Anila Hashmi^1,2,3, Stan Sidhu^3,4, Gyorgy Hutvagner²

¹NSW Health Pathology, Liverpool, NSW, Australia
²University of Technology Sydney, Sydney, NSW, Australia
³Kolling Research Institute, NSW Health, Sydney, NSW, Australia
⁴Endocrine Surgery Unit, Royal North Shore Hospital, Northern Sydney Local Health District, St Leonards, NSW, Australia

Background: Adrenocortical carcinoma (ACC) is an aggressive malignancy with high recurrence rate, even after complete surgical resection. Currently biomarkers are limited in their ability to assist diagnosis or prognostication1. While secretion of cortisol and androgens is associated with poorer outcomes2, biomarkers with greater prognostic accuracy would be desirable. AGO2, a key regulator of miRNA function, plays a significant role in tumour biology3. This study evaluated AGO2’s prognostic potential in ACC by correlating its expression with clinicopathological features, including hormone status, and overall survival.

Methods: Clinicopathological parameters, including age, sex, Weiss score, pathological stage, molecular subtype, hormone production, and overall survival, were analysed for associations with AGO2 gene expression (TCGA-79-ACC) and protein levels (Kolling Tumour bank-15- ACC)3.  P-values <0.05 were considered significant.

Results: Elevated AGO2 expression was significantly associated with poor survival (p <0.001), high Weiss scores, and advanced tumour stage. Notably, increased AGO2 expression was linked with poor outcomes after controlling for hormone status.

Conclusion: AGO2 shows potential as an ACC biomarker, offering independent prognostic information. These findings support the value of further investigation into the role AGO2 could play in supporting the diagnosis and prognosis of ACC.

Statement of Originality: This study is the first to evaluate AGO2 expression in relation to clinicopathological factors, including hormone status, in ACC, providing original analyses into AGO2’s potential role as a prognostic biomarker in this malignancy.

References: 

1) Fassnacht, M., et al. (2018). ESE Clinical Practice Guidelines on adrenocortical carcinoma. Eur J Endocrinol, 179(4), G1–G46. https://doi.org/10.1530/EJE-18-0608

2) Nastos, C., et al. (2024). Hormone-secreting ACC and survival: A meta-analysis. Langenbecks Arch Surg, 409(1), 316. https://doi.org/10.1007/s00423-024-03507-5

3) Hashmi, A., et al. (2024). AGO2 as a biomarker in adrenocortical carcinoma. Endocr Relat Cancer. https://doi.org/10.1530/ERC-24-0061

Jack Hywood¹, Jeremy VanderDoes², Gregory Rice^2
1Department of Anatomical Pathology, Royal Children's Hospital, Parkville, Australia
²Department of Statistics and Actuarial Science, University of Waterloo, Canada

The tumour microenvironment, and in particular the dynamics between tumour cells and immune cells, has increasingly been shown to contain prognostic and potentially predictive information relevant to patient care. A common technique for analysing the tumour microenvironment is the use of multiplex immunostaining, in which multiple protein antigens are detected simultaneously within a single area of tissue. This allows for accurate localisation and phenotyping of the imaged cells. There is increasing need to develop statistical techniques for the analysis of this type of data. We present here a novel statistical method for comparing cell interactions in the tumour microenvironment between two different groups of patients (e.g. responder vs. non-responders to a therapy). The technique can also be used to predict which group a new patient belongs to based on the data from their tumour.
 
Author contribution statement:
J Hywood, G Rice: Conceptualisation, methodology.
J VanderDoes: Methodology, software.

Clara Macindoe¹, Charlie Roache², Brigitte Papa¹, Beena Kumar^1,2,Samar Ojaimi^1,2

¹Monash Health Pathology, Monash Health, Clayton, Australia

²Monash University, Clayton, Australia  

 

Introduction: 
Tonsillar tissue is frequently sent for flow cytometric analysis for exclusion of haematological malignancies, often without clear clinical indication. The lack of definitive guidelines for handling tonsillar specimens may lead to inefficient use of flow cytometry. This audit assesses the diagnostic yield of routine flow cytometry on tonsillar tissue, to inform more efficient resource utilization.

Methods: 
Data were collected from 165 patients over two years for all samples sent for histological assessment. Specimens submitted for flow cytometry were further analysed based on clinical history, histology findings, and flow cytometric results.

Results: 
The median patient age was 48 (1-87) years, with a predominance of male patients (57.6%). Among 417 specimens, flow cytometry was requested for 142. Of these, only 7.7% of tonsillar specimens with suspected haematological malignancy showed monoclonal populations. No monoclonal populations were detected in cases with suspected solid malignancy or tonsillar asymmetry. Tonsillar histopathology diagnosis correlated with flow cytometry 96.1% of the time.

Discussion: 
This audit suggests that routine flow cytometry for tonsillar specimens has limited diagnostic value except in suspected haematological malignancies. Histopathology alone is often adequate, supporting a rationalized approach to reduce unnecessary flow cytometry requests, thereby conserving resources. Future analysis will evaluate the economic and environmental impacts of these findings.

Author contributions:
Clara Macindoe - Data collection, statistical analysis, write up
Charlie Roache - Data collection and statistical analysis
Brigitte Papa - Data collection
Been Kumar - Program Director of Pathology, Monash Health
Samar Ojaimi - Project lead and head of Immunopathology, Monash Health

Samuel Baumgart¹, Ruvanika Rajapakse¹, Kartik Naidu¹, Juliette Holland^1
1Laverty Pathology, 60 Waterloo Road, North Ryde, Australia

Introduction: Interferon gamma-release assays (IGRA) screen for exposure to Mycobacterium tuberculosis. The QuantiFERON®-TB Gold Plus assay separates results qualitatively into positive, indeterminate and negative categories, however various ‘borderline’ ranges have been employed1,2. Laverty Pathology’s low positive criteria is TB1 and TB2 antigen both ≥0.35 and <2.00.  

Aim: Ascertain proportions of results in each category, the reasons for indeterminate results, and whether our low positive range is appropriate.

Method: Audit of all IGRA tests performed in 2023 including subsequent results. 

Results: 26157 samples were analysed. 1461 (5.59%) were low positive and 1218 (4.66%) positive. All subsequent reversions to negative results occurred only in those with prior low positive results. Indeterminate rates were low (195/26157, 0.75%) and mainly due to inadequate mitogen response. Follow-up samples were only received in 27/195 (13.9%) indeterminate cases with the majority (17/27, 63%) becoming negative. 

Conclusions: A cutoff value of 2.00 for TB1 and TB2 antigens separated ‘low positive’ results that are more likely to revert to negative from true positive results, however further research needs to be performed to delineate the ideal borderline range. Low rates of indeterminate results suggest good pre-analytical and analytical processes however obtaining follow-up samples may be an area for quality improvement. 

References:
1. Wikell A, Jonsson J, Dyrdak R, Henningsson AJ, Eringfalt A, Kjerstadius T, et al. Latent Tuberculosis Screening under Routine Conditions in a Low-Endemicity Setting. J Clin Microbiol. 2021;59:e01370-21. doi: https://doi.org/10.1128/JCM.01370-21 

2. Wang MS, Li-Hunnam J, Chen YL, Gilmour B, Alene KA, Zhang YA, Nicol MP. Conversion or reversion of interferon gamma release assays for Mycobacterium tuberculosis infection: a systematic review and meta-analysis. Clin Infect Dis. 2024;ciae357. doi: 10.1093/cid/ciae357 

Notes:
1. This work was the original idea of the authors, and oversight was provided by JH, RR and KN. SB performed the data collection and analysis. 

2. Generative AI was NOT used at any stage in the research process including the writing process.

Katherine Fraser¹ , Joanne Moses¹

¹Department of Anatomical Pathology, LabPlus, Auckland, New Zealand

 

Background:
Endometrial cancer (EC) is the most common gynaecological malignancy in New Zealand1. The Cancer Genome Atlas has identified four molecular subgroups of EC, each with distinct prognostic and therapeutic implications: POLE-ultramutated, mismatch repair-deficient (dMMR), p53-mutant (p53abn), and no specific molecular profile (NSMP)2. Targeted POLE sequencing has recently been implemented in Northern and Midland Gynaecological Services in selected high-grade EC.

Objective:
This audit evaluates the first 70 cases of EC sent for targeted molecular analysis to assess the prevalence of molecular subgroups and selected pathological correlations.

Methods:
We identified the first 70 EC cases (01/2023–09/2024) sequenced using the SONIC FIND-IT endometrial panel, which covers  POLE, TP53, PIK3CA, and CTNNB1 mutations. We then collected and analysed selected clinicopathological and molecular data.

Results:
Of the 70 cases, 13% were POLE-mutated, 31% dMMR, 40% p53-abnormal, and 13% NSMP. Thirteen percent had multiple molecular classifiers, with p53 as the second classifier in all cases. Nine cases showed discordance between p53 immunohistochemistry and TP53 sequencing.

Conclusion:
This audit highlights the promising integration of molecular classification in Northern and Midland regions of New Zealand and emphasizes the need to address discordances between IHC and sequencing results for more accurate EC diagnostics.

Statement of originality
My role in this audit was to collect and analyse the data from each case. As trends emerged, I added additional data points. I summarized the results and contributed to the conclusions. I also designed and developed the poster with supervision and guidance from Dr. Joanne Moses.