ARM Cell Potency Project & Developing a Bioassay for an Allogeneic Cell Therapy Product

Transcription

1 ARM Cell Potency Project & Developing a Bioassay for an Allogeneic Cell Therapy Product Dr. Gil Van Bokkelen CASSS BioAssays 2013 March 4 th, 2013

2 ARM - Potency Project Overview Project goal: To identify and evaluate existing cell potency assays, recommend best practices, and assess the feasibility of developing standards for assessment of potency of cell therapy / regenerative medicine products Working Groups: WG 1 Methods Registry and Survey Identify best practices to inform assay selection and development (ongoing) WG 2 Reference Standards and Materials Elucidate tools/road maps to measure variability for common cell types (completed) WG 3 Assay development roadmap and validation Map assay requirements to specific product types and validate (completed) WG 4 Gap analysis of current inventory To ensure analytical capabilities meet measurement requirements (Initiated April 2012) 2

3 ARM Potency Project Output - Working Group 3 Potency Assay - Method Validation Roadmap Categories: Detection of Proteins Differentiation Engraftment Genetic Testing Immunotherapy Cell-based assays Viability

4 WG#3 Assay-specific Development Roadmap and Validation Goal: To provide for validated Analytical Methodologies that can be used in characterization and potency assessment, specifically: a) Survey existing Analytical Methodologies available and used for characterization & potency assessment (based on output from WG#1) b) Select sub-set of Assay Methodologies commonly used that are commercially available c) Identify VENDORS that provide commercially available solutions to these Analytical Methodologies d) Qualify and Validate selected Analytical Methodologies to provide guidance on assay variability and provide as compendium / monographs to the Developer community and other stakeholders e) Conduct forward-looking exercise to identify potential new Analytical Methodologies that could be applied toward characterization & potency assessment. 6 4

5 Recommended Approach for the Developer 1. Obtain the road map template (WG#2 output) for your cell type / product type of interest 2. Search database/registry and survey results (WG#1 output) to understand previous attempts at characterization with your specific cell type / product type 3. Search inventory of available (validated) analytical methodologies and tools that are available for purposes of measuring various cell type / product type attributes required for complete characterization of your specific product based on potency road map 4. Use this information to create a customized characterization road map for your specific product of interest 5. Reference the gap analysis (WG#4 output) to refine road map This road map should track with specific regulatory requirements of product characterization. 6 5

6 ARM ISCT Joint White Paper Project Objective: Develop Better Understanding of Current Tests for Product Quality and Consistency for Cell Therapy Products Four White Papers: White Paper 1 Cell Content (Quantity) and Viability White Paper 2 Identity and Purity Tests White Paper 3 Angiogenesis White Paper 4 Immunomodulation

7 Key Sections of White Papers 1) Scope Product Type (i.e. allogeneic, autologous) and why assays are being used 2) Assay descriptions advantages and disadvantages, and sensitivity 3) Assay qualification and validation reference materials if applicable 4) Considerations for Potency Determination 5) Relevance to MoA (not included in each white paper) 6) Case Studies 7) Comparison to Content Assays for Other Biologics 8) Regulatory Considerations 9) Future Considerations

8 Some Considerations for Potency Assay Development Potency definition and features - Quantitative, bioactivity, etc. Relationship to MOA(s) Potency surrogates Reference materials Cell content/ viability not sufficient for potency, but can be incorporated into potency assay matrix

9 Development of a BioAssay for an Allogeneic Cell Therapy Product NASDAQ: ATHX

10 Forward Looking Statements The statements and discussions contained in this presentation that are not historical facts constitute forward-looking statements, which can be identified by the use of forward-looking words such as believes, expects, may, intends, anticipates, plans, estimates and analogous or similar expressions intended to identify forward-looking statements. These forward-looking statements and estimates as to future performance, estimates as to future valuations and other statements contained herein regarding matters that are not historical facts, are only predictions, and that actual events or results may differ materially. We cannot assure or guarantee you that any future results described in this presentation will be achieved, and actual results could vary materially from those reflected in such forward-looking statements. Information contained in this presentation has been compiled from sources believed to be credible and reliable. However, we cannot guarantee such credibility and reliability. The forecasts and projections of events contained herein are based upon subjective valuations, analyses and personal opinions. This presentation shall not constitute an offer to sell or the solicitation of an offer to buy any securities. Such an offer or solicitation, if made, will only be made pursuant to an offering memorandum and definitive subscription documents.

11 Development Status of Key MultiStem Programs Inflammatory & Immune Inflammatory Bowel Disease* HSC Transplant / GVHD Solid Organ Transplant Diabetes Cardiovascular Acute Myocardial Infarction PVD/PAD/CLI Congestive Heart Failure Neurological Ischemic Stroke Traumatic Brain Injury Multiple Sclerosis Spinal Cord Injury Preclinical Phase I Phase II Phase III BLA Orphan Status Granted Recent Data Recent Data Recent Data Enrolling (U.S., Canada & Europe) Results announced Feb, 2012 Phase 2/3 Trial design under FDA review Trial Authorized in Germany Results published Nov, 2011 Phase 2 Authorized by FDA Enrolling (U.S.) * In partnership with Currently 5 Clinical Programs (4 in/at mid-stage Clinical Development)

12 MultiStem Cell Therapy: A Multimodal Biologic Product MultiStem Cell Therapy Expanded, banked product Obtained from healthy, consenting adult donors Frozen storage Administered systemically (i.v.) or locally (catheter, injection, matrix or implant) MultiStem Product Profile Cell therapy product based on patented (MAPC ) technology Developing for off-the-shelf administration no tissue matching needed Expanded product with high production yield (e.g., millions of doses from each donor) Long storage life can be kept frozen for years Consistent safety profile Promotes healing and tissue repair through multiple mechanisms of action Not a permanent transplant cells cleared from the body over time (like a drug) 12

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14 The Necessity of a Validated Potency Assay Regulatory guidelines stipulate that biological products should be safe, pure, potent, stable and effective for human use. Potency testing is measurement of the specific ability or capacity of the product to effect a given result, as indicated by appropriate laboratory tests or by adequately controlled clinical data through the administration of the product in the manner intended A well defined potency test should: Indicate biological activity specific to the product Provide test results for release of the product Provide quantitative data Meet predefined acceptance/rejection criteria Include appropriate reference materials, standards and/or controls

15 Potency Assays For Cell Therapeutics: Contrast to Biologics Drug, Biologic Potency Single target of action Single active ingredient Stable PK/PD profile Defined reference standards Cell Therapeutic Potency Multiple parallel mechanistic pathways May have multiple active ingredients Dynamic response to injury environment Difficult to define reference standards

16 Challenges in Cardiovascular Disease Approximately 785,000 first time heart attacks occurring annually in U.S., + 470,000 recurrent myocardial infarctions (MI s) and over 7.9 Million heart attack survivors Over 5.9 Million individuals with Congestive Heart Failure (CHF) in U.S. high morbidity & mortality; very high cost of care Over 8 Million individuals with peripheral arterial or vascular disease (PAD/PVD) in U.S more than 2 Million at risk of Critical Limb Ischemia (CLI) Published studies with independent labs show MultiStem could provide multiple benefits to treat damage in acute or chronic cardiovascular disease According to AHA Estimates, direct costs from cardiovascular disease will grow from $273 Billion annually (2010) to more than $818 Billion annually (2030) as a result of an aging and increasingly obese population 16

17 MultiStem : Acute Myocardial Infarction Administration of MultiStem following a heart attack Opportunity to improve outcomes & reduce risk of progression to CHF Blockage causing Angioplasty to ischemia clear blockage Rapid, efficient delivery of cells to ischemic area Retention of cells in relevant region of heart Multiple mechanisms of action, safe & well tolerated Delivery of MultiStem 5 sec 30 sec 60 sec Vessel patency Imaging data obtained from first patient enrolled in Phase 1 clinical trial treated at Cleveland Clinic 17

18 DLVSV Highlights from AMI Clinical Trial at One Year Design: Open label, Ph 1 dose escalation study, involving 25 patients (3 dose levels, registry) Results: Excellent safety profile observed through one year Efficacy: Meaningful improvements seen in LVEF, stroke volume, wall motion Detailed analysis of one month data also suggests trend toward reduced tachycardia Stroke Volume Ejection Fraction 40 4 month 12 month Reg 20M 50M 100M Evaluation of target patient population (baseline LVEF of >30 <45 at time of MultiStem administration) See published Results from Phase 1 Clinical Study (Circ Research Jan. 2012, Penn et al) 18

19 Administration of MultiStem Promotes Increased Ejection Fraction and Vascular Density in Pig AMI Model Wang et al, Circulation

20 Cell Treatment Results in Increased Microvasculature in Ischemic Hind Limb Increased Vasculature and Perfusion Cell Distribution to Neurovascular Bundles Only in Injured Limb uninjured Wragg, A et al J Mol Med 2008

21 Enhancing Repair in Vascular Injury & Chronic Heart Ischemia Models Figure 1: Administration of human MAPCs in a rodent model of peripheral vascular ischemia results in significantly improved blood flow in the hind limb region by day 30 relative to control animals treated with PBS. Images taken from Aaranguren et al Cell Transplant Note: This data was generated independently (i.e. not an Athersys sponsored study), but is consistent with previous published studies conducted by the company and our collaborators. Figure 2: Treatment with MultiStem increased vessel density in the border region of the infarct zone in a rat model of heart failure at 4 weeks compared to control animals. Vessel density was determined by the number of VWF staining vessels (in green) per mm 2 21

22 Angiogenic Factor Screening - Comparing MultiStem to MSC (when derived from same donors) Donor 1 Donor 2 Donor 3 Angiogenic Factor Immunoblot Capture Assay using 60 angiogenic factor mab panel Comparison of activity from MSC (top) and MultiStem (bottom) MSC / MultiStem from same donor bone marrows Experiment repeated across three distinct donors MSC147 P4 MSC149 P6 MSC153 P6 MultiStem expresses a broader range of angiogenic factors than MSC MultiStem147 P4 MultiStem149 P6 MultiStem153 P6

23 Steps to Designing a Surrogate Angiogenic Potency Assay Confirm angiogenesis using in vivo pre-clinical model Test angiogenic potential using an in vitro angiogenesis assay Identify necessary angiogenic factor(s) Immuno-deplete angiogenic factors Add back and identify minimum levels required for angiogenic activity Correlate to levels in production runs. Define threshold as lot release criteria Set reference standards (including cell line controls) and qualify assay

24 MultiStem Induces Angiogenesis in Surrogate in vivo Assays vessels/hpf In vivo Matrigel plug assay Reduced factor matrigel +: PBS FGF2 Cells SMA + vessels ** Isolate Matrigel plugs 0 PBS FGF2 MultiStem ** p<0.05 PBS FGF2 MultiStem Tri-chrome SMA * * * * * *

25 MultiStem Induction of Angiogenesis using an in vitro Assay Collect and test serum free MultiStem conditioned media 24 hrs Analyze for angiogenic factors Plate cells in normal media Change to serum free media Collect serum free MultiStem conditioned media Analyze MultiStem conditioned media in an in vitro angiogenesis assay HUVECS on reduced growth factor Matrigel +/- CM 18 hours Count tube formation per high powered field See: Development of a surrogate angiogenic potency assay for clinical-grade stem cell production. Cytotherapy 2012 Sep;14(8):

26 Avg. Concentration [pg/ml] Standardized to Dose Avg Number Tubes/field MultiStem Secretes Factors That Promote Angiogenesis in vitro EGM EBM MultiStem Day4 CM MultiStem Basal Media VEGF CXCL Interleukin 8 0 Lot 1 Lot 2 Lot 3 Lot 4 0 Lot 1 Lot 2 Lot 3 Lot 4 0 Lot 1 Lot 2 Lot 3 Lot 4 Measured Levels of Angiogenic Factors in Clinical Production Campaigns

27 Ave. number of Tube/Field Knock Down of VEGF,CXCL5 and IL-8 Results in Sub-Optimal Vascular Tube Formation EGM BASAL 30 VEGF 25 CXCL5 IL-8 20 CM CM (-VEGF) CM (-IL8) CM (-CXCL5) e 5 0 EGM Multistem Basal Media MulitStem CM MultiStem CM + IgG MultiStem CM (- VEGF, CXCL5 or IL-8) Knock-Down Vascular tube formation requires action of multiple factors Angiogeneic pathways are redundant complicates reductionist approach

28 Ave # of Tubes Formed Per Field Ave # of Tubes Formed Per Field Minimum Levels of VEGF, CXCL5 and IL-8 Required for Induction of Angiogenesis VEGF CXCL5 IL-8 0 pg/ml 0 pg/ml 0 pg/ml pg/ml 250 pg/ml 20 pg/ml pg/ml 1000 pg/ml 60 pg/ml 0 EBM EGM SFM SFCM ISO 0 pg/ml IL8 20 pg/ml IL8 60 pg/ml IL8 80 pg/ml IL8 100 pg/ml IL pg/ml 2000 pg/ml 80 pg/ml Pass-Fail Criteria: Knockdown and Add-back Sets Critical Threshold Required for Activity

29 Multiple Angiogenic Factors Required for Patent Vessel Formation

30 Clinical Manufacturing Lots Show Consistent Cytokine Expression Requires qualification of ELISAs for use as a potency assay Potency Assay Provides Consistency and Comparability Guidance in Process Improvements for Phase III Potency Assay Ensures Angiogenic Factor Production is Above Critical Pass/Fail Threshold

31 Potency Reflects the Composite PK/PD Profile of a Cell Product Cell Size & Other Factors Impact Biodistribution to Lung, Liver, Spleen Reservoir

32 Nothing is Easy Reductionist approach to correlate angiogenic knockdown in preclinical ischemic model may not be possible or fully informative - Parallel repair pathways (cytoprotection, anti-inflammatory injury, recruitment of host progenitors) may obscure angiogenic response Biodistribution of product may be key and impact pre-clinical, clinical potency independent of factor production - Effective dose of product influenced by cell size, tissue targeting receptors influenced by culture conditions Question for tomorrow: How many composite properties are required for a comprehensive potency assessment?

33 Thanks! Athersys Cardiovascular Team Nick Lehman Tony Ting Julie Woda Cardiovascular Collaborators Johnathan Lindner (Oregon Health Sciences Univ) Aernout Lutton (KU Leuven BE) Marc Penn (CCF, Summa Health Andrew Wragg (NHLBI)