Specialized Workshop F2Friday September 30, 2022: 7am - 5:30pm - Topic List

"Latest Advancements in Gene Therapy, Cell Therapy, and Vaccine Bioanalysis"

Immunogenicity & Risk Assessment of Gene, Cell Therapies & Vaccines; LNP & Viral Vectors for Oligonucleotide-based Therapy/Vaccines; qPCR/ddPCR Method Development & Validation Strategies; NGS & nanoString New and Specific Developments; Current Regulatory Expectations

(You can scroll down to see the details of each topic)

Part 1: Immunogenicity & Risk Assessment for Gene Therapy, Cell Therapy and Vaccines

  • Topic 1:

    Immunogenicity on Novel Drug Modalities & Expectations for Immunogenicity Risk Assessment for Gene Therapy programs: Lesson Learnt based on Novel Data from Non-clinical and Clinical Studies
    Dr. Yuanxin Xu, Vice President Early Development and Translational Medicine, Intellia

  • Topic 2:

    Current Development/Validation challenges of in vitro Human PBMC Cytokine Release Assays for Immunogenicity Risk Assessment: How can these assays be Applied across Multiple Modalities to Justify Bioanalytical Strategy?
    Dr. Johanna Mora, Scientific Director, Bristol Myers Squibb

  • Topic 3:

    Recent Developments in Vaccine Immunogenicity Strategies: Harmonization Process, China NMPA Draft Guidelines for Vaccine Immunogenicity and Novel Case studies
    Mr. Francis Dessy, Clinical Assay Senior Advisor, GlaxoSmithKline

Part 2: LNP and Viral Vectors Immunogenicity for Oligonucleotide-based Therapies & Vaccines

  • Topic 4:

    "The mRNA-based Therapies & Vaccines Era”: Clinical bioanalytical strategy, PK, PD and safety/disease biomarkers
    Dr. Yuling Wu, Head of Discovery Bioanalysis, AstraZeneca

  • Topic 5:

    Immunogenicity Assessment for Oligonucleotide-based Therapeutics: Lesson Learned based on Novel Data from Non-clinical and Clinical Studies
    Dr. Susovan Mohapatra, Associate Director DMPK, Stoke

  • Topic 6:

    Are LNPs used for Oligonucleotide-based Therapies & Vaccines Immunogenic? State-of-art Industry Experience and Novel Data in this field
    Dr. Adam Williams, Senior Scientist, AstraZeneca

  • Topic 7:

    Comparing & Contrasting Cell-based vs LBA for pre-existing anti-AAV TAbs and NAbs Assays: How to select a suitable assay platform?
    Dr. Lin Yang, Director of Bioanalytical Research, REGENXBIO

Part 3: qPCR/ddPCR Infectivity, Vaccine & Biodistribution Assays

  • Topic 8:

    Recent and Novel Developments on Universal Genomic Assay Strategy to Characterize the Biodistribution in Gene Therapy by qPCR/ddPCR
    Dr. Hiroshi Sugimoto, Associate Director, Takeda

  • Topic 9:

    Updated Strategies for qPCR Infectivity Assays Development/Validation for Gene Therapy and Oncolytic Virus
    Mr. Justin Smith, Associate Director & Biomarker Clinical Assay Lead, Pfizer

  • Topic 10:

    Advanced approaches in Molecular Assays to Support Vaccines Studies: qPCR, ddPCR and NGS Applications
    Dr. Ludovic Malvaux, Director & Head of Applied Microbiology Platform, GlaxoSmithKline

Part 4: NGS and NanoString Assays

  • Topic 11:

    Next Generation Sequencing (NGS) Bioanalytical Applications in Genetics & Genomics
    Dr. Jessica Seitzer, Vice President, Head of Genomic Operations, Intellia

  • Topic 12:

    Customizing NGS Bioanalytical Applications to Different Phases of Drug Development: What to consider in NGS use in Discovery vs Clinical Monitoring
    Dr. Sean Downing, Senior Director Clinical Biomarker Innovation & Development, Takeda

  • Topic 13:

    Recommendations for NGS Assay Fit for Purpose Validation
    Mr. Serge Picard, Senior Manager Molecular Assay Development, Bristol Myers Squibb

  • Topic 14:

    Recent Developments in Using NanoString Technology as an Alternative to qPCR: Understanding the Technology and its Potential Application in Bioanalysis
    Dr. David Bettoun, Vice President Discovery and non-clinical development, Larimar

Part 5: White Paper in Bioanalysis

  • 2022 White Paper on Gene Therapy, Cell Therapy and Vaccine Topics:

    Consensus & Conclusions on Gene Therapy, Cell Therapy and Vaccine Topics for 2022 White Paper

Finale: ASK THE REGULATORS!

  • Panel Discussion with Regulators:

    Regulatory Feedbacks on Submitted studies and Inspections/audits Outcomes on Gene Therapy, Cell Therapy & Vaccines

    • Dr. Joshua Xu (US FDA NCTR)
    • Dr. Nirjal Bhattarai (US FDA CBER)
    • Dr. Zuben Sauna (US FDA CBER)
    • Dr. Heba Degheidy (US FDA CBER)
    • Dr. Vijaya Simhadri (US FDA CBER)
    • Dr. Leslie Wagner (US FDA CBER)
    • Dr. Therese Solstad (Norway NoMA/EU EMA)
    • Dr. Shirley Hopper (UK MHRA)
    • Dr. Omar Tounekti (Health Canada)
    • Dr. Chad Irwin (Health Canada)
    • Dr. Julie Joseph (Health Canada)
    • Dr. Lucia Zhang (Health Canada)
    • Dr. Takenori Yamamoto (Japan MHLW)

 

Topic DETAILS of F2

Part 1: Immunogenicity & Risk Assessment for Gene Therapy, Cell Therapy and Vaccines
  • Topic 1

    Immunogenicity on Novel Drug Modalities & Expectations for Immunogenicity Risk Assessment for Gene Therapy programs: Lesson Learnt based on Novel Data from Non-clinical and Clinical Studies
    Dr. Yuanxin Xu, Vice President Early Development and Translational Medicine, Intellia

    • Understanding the Immunogenicity Risk Assessment for Novel Drug Modalities
      • Experience, evidence, and established approaches for assessing immunogenicity risk
      • Similarities & Differences of Immunogenicity for Gene Therapy vs Therapeutic Proteins
      • Is the Traditional Tiered Approach used for biotherapeutics suitable for Gene Therapy?
      • Evolving Industry standard and Lesson Learned for approved Gene Therapies
    • Current Regulatory expectations of immunogenicity of Gene Therapy
      • Regulatory Guidance/Guideline on follow up of patients administered with Gene therapy
      • 2009 EU EMA Guideline & 2018 US FDA Guidance
    • Factors to consider for Risk Assessment for Gene Therapy
      • Route of Administration & Vector used: Non-Viral (LNPs) and Viral Vectors
      • Characteristics of the Transgene
      • Immunogenicity of Nucleic Acids due to DAMPs, PAMPs, B Cells direct activation, production of anti-dsDNA ab
    • Present focus on new drug modalities risk assessment
      • CRISPR/Cas9 genome editing technology for human therapeutic
    • in vivo and ex vivo recent/ongoing studies
      • Cells edited in the body (in vivo)
      • Cells edited outside the body (ex vivo) from a patient or donor
    • Importance of Non-clinical NHP studies to evaluate immunogenicity
      • Immunogenicity Risk Assessment
    • Interim Clinical Data
      • What conclusions can be drawn from interpretation of recent immunogenicity clinical data
    • Case Studies: Lesson learned based on novel data from non-clinical and clinical studies
  •  

  • Topic 2

    Current Development/Validation challenges of in vitro Human PBMC Cytokine Release Assays for Immunogenicity Risk Assessment: How can these assays be Applied across Multiple Modalities to Justify Bioanalytical Strategy?
    Dr. Johanna Mora, Scientific Director, Bristol Myers Squibb

    • Importance of PBCM assay to assess if there is T cell activation through the cytokine Multiplex readout
      • Validation strategy for in vitro human PBMC cytokine release assay used to assess immunogenicity risk from Cell Therapy product residuals
    • Luminex readout
      • Traditional approach of looking at a single cytokine readout
      • Novel approach of using a full Luminex panel with a priori defined criteria on which cytokines to focus on for innate and adaptive immune responses
    • Applications to Cell Therapy
      • Issues with the assay
      • Solutions to data interpretation
    • Flow Cytometry
      • Understanding how in vitro human PBMC cytokine release assay validation strategy can be applied to a Flow Cytometry Assay used for T cell proliferation
    • Case Studies: Development/Validation strategies for of in vitro Human PBMC Cytokine Release Assays for Immunogenicity Risk Assessment
  •  

  • Topic 3:

    Recent Developments in Vaccine Immunogenicity Strategies: Harmonization Process, China NMPA Draft Guidelines for Vaccine Immunogenicity and Novel Case studies
    Mr. Francis Dessy, Clinical Assay Senior Advisor, GlaxoSmithKline

    • Importance of Immunogenicity Assessments in Vaccine Efficacy evaluation
      • Preclinical to clinical study and post-marketing studies
      • Recommendations highlighting the need for Vaccine specific guidelines
    • Unique characteristics of Vaccine Immunogenicity Assays
      • Specificities of Vaccine Immunogenicity Assays vs ADA Assays
    • Status of the vaccine immunogenicity assays development and validation initiative
      • Current Harmonization Process
      • Sensitivity limits (LOB, LOD, LLOQ) verification in validation
      • Clarification on Definitions and interpretation
      • Importance to make the link with the assay intended use
      • Verification vs re-assessment
      • Impact of imputation of non-measurable values
    • The China NMPA Draft Guidelines for Vaccine Immunogenicity
      • First concrete attempt for a vaccine specific guideline
      • Expectation to have similar Guidelines from other Regulatory Agencies
    • Case Studies: Procedures for vaccine immunogenicity assays development/validation and harmonization,

Part 2: LNP and Viral Vectors Immunogenicity for Oligonucleotide-based Therapies & Vaccines
  • Topic 4:

    The mRNA-based Therapies & Vaccines Era”: Clinical bioanalytical strategy, PK, PD and safety/disease biomarkers
    Dr. Yuling Wu, Head of Discovery Bioanalysis, AstraZeneca

    • Messenger ribonucleic acid (mRNA)-based Therapies & Vaccines
      • Widely proven as a promising treatment strategy in immune therapeutics
      • Numerous ongoing pre-clinical and clinical trials against various infectious diseases and cancers
    • Many advantages associated with mRNA drugs/vaccines
      • High efficacy
      • Relatively low severity of side effects
      • Low realization costs
      • Recent technological advancements
      • Better understanding of mRNA immunogenicity
    • Intratumorally injected mRNA- based therapy:
      • Lipid nanoparticle (LNP)-formulated mRNA encoding a key cytokine
      • Evaluation of cytokine production and enhances antitumor immune response
    • Bioanalytical strategies and challenges to support clinical trials
      • PK Assays
      • PD and safety/disease biomarkers
    • Bioanalytical tools
      • Molecular-based technique (PK)
      • Ultra-sensitive LBA technology (target PD)
      • Multiplex cytokines platform (safety/disease biomarkers)
    • Case Studies: Implementation of PK and Biomarker Assays to support clinical trials for IT administration of mRNA-based cytokine

  • Topic 5:

    Immunogenicity Assessment for Oligonucleotide-based Therapeutics: Lesson Learnt based on Novel Data from Non-clinical and Clinical Studies
    Dr. Susovan Mohapatra, Associate Director DMPK, Stoke

    • Oligonucleotide-based Therapeutics
      • Recent developments
      • Future perspective
    • Understanding the challenges associated with ADA assay development for Oligonucleotides
        Current experience in overcoming those challenges
    • Immunogenicity Risk Assessment for Oligonucleotide-based therapeutics in
      • Non-clinical Development
      • Clinical studies
    • Understanding Regulatory Expectations for Oligonucleotides Immunogenicity
      • Regulatory feedback on approved Oligonucleotides
      • Recommendations for Immunogenicity Assessment Strategy for oligonucleotide-based therapeutics
    • Case Studies: Challenges in ADA assay development for Oligonucleotide-based Therapeutics

  • Topic 6:

    Are LNPs used for Oligonucleotide-based Therapies & Vaccines Immunogenic? - State-of-art Industry Experience and Novel Data in this field
    Dr. Adam Williams, Senior Scientist, AstraZeneca

    • LNP-mRNA-based Therapies & Vaccines
      • Often formulated within lipid nanoparticle (LNP) encapsulation
      • LNP potential issues with immunogenicity that must be evaluated during clinical trials
    • LNPs are made up of multiple components of these drug products
      • Polyethylene glycol (PEG) that may contribute towards generation of anti-drug antibodies (ADA)
    • Pre-existing ADA against PEG
      • May complicate these immunogenicity assessments
      • Need to differentiate between pre-existing and drug product-initiated ADA.
    • LNP anti-PEG detection
      • Standard homogenous assay format regularly used in bioanalysis for ADA assessments is not feasible for LNP anti-PEG
      • Challenges in producing valid assays that will meet regulatory expectations
      • Development/Validation of LNP anti-PEG ADA method
    • How is the Industry/Regulators dealing with pre-existing antibodies against LNP based drugs/vaccines, given that a high proportion of the worlds population have now been exposed to LNP therapeutics via COVID vaccines?
      • Observations of incidence of ADA development against the LNP in the early clinical setting
      • Pre-clinical data demonstrates LNP exposure initiates immune responses and generation of anti-PEG antibodies
    • Case Studies: Development/Validation of LNP anti-PEG ADA methods for LNP-mRNA-based Therapies & Vaccines

  • Topic 7:

    Comparing & Contrasting Cell-based vs LBA for pre-existing anti-AAV TAbs and NAbs Assays: How to select a suitable assay platform?
    Dr. Lin Yang, Director of Bioanalytical Research, REGENXBIO

    • Evaluation of Pre-existing anti-AAV TAbs and NAbs for subject eligibility in enrollment in AAV based gene therapy
      • Cell-based neutralizing (NAbs)
      • Bridging LBA based total antibodies (TAbs)
    • Selection of a suitable method for detection of anti-AAV antibodies
      • Critical to determine subject to enroll in Gene Therapy studies
      • To be further developed as companion diagnostic (CDx)
    • Comparing NAbs and TAbs assays
      • Developed and validated from different labs using a set of commercial and study samples
      • NAb/TI assays using the same cell line and reagents but with different MOI
      • TAb assays using the same set of samples and same ECL assay
    • Advantages of the bridging ECL based TAb assay over NAb assay
      • Sensitivity
      • Reproducibility/robustness,
      • Standardization and throughput
    • Correlation between TAb and NAb levels
      • How to compare the results between the labs and what criteria should be established for patient enrollment?
      • Overall concordance between NAb and TAb
      • AAV transduction efficiencies with varying levels of anti-AAV NAb and TAb titers
    • Case Studies: Statistical assessment of NAb and TAb results and determination of the most appropriate assay for developing a CDx for clinical trials

 

Part 3: qPCR/ddPCR Infectivity, Vaccine & Biodistribution Assays
  • Topic 8:

    Recent and Novel Developments on Universal Genomic Assay Strategy to Characterize the Biodistribution in Gene Therapy by qPCR/ddPCR
    Dr. Hiroshi Sugimoto, Associate Director, Takeda

    • State-of-the-art qPCR/ddPCR Bioanalysis & Biomarker Strategy for Gene Therapy
      • Selection of bioanalytical platforms qPCR vs ddPCR
      • ddPCR Emerging Technology vs. conventional qPCR
    • Genomic Assay workflow to characterize the Biodistribution
      • Pros/Cons of absolute quantification by ddPCR
      • Partitioning the target gene analyte into nano droplets
      • Opportunities and challenges of ddPCR biodistribution assay
      • Understanding and overcoming ddPCR limitations
    • Expectations for ddPCR biodistribution assay Bioanalytical Performance
      • Characterization of critical assay parameters
      • Sensitivity, selectivity, and Reproducibility
      • Limiting dilutions and Poisson statistics
    • Case Studies: Current industry best practice on Genomic Assay for Gene Therapy Biodistribution

  • Topic 9:

    Updated Strategies for qPCR Infectivity Assays Development/Validation for Gene Therapy and Oncolytic Virus
    Mr. Justin Smith, Associate Director & Biomarker Clinical Assay Lead, Pfizer

    • Current challenges regarding implementation of conventional Infectivity Assay platforms in the clinic
      • Importance of ensuring the viral safety of gene therapy products,
      • Development of qPCR assay for the detection of replication competent & contaminating viruses
    • Investigation of a Novel Platforms to assess infectivity in a replication competent virus
      • Cross comparison between the Novel Platforms vs. traditional infectivity assay
    • Fit for Purpose Validation of Infectivity Assays
      • Highlighting the key parameters
      • Outcome of the validations
    • Regulatory Requirements for Gene Therapy and Oncolytic Virus Infectivity Assays
      • The 2015 US FDA Guidance on “Design and Analysis of Shedding Studies for Virus or Bacteria-Based Gene Therapy and Oncolytic Products”
    • Developing the right strategy for assay implementation in clinical testing
    • Case Studies: qPCR infectivity assays development/validation for gene therapy and oncolytic virus

  • Topic 10:

    Advanced approaches in Molecular Assays to Support Vaccines Studies: qPCR, ddPCR and NGS Applications
    Dr. Ludovic Malvaux, Director & Head of Applied Microbiology Platform, GlaxoSmithKline

    • Real-Time qPCR in Vaccines Clinical Trials
      • Recent recommendations on the use of qPCR for Vaccines
      • To demonstrate the efficacy of the investigational product on the presence of the causal pathogen
      • To appropriately define the serotype of specific pathogens (e.g., Streptoccocus pneumoniae)
      • To differentiate closely related bacteria/viruses (e.g., Flu A/B discrimination).
    • qPCR Validation
      • Extensive characterization and validation of real-time qPCR assays
      • Validation package required to support the required intended use (CoU)
      • Limit of detection (LOD) characterization and validation
    • Monitoring of real-time qPCR assay over time
      • Considerations on the appropriate monitoring of qPCR assay
      • Critical importance to ensure stability over time.
    • ddPCR
      • Implementation of ddPCR in vaccines
      • Help to better characterize assays standards and controls.
    • NGS applications for Vaccines
      • Implementation of Whole Exome Sequencing (WGS) and Deep Sequencing in clinical trials
    • NGS Support for Epidemiological and exploratory endpoints
      • Lineage identification
      • Identification of point mutation in antigenic site
      • Microbiome evaluation
    • Case Studies: Approaches in Molecular Assays to Support Vaccines Studies

 

Part 4: NGS and NanoString Assays
  • Topic 11:

    Next Generation Sequencing (NGS) Bioanalytical Applications in Genetics & Genomics
    Dr. Jessica Seitzer, Vice President, Head of Genomic Operations, Intellia

    • Current NGS Bioanalytical methods in in Genetics & Genomics
      • NGS direct impact in the development of novel Gene & Cell Therapies
      • Need for building out relevant technologies to evaluate efficacy and safety
    • De-risking of Gene and Cell therapy products
      • Critical NGS methods
    • Characterization of Gene and Cell therapy products
      • NGS Methods to characterize these products
      • Identification and analysis of intended and potential unintended genomic changes that can occur due to cell engineering or genome editing
    • Assay Development
      • Important factors to consider in developing NGS technologies in support of gene and cell therapy products
    • Case Studies: NGS Bioanalytical Applications in Genetics & Genomics

  • Topic 12:

    Recommendations for NGS Assay Fit for Purpose Validation
    Dr. Sean Downing, Senior Director Clinical Biomarker Innovation & Development, Takeda

    • When to use NGS vs PCR for DNA and extrapolated to RNA
      • General survey vs specific gene of interest
      • Hypothesis generating vs hypothesis testing
      • Application to Early-stage vs Late-stage trials
    • Disease State and need for depth of coverage
      • Somatic mutations vs Germline SNPs (Allelic frequency and Need for depth of coverage)
      • Oncology vs Rest of therapeutic areas
    • FFPE vs Blood
      • Oncology (somatic mutations) vs Other Therapeutics areas (germline SNPs)
      • “Quality” of matrix (Fragmented vs full length, Stochastic process of fixation and random fragmentation overcomes, for germline SNPs, blood will be the preferred matrix)
    • Fit for Purpose Assay Validation
      • Reproducibility and repeatability
      • Sample matrix
      • Variant types
      • Sensitivity (Lower Limit of Detection, Priori)
    • Whole genome vs targeted panel (includes WES)
      • Potential problem areas
      • Number of samples and runs
    • CRO vs In-house
      • CROs should be able and willing to provide “raw” validation data
      • Do you need to “validate” every possible variation?
    • Case Studies: Application of NGS in Oncology and other therapeutic areas and Assay Fit for Purpose Validation

  • Topic 13:

    Customizing NGS Bioanalytical Applications to Different Phases of Drug Development: What to consider in NGS use in Discovery vs Clinical Monitoring
    Mr. Serge Picard, Senior Manager Molecular Assay Development, Bristol Myers Squibb

    • Current best practices in NGS Applications in Discovery vs Clinical Monitoring
      • Quality framework across labs
    • Specific consideration for Assay Validation
      • Proficiency testing
      • Qualification vs Validation
    • Importance of Automation
      • Manual processes vs Automation
      • Control strategy
    • RNA-sequencing (RNA-seq)
      • Variety of applications
      • Workflow implementation
      • Major steps in RNA-seq data analysis
      • Quality control of raw reads
      • Read alignment
      • Quantification of gene and transcript levels differential gene expression,
      • Functional profiling and advanced analysis
    • Case Studies: NGS applications to different Phases of Drug Development

  • Topic 14:

    Recent Developments in Using NanoString Technology as an Alternative to qPCR: Understanding the Technology and its Potential Application in Bioanalysis
    Dr. David Bettoun, Vice President Discovery and non-clinical development, Larimar

    • What is NanoString and Why is there an increased interested in its applications?
      • Nanostring Technology
    • Applications in Gene Expression
      • Comparison to RT-qPCR
      • NanoString as accurate method of quantifying gene expression levels
      • Consistent quantitation of gene expression level regardless of codeset composition
    • Nanostring quantitation
      • High precision
      • Impact of level of expression of genes of interest (GOI)
    • Importance of proper selection of housekeeping genes
      • Accurate gene expression normalization across a large number of samples
    • Uniform timing of samples collection
      • Impact of the circadian regulation of gene expression
    • Case Studies: Use of NanoString as alternative to qPCR in gene expression




Agenda at a Glance Agenda at a Glance