Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT03343834 |
| Other study ID # |
NI15023 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
N/A
|
| First received |
|
| Last updated |
|
| Start date |
December 21, 2017 |
| Est. completion date |
August 5, 2021 |
Study information
| Verified date |
October 2021 |
| Source |
Assistance Publique - Hôpitaux de Paris |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Interventional
|
Clinical Trial Summary
Scientific context
Epstein-Barr virus has a causal role in the pathogenesis of multiple distinct lymphomas.
Post-transplant lymphoproliferative diseases (PTLD) are the most frequent EBV-induced
proliferations. PTLD after allogeneic stem cell transplantation has an incidence lower than
5% but may increase up to 10-20% in patients with established risk factors. EBV-DNAemia is
predictive of EBV-PTLD and is routinely performed using qPCR on whole blood. Preemptive
therapeutic strategies with anti-CD20 antibody are used when patients are above a defined
EBV-DNAemia threshold. This approach remains limited since it does not discriminate between
an EBV-induced lymphoproliferation (latent cycle) and/or a replicating virus (replicative
cycle).
Epigenetic modifications plays a central role in regulating the switch from latent to lytic
gene expression. Specific DNA modifications can be regarded as molecular signatures for EBV
genomes associated with the status of the viral infection (latent vs lytic). Accordingly,
these signatures may be envisioned as a potent tool to characterize the state of the viral
infection in vivo.
Description of the project Our primary objective is to estimate the respective percentages of
EBV-lytic and EBV-latent genomes (proliferating cells) in patients presenting with a high
EBV-DNAemia after allogeneic stem cell transplantation HSCT by analysing the epigenetic
modifications of EBV genome on specific sites. Our secondary objectives are i) to determine
risk factors associated with each "latent versus lytic EBV" profiles and ii) to correlate the
"latent versus lytic EBV" profiles with response to rituximab infusion and patient outcomes.
For this purpose, a retrospective study (n=80) and a prospective study (estimation n=58) will
be established. The different steps of this project are:
1. To study epigenetic modifications. The laboratory is developing a new approach to
distinguish between latent and lytic genomes.
2. To realize quantitative analysis by RT-PCR of different EBV transcripts specific of the
latent or of the lytic phase of the virus This method will be applied on RNA extracted
from patient blood samples with elevated EBV viral load, under condition preserving RNA
integrity. The results will be validated on a prospective cohort of HSCT patients (n=58)
(Saint-Antoine Hospital and La Pitié-Salpêtrière Hospital).
3. To perform quantitative analysis of EBV genomes in plasma, saliva and total blood
samples by current routine procedures In addition to total blood samples, plasma and
saliva will be collected since free viral particles are known to accumulate in these
biological fluids upon EBV reactivation. These samples will be treated by normalized
procedures that are routinely used in the medical virology laboratory to quantify EBV in
human samples.
Expected results By establishing a simple method for studying epigenetic modifications of EBV
genomes, we expect to understand the significance of high EBV viral load and the
pathophysiology of post-HSCT PTLD. We aim to distinguish between the latent / lytic profiles
of HSCT patients and to correlate their respective risks for developing PTLD. Establishing
the epigenetic EBV profile in the post-HSCT setting when facing increase viral load and PTLD
will improve our understanding of the biological mechanisms determining EBV-status in
post-HSCT. This should improve major medical and economical issues. These results could have
a major therapeutic
Description:
Study duration :
Total study duration: 37 months Duration of recruitment: 24 months Duration of participation
for each patient: 13 months
Inclusion criteria:
Retrospective study: Patient who underwent HSCT, in Saint-Antoine hospital, between
2010-2015, treated by rituximab for high level EBV-DNAemia (above 3.3 log copies/mL).
Prospective study: Patients who underwent HSCT, in Saint-Antoine hospital and la
Pitié-Salpêtrière, in 2016-2017, treated by rituximab for high level EBV-DNAemia (above 10
000c/mL), And/or having post-transplant lymphoproliferative diseases(PTLD) Concerned
population Allogeneic hematopoietic stem cell transplantation (allo-HSCT) patients with a
high Epstein-Barr virus (EBV) viral load
Study endpoints:
Primary endpoint (linked to the primary objective) Respective percentages of latent versus
replicative EBV profiles, defined by the methylation level of specific sites of EBV genome,
in a cohort of HSCT patients with a high EBV viral load (above 3.3 log copies/ml) before the
initiation of treatment per rituximab. A methylation index will be calculated from the
respective differences of the "Cycle thresholds" of the methyl-qPCR performed on patients
blood samples Secondary endpoints (linked to the secondary objectives)
1. Characteristics of the patients according to the EBV q-PCR results
2. Patients' outcome according to the EBV profile defined by the methyl-qPCR :
- Response to Rituximab infusion (EBV DNA-emia < 3.3 log copies/mL) after a maximum
of 4 infusions)
- Disease-free survival (DFS) at 12 months defined as survival without relapse
- Overall survival (OS) at 12 months
- Relapse incidence (RI) at 12 months
- Non-relapse mortality (NRM) at 12 months
- Number of post-HSCT infections within 12 months
Statistical Analysis :
1. Comparison between characteristics of latent and lytic genomes will be performed using
parametric (Student t-test) or non-parametric (Mann Whitney test) when appropriate for
continuous variables, Chi-Square test for qualitative variables.
Multivariate analysis for determination of risk factors associated with "lytic EBV"
profiles will be performed using logistic regression.
2. Correlation between "latent versus lytic EBV" profiles" and response to rituximab
infusion will be performed using logistic regression including in the model all other
factors potentially associated to the response to rituximab. OS and DFS will be
estimated by using the Kaplan-Meier method. Cumulative incidences of RI and NRM will be
calculated from the date of inclusion to the date of relapse or death in remission,
respectively, with the other event being the competing risk. Death or progression will
be considered as a competing events for estimating the incidence of acute GVHD.
Multivariate analyses for survival endpoints will be performed using the Cox
proportional hazards model. The type I error rate is fixed at 0.05 for determination of
factors associated with time to event outcomes. All statistical analyses will be
performed with R software packages (R Foundation for Statistical Computing, Vienna,
Austria)
