A Case Report on Longitudinal Collection of Tumour Biopsies for Gene Expression-Based Tumour Microenvironment Analysis from Pancreatic Cancer Patients Treated with Endoscopic Ultrasound Guided Radiofrequency Ablation

Patrick V Lawrence; Krisha Desai; Christopher Wadsworth; Nagina Mangal; Hemant M Kocher; Nagy Habib; Anguraj Sadanandam; Mikael H Sodergren

doi:10.3390/curroncol29100531

A Case Report on Longitudinal Collection of Tumour Biopsies for Gene Expression-Based Tumour Microenvironment Analysis from Pancreatic Cancer Patients Treated with Endoscopic Ultrasound Guided Radiofrequency Ablation

Curr Oncol. 2022 Sep 21;29(10):6754-6763. doi: 10.3390/curroncol29100531.

Authors

Patrick V Lawrence¹, Krisha Desai¹, Christopher Wadsworth², Nagina Mangal^{1

2}, Hemant M Kocher^{3

4

5}, Nagy Habib², Anguraj Sadanandam^{1

6}, Mikael H Sodergren²

Affiliations

¹ Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.
² Department of Surgery and Cancer, Imperial College London, London W12 0HS, UK.
³ Centre for Tumour Biology, Barts Cancer Institute-A CRUK Centre of Excellence, Queen Mary University of London, London EC1M 5PZ, UK.
⁴ Barts and the London HPB Centre, The Royal London Hospital, Barts Health NHS Trust, London EC1M 5PZ, UK.
⁵ Barts Pancreas Tissue Bank, Barts Cancer Institute-A CRUK Centre of Excellence, Queen Mary University London, London EC1M 5PZ, UK.
⁶ Centre for Global Oncology, Division of Molecular Pathology, The Institute of Cancer Research, London SM2 5NG, UK.

Abstract

Background: Most patients with pancreatic ductal adenocarcinoma (PDAC) are metastatic at presentation with dismal prognosis warranting improved systemic therapy options. Longitudinal sampling for the assessment of treatment response poses a challenge for validating novel therapies. In this case study, we evaluate the feasibility of collecting endoscopic ultrasound (EUS)-guided longitudinal fine-needle aspiration biopsies (FNABs) from two PDAC patients and conduct gene expression studies associated with tumour microenvironment changes associated with radiofrequency ablation (RFA).

Methods: EUS-guided serial/longitudinal FNABs of tumour were collected before and after treatment from two stage III inoperable gemcitabine-treated PDAC patients treated with targeted RFA three times. Biopsies were analysed using a custom NanoString panel (144 genes) consisting of cancer and cancer-associated fibroblast (CAFs) subtypes and immune changes. CAF culture was established from one FNAB and characterised by immunofluorescence and immunoblotting.

Results: Two-course RFA led to the upregulation of the CD1E gene (involved in antigen presentation) in both patients 1 and 2 (4.5 and 3.9-fold changes) compared to baseline. Patient 1 showed increased T cell genes (CD4-8.7-fold change, CD8-35.7-fold change), cytolytic function (6.4-fold change) and inflammatory response (8-fold change). A greater than 2-fold upregulation of immune checkpoint genes was observed post-second RFA in both patients. Further, two-course RFA led to increased PDGFRα (4.5-fold change) and CAF subtypes B and C genes in patient 1 and subtypes A, B and D genes in patient 2. Patient 2-derived CAFs post-first RFA showed expression of PDGFRα, POSTN and MYH11 proteins. Finally, RFA led to the downregulation of classical PDAC subtype-specific genes in both patients.

Conclusions: This case study suggests longitudinal EUS-FNAB as a potential resource to study tumour and microenvironmental changes associated with RFA treatment. A large sample size is required in the future to assess the efficacy and safety of the treatment and perform comprehensive statistical analysis of EUS-RFA-based molecular changes in PDAC.

Keywords: fine-needle aspiration biopsy; immune checkpoint genes; pancreatic ductal adenocarcinoma; radiofrequency ablation; tumour microenvironment.

Publication types

Case Reports
Research Support, Non-U.S. Gov't

MeSH terms

Biopsy
Carcinoma, Pancreatic Ductal* / genetics
Carcinoma, Pancreatic Ductal* / surgery
Gene Expression
Humans
Pancreatic Neoplasms* / genetics
Pancreatic Neoplasms* / pathology
Pancreatic Neoplasms* / surgery
Radiofrequency Ablation*
Receptor, Platelet-Derived Growth Factor alpha / genetics
Tumor Microenvironment
Ultrasonography, Interventional

Substances

Receptor, Platelet-Derived Growth Factor alpha

Grants and funding

Funding, in part, was secured from EMcision (now acquired by Boston Scientific) and badged by the National Institute for Health Research (NIHR) for conducting the trial. No other funding source to be declared.