search
Back to results

Methyl-qPCR : a New Predictive Marker for Epstein-Barr Virus-associated Lymphoproliferations During Allogeneic Hematopoietic Stem Cell Transplantation (EBVALLO)

Primary Purpose

Epstein-Barr Viraemia, Hematologic Diseases, Allogeneic Disease

Status
Completed
Phase
Not Applicable
Locations
France
Study Type
Interventional
Intervention
biological collection
Sponsored by
Assistance Publique - Hôpitaux de Paris
About
Eligibility
Locations
Arms
Outcomes
Full info

About this trial

This is an interventional basic science trial for Epstein-Barr Viraemia focused on measuring Allogenic cell stem transplant (HSCT), EBV-induced lymphoproliferation, Epigenetic

Eligibility Criteria

18 Years - undefined (Adult, Older Adult)All SexesDoes not accept healthy volunteers

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 2017-2018,
  • treated by rituximab for high level EBV-DNAemia (above 10 000c/mL),
  • And/or having post-transplant lymphoproliferative diseases(PTLD)

Exclusion Criteria:

  • Refusal to sign the informed consent
  • Patient non-beneficiary of the Social Security system

Sites / Locations

  • Service d'hématologie Clinique

Arms of the Study

Arm 1

Arm Type

Other

Arm Label

EBV+ allograft population

Arm Description

Retrospective study (n=80) : 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 (n=58) : 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

Outcomes

Primary Outcome Measures

Respective percentages of latent versus replicative EBV profiles
Defined by epigenetic modifications of EBV genome, in a cohort of HSCT patients with a high EBV viral load (above 10 000 c/ml) before the initiation of treatment per rituximab.

Secondary Outcome Measures

Characteristics of the patients according to the EBV results
To determine the risk factors associated with latent versus replicative EBV profiles, in allo-HSCT with a high viral load.
Patients' outcome according to the EBV profile
- Response to Rituximab infusion (EBV DNA-emia < 3 log copies/mL after a maximum of 4 infusions)

Full Information

First Posted
June 19, 2017
Last Updated
October 22, 2021
Sponsor
Assistance Publique - Hôpitaux de Paris
Collaborators
Institut National de la Santé Et de la Recherche Médicale, France
search

1. Study Identification

Unique Protocol Identification Number
NCT03343834
Brief Title
Methyl-qPCR : a New Predictive Marker for Epstein-Barr Virus-associated Lymphoproliferations During Allogeneic Hematopoietic Stem Cell Transplantation
Acronym
EBVALLO
Official Title
Methyl-qPCR : a New Predictive Marker for Epstein-Barr Virus-associated Lymphoproliferations During Allogeneic Hematopoietic Stem Cell Transplantation
Study Type
Interventional

2. Study Status

Record Verification Date
October 2021
Overall Recruitment Status
Completed
Study Start Date
December 21, 2017 (Actual)
Primary Completion Date
August 5, 2021 (Actual)
Study Completion Date
August 5, 2021 (Actual)

3. Sponsor/Collaborators

Responsible Party, by Official Title
Sponsor
Name of the Sponsor
Assistance Publique - Hôpitaux de Paris
Collaborators
Institut National de la Santé Et de la Recherche Médicale, France

4. Oversight

Studies a U.S. FDA-regulated Drug Product
No
Studies a U.S. FDA-regulated Device Product
No
Data Monitoring Committee
Yes

5. Study Description

Brief 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: To study epigenetic modifications. The laboratory is developing a new approach to distinguish between latent and lytic genomes. 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). 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
Detailed 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) Characteristics of the patients according to the EBV q-PCR results 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 : 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. 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)

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Epstein-Barr Viraemia, Hematologic Diseases, Allogeneic Disease
Keywords
Allogenic cell stem transplant (HSCT), EBV-induced lymphoproliferation, Epigenetic

7. Study Design

Primary Purpose
Basic Science
Study Phase
Not Applicable
Interventional Study Model
Single Group Assignment
Masking
None (Open Label)
Masking Description
1 population divided in 2 groups: Retrospective study (n=80): 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 (n=58): 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
Allocation
N/A
Enrollment
138 (Actual)

8. Arms, Groups, and Interventions

Arm Title
EBV+ allograft population
Arm Type
Other
Arm Description
Retrospective study (n=80) : 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 (n=58) : 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
Intervention Type
Biological
Intervention Name(s)
biological collection
Intervention Description
At D0: blood samples saliva sample At D8: blood sample the results will be compared to the analysis of EBV transcripts that are specific for the latent or replicative phase of the virus by reverse transcription-quantitative PCR (RT-qPCR), and to the detection of free virus in the saliva, which is associated with viral reactivation
Primary Outcome Measure Information:
Title
Respective percentages of latent versus replicative EBV profiles
Description
Defined by epigenetic modifications of EBV genome, in a cohort of HSCT patients with a high EBV viral load (above 10 000 c/ml) before the initiation of treatment per rituximab.
Time Frame
12 months
Secondary Outcome Measure Information:
Title
Characteristics of the patients according to the EBV results
Description
To determine the risk factors associated with latent versus replicative EBV profiles, in allo-HSCT with a high viral load.
Time Frame
12 months
Title
Patients' outcome according to the EBV profile
Description
- Response to Rituximab infusion (EBV DNA-emia < 3 log copies/mL after a maximum of 4 infusions)
Time Frame
12 months

10. Eligibility

Sex
All
Minimum Age & Unit of Time
18 Years
Accepts Healthy Volunteers
No
Eligibility Criteria
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 2017-2018, treated by rituximab for high level EBV-DNAemia (above 10 000c/mL), And/or having post-transplant lymphoproliferative diseases(PTLD) Exclusion Criteria: Refusal to sign the informed consent Patient non-beneficiary of the Social Security system
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
Eolia Brissot, Associate Professor
Organizational Affiliation
Assistance Publique - Hôpitaux de Paris
Official's Role
Principal Investigator
Facility Information:
Facility Name
Service d'hématologie Clinique
City
Paris
ZIP/Postal Code
75012
Country
France

12. IPD Sharing Statement

Plan to Share IPD
Undecided

Learn more about this trial

Methyl-qPCR : a New Predictive Marker for Epstein-Barr Virus-associated Lymphoproliferations During Allogeneic Hematopoietic Stem Cell Transplantation

We'll reach out to this number within 24 hrs