexity parameter. The lasers in this instrument are 488nm (Blue), 633nm (Red) and 405nm (Violet). It also features a High Throughput Sampler (96 well Plate Loader). These instruments operate on a PC-based platform using FACSDiva (version 8) software.
- AutoMACS Pro Separatoris a benchtop instrument for high-speed magnetic cell sorting of multiple samples. Employing MACS® Technology, it is designed for cell isolation in a fully automated, walk-away fashion. It can be used as a pre-sorter for speeding up flow sorting.
- Data Analysis Software – FlowJo– is a comprehensive analysis package specifically designed for handling list mode data generated by cytometers. ONJCRI provides several Flow Jo dongles, which are available to Institute members at no cost. Users can download the latest FlowJo version for Windows or Mac by contacting Institute’s IT Department.
Contact
If you would like to discuss opportunities to use our platform technologies and services, please contact:
Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518
Mammalian Protein Expression Facility
Mammalian Protein Expression Facility
This unique facility is located in a dedicated suite of cleanrooms and can produce small to large amounts of high quality recombinant proteins and antibodies for use in medical research.
Cell Line Development
Transient expression, stable expression, and isolation/enrichment of high producing clones can be performed.
Production Systems
A range of production systems are available depending on the scale of production required. Perfusion systems are able to produce material every two days ranging from tens of milligrams to several hundreds of milligrams per harvest. A proprietary biphasic approach to production is used during protein production in stirred tank bioreactors. Yields up to 4g/L of mAb in a GS CHO cell line can be achieved.
Purification and Product Characterisation
Production material is purified using a variety of chromatography techniques such as affinity, size exclusion and ion exchange. Operating procedures are in place to allow for flexibility to process materials ranging from milligrams to several grams of material.
Quality assessments are conducted through a combination of SDS-PAGE, size exclusion chromatography using HPLC, and binding kinetics using BIAcore.
Contact
If you would like to discuss opportunities to use our platform technologies and services, please contact:
Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518
VECTRA Multi-Spectral Imaging Platform
VECTRA Multi-Spectral Imaging Platform
The Vectra multi-spectral immuno-histochemistry platform is the first of its kind in Australia. It allows researchers to more accurately define the immune microenvironment, and helps detect mechanisms a tumour may use to evade the immune system.Its acquisition was supported by a grant from the Ian Potter Foundation in collaboration with La Trobe University and the Peter MacCallum Cancer Centre.
The Vectra platform and its analysis software allows researchers to quantify cells which are positive for one or both of the molecules and to analyse their localisation within the cells (for example, in the membrane, cytoplasma or nucleus).
Contact
If you would like to discuss opportunities to use our platform technologies and services, please contact:
Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518
Functionalities
The Vectra platform can simultaneously detect up to seven different proteins of interest on one FFPE-tissue slide using Opal chemistry and spectral un-mixing.
It has an integrated automated slide-loader for up to 200 slides, which can be loaded and scanned, fully automated by user-defined protocols. Pre-scans of whole tissue slides will be performed at 4x or 10x magnification (RGB) followed by high-power-field (HPF) imaging of regions of interest in the multispectral mode (20x or 40x magnification).
The multiplex IHC images lead to a comprehensive understanding of complex cellular interactions which is not accessible by other methods. Opal follows the standard IHC workflow using unlabelled primary antibodies, followed by the addition of anti-species-HRP conjugate and detection substrate. Opal fluorescent detection substrates bind covalently near the epitope, allowing subsequent removal of antibodies to clear the tissue for detection of the next target. The signal remains intact after antibody removal.
Localisation studies of proteins in cellular compartments, co-localisation of proteins and quantification of cells expressing single markers or marker combinations can be subsequently performed with specialised software available on several work stations at the ONJCRI.
Spatial relationships of cell types to each other and within the tissue context can be analysed using Spotfire software. Scanning of the slides and selection of tissue sections and areas of interest can be automated on the Vectra system and allows for review by a pathologist before analysis. Due to the TSA based signal amplification, primary antibody concentration can be reduced up to 100X and all markers of interest can be detected with antibodies raised in the same species.
Aperio AT2 Whole Slide Scanner
The Aperio AT2 is a high-volume microscope capable of digitalising histochemical stained slides. The Aperio AT2 has a capacity for up to 400 slides that can be scanned with a sustained rate of 50 slides per hour at 20 X. With high first scan success rate, your images will be uploaded to our microscopy server, allowing remote access for collaborative research teams worldwide.
Analysis can be performed using the Aperio ImageScope – Pathology Slide Viewing Software. This software contains integrated macros and algorithms for the analysis of many common pipelines for investigating your histochemical stained tissues. Image export is easily achieved, allowing free access to your datasets in your analysis software of choice.
BOND RX Fully Automated Research Stainer
The BondRX allows you to fully automate IHC, ISH, FISH, CTC and multiplexing staining experiments. Reduce your manual work to increase your efficiency and consistence with your staining experiments. The BondRX allows you to customise all steps in your staining protocol from baking and dewaxing, antigen retrieval to the length and temperature of your staining approach. The BondRX has a 30-slide capacity, with finished trays of 10 slides being capable of replaced continuously. Up to three separate staining protocols can be run simultaneously.
The BondRX allows you to leverage established protocols and reagents or develop completely custom and reproducible staining experiments with your own novel agents. The BondRX dramatically reduces the time required to perform your experiment. A bench run seven-plex experiment would require approximately 5 days of hands-on time manual time but with the BondRX this is reduced to a reproducible and consistent ~17 hours.
The ACRF Centre for Translational Cancer Therapeutics and Imaging
The ACRF Centre for Translational Cancer Therapeutics and Imaging
This Centre was established for medical research and preclinical investigations and houses a range of advanced imaging technologies and platforms including:

Contact
If you would like to discuss opportunities to use our platform technologies and services, please contact:
Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518
PET MRI imaging
By combining a high-performance PET system and compact MRI technology, NanoScan® PM PET/MRI provides preclinical whole body soft tissue images with detailed quantitative imaging data within just one study. The PET camera offers quantitative 3D spatial resolution at 700 µm combined with a uniquely large field-of-view. The 1 Tesla permanent magnet for MRI provides 100 µm resolution with advanced sequences and ensures robust imaging across a broad range of biological applications including:
- Oncology
- Tumour biology
- Stem cell investigations
- Regenerative medicine
- Neuroscience and receptor studies
- Cardiology
- Immunology and inflammation
- Multimodal contrast agent development
- Animal model development and phenotyping
- Nephrology
- Pharmacokinetics
- PET development of radiotracers
SPECT CT imaging
The NanoSPECT/CTTM is an in-vivo molecular imaging system suitable for use with small animals and unifies functional (SPECT) and anatomical (CT) imaging procedures for preclinical investigations. With 250μm 3D SPECT and 30μm CT spatial resolution, exceptional quantification is permitted with an accuracy over 97%.
The system enables the examination of bio-chemical processes in healthy and disease models (including cancer, diabetes and stroke). It determines the localisation of radio-labelled compounds used as probes for the disease state, and monitors the efficacy of interventions or therapies. In living subjects, the effect of drugs in development can be monitored and compared in real time and at multiple time points within the one subject. Drug localisation as well as uptake can be quantified and visualised from the data collected. The system is also suitable for monitoring genetic modifications and gene-therapeutic healing procedures using appropriate preclinical models.
IVIS spectrum imaging
The IVIS® Spectrum is a versatile and advanced in vivo imaging system, which uses a novel patented optical imaging technology to allow non-invasive longitudinal monitoring of disease progression, cell trafficking and gene expression patterns in living animals. High efficiency filters and spectral un-mixing algorithms take full advantage of bioluminescent and fluorescent reporters across blue to near-infrared wavelengths. It also offers single-view 3D tomography for both fluorescent and bioluminescent reporters, which can be analysed in an anatomical context using a Digital Mouse Atlas or registered with the IVIS multimodality module to other tomographic technologies such as MR, CT or PET.
For advanced fluorescence pre-clinical imaging, the IVIS Spectrum can use either trans-illumination (from the bottom) or epi-illumination (from the top) to illuminate in vivo fluorescent sources. 3D diffuse fluorescence tomography can be used to determine source localisation and concentration using the combination of structured light and trans illumination fluorescent images. The instrument is equipped with 10 narrow band excitation filters (30nm bandwidth) and 18 narrow band emission filters (20nm bandwidth), which assist in significantly reducing autofluorescence, via the spectral scanning of filters and the use of spectral unmixing algorithms. In addition, the spectral unmixing tools allow the researcher to separate signals from multiple fluorescent reporters in the same animal.
Research Platforms
Our research services
All research activities at the Olivia Newton-John Cancer Research Institute are enhanced and supported by outstanding platform technologies, facilities and technical expertise.
We also have a number of platform technologies and services which can be utilised by other Institutions and organisations. These include:
- PET MRI Imaging
- SPECT CT imaging
- IVIS spectrum imaging
- Inverted confocal microscopy
- Multiphoton microscopy
- NanoString
- VECTRA imaging
- Aperio
- BondRX
- Mammalian protein expression
- Flow cytometry
Contact
If you would like to discuss opportunities to use our platform technologies and services, please contact:
Mark Frewin
Laboratory and Facilities Manager
P +61 3 9496 5299
M +61 434 242 518
Receptor Biology Laboratory
Our research focus
Eph receptors
Eph receptors are cell surface proteins that guide cell migration by binding to other cell-bound proteins (ephrins) on adjacent cells, thereby controlling cell-cell adhesion. Ephs coordinate cell movement during normal development of tissue and organ boundaries, and the vascular and neural networks. They are generally scarce in adults but reappear in cancers, where they are often on early ‘progenitor’ cell types, associated with blood vessel formation, and tumour cell invasion and spread.
EphA3 is a particular focus, which we investigate in tumour models, using ‘knock-down’ mice or by treatment with a specific antibody we helped develop, with the aid of drug payloads to specifically target the tumour microenvironment.
ADAM metalloproteases
ADAM metalloproteases (or ADAMs) are cell membrane-bound proteases that shed a range of other membrane proteins, regulating the activity of diverse cell surface receptors. These include Ephs and other receptors controlling cancer cell growth, drug resistance, and invasion and spread to other tissues. ADAMs also play an important role in the tumour microenvironment and in inflammation.
ADAM10 and 17 are of particular interest and we are investigating their function in tumours, as well as developing antibodies and antibody-drug conjugates as potential new therapies.
Fast facts
Cell surface (or membrane) receptors are proteins attached to a cell’s exterior which can receive external signals, usually by binding with another protein. The bound receptors then send signals into the cell to modify its behaviour, including movement, proliferation and survival.
They are distinctive in that they bind to proteins attached to adjacent cells. This allows them to control cell adhesion, migration and invasion. They are important in normal embryonic development but reappear in certain cell types in tumours and their surrounding environment, including new tumour blood vessels, which support tumour growth and spread.
A protease is a protein which cuts other proteins in a very controlled manner. ADAMs are a type of cell surface metalloprotease (‘metallo’ refers to their dependence on metal ions). ADAM10 and 17 control the activity of various cell surface receptors and are essential in normal cellular development. However they become overly active in tumours and their surrounding environment by supporting tumour growth, survival and drug resistance.
Recent publications
CANCERS
Preferential Antibody and Drug Conjugate Targeting of the ADAM10 Metalloprotease in Tumours
DOI: 10.3390/cancers14133171
28 June 2022
Biochemical Journal
The intracellular domains of the EphB6 and EphA10 receptor tyrosine pseudokinases function as dynamic signalling hubs
DOI: 10.1042/BCJ20210572
14 September 2021
Cancer Research
Eph Receptors in the Immunosuppressive Tumor Microenvironment
DOI: 10.1158/0008-5472.CAN-20-3047
15 Feb 2021
Our team
Meet our researchers
- A/Prof Peter Janes - Head, Receptor Biology Laboratory Publications
- Mary Vail - Senior Research Scientist Publications
- Stacey Allen - Senior Research Assistant
- Hengkang Yan - Honorary
- Sakshi Arora - PhD Student