Slide 1: Nonclinical considerations for cell and gene therapies
2023 Nonclinical considerations for cell and gene therapies Brian E. McIntosh, PhD ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 518817_DX_PRES_Nonclinical Safety Cell and Gene Therapy Webinar_0923
Slide 2: About us
About us Brian E. McIntosh, PhD Global Preclinical Cell & Gene Therapy Lead, and Head of Toxicology Subject Matter Experts, Safety Assessment/Toxicology, Labcorp • 25+ years combined industry and academic experience within regenerative biology, oncology, toxicology, and gene therapy • Responsible fo
Slide 3: Today you'll hear about
Today you’ll hear about • How advanced therapies are defined • Preclinical study and regulatory/safety considerations and strategies when assessing advance therapies • Analytical design and validation strategies • Biodistribution studies and data integrity • Titre assays • CMC considerations • Phase
Slide 4: What type of advanced therapy are you interested in advancing?
“What type of advanced therapy are you interested in advancing and where does your product fit?” ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 4
Slide 5: Advanced therapies: cell vs. gene
N O N C L I N I C A L D E V E L O P M E N T C O N S I D E R A T I O N S F O R C E L L A N D G E N E T H E R A P I E S Advanced therapies: cell vs. gene Cell-Based Therapies Gene Therapies (Regenerative Medicinal Products) • Process of replacing, engineering or • Technique for correcting defective ge
Slide 6: Preclinical program goals for advanced therapeutics
Preclinical program goals for advanced therapeutics • Recognize data necessary to support development depend on the characteristics of the product • Recognize preclinical studies are designed to support use of a specific product for a specific clinical indication • Identify biologically relevant dos
Slide 7: Preclinical program goals for advanced therapeutics (cont.)
Preclinical program goals for advanced therapeutics (cont.) • Recognize test system must be appropriate • Species, disease models, immunodeficient animals • Autologous, allogeneic or xenogenic • Recognize delivery must represent clinical route of administration • May include device compatibility • D
Slide 8: Fast track and breakthrough designation programs
E X P E D I T E D P R O G R A M S F O R R E G E N E R A T I V E M E D I C I N E T H E R A P I E S F O R S E R I O U S C O N D I T I O N S Fast track and breakthrough designation programs Fast Track (Designation): for therapeutics intended to treat a serious condition where data demonstrate the poten
Slide 9: Issues to overcome for in vivo viral delivery
G E N E T H E R A P Y P R O D U C T S Issues to overcome for in vivo viral delivery • Short-lived • Difficult to rapidly integrate therapeutic DNA into genome • Rapidly dividing characteristic of certain cells may prevent long-term efficacy • May require multiple rounds of therapy • Immune response
Slide 10: Refresher preclinical study considerations
G E N E T H E R A P Y P R O D U C T S Refresher preclinical study considerations • The test system must be appropriate and may require a prescreen for neutralizing antibodies • Delivery must represent a clinical route of administration • The test system may require immune suppression • Study length
Slide 11: Preclinical regulatory considerations
C E L L T H E R A P Y D E R I V E D E X V I V O Preclinical regulatory considerations In vitro properties In vivo concerns • Safety: acute, tumorigenicity, biodistribution, efficacy • Open dialogue with the FDA-OTAT, EMA, PMDA… Source: Fink DW. FDA regulation of stem cell-based products. Science. 20
Slide 12: Perspective shifts in early development
C A R - T T H E R A P Y P R O D U C T S Perspective shifts in early development Safety is primarily shifted to the manufacturing phase (CMC) • Personalized or individualized medicine • Variation in cell source • Limited production and small batch size • Establishment of minimal criteria to ensure sa
Slide 13: Preclinical study considerations
C A R - T T H E R A P Y P R O D U C T S Preclinical study considerations A scientific discussion dictates in vivo safety designs • Controversy: definitive therapeutic is generally short-lived—beware the rise of the clone! • Safety studies generally focus on long-term tumorigenicity (three to six mon
Slide 14: Preclinical study considerations (cont.)
C E L L T H E R A P Y P R O D U C T S Preclinical study considerations (cont.) • In vivo tumorigenicity • Derived CBT/RMP dosed at varied concentrations to an ideally confined location in the animal • One group may be required for a positive parental cell line or tumorigenic line • e.g., iPSC, MCF7,
Slide 15: Preclinical study considerations (cont.)
C E L L T H E R A P Y P R O D U C T S Preclinical study considerations (cont.) • Acute/efficacy/dose response • Employ disease or injury model test system • Adverse effects with respect to dose, onset and duration • Dependent on cellular response and physiology to detect signal and time course • Com
Slide 16: Closing remarks
Closing remarks Cell and gene therapy study designs • Changing SA landscape with Cell and Gene Therapeutics • Therapeutic dictates the study design • Seek advice from a regulatory body and knowledgeable source prior to initiating work • The process is the product ©2023 Laboratory Corporation of Amer
Slide 17: Design considerations for bioanalytical assays to support biodistribution studies
“What are the design considerations for bioanalytical assays to support biodistribution studies?” ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 17
Slide 18: Biodistribution studies
Q P C R / R T - Q P C R A S S A Y D E S I G N C O N S I D E R A T I O N S Biodistribution studies Assay design is critical • Span the junction of modifications • Theoretical specificity • Feasibility • Multiple applications Biodistribution • CMC • Clinical Toxicology Validation considerations • LOQ,
Slide 19: Biodistribution studies
Q P C R / R T - Q P C R A S S A Y D E S I G N C O N S I D E R A T I O N S Biodistribution studies Sensitivity is critical • LOQ: 50 copies/µg of genomic DNA • Chemistry is important (e.g., TaqMan or SYBR Green) Specificity • Importance of assay design Biodistribution Extraction considerations • Chal
Slide 20: Key analytical and design considerations for preclinical biodistribution studies
“What are the key analytical and design considerations for preclinical biodistribution studies?” ©2022 Laboratory Corporation of America® Holdings All rights reserved. ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 20
Slide 21: Biodistribution studies
Q P C R / R T - Q P C R A S S A Y D E S I G N C O N S I D E R A T I O N S Biodistribution studies • Combined with toxicology study: separate biodistribution groups/cohorts • Route of administration: dependent on therapy • Time points: one week, one month and three months • Assessment of distribution
Slide 22: Biodistribution studies
Biodistribution studies Sterile, biological handling Dispensary Specific procedures and aliquots Animal Specialized dosing room Handling, separation and housing Biodistribution Precautions and procedures Necropsy Disposable equipment and sample collection Toxicology Purpose-built laboratories Extrac
Slide 23: Biodistribution studies: responsibilities
Biodistribution studies: responsibilities SME SD PI Sterile, biological handling Dispensary Specific procedures and aliquots Animal Specialized dosing room Handling, separation and housing Biodistribution Precautions and procedures Necropsy Disposable equipment and sample collection Toxicology Purpo
Slide 24: Biodistribution studies
C O N F I D E NC E I N D A T A Biodistribution studies Significant risks for compromised studies • Cost of repeats • Delays to submissions • Impact to clinical trial design Important to have alignment among in-life, necropsy Biodistribution and analytical • Sample collection • Guided by analytical t
Slide 25: Biodistribution Studies
T I T R E / D O S E D E T E R M I N A T I O N Biodistribution Studies CMC information for human gene therapy investigational INDs: FDA draft guidance for industry, July 2018 • Ensure ability to compare the preclinical dose to the clinical dose • Use the same qualified method to quantitate preclinica
Slide 26: Chemistry, Manufacturing and Controls
Chemistry, Manufacturing and Controls “What questions do you need to address to ensure safety, efficacy and consistency for IND/IMPD submissions?” ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 26
Slide 27: Biodistribution studies
C M C Biodistribution studies • How much do you have? • How potent is it? • How safe is it? CMC • How pure is your product? • Is it what you think it is? Clinical • Is it stable? ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 27
Slide 28: Biodistribution studies
C R I T I C A L Q U A L I T Y A T T R I B U T E S Biodistribution studies Quantity and titre AAV Vectors CAR-T Part 1 Part 2 CMC Genomic titre (qPCR) Vector titre (qPCR) Viability (flow) Clinical Capsid titre (ELISA) Cell count (flow) Part 1: retroviral/lentiviral vector Part 2: active substance (CA
Slide 29: How potent is it?
C R I T I C A L Q U A L I T Y A T T R I B U T E S How potent is it? Biological Activity, Transduction and Potency AAV Vectors CAR-T Part 1 Part 2 Cell-based assay Cell-based assay with PBMC transduction with ELISA CMC ELISA endpoint endpoint Clinical Infectious titre (cell- Transduction based qPCR e
Slide 30: How safe is it?
C R I T I C A L Q U A L I T Y A T T R I B U T E S How safe is it? Safety Testing AAV Vectors CAR-T Part 1 Part 2 CMC Safety testing ICH Safety testing ICH Q5A Sterility Q5A Clinical rcAAV RCR/RCL Mycoplasma Endotoxin ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 30
Slide 31: How pure is it?
C R I T I C A L Q U A L I T Y A T T R I B U T E S How pure is it? Purity and Impurities AAV Vectors CAR-T Part 1 Part 2 Host cell proteins and HCP and HCDNA Vector (qPCR) DNA (ELISA and qPCR) (ELISA and qPCR) CMC Beads BSA, Benzonase (ELISA) RCR/RCL (microscopy) Clinical Capsid purity (CE) Empty vs.
Slide 32: Is it what you think it is?
C R I T I C A L Q U A L I T Y A T T R I B U T E S Is it what you think it is? Identity AAV Vectors CAR-T Part 1 Part 2 CMC Sequencing, mass Sequencing qPCR spectrometry Clinical ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 32
Slide 33: Phase-appropriate validation plan
“How can I rely on the data for submission, and what does a phase- appropriate validation plan look like?” ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 33
Slide 34: Regulatory perspective
V A L I D A T I O N S Regulatory perspective CMC information for human gene therapy investigational INDs: FDA draft guidance for industry, July 2018 • Validation not required for IND Phase I • Appropriately controlled • Complete validation before BLA Biodistribution • Evaluate performance through de
Slide 35: Development pathway
Development pathway CMC: Validation Commercial Phase III CMC Phase II Preclinical Phase I IND/IMPD BLA/MAA Product Understanding ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 35
Slide 36: Assay validation
D E F I N I T I O N S Assay validation Fit for purpose/controlled/suitability • Compendial • One run to qualify (additional for small volumes) • Identity/purity (qPCR, ELISA, CE-SDS, MS, FACS) CMC • Two to three runs • Titre (qPCR, ELISA) • Three to four runs • Potency (cell-based) • 15 runs ©2023 L
Slide 37: Assay validation
D E F I N I T I O N S Assay validation Limited validation • Identity/purity/titre (qPCR, ELISA, CE-SDS, MS) • Seven to eight runs • Potency (cell-based) CMC • Nine runs Full validation (robustness) • Design of experiment (DOE) • Eight runs • qPCR included with limited validation ©2023 Laboratory Cor
Slide 38: Assay validation
D E F I N I T I O N S Assay validation Validation Requirement Commercial Phase III Phase II Fit for CMC Purpose Phase I Compendial ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 38
Slide 39: Assay validation
D E F I N I T I O N S Assay validation Validation Requirement Commercial Phase III Phase II Fit for CMC Purpose Phase I Identity/Purity/Potency Fit for Purpose Limited Validation Full Validation Limited Validation Full Validation ©2023 Laboratory Corporation of America® Holdings. All rights reserved
Slide 40: Assay validation
D E F I N I T I O N S Assay validation Validation Requirement Commercial Phase III Phase II Fit for CMC Purpose Phase I Titre Assays Fit for Purpose Limited Validation Full Validation Limited Validation Preclinical ©2023 Laboratory Corporation of America® Holdings. All rights reserved. 40
Slide 41: Thank you
Thank you ©2023 Laboratory Corporation of America® Holdings. All rights reserved.