Immune Mechanisms After Radiofrequency Ablation of Pulmonary Metastases From Colorectal Cancer Origin- ARFIM Study
Phase: NA
Conditions: Immune Evasion, Tumor, Neoplastic Cells, Circulating, Circulating Tumor Cell, Pulmonary Metastasis, Colo-rectal Cancer
Sponsor: Institut Bergonié

EVALUATION OF IMMUNE REACTION AFTER 
RADIOFREQUENCY TREATMENT OF PULMONARY 
METASTASES IN COLORECTAL CANCER 
ARFIM STUDY 
 
Category 2 non-interventional research involving humans with minimal risks and 
constraints (RIPH 2) 
 
IB 2018-03 
NCT03960021 
Protocol version No. 4 - 10/06/2021 
 
This research is supported by the Groupement Interrégional de Recherche Clinique et 
d'Innovation Sud-Ouest Outre-Mer Hospitalier (Southwest-Overseas Interregional Hospital 
Group for Clinical Research and Innovation - GIRCI SOHO) 
 
 
Coordinator 
Dr Jean PALUSSIÈRE, Coordinator, Radiologist 
 
Department of Medical Radiology 
Institut Bergonié 
33076 Bordeaux Cedex 
 
 
Clinical Research and Epidemiology Unit 
Prof. Simone MATHOULIN-PELISSIER, Unit Head 
 Institut Bergonié 
Caroline LALET, Clinical studies manager 
Institut Bergonié 
Valérie DUMAS, Clinical Research Associate,  
Institut Bergonié 
Carine BELLERA, Biostatistician 
Institut Bergonié 
Fanny BOUTEILLER, Biostatistician  
Institut Bergonié 
 
 
Molecular Pathology Unit 
Dr Isabelle SOUBEYRAN, Unit head 
 
 
 
                      Institut Bergonié 
Dr Benjamin BONHOMME, Biopathologist 
 
 
 
                      Institut Bergonié 
Céline AUZANNEAU, Biological engineer  
 
 
 
 
         Institut Bergonié 
 
 
SPONSOR 
INSTITUT BERGONIÉ 
229 cours de l’Argonne 33076 BORDEAUX Cedex 
Tel: +33 (0)5.56.33.33.33 – Email: drci@bordeaux.unicancer.fr 
 
 

SPONSOR 
Institut Bergonié 
COORDINATING/PRINCIPAL 
INVESTIGATOR 
Dr Jean PALUSSIÈRE 
SCIENTIFIC MANAGER 
Nicolas PANGON, radiology intern 
TITLE 
Evaluation of the immune response after radiofrequency 
treatment of lung metastases of colorectal cancer. ARFIM 
study. 
RATIONALE / BACKGROUND 
Radiofrequency (RF) is now widely used in the percutaneous 
treatment of lung metastases of colorectal cancers. 
The pathophysiology of reactions induced locally by this ablation 
technique at metastatic sites is poorly known, particularly in terms 
of immunology. Some studies have shown that RF ablation may 
alter the environment and immunological mechanisms, not only at 
treated sites, but also at distant sites. However, this RF-induced 
immune response is insufficient to cause the regression of 
metastases at a distance from the treated site. 
From an immunological point of view, the importance of the PD-
1/PD-L1 axis has been shown in tumour mechanisms of 
resistance to the immune system, in particular by inhibiting 
lymphocyte action. Combining RF with targeted anti-PD-1/PDL-1 
therapies could therefore be an interesting therapeutic option to 
control metastases distant from those treated with RF. 
OBJECTIVES 
Primary objective: To evaluate the RF-induced immune response 
to lung metastases of colorectal cancers (CRC). The 
radiofrequency-induced immune response will be jointly evaluated 
in 3 ways: by measuring changes to lymphocyte infiltration of 
metastasis tumour stroma, by quantifying the PD-1 and PD-L1 
interaction by immune Förster Resonance Energy Transfer 
(iFRET), and by assaying blood lymphocyte sub-populations.  
Secondary objectives:  
 To evaluate the impact of RF on tumour activity by measuring 
the level of circulating tumour cells and circulating DNA, along 
with as the expression of the PDL-1 ligand on tumour cells.  
 Clinical evaluation with monitoring of metastatic disease and 
especially RF-treated sites 
 Evaluation of procedure-related complications (RF and 
biopsies) 

ENDPOINTS 
Primary endpoint:  
Lymphocyte infiltration rate on histological analysis of metastases. 
Biopsy samples will be taken before RF and 4-6 weeks after the 
first RF in a second procedure on another initially untreated 
metastasis. A lymphocyte infiltration rate of 20% or more is 
considered significant. A comparative measurement of lymphocyte 
infiltration rates will be performed. 
A blood assay of NK, CD4, CD8 T cells and their HLADR+ 
activated sub-population, CD25+, CD38+, will be performed 
before, then 30 minutes after each RF, in order to evaluate 
distribution changes.  
Secondary endpoints:  
 Rate of circulating tumour cells and of circulating DNA before 
and after each RF (before RF, at 30 minutes and on D1 after 
RF). Quantification of PDL-1 ligand expression on tumour cells 
before and after RF from histological samples 
 Assessment of the progress of treated sites and of any distant 
metastatic disease by scans at 3, 6, 9 and 12 months, as part 
of standard follow-up.  
 Complications will be graded (1 to 5) according to version 5 of 
the NCI-CTAE scale. 
STUDY DESIGN 
Prospective single-centre pathophysiology study 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
INCLUSION CRITERIA 
1. Patient aged 18 years or over. 
2. WHO performance status ≤ 2. 
3. Histologically confirmed colorectal cancer. 
4. Resected primary tumour. 
5. Lung metastases 
a. bilateral (or unilateral if: number ≥ 5 and requiring two 
interventions), 
b. maximum diameter ≤ 4 cm, 
c. stable or slowly progressive (radiologically defined), with or 
without chemotherapy, 
d. eligible for radiofrequency therapy. 
6. Thorax-abdomen-pelvis CT scan and/or PET scan: 
a. performed within 8 weeks prior to inclusion, 
b. finding lung metastases of which no more than 10 can be 
treated with radiofrequency during both times (supplemental 
metastases being considered untreatable with radiofrequency 
due to their small size). 
7. Inclusion criterion No. 7 was deleted during MS2. 
8. Decision on local treatment taken during the multidisciplinary 
meeting or during an Interventional Radiology opinion meeting 
(no contraindication). 
9. Life expectancy ≥ 3 months. 
10. Free, informed and written consent signed by the participant 
and the investigator (on the day of inclusion at the latest and 
before any tests required by the study).  
11. Patient with social security cover in accordance with Article 
1121‐11 of the French public health code. 

EXCLUSION CRITERIA 
1. Non-lung or liver metastases. 
2. Contraindication to general anaesthesia. 
3. Contraindication to radiofrequency treatment tumour 
localization (< 1 cm of the hilus), respiratory failure (FEV1 < 1 
l), 
4. Pregnant or breastfeeding woman. 
5. Concomitant inclusion in another interventional radiology 
study.   
6. Patient incapable of following and adhering to research 
procedures for geographical, social or psychological reasons. 
7. Patient deprived of freedom or subject to a legal protective 
measure. 
8. Previous treatment with hormone therapy. 
STUDY 
TREATMENTS/STRATEGIES/PROCEDU
RES 
Lung radiofrequency treatment of lung metastases of colorectal 
cancer. 
The procedure is performed under general anaesthesia. 
A RF needle is deployed, under strict asepsis, into the metastasis 
under imaging guidance. Once deployed, heating is performed to 
destroy the lesion. Each patient will undergo 2 procedures, to treat 
all metastases 
Biopsy samples are taken from a location before the heating 
phase. 
STUDY SIZE 
20 patients 
STUDY DURATION 
Duration of the inclusion period: 24 months 
Duration of participation of each patient: 12 months 
Total study duration: 36 months  
DATA STATISTICAL ANALYSIS 
Number of subjects required 
This exploratory study is not based on a statistical hypothesis but 
on feasibility criteria in terms of recruitment. 20 patients will be 
included in this study  
Statistical analyses 
Patient characteristics will be described for the eligible and eligible 
evaluable populations:  
 Compliance with eligibility criteria, 
 Socio-demographic characteristics, 
 Clinical and tumour characteristics,  
 Performing procedures (RF, radiological examinations). 
Analysis of primary and secondary endpoints:  
 will concern the eligible and evaluable population. 
 Quantitative variables will be described from mean values (+/- 
sd) if the normality hypothesis is met, and otherwise from non-
parametric statistics (median, range, quartiles). 
 Qualitative variables will be described based on frequency, 
percentage and 95% confidence interval (binomial law). 
EXPECTED BENEFITS 
 Better understanding of radiofrequency-induced 
pathophysiological and immunological mechanisms of lung 
metastases 
 with the possibility of developing combined RF/immunotherapy 
treatments 
 an evaluation of metastatic disease activity with a new 
endpoint consisting of the circulating tumour cell (CTC) level