Sequential Analysis Before and After Treatment Initiation to Unravel the Role of Naturally Occurring Extracellular Vesicles in HIV Infection

Recruiting

• Conditions: Hiv

• Registration Number: NCT04653610
• Lead Sponsor: University Hospital, Ghent

Brief Summary

This study aims to evaluate the role of extracellular vesicles in HIV-infection, by determining the expression profile and content of EVs before and after treatment initiation in HIV-infected patients, through extensive blood and tissue sampling (leukapheresis, stool sampling and colon biopsies). A one-time sampling (blood, stool, colon biopsies) will also be performed in HIV-seronegative healthy volunteers to confirm that results found in HIV-infected patients are related to the disease.

Detailed Description

Not available

Study Timeline

• Study First Submitted: 2020-10-23
• Study First Submitted QC: 2020-11-26
• Study First Posted: 2020-12-04
• Study Start: 2021-01-27
• Status Verified: 2024-02
• Last Update Submitted: 2024-02-06
• Last Update Posted: 2024-02-08
• Primary Completion: 2027-01-01
• Study Completion: 2027-01-01

Recruitment & Eligibility

• Status: RECRUITING
• Sex: All
• Target Recruitment: 105

Inclusion Criteria

• Documented untreated HIV-1 infection defined as followed: HIV-1-specific antibody+(western blot); p31+ (ELISA)
• CD4 T cell count will be determined standard of care (SOC). A minimum of 16 patients will be included with a CD4 T cell count lower than 350 cells/µl and a minimum of 16 patients with a CD4 T cell count higher than 350 cells/µl
• Able and willing to provide written informed consent
• Age ≥ 18 years and ≤ 65 years
• Ability to attend the complete schedule of assessments and patient visits as described in the schedule below
• Ability and willingness to have blood, stool and colon samples collected and stored for 20 years after finalizing the study, and used for various research purposes

A.2.

Exclusion Criteria

• Recent HIV-infection, early diagnosis

• Previous or current history of opportunistic infection (AIDS defining events as defined in category C of the CDC clinical classification), consisting of chronic HIV-1 infection

• Evidence of active HBV infection (Hepatitis B surface antigen positive or HBV viral load positive in the past and no evidence of subsequent seroconversion (=HBV antigen or viral load negative and positive HBV surface antibody))

• Evidence of active HCV infection: HCV antibody positive result within 60 days prior to study entry with positive HCV viral load or, if the HCV antibody result is negative, a positive HCV RNA result within 60 days prior to study entry

• Current or known history of cardiomyopathy or significant ischemic or cerebrovascular disease

• Current or known history of cancer

• Pregnancy or breastfeeding

• Any conditions, including preexisting psychiatric and psychological disorders, which will in the opinion of the investigator interfere with the trial conduct or safety of the participant. An initial psychiatric assessment will be made by the treating physician. Since making a correct psychological assessment at the time of diagnosis can be difficult, a visit with a psychologist is planned for patients included in the study, for a second evaluation. This will be planned within the first month after diagnosis. In consultation with the psychologist, further sampling will be planned or the patient will be excluded from further sampling.

• Previous participation in a trial evaluating an immune modulating agent

• Abnormal laboratory tests results at screening:

  1. Confirmed hemoglobin <11g/dl for women and <12 g/dl for men
  2. Confirmed platelet count < 100 000/µl
  3. Confirmed neutrophil count <1000/μl
  4. Confirmed AST and/or ALT > 10x ULN

• Active drug or alcohol use or dependence that, in the opinion of the site investigator, would interfere with adherence to study requirements

• Acute or serious illness, in the opinion of the site investigator, requiring systemic treatment and/or hospitalization within 60 days prior to entry

• Known inflammatory bowel disease (Crohn's disease or ulcerative colitis)

B. Healthy Volunteers

B.1. Inclusion Criteria:

• Able and willing to provide written informed consent
• Age ≥ 18 years and ≤ 65 years
• Ability to attend the complete sampling schedule, as described below
• Ability and willingness to have blood, stool and colon samples collected and stored for 20 years and used for various research purposes

B.2. Exclusion Criteria:

• HIV-infection

• Evidence of active HBV infection (Hepatitis B surface antigen positive or HBV viral load positive in the past and no evidence of subsequent seroconversion (=HBV antigen or viral load negative and positive HBV surface antibody))

• Evidence of active HCV infection: HCV antibody positive result within 60 days prior to study entry with positive HCV viral load or, if the HCV antibody result is negative, a positive HCV RNA result within 60 days prior to study entry

• Current or known history of cardiomyopathy or significant ischemic or cerebrovascular disease

• Current or known history of cancer

• Pregnancy or breastfeeding

• Any conditions, including preexisting psychiatric and psychological disorders, which will in the opinion of the investigator interfere with the trial conduct or safety of the participant

• Previous participation in a trial evaluating an immune modulating agent

• Abnormal laboratory tests results at screening:

  1. Confirmed hemoglobin <11g/dl for women and <12 g/dl for men
  2. Confirmed platelet count < 100 000/µl
  3. Confirmed neutrophil count <1000/μl
  4. Confirmed AST and/or ALT > 10xULN

• Active drug or alcohol use or dependence that, in the opinion of the site investigator, would interfere with adherence to study requirements

• Acute or serious illness, in the opinion of the site investigator, requiring systemic treatment and/or hospitalization within 60 days prior to entry

• Known inflammatory bowel disease (Crohn's disease or ulcerative colitis)

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Primary Outcome Measures

Name	Time	Method
Extracellular Vesicles analysis-RNAsequencing	6 years	The isolated EVs will be further characterized through RNA sequencing.
Quantification of HIV DNA and RNA	6 years	Digital PCR
Extracellular Vesicles analysis-NTA	6 years	Extracellular vesicles (EV) will be isolated through a combination of size-exclusion chromatography (SEC) and Optiprep density gradient (ODG). Nanoparticle Tracking Analysis (NTA) will be performed to obtain the concentration and size distribution of EVs in the samples.
Immunological analysis-ELISA	6 years	Immunophenotyping by flow cytometric assays will be performed of different cells to assess the phenotype of innate immune cells, using ELISA.
Virological analysis-FLIPS	6 years	HIV will be characterized by Full Length Individual Proviral Sequencing (FLIPS).
Extracellular Vesicles analysis-western blot	6 years	The isolated EVs will be further characterized through western blot.
Gene expression analysis/transcriptomics	6 years
Microbiome monitoring	6 years	Gut microbiome will be analyzed in stool and colon biopsies using next-generation sequencing (NGS) of rRNA gene amplicons to identify bacteria at genus/species level
Virological analysis-integration site	6 years	HIV will be characterized by integration site analysis.
Extracellular Vesicles analysis-PCR	6 years	The isolated EVs will be further characterized through PCR.
Extracellular Vesicles analysis-microscopy	6 years	The isolated EVs will be further visualized by (electron) microscopy.
Extracellular Vesicles analysis-reporter assays	6 years	Reporter assays will be performed to quantitatively measure bacterial EV-associated lipopolysaccharide (LPS).
Extracellular Vesicles analysis-proteomics	6 years	The isolated EVs will be further characterized through proteomic analysis.
Immunological analysis-FACS	6 years	Immunophenotyping by flow cytometric assays will be performed of different cells to assess the phenotype of innate immune cells, using FACS analysis.

Trial Locations

Locations (1)

Ghent University Hospital; 🇧🇪 Ghent, Oost-Vlaanderen, Belgium