role of mutations outside the rt domain of hbv polymerase
play

Role of mutations outside the RT-domain of HBV polymerase on - PowerPoint PPT Presentation

Dec 07, 2022 •271 likes •602 views

AREVIR-GenaFor-Meeting, Bonn, 05. May 2010 Role of mutations outside the RT-domain of HBV polymerase on antiviral resistance PD Dr. Dieter Glebe Institut fr Medizinische Virologie Justus-Liebig-Universitt Gieen HBV Polymerase

  1. “AREVIR-GenaFor-Meeting”, Bonn, 05. May 2010 Role of mutations outside the RT-domain of HBV polymerase on antiviral resistance PD Dr. Dieter Glebe Institut für Medizinische Virologie Justus-Liebig-Universität Gießen

  2. HBV Polymerase Secondary- Primary Resistance- resistance mutations mutations Zoulim & Locarnini, HBV resistance to nucleos(t)ide analogues . Gastroenterology, 2009, modified

  3. HBV genotypic assay Adapt treatment based on genotypic information Zoulim & Locarnini, HBV resistance to nucleos(t)ide analogues . Gastroenterology, 2009; 137:1593-1608

  4. Look at www.genafor.org g2p[hbv] – Input Page

  5. Look at www.genafor.org g2p[hbv] – Result Page

  6. HBV resistance: from genotype to phenotype BMBF project (2009-2012): H epatitis B therapies o ptimized by p henotypic e valuation (HOPE) Goals: – Validate resistance profile of HBV polymerase as a biomarker – Rapid rating of HBV (antiviral)-mutants ( Geno2pheno ) – improved clinical outcome of antiviral therapy • Involved groups: • Helmholtz Zentrum München (HMGU) (Protzer), • Justus-Liebig-Universität-Giessen (JLU) (Glebe, Gerlich), • Universität Duisburg-Essen University (Roggendorf, Lu) • Universität zu Köln (Kaiser), • Humboldt Universität Berlin/ Charite (van Boemmel), • Medizinische Hochschule Hannover (MHH) (Manns, Wursthorn), • Max-Planck-Institut Saarbrücken (MPI) (Lengauer, Beggel) • Dade Behring/Siemens Medical Solutions Diagnostics GmbH Deutschland Leverkusen,

  7. HBV resistance: from genotype to phenotype Phenotypic in vi vitr tro o resistance assay Expression plasmid with HBV-Insert Pol-frame amplicon from patients serum Wildtype pCH9-3091 pCH9-3091

  8. HBV resistance: from genotype to phenotype Wildtype Wildtype Wildtype Transfection of HuH7-cells pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 Wildtype Wildtype Wildtype pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 pCH9-3091 - incubation for 3 days - incubate over night -purify DNA from supernatant - wash cells - quantify DNA with Real time PCR

  9. HBV resistance: from genotype to phenotype Adefovir Entecavir Lamivudin Tenofovir no drug - 72 datapoints for every clinical isolate

  10. HBV resistance: from genotype to phenotype WT-Adefovir WT-Entecavir 1,4 1,2 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 Adefovir [nM] Entecavir [nM] WT-Lamivudin WT-Tenofovir 1,2 1,4 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Lamivudin [nM] Tenofovir [nM]

  11. HBV resistance: from genotype to phenotype WT-Adefovir WT-Entecavir 1,4 1,2 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 Adefovir [nM] Entecavir [nM] WT-Lamivudin WT-Tenofovir 1,4 1,4 1,2 1,2 relative replication relative replication 1 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Lamivudin [nM] Tenofovir [nM]

  12. HBV resistance: from genotype to phenotype Nucleoside analogs Nucleotide analogs Lamivudin 100 mg/d Adefovir dipivoxil 10 mg/d Entecavir 0,5 mg/d Tenofovir disoproxil 245 mg/d Telbivudin 1,0 mg/d aus: Schaefer, Glebe, Gerlich. Hepatitis B Virus ; in: Doerr & Gerlich, Medizinische Virologie HBV-Polymerase rtM204I Zoulim & Locarnini, HBV resistance to nucleos(t)ide S: sensitive; R: resistance; I: intermediate analogues . Gastroenterology, 2009; 137:1593-1608

  13. HBV resistance: from genotype to phenotype Adefovir Entecavir 1,4 1,2 1 24 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 Adefovir [nM] Entecavir [nM] HBV-Polymerase Lamivudin Tenofovir rtM204I 2 1,4 >10000 3 1,8 1,2 1,6 relative replication relative replication 1 1,4 1,2 0,8 1 0,6 0,8 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Lamivudin [nM] Tenofovir [nM]

  14. Phenotypic in vitro resistance assay Adefovir Entecavir 1,4 1,2 2,5 21,5 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 Adefovir [nM] Entecavir [nM] HBV-Polymerase rtL180M,rtM204V Lamivudin Tenofovir 1,8 1,4 >10000 3,3 1,6 1,2 1,4 relative replication relative replication 1 1,2 0,8 1 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Tenofvir [nM] Lamivudin [nM]

  15. HBV resistance: from genotype to phenotype Nukleoside analogs Nukleotide analogs Lamivudin 100 mg/d Adefovir dipivoxil 10 mg/d Entecavir 0,5 mg/d Tenofovir disoproxil 245 mg/d Telbivudin 1,0 mg/d aus: Schaefer, Glebe, Gerlich. Hepatitis B Virus ; in: Doerr & Gerlich, Medizinische Virologie HBV-Polymerase rtL180M,rtT184A,rtM204V Zoulim & Locarnini, HBV resistance to nucleos(t)ide S: sensitive; R: resistance; I: intermediate analogues . Gastroenterology, 2009; 137:1593-1608

  16. HBV resistance: from genotype to phenotype Adefovir Entecavir 1,4 1,2 2,5 153 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 Adefovir [nM] Entecavir [nM] HBV-Polymerase rtL180M,rtT184A,rtM204V Lamivudin Tenofovir 1,6 1,4 >10000 3,2 1,4 1,2 relative replication relative replication 1,2 1 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Lamivudin [nM] Tenofvir [nM]

  17. Phenotypic in vitro resistance assay Adefovir Entecavir 1,2 1,4 4 1 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 0,01 0,1 1 10 100 1000 10000 100000 Entecavir [nM] Adefovir [nM] HBV-Polymerase Tenofovir Lamivudin rtN236T 1,6 1,6 6 2,3 1,4 1,4 relative replication 1,2 relative replication 1,2 1 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Tenofovir [nM] Lamivudin [nM]

  18. HBV resistance: from genotype to phenotype Nukleoside analogs Nukleotide analogs Lamivudin 100 mg/d Adefovir dipivoxil 10 mg/d Entecavir 0,5 mg/d Tenofovir disoproxil 245 mg/d Telbivudin 1,0 mg/d aus: Schaefer, Glebe, Gerlich. Hepatitis B Virus ; in: Doerr & Gerlich, Medizinische Virologie HBV-Polymerase rtA181V Zoulim & Locarnini, HBV resistance to nucleos(t)ide S: sensitive; R: resistance; I: intermediate analogues . Gastroenterology, 2009; 137:1593-1608

  19. HBV resistance: from genotype to phenotype Adefovir Entecavir 1,4 1,6 1,5 4,5 1,4 1,2 1,2 relative replication relative replication 1 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 0,01 0,1 1 10 100 1000 10000 100000 Entecavir [nM] Adefovir [nM] HBV-Polymerase rtA181V Tenofovir Lamivudin 1,4 2 18 4 1,8 1,2 1,6 relative replication relative replication 1 1,4 1,2 0,8 1 0,6 0,8 0,4 0,6 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Tenofovir [nM] Lamivudin [nM]

  20. HBV resistance: from genotype to phenotype Nukleoside analogs Nukleotide analogs Lamivudin 100 mg/d Adefovir dipivoxil 10 mg/d Entecavir 0,5 mg/d Tenofovir disoproxil 245 mg/d Telbivudin 1,0 mg/d aus: Schaefer, Glebe, Gerlich. Hepatitis B Virus ; in: Doerr & Gerlich, Medizinische Virologie HBV-Polymerase rtA181V,rtN236T Zoulim & Locarnini, HBV resistance to nucleos(t)ide S: sensitive; R: resistance; I: intermediate analogues . Gastroenterology, 2009; 137:1593-1608

  21. HBV resistance: from genotype to phenotype Adefovir Entecavir 1,4 1,2 13,5 1 1,2 1 relative replication relative replication 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 Adefovir [nM] Entecavir [nM] HBV-Polymerase Tenofovir Lamivudin rtA181V,rtN236T 1,4 1,4 17,5 15 1,2 1,2 relative replication relative replication 1 1 0,8 0,8 0,6 0,6 0,4 0,4 0,2 0,2 0 0 0,01 0,1 1 10 100 1000 10000 100000 0,01 0,1 1 10 100 1000 10000 100000 Lamivudin [nM] Tenofovir [nM]

  22. Phenotypic in vitro resistance assay Nukleoside analogs Nukleotide analogs Lamivudin 100 mg/d Adefovir dipivoxil 10 mg/d Entecavir 0,5 mg/d Tenofovir disoproxil 245 mg/d Telbivudin 1,0 mg/d aus: Schaefer, Glebe, Gerlich. Hepatitis B Virus ; in: Doerr & Gerlich, Medizinische Virologie HBV-Polymerase rtA181V,rtN236T… Zoulim & Locarnini, HBV resistance to nucleos(t)ide S: sensitive; R: resistance; I: intermediate analogues . Gastroenterology, 2009; 137:1593-1608

Recommend

DNA polymerase kappa produces interrupted mutations and displays polar pausing within

DNA polymerase kappa produces interrupted mutations and displays polar pausing within

688696 Nucleic Acids Research, 2008, Vol. 36, No. 2 Published online 13 December 2007 doi:10.1093/nar/gkm1089 DNA polymerase kappa produces interrupted mutations and displays polar pausing within mononucleotide microsatellite sequences

132 views • 9 slides
Novel rtM204 Mutations in HBV Polymerase Confer Reduced Susceptibility to Adefovir and Tenofovir

Novel rtM204 Mutations in HBV Polymerase Confer Reduced Susceptibility to Adefovir and Tenofovir

Novel rtM204 Mutations in HBV Polymerase Confer Reduced Susceptibility to Adefovir and Tenofovir Ke Zhang, Christian Bach, Yuchen Xia, Oliver Schildgen, Ulrike Protzer Institute of Virology Technische Universitt Mnchen / Helmholtz Zentrum

277 views • 26 slides
Protein Domain-Centric Approach to Study Cancer Somatic Mutations from High-throughput Sequencing

Protein Domain-Centric Approach to Study Cancer Somatic Mutations from High-throughput Sequencing

Protein Domain-Centric Approach to Study Cancer Somatic Mutations from High-throughput Sequencing Studies Dr. Maricel G. Kann Assistant Professor Dept of Biological Sciences UMBC The term protein domain (or domain ) refers to a region of the

809 views • 33 slides
Lessons learned during the HOPE-Project - which role have HBV mutations in the times of

Lessons learned during the HOPE-Project - which role have HBV mutations in the times of

11.04.2013 AREVIR Meeting Kln Lessons learned during the HOPE-Project - which role have HBV mutations in the times of tenofovir and entecavir? Florian van Bmmel University Hospital Leipzig Hepatology Section Germany Current

361 views • 33 slides
Mutations, the molecular clock, and models of sequence evolution Why are mutations important?

Mutations, the molecular clock, and models of sequence evolution Why are mutations important?

Mutations, the molecular clock, and models of sequence evolution Why are mutations important? Mutations can Mutations drive be deleterious evolution Replicative proofreading and DNA repair constrain mutation rate UV damage to DNA UV

634 views • 21 slides
Mutations What is a mutation? ANY change in the genetic material (DNA) Mutations may be

Mutations What is a mutation? ANY change in the genetic material (DNA) Mutations may be

Mutations What is a mutation? ANY change in the genetic material (DNA) Mutations may be spontaneous or caused by mutagens (physical and chemical agents) Mutations are the primary source of genetic variation (not all are harmful)

644 views • 27 slides
Minimal Domain Expertise in a Financial Domain Mayank Kejriwal Role Relevance Problem In the

Minimal Domain Expertise in a Financial Domain Mayank Kejriwal Role Relevance Problem In the

Predicting Role Relevance with Minimal Domain Expertise in a Financial Domain Mayank Kejriwal Role Relevance Problem In the context of a text fragment, how relevant are the triples? "For the year ended December 31, 2015, billed business

478 views • 11 slides
DNA mutations http://www.ncbi.nlm.nih.gov/books/NBK2 1897/ 1 Types of mutations

DNA mutations http://www.ncbi.nlm.nih.gov/books/NBK2 1897/ 1 Types of mutations

DNA mutations http://www.ncbi.nlm.nih.gov/books/NBK2 1897/ 1 Types of mutations Micromutation that involve small regions of the DNA Macromutations that involve the chromosomes as a whole 2 DNA micromutations Single-base mutations

1.5k views • 135 slides
Polymerase Chain Reaction Dr. Lalani Yatawara Department of MLS, FAHS Introduction PCR,

Polymerase Chain Reaction Dr. Lalani Yatawara Department of MLS, FAHS Introduction PCR,

Polymerase Chain Reaction Dr. Lalani Yatawara Department of MLS, FAHS Introduction PCR, polymerase chain reaction, is an in-vitro technique for amplification of a region of DNA whose sequence is known or which lies between two regions of

243 views • 23 slides
mRNA Library Prep: cDNA clean up through adapter ligation Deoxynucleotide Mix T4 DNA

mRNA Library Prep: cDNA clean up through adapter ligation Deoxynucleotide Mix T4 DNA

mRNA Library Prep: cDNA clean up through adapter ligation Deoxynucleotide Mix T4 DNA polymerase and Klenow enzyme converts overhangs into blunt ends T4 DNA Polymerase T4 is a bacteriophage of E. coli. E. coli DNA Polymerase

195 views • 4 slides
Something very different - We can use this graph to fi nd mutations in new cases - Perhaps

Something very different - We can use this graph to fi nd mutations in new cases - Perhaps

Covid-19 genome variation graph Something very different - We can use this graph to fi nd mutations in new cases - Perhaps mutations which increase virulence Covid-19 phylogeny De Brujn graph - Genome assembly - Inferred using post order

445 views • 10 slides
Regulatory Motifs Gene Regulation Promoter Gene -35 -10 RNA polymerase Negative Positive

Regulatory Motifs Gene Regulation Promoter Gene -35 -10 RNA polymerase Negative Positive

Regulatory Motifs Gene Regulation Promoter Gene -35 -10 RNA polymerase Negative Positive Regulation Rep Regulation Act RNA polymerase 1 What if we believed that a number of genes were regulated by the same transcription factor? TF

506 views • 13 slides
A C Close se-Up L Look ook a at PCR Polymerase Chain Reaction (PCR) and cellular DNA

A C Close se-Up L Look ook a at PCR Polymerase Chain Reaction (PCR) and cellular DNA

A C Close se-Up L Look ook a at PCR Polymerase Chain Reaction (PCR) and cellular DNA replication both result in synthesis of new DNA molecules Rev eview ew: DNA NA Rep eplic lication DNA r repl plication i is a semiconser

123 views • 9 slides
Reframing the situation Implementing penalties for frameshifting mutations in a sequence

Reframing the situation Implementing penalties for frameshifting mutations in a sequence

Reframing the situation Implementing penalties for frameshifting mutations in a sequence alignment function Frameshift mutations Esciencecentral.org How do you do it? Movement Score Indel (1) -1 Indel (3) 0 Mismatch 0 Match 1

197 views • 8 slides
} { 64 MB Server Server 64 bit unique handle Many Client Many Triple

} { 64 MB Server Server 64 bit unique handle Many Client Many Triple

The Need Component failures normal Due to clustered computing Files are huge Google File System By traditional standards (many TB) Most mutations are mutations Not random access overwrite CSE 454 Co-Designing

321 views • 5 slides
EXPLORING THE ROLE FOR ETHICS AND EMPATHY IN THE CYBER DOMAIN By Gloria Barron,

EXPLORING THE ROLE FOR ETHICS AND EMPATHY IN THE CYBER DOMAIN By Gloria Barron,

EXPLORING THE ROLE FOR ETHICS AND EMPATHY IN THE CYBER DOMAIN By Gloria Barron, M.Ed. Computer Concepts, Computer Applications Montgomery College, Rockville Global Humanities Institute Many Voices, One Campus (MVOC) Fellowship

482 views • 25 slides
Phenotypic evaluation of resistance profiles of HBV polymerase against Lamivudine reveals an

Phenotypic evaluation of resistance profiles of HBV polymerase against Lamivudine reveals an

AREVIR-GenaFor-Meeting, Kln, 11.04.2013 Phenotypic evaluation of resistance profiles of HBV polymerase against Lamivudine reveals an unexpected resistance against Adefovir and Entecavir Dieter Glebe Institute for Medical Virology

592 views • 22 slides
Exploring The Role of Print 21H.343 J / CC.120J Spring 2016 1 The Role of Print in an Age of

Exploring The Role of Print 21H.343 J / CC.120J Spring 2016 1 The Role of Print in an Age of

Renaissance Religion and Cartography: Exploring The Role of Print 21H.343 J / CC.120J Spring 2016 1 The Role of Print in an Age of Religious Reformation Th e s e image s are in the public domain. Erasmus, Novum Instrumentum , 1516 edition,

374 views • 21 slides
Role of Domain Ignorance in Software Development Gaurav Mehrotra Masters Thesis Presentation

Role of Domain Ignorance in Software Development Gaurav Mehrotra Masters Thesis Presentation

Role of Domain Ignorance in Software Development Gaurav Mehrotra Masters Thesis Presentation Daniel M. Berry, Supervisor Outline Motivation Related Work Problem Statement Empirical Study Design Results Applications

901 views • 55 slides
Pollution-Induced Germ-line Mutations in Gull and Mouse Tandem Repeat DNA Dr. James S. Quinn

Pollution-Induced Germ-line Mutations in Gull and Mouse Tandem Repeat DNA Dr. James S. Quinn

Pollution-Induced Germ-line Mutations in Gull and Mouse Tandem Repeat DNA Dr. James S. Quinn Biology Department McMaster University DNA Damaging Agents In the Environment: - heritable mutations? Hamilton Harbour Remedial Action Plan:

840 views • 60 slides
Multi-domain Interventions to Prevent Cognitive Impairment and Alzheimers Disease : The Role of

Multi-domain Interventions to Prevent Cognitive Impairment and Alzheimers Disease : The Role of

Multi-domain Interventions to Prevent Cognitive Impairment and Alzheimers Disease : The Role of the Gut Microbiome Prof. Miia Kivipelto, MD, PhD Director of Research and Development Karolinska Institutet, NVS, Center for Alzheimer Research

526 views • 25 slides
Arevir University of Cologne Institute of Virology Analysis of resistance mutations of HI-Virus

Arevir University of Cologne Institute of Virology Analysis of resistance mutations of HI-Virus

Arevir University of Cologne Institute of Virology Analysis of resistance mutations of HI-Virus Bioinformatics analysis of relations between mutations of the HIV genome and phenotypical drug resistance for the optimization of anti-retroviral

262 views • 22 slides
The Role of the Interval Domain in Modern Exact Real Airthmetic Andrej Bauer Iztok Kavkler

The Role of the Interval Domain in Modern Exact Real Airthmetic Andrej Bauer Iztok Kavkler

The Role of the Interval Domain in Modern Exact Real Airthmetic Andrej Bauer Iztok Kavkler Faculty of Mathematics and Physics University of Ljubljana, Slovenia Domains VIII & Computability over Continuous Data Types Novosibirsk,

539 views • 15 slides
Specific Learning Disabilities: The Role of Working Memory and Other Domain-specific Deficits

Specific Learning Disabilities: The Role of Working Memory and Other Domain-specific Deficits

Specific Learning Disabilities: The Role of Working Memory and Other Domain-specific Deficits Lisa Archibald, PhD Western University, Canada Cogmed Conference 2014 Specific Learning Disabilities Appear to have typical potential to learn

562 views • 55 slides

More recommend