04/10/2008©Novocell 2008 Developing a Safe hESC-Product for Diabetes FDA Advisory Committee Meeting April 10, 2008 Melissa Carpenter, Ph.D.

Slides:



Advertisements
Similar presentations
Regulatory Framework Leigh Shaw, Director.
Advertisements

Basics of stem cell culture Dr Shafaei. Definition of cell culture Cell culture refers to the removal of cells from an animal or plant and their subsequent.
rhBMP-2: origin, biology and preclinical safety
4th International Symposium on Stem Cell Therapy Madrid, April 26-27, 2007 Stem Cell Thertapy for STEMI. Is is Time for a Large Scale Clinical Trial ?
Stem Cell Basics Introduction to Embryonic and Adult Stem Cells.
1 Cellular, Tissue and Gene Therapies Advisory Committee Meeting azfibrocel-T (previously Isolagen Therapy (IT)) BLA Proposed Indication: Treatment.
From Stem Cells to Beta Cells: Possible Cure for Diabetes Mellitus By Ryan Scavinski.
Dan Takefman, Ph.D. Chief, Gene Therapy Branch
COMPARABILITY PROTOCOLS ACPS March 12-13, 2003 Stephen K. Moore, Ph.D. Chemistry Team Leader CDER/Office of New Drug Chemistry Co-Chair, Comparability.
Human Induced Pluripotent Stem Cells (hiPS Cells)
Processing and Product Quality Issues Keith Wonnacott Ph.D. Office of Cellular, Tissue, and Gene Therapies E BC R Moving from Investigational to Licensed.
MARIE CSETE MD, PhD CHIEF SCIENTIFIC OFFICER
Stem Cells and Society: Ethics and Advances
The demand for islet cells for treatment of diabetes and lack of pancreata and the problem of immunosuppression with the allogenic transplantation directed.
Denise K. Gavin, Ph.D. Division of Cellular and Gene Therapies
Long-Term Follow-Up of Subjects in Gene Transfer Clinical Protocols Vector Classes with Potential for Long-Term Risks Carolyn A. Wilson, Ph.D. Division.
Cancer stem cells IOSI Journal Club Giulia Poretti January 19, 2007.
What are they and what can we do with them?
Food and Drug Administration Food and Drug Administration Center for Biologics Evaluation and Research Biological Response Modifiers Advisory Committee.
Food and Drug Administration Center for Biologics Evaluation and Research The Office of Cellular, Tissue, and Gene Therapies Web Seminar Series presents:
Ensuring Product Quality in Gene Transfer Clinical Trials
Nonclinical Studies Subcommittee Advisory Committee for Pharmaceutical Science CMC Issues for Screening INDs Eric B. Sheinin, Ph.D. Acting Deputy Director.
Altogen Labs, 4020 S Industrial Dr, Suite 130, Austin TX 78744, USA Contact us at: Provider of Global.
Animal Models for Porcine Xenotransplantation Products Intended to Treat Type 1 Diabetes or Acute Liver Failure CTGTAC #47 May 14, 2009.
Human Embryonic Stem Cells: Considerations for Therapeutic Development Jane Lebkowski Ph.D. Geron Corporation Cell, Tissue and Gene Therapy Advisory Committee.
E BC R Overview of FDA Regulatory Issues Darin J. Weber, Ph.D. Office of Cellular, Tissue and Gene Therapies.
1 Safety of Cell Therapies Derived from Human Embryonic Stem Cells CTGTAC #45 April 10, 2008.
Potency Testing for an Autologous Cellular Immunotherapy Nicole Provost, PhD VP Product Development Dendreon Corporation February 9, 2006.
U.S. Food and Drug Administration CENTER FOR BIOLOGICS EVALUATION AND RESEARCH March 29, 2007 Sipuleucel-T BLA US Food and Drug Administration Center.
Dr. Abdullah Aldahmash. Cell Stem Cell – Definition A cell which has the ability to continuously divide and differentiate into various other kinds of.
Stem cells Helena Fulkova Institute of Animal Science
Cellular Therapies for Repair and Regeneration of Joint Surfaces— Product Characterization and Testing Malcolm Moos Jr., M.D., Ph.D. Medical Officer
National Institute for Biological Standards and Control Assuring the quality of biological medicines Bank Update II: focusing on contributions to R&D Lyn.
Stem cell biology basics By the end of this lecture you will: 1.Understand where stem cells come from 2.Understand the different types of stem cells 3.Understand.
Human embryonic stem cells (hES cells) and human induced-pluripotent stem cells (iPS cells) are uniquely defined by their pluripotent differentiation potential.
Altogen Biosystems offers the Transfection Reagent for PANC-1 Cells Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection.
Products > B16-F10 Transfection Reagent (Mouse Melanoma Cells) Altogen Biosystems offers the B16-F10 Transfection Reagent among a host of 100+ cell line.
Altogen Biosystems offers the J774A Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. The J774A Transfection Reagent.
Identification of Cancer Stem Cells using Flow Cytometry Analysis تعيين الخلايا الجذعية السرطانية باستخدام تحليل التدفق الخلوي Dr. Ayat Al-Ghafari Biochemistry.
정책이력 일반 현황 Telomerase reverse transcriptase (TERT) related with telomerase activity regulates tumorigenic potential of mouse embryonic stem cells Seung.
Altogen Biosystems offers the 786-O Transfection Reagent among a host of 100+ cell line specific In Vitro Transfection Kits. 786-O Transfection Reagent.
Products > Transfection Reagent for C6 Cells (Glioma Cells, CCL-107) Altogen Biosystems offers the C6 Transfection Reagent among a host of 100+ cell line.
Products > 3T3-L1 Transfection Reagent (Embryonic Fibroblast Cells, CL-173) Altogen Biosystems offers the 3T3-L1 Transfection Reagent among a host of 100+
Products > T98G Transfection Reagent (Glioblastoma Cells, CRL-1690)
Olga Genbacev, Ph. D. , Ana Krtolica, Ph. D. , Tamara Zdravkovic, M. S
Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products.
Products > NIH3T3 Transfection Reagent (Swiss Mouse Fibroblasts)
The Lifecycle of Pharmaceutical products
Supplementary Figure S1
Products > Weri-Rb-1 Transfection Reagent (Retinoblastoma Cells)
The Generation of Six Clinical-Grade Human Embryonic Stem Cell Lines
Products > A375 Transfection Reagent (Melanoma Cells, CRL-1619)
Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products.
Establishment of Endoderm Progenitors by SOX Transcription Factor Expression in Human Embryonic Stem Cells  Cheryle A. Séguin, Jonathan S. Draper, Andras.
Results of Phase 1 Trial of Treg Adoptive Cell Transfer in Living Donor Kidney Transplant Recipients TRACT Therapeutics™
Development of humanized culture medium with plant-derived serum replacement for human pluripotent stem cells  Michaela Kunova, Kamil Matulka, Livia Eiselleova,
Volume 5, Issue 4, Pages (October 2015)
Meeyoung Cho, Ph. D. , Eun Ju Lee, Ph. D. , Hyun Nam, Ph. D
Olga Genbacev, Ph. D. , Ana Krtolica, Ph. D. , Tamara Zdravkovic, M. S
Volume 10, Issue 4, Pages (April 2012)
Transduction of Human Embryonic Stem Cells by Foamy Virus Vectors
Products > IMR-90 Transfection Reagent (Lung IMR-90, CCL186)
Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products.
Volume 6, Issue 3, Pages (March 2016)
Wei Jiang, Yuting Liu, Rui Liu, Kun Zhang, Yi Zhang  Cell Reports 
Volume 4, Issue 1, Pages (January 2015)
Volume 4, Issue 4, Pages (April 2015)
Products > CLBPEC Transfection Reagent (Neuroblastoma Cells)
Volume 11, Issue 6, Pages (December 2018)
Presentation transcript:

04/10/2008©Novocell 2008 Developing a Safe hESC-Product for Diabetes FDA Advisory Committee Meeting April 10, 2008 Melissa Carpenter, Ph.D.

04/10/2008©Novocell 2008 Cell Therapy for Diabetes: The Edmonton Protocol Adapted from J. Shapiro

04/10/2008©Novocell 2008 Development of a Safe hESC-Derived Product Characterize the Cell Product In Vitro Evaluate Cell Product In Vivo in appropriate animal model –Efficacy –Stability –Toxicity –Tumorigenicity

04/10/2008©Novocell 2008 Steps in Characterizing a Cell Product Starting Material Cell Product Definitive Endoderm Foregut Endoderm Pancreatic Endoderm hESC Insulin Producing Cell Starting cellular material –hESCs Intermediate populations Final cellular product –Insulin producing cell population

04/10/2008©Novocell 2008 Established on GMP-compliant human fibroblast feeders PTC testing completed and passed All processing reagents compliant for clinical manufacturing Initial characterization of cell line complete (surface markers, karyotype, differentiation) MCB & WCB established PTC testing complete Starting Material: Derivation Under Clinical Manufacturing Conditions CyT49

04/10/2008©Novocell 2008 Assays for In Vitro Characterization of Starting Material (hESCs) 1.Genetic identity 2.Markers Flow cytometry Q-PCR Immunocytochemistry 3.Karyotype 4.Stability 5.Differentiation

04/10/2008©Novocell 2008 In Vitro Characterization of Starting Material: 1) Genetic Identity Cell line identity by STR genotyping PowerPlex® 1.1 Plus 2.1 System (14 STR loci) Power of 1 in 1x10 16

Nanog SSEA-4 Oct3/4 TRA-1-81 In Vitro Characterization of Starting Material: 2) Markers

04/10/2008©Novocell 2008 In Vitro Characterization of Starting Material: 3) Karyotype Cell LinePassage Range CyT Normal Karyotype on Feeders Cell Line Passage # at Thaw Passage # at Karyotype CyT CyT CyT Normal Karyotype after Cryopreservation Cell Line# Passages on Feeders # Passages FF Total # Passages CyT CyT CyT Normal Karyotype Feeder-Free CyT49 p25

04/10/2008©Novocell 2008 Cytogenetic Analysis G-Banding –Allows detection of numerical abnormalities, inter- chromosomal abnormalities, intra-chromosomal abnormalities –Performed in cytogenetics lab –20 cells or more examined –Clinically correlated Spectral Karyotype (SKY) analysis –Allows detection of unknown rearrangements Correlation between aneuploidy and adverse outcome currently unclear

04/10/2008©Novocell 2008 In Vitro Characterization of Starting Material: 4) Stability Key Requirements for Stability –Stable over long term culture Expansion of cells prior to differentiation for cell product –Stable over long term cryopreservation

04/10/2008©Novocell 2008 Expansion of hESCs hESC Starting MaterialCell Product Definitive Endoderm Foregut Endoderm Pancreatic endoderm Insulin Producing Cell Viability Consistent Composition Normal Karyotype Ability to Differentiate

04/10/2008©Novocell 2008 In Vitro Characterization of Cell Product Starting Material Cell Product Enrichment of Cell Product & In-Process Testing Predictive of Outcome Definitive Endoderm Foregut Endoderm Pancreatic endoderm Endocrine cells hESC SOX17 CER FOXA2 OCT4 NANOG SOX2 ECAD HNF1B HNF1A HNF4A NKX6.1 NGN3 PAX4 NKX2.2 INS C-PEP PAX6 GCG GHRL SST PP

04/10/2008©Novocell 2008 Recommended Characterization of Cell Based Product Code of Federal Regulation for Food and Drugs (21 CFR 600 – Biologics) –Sterility –Purity –Potency –Identity –Stability –Safety –Efficacy “Active component/cell”, accessory cells, carrier solution, inappropriate components

04/10/2008©Novocell 2008 Identity Analysis Includes Assessment of Different Populations in Product Cell Product might be a heterogeneous population Cell Product assessment will include: –“Functional” cell Cell population capable of producing insulin –Accessory cells Other endocrine cells –Inappropriate cells Undifferentiated cells Cytotoxic cells –“Bystander” cells

04/10/2008©Novocell 2008 In Vitro Assays for Identity Analysis hESC- insulin QPCR Flow Cytometry Immunocytochemistry Sensitivity

04/10/2008©Novocell 2008 Detection of Rare ESCs Using Real- time PCR for OCT4 0.04% ESCs detected = 8 ESCs in 20,000 HEFs results in signal detectable above HEF expression Decreasing numbers of ESCs mixed in with 20,000 HEFs

04/10/2008©Novocell 2008 side (90  ) scatter controlOct3/4 50% 5% 0.5% hESC + HEF controlOct3/4 side (90  ) scatter hESC only Flow Cytometric Analysis for Detection of hESCs

04/10/2008©Novocell 2008 Summary of In Vitro Characterization of Cell Product Assessment of Starting Material Identity and stability of cell product Assay validation required Sensitivity of assays needs to be balanced with consumption of product for QC Predictive of clinical outcome –Safety –Efficacy

04/10/2008©Novocell 2008 Safety/Tumorigenicity for Cellular Product Dosing / Toxicity Biodistribution Where do the cells go? Maintain identity if found in other tissues? Stability Glycemic control De-differentiated cells? Tumorigenicity In vivo In vitro

04/10/2008©Novocell 2008 What is a Relevant Animal Model? Many cell based products are species-specific Will large animal studies be meaningful? –Is there a suitable large animal model? ??

04/10/2008©Novocell 2008 Teratoma vs Teratocarcinoma Teratoma = benign tumor Teratocarcinoma = malignant tumor Risk of teratoma formation will be balanced with patient population and implant site

04/10/2008©Novocell 2008 Tumorigenicity: What is the appropriate assay? How many ES cells does it take to make a teratoma? –Is there an absolute number of cells required? –Is there a frequency required (percentage of cells)? –Needs to be measured for each cell line, each product? What is the effect of implant site on teratoma formation? –Are some sites more permissive? –Do the neighboring cells (from graft or from implant site) influence teratoma formation? Are other cell types tumorigenic? Does the immune status of the recipient affect teratoma formation? What does a negative result mean?

04/10/2008©Novocell 2008 All ES Cells are NOT Equal: Origin May Influence Tumorigenicity Human does not equal mouse –Single cell cloning –Requirements for self-renewal are different MouseHuman Morphological Character Rounded colonies Flat colonies Growth Requirements LIF, BMPbFGF, activin Marker Expression SSEA-1SSEA-4 Spontaneous Trophoblast Differentiation noyes

04/10/2008©Novocell 2008 Developing a Safe hESC-derived Product: Summary Characterize the cell product Demonstrate safety and efficacy of cell product Sensitive and specific assays required Assays which are predictive of clinical outcome required

04/10/2008Copyright Novocell 2008 Developing a Cell Product for Diabetes Edmonton protocol and others successfully implanted autologous islets under immunosuppression hESCs provide a cell source for Diabetic therapies Novocell has developed encapsulation technology –Preliminary clinical evaluation