Skip to Main content Skip to Navigation
Theses

De la détection de l'ADNccc par de nouvelles technologies à la preuve de concept de sa dégradation à visée thérapeutique dans des modèles d'infection par le virus de l'hépatite B

Abstract : Hepatitis B virus {HBV) is a major health problem with 250 million chronic carriers, despite the existence of a preventive vaccine. Currently the treatments used are nucleos{t)ide analogues and / or interferon a. Although they efficiently reach a decrease of the viral load, they do not allow the eradication the disease due to the persistence of the cccDNA, the minichromosome of the hepatitis B. This DNA serves as a template for the viral transcription and only a single copy suffice for the infection rebound. However, the techniques used routinely for the quantification of the cccDNA are not sensitive enough to be able to detect low concentrations of this DNA, in particular in biopsies of patients chronically infected and long term treated. ln addition, it is necessary to develop new strategies to target the cccDNA in order to clear the infection. Thus, my thesis work is based on the development of a new technology: the Droplet Digital PCR {ddPCR) to allow the quantification of cccDNA in patient biopsies. This technique allows a gain of 2 log in sensitivity compared to the qPCR technique currently used in routine. lt allowed us to see the presence of this DNA in long-term treated patients even in the presence of interferon. The presence of pgRNA and ChlP experiments also confirmed that the cccDNA was still transcriptionally active.These results confirm the requirement to develop new therapeutics to allow the inactivation or even the elimination of the cccDNA. One of the strategies envisaged is the CRlSPR / Case 9 system. Thus, the following part of this thesis was to develop this system in hepatitis B virus infection models. To reduce off-target effect we tested 8 different guide RNAs incorporated via ribonucleoproteins into HepG2- NTCP cells. Preliminary results have shown that this system can reduce the pool of cccDNA in these cells and open up the possibilities to test this model on PHH and opens interesting perspectives for the development of new treatments
Document type :
Theses
Complete list of metadatas

Cited literature [29 references]  Display  Hide  Download

https://tel.archives-ouvertes.fr/tel-02021137
Contributor : Abes Star :  Contact
Submitted on : Friday, February 15, 2019 - 4:32:08 PM
Last modification on : Wednesday, June 10, 2020 - 6:42:06 PM
Long-term archiving on: : Thursday, May 16, 2019 - 6:54:41 PM

File

TH2018INCHAUSPEAURORE.pdf
Version validated by the jury (STAR)

Identifiers

  • HAL Id : tel-02021137, version 1

Collections

Citation

Aurore Inchauspe. De la détection de l'ADNccc par de nouvelles technologies à la preuve de concept de sa dégradation à visée thérapeutique dans des modèles d'infection par le virus de l'hépatite B. Virologie. Université de Lyon, 2018. Français. ⟨NNT : 2018LYSE1218⟩. ⟨tel-02021137⟩

Share

Metrics

Record views

260

Files downloads

218