A translational study of the role of Fusobacterium nucleatum in oral cancer treatment efficacy using tumour-on-chip models

Primary supervisor: Miguel Reis Ferreira, King’s College London

Secondary supervisor: Oliver Pearce, Queen Mary University of London

Tertiary supervisor: Stefaan Verbruggen, Queen Mary University of London

Project

Oral squamous cell carcinoma (OSCC) is devastating. Patients with high-risk OSCC undergo mutilating surgery and post-operative radiotherapy (+- chemotherapy), often resulting in life-long inability to swallow and/or communicate. Despite this, recurrence rates are high [1]. Relapsed OSCC is notoriously difficult to treat. There is a clinical unmet need to improve OSCC treatment, which has not significantly changed in the last decade.

Tumour-associated microbiota may modulate treatment response, particularly in organ-preserving therapies such as radiotherapy or immunotherapy [2]. The Reis Ferreira (MRF) group has identified that patients with detectable and/or higher abundance of Fusobacterium nucleatum (Fnuc), a Gram-negative anaerobic oral commensal bacterium, in their OSCC have better survival and that this bacterium promotes OSCC death in 2D cell culture models. Previous research also indicates that Fnuc impacts on cancer-associated immunity, although these relationships are under-researched in OSCC [3,4].

Herein, we will test the hypothesis that tumour-associated Fnuc improves the effectiveness of non-surgical OSCC treatment.

The candidate will initially develop innovative 2D and 3D tumour-on-chip microfluidic models of OSCC, immune cells and bacteria at the Pearce and Verbruggen labs, which specialise in tumour microenvironment and in modelling microenvironments on in vitro microfluidics and chip devices respectively [5]. The FDA Modernization Act 2.0 approved these systems to replace animal models for pre-clinical drug safety and efficacy testing, following the European Directive 2010/63/EU to reduce or replace animals in science and medicine. OSCC 3D cultures will leverage experience from Prof Tony Ng’s and Dr Anthony Kong’s labs (King’s College London). Tumour tissue and PBMC will be obtained from the KHP head and neck cancer biobank. Commercially-available Fnuc strains will be used, but a culture collection of patient-derived Fnuc matching obtained tumour tissue will also be created. In collaboration with the radiotherapy department (Guys), a protocol for irradiation of these models using clinically-active linear accelerators will be developed leveraging previous experience of MRF’s group.

Using tissue from patients treated for stage III-IV OSCC at Guys, intra-tumoural Fnuc will be quantified using fluorescence in situ hybridisation in fixed OSCC and deep sequencing in fresh aseptically-collected matched samples. The radio, chemo and immunotherapy sensitivity of patient-derived organoids in microfluidic models will be ascertained and the effect of Fnuc co-culture in treatment effectiveness will be assessed in both Fnuc-high and Fnuc-low tumours, defining whether Fnuc has a direct impact in OSCC treatment resistance. RNAseq will delineate differential expression in OSCC treated with Fnuc.

RNAseq will assess the effect of Fnuc in PBMCs in isolation to determine immune cell populations of interest. The concomitant effect of immune and non-immune mediated effects of Fnuc in treatment response will then be tested using tri-culture microfluidic models.

Candidate background

This project would suit candidates with a background in biology and an interest in cancer and/or radiotherapy research, 3D and organ-on-chip models.

Potential Research Placements

1. Rebecca Carter, UCL Preclinical Radiotherapy Core Facility; David Eaton, Radiotherapy Physics Guys and St Thomas/ King’s College London
2. David Moyes, Centre for Host-Microbiome Interactions, King’s College London
3. Ann Sandison, Head and neck pathology department, Guys and St Thomas NHS Foundation Trust

References

1. Sambasivan K, Sassoon I, Thavaraj S, Kennedy R, Doss G, Michaelidou A, Odell E, Sandison A, Hall G, Morgan P, Collins LHC, Lyons A, Cascarini L, Fry A, Oakley R, Simo R, Jeannon JP, Petkar I, Reis Ferreira M, Kong A, Lei M, Guerrero Urbano T. TNM 8 staging is a better prognosticator than TNM 7 for patients with locally advanced oral cavity squamous cell carcinoma treated with surgery and post-operative radiotherapy. Radiother Oncol. 2021;160:54-60.
2. Reis Ferreira M, Pasto A, Ng T, Patel V, Guerrero Urbano T, Sears C, Wade W. The microbiota and radiotherapy for head and neck cancer: what should clinical oncologists know? Cancer Treat Rev. 2022 Aug;102442.
3. Michikawa C, Gopalakrishnan V, Harrandah AM, Karpinets T V, Garg RR, Chu RA, Park YP, Chukkapallia SS, Yadlapalli N, Erikson-Carter KC, Gleber-Netto FO, Sayour E, Progulske-Fox A, Chan Edward K.L., Wu X, Zhang J, Jobin C, Wargo JA, Pickering CR, Myers JN, Silver N. Fusobacterium is enriched in oral cancer and promotes induction of programmed death-ligand 1 (PD-L1). Neoplasia. 2022 Sep;31:100813.
4. Gur C, Ibrahim Y, Isaacson B, Yamin R, Abed J, Gamliel M, Enk J, Bar-On Y, Stanietsky-Kaynan N, Coppenhagen-Glazer S, Shussman N, Almogy G, Cuapio A, Hofer E, Mevorach D, Tabib A, Ortenberg R, Markel G, Miklić K, Jonjic S, Brennan CA, Garrett WS, Bachrach G, Mandelboim O. Binding of the Fap2 protein of fusobacterium nucleatum to human inhibitory receptor TIGIT protects tumors from immune cell attack. Immunity. 2015;
5. Nolan J, Pearce OMT, Screen HRC, Knight MM, Verbruggen SW. Organ-on-a-Chip and Microfluidic Platforms for Oncology in the UK. Cancers (Basel). 2023 Jan 19;15(3):635.