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Dégradation de l’ADNccc 2.3

2 Matériels et Méthodes

Données sur les cohortes

Cohorte PEGAN\

&@=A.&7A@.7"5B@%7@"&A&@@.9BF&?AZ"97A?:5(&A?7%96.@((A.&7A ,(@%&v} |z 7@ &A AA&.7A@ %_B7& -(=A.A&  "-?97.>B& F&" %&@ 7A.,'7&@ & 7(,A.+@] 5@ =?(@&7A.&7A B7& "-?,& F.?5& 7(,A.F& =9B? 5_ %&  &A 97A (A( A?.A(@ =&7%7A B 69.7@ B7 7 =? 759,B&@ 7B"5(9@jAk.%.>B&@ %7@ A?9.@ @&?F."&@ %& @9.7@ A&?A..?&@ &7 -(=A959,.&&7?7"&]&@"?.A'?&@%_.7"5B@.97(A.&7A\

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Cohorte de transplantés hépatiques de l’hôpital de Beaujon :

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Extraction d’ADN et ARN des échantillons de biopsie

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Quantification ADN et ARN par qPCR

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Quantification ADN et ARN par ddPCR

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Chromatine immunoprécipitation (µChIP)

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3 Résultats

Evaluation de l’ADNccc et de son activité transcriptionnelle dans la cohorte PEGAN :

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Evaluation de l’ADNccc et de son activité transcriptionnelle dans la cohorte de patients traités par la Telbivudine

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Fifty-six patients (46 males and 10 females) from the clinical trial NV-02B-015 (Novartis Pharmaceuticals) were prospectively enrolled for the CLDT600ACN04E1 study, an extension of CLDT600ACN04E study assessing the efficacy and safety in fourth to sixth year of Telbivudine treatment in patients with Chronic Hepatitis B (CHB) (Novartis Pharmaceuticals). New consent was given and the study was approved by the competent ethical committee.

=A.&7A@-%"96=5&A&%xA9zI&?@9+&5!.FB%.7&A-&?=I""9?%.7,A9A-&NV-02B-015 and CLDT600ACN04E1 clinical trials 7% A-& A?&A6&7A G@ "97A.7B&%] 5.7."5 7% F.?959,."5%AG&?&"955&"A&%!&+9?&!&,.77.7,9+A?&A6&7Aj!@&5.7&jk9+NV-02B-015 studyk 7% A A-& &7% 9+ +9559G B= j&7% =9.7A jk 9+ CLDT600ACN04E1 studyk jB==5&6&7A?I .,B?& vk] .@A959,I G@ @@&@@&% 97 5.F&? A.@@B& A  7% 5.F&? !.9=@I =&?+9?6&%Asnap frozen at -80°C to subsequent viral markers analysis.

For the study of cccDNA epigenetic regulation features, 7 untreated CHB patients from an historical cohort were used as comparative samples. These 7 CHB patients underwent liver biopsy in the frame of their clinical follow-up and a fragment was preserved for research purpose after patient’s consent and stored at -80°C. The protocol was approved by institutional Ethic Committees.

None of the patients from the clinical trial or the historical cohort presented Human Immunodeficiency Virus (HIV), Hepatitis C virus or Hepatitis Delta virus co-infection.

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Intrahepatic total HBV DNA, cccDNA and pgRNA quantification using qPCR method

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