Biology subjects

Shields, J. D.

Publications and source records attributed to Shields, J. D..

5 recordsLinked to original sources

bioRxiv2025

Immune Profiling the Axilla with Fine Needle Aspiration is Feasible to Risk-Stratify Breast Cancer

Whether breast cancer (BC) has metastasised to the regional axillary lymph nodes (ALN) is an important prognostic factor that directs the extent of axillary surgery. However, surgically clearing the axilla carries morbidity, so less invasive methods of risk-stratifying patients are needed. ALN fine needle aspiration (FNA) is currently used to detect BC metastases, but these samples also contain immune cells. Here we show that FNA acquires sufficient leukocytes for comprehensive immunophenotyping of reactive, patient-derived ALN. All CD4+ and CD8+ T cells subsets (naive, terminal effector, central and effector memory) and rarer (<2%) natural killer (NK) and plasmacytoid dendritic cell (pDCs) populations are represented. Importantly, the immune cell profile of one reactive ALN appears to reflect the immune status of the patients axilla. We show that FNA captures immune differences between patients with [≤]1 or [≥]2 metastatic ALN. Significantly increased numbers of naive CD4+ T cells, but significantly fewer terminal effector, central memory and effector memory subpopulations, were obtained from patients with [≥]2 metastatic ALN. Moreover, despite their sparse distribution pattern on whole-section immunohistochemistry (WSI), FNA revealed that CD56+ NK cell activation receptors were decreased in patients with [≥]2 metastatic ALN. Finally, FNA captured a significant decrease in pDCs in patients with [≥]2 metastatic ALN, despite their clustered distribution pattern on WSI. In summary, FNA is not only feasible for sampling leukocytes from reactive, patient-derived ALN, but also identifies immune cell profiles that reflect axillary tumour burden in BC. Thus, this technique could be used to risk-stratify BC patients in future.

cancer biology↗

bioRxiv2024

Angiogenic and Immune Predictors of Neoadjuvant Axitinib Response in Renal Cell Carcinoma with Venous Tumour Thrombus

Venous tumour thrombus (VTT), where the primary tumour invades the renal vein and inferior vena cava, affects 10-15% of renal cell carcinoma (RCC) patients. Curative surgery for VTT is high-risk, but neoadjuvant therapy may improve outcomes. The NAXIVA trial demonstrated a 35% VTT response rate after 8 weeks of neoadjuvant axitinib, a VEGFR-directed therapy. However, understanding non-response is critical for better treatment. We conducted a multiparametric investigation of samples collected during NAXIVA using digital pathology, flow cytometry, plasma cytokine profiling and RNA sequencing. Responders had higher baseline microvessel density and increased induction of VEGF-A and PlGF during treatment. A multi-modal machine learning model integrating features predicted response with an AUC of 0.868, improving to 0.945 when using features from week 3. Key predictive features included plasma CCL17 and IL-12. These findings may guide future treatment strategies for VTT, improving the clinical management of this challenging scenario. One Sentence SummaryA comprehensive multiparametric assessment of the effect of neoadjuvant axitinib in renal cell carcinoma patients with venous tumour thrombus was performed on tissue and peripheral blood, including an integrative machine learning model, which identified both angiogenic and immune determinants of response to therapy.

cancer biology↗

bioRxiv2020

Stromal Amyloid β drives Neutrophil extracellular trap formation to augment tumour growth

Tumors are comprised of cancer cells and a network of non-cancerous stromal cells. Cancer-associated fibroblasts (CAFs) are well known to support tumorigenesis and are emerging as immune modulators. While many leukocyte populations are well studied in cancer, neutrophils have received less attention. Neutrophils can release histone-bound nuclear DNA and cytotoxic granules as extracellular traps (NETs) in a process termed NETosis. Here, we show that CAFs induce formation of NETs both within the tumor microenvironment and at systemic levels in the blood and bone marrow. These tumor-induced NETs (t-NETs) are driven by a ROS-mediated pathway dependent on PAD4 and CD11b. Remarkably, CAF-derived Amyloid {beta} was identified as the key factor driving t-NETosis, a protein with significance in both neurodegenerative and inflammatory disorders. Therapeutic inhibition of NETs in established tumors prevented growth, skewing neutrophils to a pro-inflammatory phenotype. Reciprocally, t-NETs enhanced CAF activation phenotypes. Mirroring murine observations, NETs were detected juxtaposed to CAFs in human melanoma and pancreatic adenocarcinoma, and elevated expression of amyloid and {beta}-Secretase correlated with poor prognosis. In summary, we report the existence of cross-talk between CAFs and neutrophils within the tumour microenvironment whereby CAF-induced t-NETosis supports cancer progression, identifying Amyloid {beta} as the protagonist and potential therapeutic target. SignificanceThis study defines the existence of a pro-tumor immunomodulatory function of the stroma showing the induction of Neutrophil Extracellular Traps through CAF-derived Amyloid {beta}. We term this novel process "Tumor-induced NETosis" (t-NETosis) and propose that therapeutic inhibition of this mechanism, which we observe in human melanoma and pancreatic cancer, has the potential to improve patient outcome.

cancer biology↗

bioRxiv2018

Podoplanin interaction with caveolin-1 promotes tumour cell migration and invasion

Podoplanin, a highly O-glycosylated type-1 transmembrane glycoprotein, found in lymphatic endothelial cells, podocytes, alveolar epithelial cells and lymph node fibroblasts is also expressed by tumour cells, and is correlated with more aggressive disease. Despite numerous studies documenting podoplanin expression, the mechanisms underlying its tumour-promoting functions remain unclear. Using a murine melanoma cell line that endogenously expresses podoplanin, we demonstrate interactions with proteins necessary for cytoskeleton reorganization, adhesion and matrix degradation, and endocytosis/receptor recycling but also identify a novel interaction with caveolin-1. We generated a panel of podoplanin and caveolin-1 variants to determine the molecular interactions and functional consequences of these interactions. Complementary in vitro and in vivo systems confirmed the existence of a functional cooperation in which surface expression of both full length, signalling competent podoplanin and caveolin-1 are necessary to induce directional migration and invasion, which is executed via PAK1 and ERK1 pathways. Our findings establish that podoplanin signalling mediates the invasive properties of melanoma cells in a caveolin-1 dependent manner.\n\nSummary StatementThis manuscript describes a new interaction and functional cooperation between podoplanin and caveolin1 that drives tumour cell invasion into surrounding tissues.

cancer biology↗

bioRxiv2018

Tumors induce de novo steroid biosynthesis in T cells to evade immunity

Tumors subvert immune cell function to evade immune responses, yet the complex mechanisms driving immune evasion remain poorly understood. Here we show that tumors induce de novo steroidogenesis in T lymphocytes to evade anti-tumor immunity. Using a novel transgenic steroidogenesis-reporter mouse line we identify and characterize de novo steroidogenic immune cells. Genetic ablation of T cell steroidogenesis restricts primary tumor growth and metastatic dissemination in mouse models. Steroidogenic T cells dysregulate anti-tumor immunity, and inhibition of the steroidogenesis pathway was sufficient to restore anti-tumor immunity. This study demonstrates T cell de novo steroidogenesis as a mechanism of anti-tumor immunosuppression and a potential druggable target.

immunology↗