Invited Presentation Abstracts
Mechanobiology-Inspired Antithrombotic Strategies and Point-of-Care Microtechnologies
Lining Arnold Ju
University of Sydney
Cardiovascular diseases remain the leading cause of death globally, with thrombosis playing a central role in their pathogenesis. Current antithrombotic therapies, while effective, often carry significant bleeding risks due to their inability to differentiate between pathological and physiological blood clotting. This presentation introduces our integrated approach that combines fundamental mechanobiology with translational engineering to address critical clinical needs in cardiovascular medicine, potentially transforming how we diagnose, monitor, and treat thrombotic disorders.
First, using our single-cell biomechanical nanotools such as Biomembrane Force Probe (BFP), we present insights into thrombosis mechanobiology, particularly focusing on the role of von Willebrand Factor (VWF) and other mechanoreceptors (GPIbα, integrin αIIbβ3 and PIEZO1 ion channels) in distinguishing between “good” and “bad” mechanical forces in thrombosis. These helped uncover new therapeutic targets for force-sensitive antithrombotic strategies. Second, we demonstrate a personalized vessel-on-chip platform that recreates patient-specific blood vessel geometries and flow conditions, enabling precise evaluation of thrombotic risk and drug responses. Finally, we introduce novel point-of-care microtechnologies for rapid blood coagulation testing, including an AI-powered platform called SmartClot, which promises to revolutionize home-based coagulation monitoring. These innovations represent a significant advancement toward more effective and safer antithrombotic treatments, with potential applications ranging from preventive care to personalized medicine.
The contribution of flow cytometry to diagnostic pathology
William Sewell
SydPath, St Vincent’s Hospital Sydney and Faculty of Medicine and Health, University of NSW Sydney
This presentation evaluates the strengths and limitations of flow cytometry in the contemporary pathology laboratory. Strengths of flow cytometry include rapid provision of results and determination of comprehensive immunophenotypes. In lymphoma, flow cytometry is valuable in assessment of clonality, and our recent results in assessing T cell clonality with TRBC1 and TRBC2 will be highlighted. In the assessment of biopsies, immunophenotypic information is provided by both flow cytometry and immunohistochemistry, and the presentation compares the contributions of the two modalities. The valuable role of flow cytometry in fine needle aspirates and other paucicellular samples will be presented. Limitations of flow cytometry will be discussed, including its failure to detect certain types of haematological malignancies, and under-estimation of other types. The continuing contribution of flow cytometry in the face of rapid advances in genetics will be evaluated. Both flow cytometry and genetics are used to assess minimal residual disease, and the merits of the two systems will be compared. Flow cytometry is a standard feature of immunological research, but only a fraction of this methodology has become established in diagnostic laboratories. An example is our use of flow cytometry to enumerate CMV antigen-responsive T cells. Although there are challenges, flow cytometry is well established and will continue to make an important contribution to diagnostic pathology.
Diagnostic flow cytometry in hematologic neoplasms today and tomorrow
Wolfgang Kern
MLL Munich Leukemia Laboratory
Flow cytometry is an integral part of diagnostics in hematologic neoplasms in combination with cytomorphology, histopathology, cytogenetics, FISH and molecular genetics. Continuously increasing insights into genetic alterations causing these diseases and being target for innovative therapies causes overall increases in diagnostics. This is paralleled by innovations in diagnostic technologies. In flow cytometry higher numbers of fluorochromes can be analyzed and spectral cytometry is anticipated to significantly accelerate this evolution. Further, more scalable sample preparation devices and well as AI-based data analysis tools are more and more shaping diagnostic routine procedures. This presentation will provide a respective perspective.
Uncovering the molecular basis for treatment resistance in autoimmune disease with single cell multi-omics
Joanne Reed
Westmead Institute for Medical Research
The treatment of autoimmune disease has advanced with the introduction of rituximab, a monoclonal antibody targeting CD20 on the surface of B cells. However, a significant subset of patients exhibit treatment resistance leading to relapse of disease symptoms. Treatment resistance may arise from the recruitment of new pathogenic B cell clones or insufficient depletion of pathogenic clones due to molecular changes or tissue location. The goal of this study was to use flow cytometry to predict disease relapse and determine the molecular drivers of treatment resistance with single cell omics.
To identify treatment-resistant cells we performed flow cytometry on blood, bone marrow and nasal swabs collected from patients with autoimmune diseases undergoing rituximab treatment. Single cell RNA and DNA sequencing was performed on a patient followed through five cycles of clinical response to rituximab and relapse over 3 years.
Treatment resistance was caused by insufficient depletion of a subpopulation of pathogenic B cell clones that carried distinct mutations and lacked CD20 expression at the time of treatment. Despite depletion of B cells in the nasal passage and blood, pathogenic B cell clones re-populated the blood over time and remained in the bone marrow. These findings support the use of flow cytometry to track treatment response and single cell omics to identify more targeted treatment.
Breakthrough Technologies to Unveil and Predict the Hidden Architecture of Life
Joakim Lundeberg
KTH Royal Institute of Technology
Multicellular life is more than the sum of its cellular components: it is shaped by their intricate spatiotemporal networks that define the form and function of tissues, organs, and whole organisms. While genome sequencing revealed the molecular code of life and single-cell technologies uncovered the identity and diversity of individual cells, spatial biology represents the next technological frontier towards decoding this complexity by capturing how cells are organized, communicate, make decisions, and change within intact tissues across space and time
Over the past decade, spatial biology has emerged as a paradigm-shifting concept in life sciences. This extraordinary momentum was driven by groundbreaking technological advances collectively termed ‘spatial omics’, enabling highly multiplexed molecular profiling of RNA expression, protein abundance, genomic variation, epigenetic states, lipids, glycans, and metabolites together with their precise spatial coordinates, while preserving the intact architecture of the source cells and their microenvironments. These technologies thus reveal how cells are arranged, how they communicate with each other and their surroundings, and how these interactions collectively shape tissue function in health and disease.
Prof Joakim Lundeberg together with Prof. Jonas Frisén (Karolinska Institutet), pioneered this field by developing Spatial Transcriptomics (ST), the first method enabling transcriptome-wide mapping of mRNA directly in tissue sections (Science 2016). This foundational approach underlies the widely adopted Visium platform (10x Genomics), the most used spatial transcriptomics system worldwide, and recognized as Method of the Year in 2020 by Nature Methods. This presentation will provide an overview of the current methodology insights into these landscapes across development, health, and disease.
Mapping early dendritic cell–T cell interactions after allergen exposure
Franca Ronchese
Malaghan Institute of Medical Research
Allergic inflammation is characterised by the production of IL-4, IL-5 and IL-13 by CD4+ T helper 2 (TH2) cells. These cells respond to otherwise harmless environmental antigens, including house dust mite, pollen and animal dander, and release mediators that drive inflammation and tissue damage. However, it remains unclear how the first encounter with an allergen programs the immune system to overreact during subsequent exposures.
Allergic immune responses begin when dendritic cells in barrier tissues, such as the skin, airway and gut, take up allergens. These dendritic cells then migrate to lymph nodes, where they interact with T cells and instruct them to differentiate into TH2 cells. Using fluorescently labelled allergens, we identified the skin dendritic cells that capture allergens and characterised their gene expression by bulk and single-cell RNA sequencing to define allergen-induced gene programs. We also monitored early TH2 differentiation using flow cytometry, fluorescent cytokine reporters and single-cell transcriptomics. Together, these studies reveal early dendritic cell–T cell communication events that promote TH2 priming after allergen exposure.
Validation of a 24-colour Spectral B-Acute Lymphoblastic Leukaemia (B-ALL) assay for Measurable Residual Disease (MRD) detection
Melissa Nardi
Peter MacCallum Cancer Centre
Emerging immunotherapies present new challenges for detecting measurable residual disease (MRD) in B-acute lymphoblastic leukaemia (B-ALL) particularly in regenerating post-treatment bone marrow. Conventional B-cell markers such as CD19, CD20, and CD22 may be diminished or lost following targeted therapies, may undergo antigen escape, or may not be represented in the patient’s original immunophenotype. To overcome these limitations, we developed a 24-colour in-house spectral flow cytometry panel informed by current literature for B-ALL MRD detection using the Cytek Northern Lights. This presentation describes the validation workflow for comparing the new spectral panel with our existing 12-colour conventional B-ALL MRD panel and key considerations for implementation into routine diagnostic use.
Innovating Cytometry for the Australian context
Helen McGuire
University of Sydney
Conducting medical research in Australia comes with distinct challenges and opportunities. This talk will explore how cytometry can provide a mechanism that have engaged this paradigm. Further, this talk will invite conversation as to how further strengthening can be achieved by leveraging community efforts.
Unravelling age-dependent tumour-immune interactions driving immunotherapy responses in paediatric cancer
Omar Elaskalani
The Kids Research Institute Australia
Children’s immune systems are developmentally distinct from those of adults, yet how this shapes cancer immunotherapy responses remains poorly understood. Using our juvenile cancer mouse models alongside paediatric patient datasets, we found age-dependent responses to immune checkpoint inhibitors and bispecific T cell engagers: identical tumours were cured in adult mice but responded poorly in juveniles, despite equivalent tumour burden. Using spectral flow cytometry with specialised 30-colour T cell and macrophage panels, and RNA sequencing, we uncovered the age-specific immune landscape underlying these responses. Juvenile tumours were infiltrated by naïve-like, PD-1 low CD8⁺ T cells, with few effector, exhausted, or resident memory cells, alongside MHC-II low, M2-like immunosuppressive macrophages. In contrast, adult tumours contained functionally activated effector and exhausted CD8⁺ T cells, indicating that productive T cell exhaustion programs accompany immunotherapy response. This anti-inflammatory, macrophage-driven milieu impeded T cell activation in juveniles. Repolarising tumour-associated macrophages reprogrammed the juvenile microenvironment towards an adult-like state, restoring CD8⁺ T cell effector responses and immunotherapy efficacy. These findings reveal that immune age dictates the tumour microenvironment and define new strategies to improve immunotherapy for children with cancer.
Tracking responses to GD2-specific CAR-T therapy in patients with brain tumours
Tessa Gargett
Royal Adelaide Hospital
Adelaide University
Glioblastoma multiforme (GBM) and diffuse intrinsic pontine glioma (DIPG) are aggressive, life-threatening brain tumours in children and adults, respectively, with poor survival outcomes and high recurrence rates. Current standard treatments—surgery, radiation, and chemotherapy for GBM; radiation for DIPG—provide limited long-term benefit, highlighting the urgent need for more effective therapies. Chimeric antigen receptor (CAR)-T cell therapy has emerged as a promising immunotherapy for targeting GBM and DIPG.
We are conducting two CAR-T cell therapy clinical trials using GD2-targeting CAR-T cells for GBM (KARPOS) and DIPG (LEVI’S CATCH), with 16 patients treated to date (8 adults and 8 children). Some patients have achieved partial tumour responses with varying toxicity profiles, while others had progressive disease. However, the encouraging objective responses in some patients are of relatively short duration, suggesting resistance mechanisms. We are monitoring immunological changes in blood and cerebrospinal fluid following CAR-T infusion using cytokine bead array, high-parameter flow cytometry, targeted proteomics, and scRNAseq. We aim to use this comprehensive patient dataset to identify biomarkers and signatures associated with objective response, toxicity, and treatment failure.
Our pilot data has identified T cell signatures potentially linked with positive response, and myeloid and B cell-associated markers that may indicate treatment failure and tumour recurrence. These signatures will be investigated in a larger cohort. The ultimate goal of this correlative study is to identify therapeutic strategies to overcome the multiple inhibitory mechanisms by which tumours evade CAR-T attack. This will guide further development of CAR-T therapy designed to persist, identify, and overcome the immune-suppressive microenvironment to achieve durable disease control.
Progressing Flow Cytometric MRD in Acute Myeloid Leukemia
Sylvie Freemann
University of Birmingham
The purpose of measurable residual disease detection in acute myeloid leukemia (AML) is to provide objective and reproducible quantification of the leukemia for the measurement of deeper remission status. This requires a technical sensitivity of at least 10-4 (1 leukemic cell in 10,000 normal cells). Although flow cytometry is a widely applicable technology for MRD, inter- and intra-patient immunophenotypic heterogeneity of AML blasts with potential shifts over time and variable background from non-AML cells has made marker selection and analysis challenging – particularly for quantitation below 10-3 (Flow MRD low-level). Additionally, advances in assay technology and reagents have to be balanced by the requirement for clinical validation. In parallel the genetic complexity of AML means both molecular and flow cytometric technologies are required to optimise monitoring, according to genetic subtype and treatment context. All these factors have to be considered for the best use of flow cytometric MRD in AML management. However, there are key principles acquired from published data, expert experience and regulatory requirements that can guide to reproducible AML MRD assay results and reporting across clinical laboratories. This talk will update participants on recommendations for flow cytometric MRD in AML and current areas of development.
ACS Conference 2026 