OAR@UM Community:

Decoding human monocyte responses to damage-associated molecular patterns (DAMPs)

Thu, 01 Jan 2026 00:00:00 GMT

Title: Decoding human monocyte responses to damage-associated molecular patterns (DAMPs) Abstract: Sepsis is defined as life-threatening organ dysfunction as a result of infection. Numerous studies have attempted to understand the mechanisms underlying this aggressive inflammatory state; however, no specific treatments have yet been approved. Several clinical trials to mitigate overwhelming mortality rates due to sepsis have been unsuccessful. These failures have been ascribed to several gaps in our understanding of sepsis pathophysiology, particularly those pertaining to immune paralysis or maladaptive tolerance. Despite those failures, the current paradigm of sepsis that predominantly centres on pathogen-associated molecular patterns and their recognition have not been revised. Damage- associated molecular patterns are known to be elevated during sepsis, particularly Adenosine Triphosphate (ATP). The project hypothesises that extracellular ATP contributes to innate immune activation and tolerance. In order to test this hypothesis, the response of human primary monocytes (white blood cells that play a major role in innate immunity) against ATP was studied. Approximately 9 whole blood samples from young Maltese donors between the ages of 18 - 30 years old were obtained in heparinised (green-top) BD Vacutainer® tubes. Monocytes from each sample were purified and exposed to LPS as a positive control or ATP for 6 hours in vitro. Enzyme-linked immunosorbent assays (ELISA) targeting the prototypical cytokine Interleukin-1β (IL-1β) found in the supernatant were measured. Altogether, the results will contribute to a growing body of evidence on DAMPs establishing a maladaptive immune response. Description: B.Sc. (Hons)(Melit.)

The effect of TGF-beta on EMT in pancreatic cancer

Thu, 01 Jan 2026 00:00:00 GMT

Title: The effect of TGF-beta on EMT in pancreatic cancer Abstract: Pancreatic cancer is one of the most aggressive forms of cancer, often having a very low prognosis if not caught early. A known contributor to the aggressiveness of pancreatic cancer is the epithelial to mesenchymal transition (EMT). The EMT is a dynamic process, during which pancreatic cells begin to downregulate epithelial features, and begin to upregulate mesenchymal ones. Advanced stages of the EMT are associated with features such as increased metastatic potential and immune evasion. A known contributor to the EMT is the cytokine transforming growth factor-beta (TGF-β). The presence of TGFβ in the microenvironment of pancreatic cancer cells is known to promote the progression of the EMT. The aim of this study was to investigate how different concentrations of TGF-β over different treatment times affect the EMT in the pancreatic cancer cell line PANC-1. To achieve this, PANC-1 cells were treated with varying TGF-β doses, and gene expression was measured after 24- and 48- hours. The genes chosen were those most commonly mentioned with regard to the EMT, as well as TGF-β. These included epithelial genes CDH1, EpCAM, and SDC1. As well as mesenchymal genes VIM, ZEB1, CD44, SMAD4, NT5E, and CDH2. In the majority of cases, a clear relationship was found between the concentration and duration of TGF-β treatment, and changes in gene expression which were indicative of EMT progression. TGF-β was concluded to promote EMT progression as epithelial genes tended to be downregulated, and mesenchymal genes tended to be upregulated following treatment. Description: B.Sc. (Hons)(Melit.)

Targeting lung tumours : phage display identification of single chain antibodies against ROR1 and AXL

Thu, 01 Jan 2026 00:00:00 GMT

Title: Targeting lung tumours : phage display identification of single chain antibodies against ROR1 and AXL Abstract: Globally, lung cancer remains the leading cause of cancer-related mortality, with a poor prognosis despite the current treatment options such as surgery, chemotherapy and radiotherapy. Immunotherapy is currently emerging as a promising alternative by harnessing the immune system’s ability to identify and eliminate tumour cells through tumour-associated antigens (TAAs) with high specificity in targeting cancer cells, minimising the adverse effects on healthy tissue, unlike the aforementioned treatments. The TAAs of interest, Receptor Tyrosine Kinase-Like Orphan Receptor 1 (ROR1) and Anexelekto (AXL) are both overexpressed in non-small cell lung cancer (NSCLC), particularly in adenocarcinomas. Both contribute to tumour growth, metastasis and overall resistance to commercially available treatments. This study aimed to generate and characterise high-affinity single-chain variable fragment (scFv) antibodies against ROR1 and AXL using the Phage Display biopanning technique, an efficient and ethical alternative to hybridoma technology. Multiple rounds of biopanning were performed to enrich phage clones with increased binding affinity and specificity toward the TAAs. Selected clones were screened and validated using enzyme linked immunosorbent assay (ELISA) to assess their binding activity, followed by Sanger Sequencing to confirm clone sequence diversity and integrity. The results showed successful enrichment of antigen-specific phage clones across four consecutive biopanning rounds for the selected scFv targeting ROR1 adding to their potential as targeted immunotherapeutic agents as a more effective and targeted approach to lung cancer treatment. In contrast, biopanning against AXL did not yield clones, highlighting the challenges and needs for further optimisation and considerations in the overall procedure. Description: B.Sc. (Hons)(Melit.)

Targeting lung tumours : phage display identification of single chain antibodies against TROP2 and TIGIT

Thu, 01 Jan 2026 00:00:00 GMT

Title: Targeting lung tumours : phage display identification of single chain antibodies against TROP2 and TIGIT Abstract: Lung cancer is the most prevalent type of cancer and continues to remain a major global cause of death. Existing treatment options such as chemotherapy, radiotherapy or surgery, frequently have low specificity, leading to toxicity and severe adverse effects. Thus, for better patient prognoses and less adverse effects, novel targeted therapies must be developed. Immunotherapies may be better alternatives as they use immune cells, which can specifically target tumour associated antigens (TAAs) and immune checkpoints. In this project, phage display biopanning was used to identify and validate single chain variable fragment (scFv) antibodies against Trophoblast Cell Surface Antigen 2 (TROP2) and T cell immunoreceptor with immunoglobulin and ITIM domain (TIGIT). However, due to consistently poor binding across all biopanning rounds, the target was subsequently changed to GlucocorticoidInduced Tumour Necrosis Factor Receptor Related Protein (GITR). A total of 75 monoclones of anti-GITR scFv displaying phages were generated and picked using a phage display library. Enzyme-linked immunosorbent assay (ELISA) was used to determine the specificity of the phage clones to the target protein GITR versus non-target protein severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) trimer. An elevated on-target signal was produced by clone 64 (p-value <0.0001). Clone 64 along with 7 non-binders were validated for their specificity, affinity and diversity using Sanger sequencing. This revealed that clone 64 was a false positive result, sharing an identical Complementarity Determining Region 3 – Variable Heavy Chain (CDR3-VH) sequence with several non-binding clones. Therefore, no functional anti-GITR scFv antibodies were identified in this study. The technical limitations encountered will guide future optimisation of library diversity, monoclonal generation and integration of high‑throughput sequencing to isolate diverse, high‑affinity scFvs against GITR. Description: B.Sc. (Hons)(Melit.)