Spatial profiling of tumour-draining lymph nodes to stratify
treatment in oral cancer

Primary supervisor: Clare Schilling, UCL

Secondary supervisor: Sheeba Irshad, King’s College London

Project

Oral squamous cell cancer (OSCC) is a devastating disease affecting vital functions such as speech, eating, and appearance. Surgery is the most common treatment, but recent data from the KEYNOTE-689 trial demonstrate promising responses to immunotherapy in the neoadjuvant setting for a subset of patients. This marks a critical shift in a disease where PD-1 inhibitors have historically shown limited benefit. However, predicting which patients will derive durable benefit remains an unmet clinical need. Biomarkers are urgently required to avoid overtreatment and the immunotherapy-related toxicity in likely non-responders, while identifying those who may be spared major surgery.

This UCL/KCL partnership offers the student access to advanced spatial technologies, robust immunology pipelines, and a cross-disciplinary mentorship environment at the intersection of biomarker discovery and treatment stratification. The student will also gain international research exposure through a collaboration the Łukasiewicz – PORT Institute in Poland, accessing complementary expertise in digital pathology, H&E based machine learning, and tissue array construction.

Proposal:

This PhD will investigate the immunological landscape of tumour-draining lymph nodes (TDLNs) in early-stage oral cancer to guide treatment stratification and de-escalation strategies. The student will use both archival and prospectively collected samples from patients undergoing sentinel lymph node biopsy, focusing on immune cell phenotypes, spatial organisation, and clinical correlates of immunotherapy response. Key findings will be validated using an independent patient cohort contributed by the Łukasiewicz – PORT team, enabling assessment of reproducibility in a multi-centre international setting.

Aim 1: Define immune architecture and metastatic potential of tumour-draining lymph nodes in early oral cancer 

Haematoxylin and eosin (H&E) stained slides will be used for histopathological quality control, tumour mapping, and selection of regions of interest. Visual assessment by expert histopathologists will guide identification of key tumour and lymph node features. In parallel, computational pathology tools – including digital image analysis and machine learning-based tissue segmentation- will enhance reproducibility and enable highthroughput identification of potentially predictive histological features associated with immune activity and metastatic risk.

Aim 2: Perform high-dimensional spatial immune profiling using multiplex proteomics

Multiplexed immunofluorescence will be performed using in-house validated panels on the Phenocycler (Akoya Biosciences) to characterise the composition, activation status, and spatial distribution of immune and stromal cells in both primary tumour and lymph node compartments. Spatial topographies will be analysed in relation to clinical features including presence/absence of occult metastases, extranodal extension, and progressionfree survival. Bioinformatic support from the Irshad lab will assist in cluster analysis, neighbourhood mapping, and dimensionality reduction.

Aim 3: Develop and test a translational immune-based assay for stratifying treatment decisions

The ultimate aim of this project is to inform the development of a minimally invasive bedside assay, such as fine needle aspiration (FNA) followed by immune profiling, to assess the immunological status of tumour-draining lymph nodes. The goal is to distinguish “hot” (immune-active) from “inert” (immune-inactive) nodes, which may correlate with both metastatic potential and likelihood of immunotherapy response. This stratification could guide treatment decisions, identifying patients who may benefit from surgery-sparing immunotherapy or, conversely, those requiring more aggressive intervention.

Potential Research Placements

1. Sheeba Irshad, Breast Immunology Group, Guys Innovation Hub, King’s College London
2. Adrian Biddle, Blizzard Institute, Queen Mary University of London
3. Patrycja Gazinska, Population Diagnostics Center, Łukasiewicz Research Network – PORT Polish Center for Technology Development, Wroclaw (PL)

References

1. Uppaluri R, Haddad RI, Tao Y, Le Tourneau C, Lee NY, Westra W, Chernock R, Tahara M, Harrington KJ, Klochikhin AL, Braña I, Vasconcelos Alves G, Hughes BGM, Oliva M, Pinto Figueiredo Lima I, Ueda T, Rutkowski T, Schroeder U, Mauz PS, Fuereder T, Laban S, Oridate N, Popovtzer A, Mach N, Korobko Y, Costa DA, Hooda-Nehra A, Rodriguez CP, Bell RB, Manschot C, Benjamin K, Gumuscu B, Adkins D; KEYNOTE-689 Investigators. Neoadjuvant and Adjuvant Pembrolizuma in Locally Advanced Head and Neck Cancer. N Engl J Med. 2025 Jul 3;393(1):37-50.
2. Goode EF, Roussos Torres ET, Irshad S. Lymph Node Immune Profiles as Predictive Biomarkers for Immune Checkpoint Inhibitor Response. Front Mol Biosci. 2021 May 24;8:674558.
3. Alternatywa: Verastegui E, Morales R, Barrera JL, Müeller A, Guzman B, Meneses A, Alfaro G. Immunological approach in the evaluation of regional lymph nodes of patients with squamous cell carcinoma of the head and neck. Clin Immunol. 2002 Jan;102(1):37-47.
4. The impact of elective cervical lymph node treatment on the tumour immune response in head and neck squamous cell carcinoma: time for a change in treatment strategy? Payne K, Nenclares P, Schilling C. BJC reports vol 2 68 (2024)
5. Incidence and predictive factors for positive non-sentinel lymph nodes in completion neck dissection following a positive sentinel node biopsy in early oral cancer. Karamchandani S, Sahovaler A, Crosbie-Jones E, McGurk M, Thavaraj S, Alibhai M, Wan S, Forster M, Sassoon I, Schilling C. Oral Oncology, Dec 2024 (159)