Publications

Background: For patients with mCRPC, PSMA-targeted radioligand treatment has significantly improved the clinical outcome. A blood-based liquid biopsy assay for recognizing PSMA protein expression on circulating tumor cells may be beneficial for better informing therapeutic decision-making and identifying the patients most likely to benefit from PSMA-targeted radioligand therapy. Methods: Using high-throughput imaging and digital AI pathology algorithms, a four-color immunofluorescence assay has been developed to find PSMA protein expression on CTCs on a glass slide. Cell line cells (LNCaP/PC3s/22Rv1) spiked into healthy donor blood were used to study the precision, specificity, sensitivity, limit of detection, and overall accuracy of the assay. Clinical validation and low-pass whole-genome sequencing were performed in PSMA-PET-positive patients with high-risk mCRPC (N = 24) utilizing 3 mL of blood. Results: The PSMA CTC IF assay achieved analytical specificity, sensitivity, and overall accuracy above 99% with high precision. In the clinical validation, 76% (16/21) of the cases were PSMA positive with CTC heterogeneity, and 88% (21/24) of the patients contained at least one conventional CTC per milliliter of blood. Thirty-six low-pass-sequenced CTCs from 11 individuals with mCRPC frequently exhibited copy number increases in AR and MYC and losses in RB1, PTEN, TP53, and BRCA2 locus. Conclusions: The analytical validation utilizing Epic Sciences’ liquid biopsy CTC platform demonstrated the potential to detect PSMA protein expression in CTCs from patients with mCRPC. This assay is positioned as an effective research tool to evaluate PSMA expression, heterogeneity, and therapeutic response in many ongoing clinical studies to target tumors that express PSMA.

<p>RB1 FISH from sample p.16.0 showing RB1 (red) and reference (green) showing a mixture of: hemizygously deleted cells (white arrows) and homozygously deleted cells (orange arrows).</p>

In patients early post-autologous stem cell transplant, seroprotection rates were high for Hemophilus influenzae type B and tetanus toxoid (70%-90%) but lower for Streptococcus pneumoniae (30%-50%) including after revaccination. There were high rates of seropositivity (67%-86%) to measles, mumps, and rubella and varicella zoster virus. Durability of protection requires assessment.

<p>Supplementary Figure 3. Pano induced pancreatic tumor cell apoptosis.</p>

Adoptive T-cell immunotherapy holds great promise for the treatment of viral complications in immunocompromised patients resistant to standard anti-viral strategies. We present a retrospective analysis of 78 patients from 19 hospitals across Australia and New Zealand, treated over the last 15 years with "off-the-shelf" allogeneic T cells directed to a combination of Epstein–Barr virus (EBV), cytomegalovirus (CMV), BK polyomavirus (BKV), John Cunningham virus (JCV) and/or adenovirus (AdV) under the Australian Therapeutic Goods Administration's Special Access Scheme. Most patients had severe post-transplant viral complications, including drug-resistant end-organ CMV disease, BKV-associated haemorrhagic cystitis and EBV-driven post-transplant lymphoproliferative disorder. Adoptive immunotherapy is well tolerated with few adverse effects. Importantly, 46/71 (65%) patients show definitive clinical improvement including reduction in viral load, clinical symptoms and complete resolution of end-organ disease. In addition, seven high-risk patients remain disease free. Based on this long-term encouraging clinical experience, we propose that a dedicated nationally funded centre for anti-viral cellular therapies should be considered to provide T cell therapies for critically ill patients for compassionate use. Adoptive T-cell immunotherapy offers promise to patients who are resistant to standard anti-viral strategies. Here the authors describe clinical observations in patients with viral complications treated with adoptive immunotherapy over the last 15 years.

β-adrenergic receptors are upregulated in melanoma cells and contribute to an immunosuppressive, pro-tumorigenic microenvironment. This study investigated the prognostic and predictive value of β-adrenoreceptor blockade by β-blockers in the EORTC1325/KEYNOTE-054 randomised controlled trial.Patients with resected stage IIIA, IIIB or IIIC melanoma and regional lymphadenectomy received 200 mg of adjuvant pembrolizumab (n = 514) or placebo (n = 505) every three weeks for one year or until recurrence or unacceptable toxicity. At a median follow-up of 3 years, pembrolizumab prolonged recurrence-free survival (RFS) compared to placebo (hazard ratio (HR) 0.56, 95% confidence interval (CI) 0.47-0.68). β-blocker use was defined as oral administration of any β-blocker within 30 days of randomisation. A multivariable Cox proportional hazard model was used to estimate the HR for the association between the use of β-blockers and RFS.Ninety-nine (10%) of 1019 randomised patients used β-blockers at baseline. β-blockers had no independent prognostic effect on RFS: HR 0.96 (95% CI 0.70-1.31). The HRs of RFS associated with β-blocker use were 0.67 (95% CI 0.38-1.19) in the pembrolizumab arm and 1.15 (95% CI 0.80-1.66) in the placebo arm. The HR of RFS associated with pembrolizumab compared to placebo was 0.34 (95% CI 0.18-0.65) among β-blocker users and 0.59 (95% CI 0.48-0.71) among those not using β-blockers.This study suggests no prognostic effect of β-blockers in resected high-risk stage III melanoma. However, β-blockers may predict improved efficacy of adjuvant pembrolizumab treatment. The combination of immunotherapy with β-blockers merits further investigation. This study is registered with ClinicalTrials.gov, NCT02362594, and EudraCT, 2014-004944-37.

<p>Focal copy number alterations proximal to viral integrations in V+MCC.</p>

Abstract Merkel cell carcinoma (MCC) is a highly aggressive neuroendocrine tumor of the skin with an estimated disease‐associated mortality of 15–33%. Australia has a higher incidence of MCC compared to the rest of the world, thought to be due to a higher ultraviolet index. The Australian MCC population is distinct from the MCC population of the Northern hemisphere, characterized by a predominantly viral negative etiology with high tumor mutational burden. The optimal management of MCC and the choice of treatment modality vary significantly across the world and even between institutions within Australia. Historically, the treatment for MCC has been resection followed by radiotherapy (RT), though definitive RT is an alternative treatment used commonly in Australia. The arrival of immune checkpoint inhibitors and the mounting evidence that MCC is a highly immunogenic disease is transforming the treatment landscape for MCC. Australia is playing a key role in the further development of treatment options for MCC with two upcoming Australian/New Zealand investigator‐initiated clinical trials that will explore the interplay of RT and immunotherapy in the treatment of early and late stage MCC.

Yuquan Pill (YQP), a Chinese patent medicine for the treatment of diabetes, is widely used in the treatment of diabetes and its complications in China. However, the efficacy of YQP on type 2 diabetes mellitus (T2DM) has not been completely assessed. The aim of this study is to evaluate the efficacy and safety of YQP in the treatment of T2DM.

<p>Supplementary Figure 4. Pano did not enhance the cytotoxicity of CARaMEL cells nor oncolysis of VV-gp100.</p>

Viral screening before initiation of biologics in patients with inflammatory bowel disease during the COVID-19 outbreakWe read with interest the Comment by Ren Mao and colleagues on the implications of coronavirus disease 2019 (COVID-19) in patients with pre-existing digestive diseases, and the strategies implemented in China to restrict the risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in patients with inflammatory bowel disease. 1 We agree with the current evidence that does not support drug suspension and also with the European Crohn's and Colitis Organisation COVID-19We declare no competing interests.

256 Background: PSMA-targeted radioligand therapy has dramatically improved clinical outcomes in men with mCRPC. A blood-based liquid biopsy assay to detect PSMA expression on circulating tumor cells (CTCs) could be helpful to optimize therapeutic decisions and identify patients most likely to benefit from PSMA-targeted therapies. Here we describe the development of this PSMA blood-based assay and the application of the single cell sequencing (SCS) pipeline to identify copy number alterations and further characterize heterogeneity in late-stage disease. Methods: A 4-color immunofluorescence (IF) assay was developed to detect PSMA expression on CTCs on the Epic Sciences liquid biopsy platform. CTCs were identified as DAPI+, cytokeratin (CK)+, and CD45- with appropriate cell morphology. Cells were plated onto glass slides and subjected to IF staining followed by CTC identification using a high throughput scanning platform with digital pathology machine learning algorithms. Cell line cells (LNCaP, PC3s, 22Rv1) of varying PSMA expression levels were spiked into normal blood (CLC/WBC; 1:10000) to create laboratory derived slides. Assay specificity, sensitivity, limit of detection (LOD), precision, and overall accuracy, as well as subcellular localization of biomarker expression were reported. Clinical validation was performed in a cohort of high-risk mCRPC patients (N=24) with previously demonstrated PSMA expression by tissue biopsy, utilizing 3ml of blood. Additionally, a subset of CTCs (36 cells from 11 patients) underwent single-cell low-pass whole genome sequencing to identify copy number alterations. Results: The PSMA CTC assay (Epic Sciences) achieved analytical specificity, sensitivity, and overall accuracy of over 99%. Precision was evaluated using intra-class correlation (ICC) (0.98). The assay showed an LOD of 1 CLC/technical replicate. PSMA expression was specific to staining outside the nucleus and distinguished from CK cytoplasmic staining. Analysis of 24 mCRPC samples revealed that 21/24 (88%) had at least one conventional CTC/mL (range: 0-30 CTCs/mL), of which 16/21 (76%) were PSMA positive (range: 0-27 CTCs/mL) with considerable tumor heterogeneity. 36 sequenced CTCs (N=11 pts) exhibited genomic heterogeneity and frequent AR (3/11), MYC (7/11), and MYCN (5/11) gains or losses of PTEN (4/11), TP53 (7/11), BRCA2 (5/11), CHD1 (6/11), and/or RB1(5/11 pts). Conclusions: The PSMA CTC assay analytical results firmly support its feasibility to detect PSMA expression in CTCs from mCRPC patient samples. The assay demonstrates high sensitivity, specificity, and overall accuracy. Many ongoing trials are designed to target PSMA-positive tumors, positioning this assay as a robust research tool to identify and longitudinally monitor PSMA expression and explore heterogeneity, response, and resistance in these patient populations.