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Production Assistance for Cellular Therapies ‐ PACT

Accelerating Your Cell Therapy

Tuesday March 14, 201712:00 12:00 –– 1:15 PM ET1:15 PM ET

NHLBI – PACT history, scope and application criteria

PACT facilities ‐ Product manufacturing capabilities• David McKenna - University of Minnesota, Molecular and Cellular

Therapeutics• Aisha Khan - Interdisciplinary Stem Cell Institute Cellular

Today’s Web SeminarToday’s Web Seminar

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• Aisha Khan Interdisciplinary Stem Cell Institute CellularManufacturing Program, University of Miami

• Joseph Gold - Center for Biomedicine and Genetics, City of Hope• Linda Kelley - Moffitt Cancer Center• Adrian Gee - Center for Cell and Gene Therapy, Baylor College of

Medicine

Emmes ‐ Application Process

PACT PACT ‐‐ OverviewOverview

PACT Goal – advance cellular therapeuticsfrom the laboratory bench into clinicaltrialsEd ti l P Educational Programs

Resource Center

PACT Services

PACT Website

https://www.pactgroup.net

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PACT Program Life CyclePACT Program Life Cycle

Began September 2003• Cell product manufacturing for clinical trials• 3 cell processing facilities

Renewed January 15, 2010

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• Scope expanded to include translational services• 5 cell processing facilities

Renewed in July 2016• Revised MissionProvide assistance for cellular therapy translational research and the manufacture of cellular therapy products

PACT Organizational StructurePACT Organizational Structure

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PACTPACT‐‐ Requests for Services Requests for Services

PACT will review requests for translational services inthe context of what is required in a product development plan and with an end goal of an eventual IND submission.

PACT Cell Processing Facilities provide the requestedproduct development services and cellmanufacturing for approved projects.

PACT does not provide direct funding to investigators.

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PACTPACT

Cell products that aid in the repair and regeneration ofdamaged and diseased tissues, organs, and biologic systems

Preclinical studies including basic and translational work

Beginning after preclinical therapeutic proof‐of‐concept studies though to validation studies immediately prior toPhase I clinical trial

Products and Services of programmatic interest to NHLBIor with support from other NIH Institutes

Proposals possessing procedural advancements tofurther foster and standardize cell therapies

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Cardiac repair and disease

Lung repair and disease

Hematologic repair and disease (non‐malignant)

H t i ti ll t l t ti i l di ft

NHLBI Scientific InterestsNHLBI Scientific Interests

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Hematopoietic cell transplantation; including graft manipulation and complications (GVHD; graft failure,infections)

Red blood cell, platelet, or white blood cell productsfor transfusion

For more information on PACT Application: Scope Criteria visit www.pactgroup.net

Other NIH Institutes and Federal partners have theability to support investigators through the PACT program

Determined on a case‐by‐case basis

Projects outside of NHLBI InterestProjects outside of NHLBI Interest

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Determined on a case by case basis

Investigators with projects that are outside of NHLBI scientific interests are strongly encouraged to discuss the possible use of PACT with their NIH or Federal Agency program officer and with the PACT NHLBI Program Director prior to starting an application

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ApplicationsApplications

Products and Services

• Supporting activities that will bring proof ofconcept discoveries into the translational development stage.

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p g• Scale‐up

• Optimizing manufacturing processes

• Pre‐clinical process development

• SOP development

• CMC section development

• Cell product manufacturing for relevant animal model studies and other IND‐enabling studies.

Also see https://www.pactgroup.net/content/pact‐services

NHLBI Contact InformationNHLBI Contact Information

Lisbeth Welniak or Traci Heath MondoroPhone: 301‐435‐0065Email: [email protected]

[email protected]

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@ g

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Molecular & Cellular Therapeutics

David H. McKenna, M.D.PACT WebinarMarch 14, 2017

Molecular & Cellular Therapeutics

www.mct.umn.edu

• Class 10,000 clean suites

• Single pass air, isolated differential air pressure rooms, negatively pressured to adjoining rooms (can be adjusted)

• Rooms terminal HEPA‐filtered air tested at1 000 10 000

MCT ‐ Production Space

www.mct.umn.edu

1,000‐10,000 particles/ft3, 20‐70 airchanges per hour

• Production rooms are constructed with epoxy liquid flooring coved base, reinforced resinous wall surfacing, and epoxy painted veneer plaster ceilings

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Recent & Current CT ProjectsProduct Type Procedure Technology

Allo NK (PB) Short-term cx/long-term cx

IM selection, IL-2/IL-15 activation, expansion

Allo Treg (UCB) Long-term cx IM selection, 3/28 or K562 (KT64.86) expansion

Allo MSC (BM) Long-term cx MNC (density gradient), cell factories

Allo UCB expansion Long-term cx IM selection, expansion

Allo UCB homing Incubation Short-term C3a incubation

Auto marrow MNC MNC (automated) MNC (density gradient)

Dendritic cell vaccine [glioblastoma l if (GBM)]

Mid-term cx IM selection, flasksmultiforme (GBM)]

Dendritic cell/tumor cell hybrid vaccine (multiple myeloma)

Mid-term cx PEG-driven fusion

GBM vaccine (lysate) Long-term cx, nebulization

MCB expansion

Allo iTreg (PB) Long-term cx IM selection, KT64.86 expansion

iPS cell banking platform Long-term cx Sendai virus

Virus-specific T cells (CMV, EBV, Adeno, etc.)

Cytokine-capture technology

IM selection

Right ventricular outflow tract (decellularized tissue tube)

Long-term cx Fibrin-based matrix, tube mold

MethodsPB‐Derived NK Cells

anti‐CD3/19

www.mct.umn.edu

Overnight:• XVIVO-15• IL-2• AB serumLot release testing

•CD3-/CD56+•CD3+•Viability•Endotoxin•Gram stain

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MethodsUCB‐Derived Treg Cells

anti‐CD25

www.mct.umn.edu

Day 0:1 x 106 cells/mL in:X-VIVO 15 10% AB serumL-glutaminen-acetylcysteineanti-CD3/CD28 beads (UPenn)

Day 3:IL-2 at 300 units/mL

Day 5-14:0.5 x 106 cells/mL

Day 15-18(+/-1):1 x 106 cells/mL

•Bead removal•Lot release testing

MSC Manufacturing

www.mct.umn.edu

• Trilineage assays• KGF assay

Manufacturing of Induced Pluripotent stem cells (iPSCs)

www.mct.umn.edu

James Dutton, PhD, Jakub Tolar, MD

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Tissue Engineering

• Decellularized tissuetube from humanneonatal fibroblasts forpediatric rightventricular outflow

Tubular Tissue Engineered Valve

www.mct.umn.edu

ventricular outflowreconstruction (RVOT).

• Cells are entrapped onfibrin gel tube

Physiological compliance

High burst strength

Completely biological

Allograft (acelluar)

Syedain et al 2014, Tissue Engr AMeier et al 2014, J Cardiovas TranSyedain et al 2013, Ann Biomed EngrSyedain et al 2010, Biomaterials

Final product is a decellularized off-the-shelftissue-engineered valve

R Tranquillo, Z Syedain

Collaborators/PIs:Bruce Blazar Jeff MillerClaudio Brunstein Zeeshan SyedainSarah Cooley Jakub TolarJames Dutton Bob TranquilloMichael Matthay John Wagner

Thank you!

CT Lab:Diane Kadidlo

Kristen ReynaMichelle Lucio

www.mct.umn.edu

Darin SumstadLisa VanOrsowNancy BostromSheryl AdamsNancy ColeyCindy StanawayLien LeMolly GroweAnh DoStacy LinnAlemayu Emiru

Michelle LucioJulie LaTour

MCT QA:Fran RabeAlison JakubekMaria Opitz& Materials Management/Administrative & Support Staff

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Interdisciplinary Stem Cell InstituteUniversity of Miami

Cell Processing Facility (CPF)

03/14/2017

Aisha Khan, M.Sc, M.B.A

Executive Director of Laboratory Operations

Interdisciplinary Stem Cell Institute

The Miller School of Medicine, University of Miami

Interdisciplinary Stem Cell Institute

• Serves as a manufacturing resource for the various clinical programs

• Facilitates translational research

• Provides regulatory and scale-up expertise to bridge the gap between basic and clinical research applications

• Provides services throughout the IND application

Cell Processing FacilityIntroductionIntroduction

development process, from the proof-of-principle and pre-clinical testing to clinical trial phase.

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Services regional and national research efforts in reparative/regenerative medicine

Equipped to do Phase I, II and III clinical trials

Facility CredentialsFacility Credentials

TRANSLATIONAL CELL

PROCESSING

FDA registered

Serves as a national resource for translational research in the area of cellular therapy

Equipped to manufacture licensed cellular products for new therapies and tissue‐engineered end‐products.

Biomedical Research Building, 9th floor(12,931.28 sq. ft., 85 rooms). Red square indicates location of the cell processing Facility

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950

949 937

951

952

945

Floor Plan of the CPF9TH floor BRB building

Legend950 – General Area of Development Lab951-Tissue Culture Room952-Tissue Culture Room

Cleanroom Facility937 – Gowning in of the Cleanroom939 – Clean Storage Room

G

946

947

948 938939

941942

944

945-General Area942- Cryogenic Freezer Room944-Gowning out of the Cleanroom946 – Tissue Culture Room947 – Tissue Culture Room948 – Tissue Culture Room949 – Tissue Culture Room

Ancillary Areas938 –Clean Storage Room941 – Cryogenic Freezer storage Room

Products• MSCs – Human

• MSCs – Rat

• MSCs – Porcine

• MSCs – Canine

• c-Kit+ Cardiac Progenitor Stem Cells – Human

• c-Kit+ Cardiac Progenitor Stem Cells – Porcine

• c-Kit+ Cardiac Progenitor Stem Cells – Rat

• c Kit+ Kidney Progenitor Stem Cells Porcine

• Cord Tissue Derived MSCs• Cord Blood Derived MSCs• Placental MSCs• Exosomes• Human and Porcine iPSC• Human T- Cell • Human T-regulatory Cells• Human NK Cells• Amniotic Fluid Stem Cells

• c-Kit+ Kidney Progenitor Stem Cells – Porcine

• Human Dendritic Cells

• Human Tumor Lysate

• Human Schwann Cells

• Rat, Porcine, & Primate Schwann Cells

• gene-modified CD34+ hematopoietic cells

• Human Dental Pulp Stem Cells

Clinical Products

Products being used for clinical trials:

• MSCs

• Bone Marrow Mononuclear Cells

• Schwann Cells

• Dendritic Cells

• Tumor Lysate

• Cord Tissue Derived MSC

• CD34+ Cells

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Mesenchymal Stem Cell Manufacturing

• GLP Manufacturing

• GMP Manufacturing

• Enabled 15 phase I/II INDs

Manufacturing Steps of MSCs

Bone marrow aspira on

Bone marrow

Ini al seeding

Ficoll gradient

MNCs

Expansion

MSCs

CD105: 99.9%

Back to

human

Manufacturing Steps of Schwann Cells • GLP

Manufacturing



Dendritic Cell Vaccine for Malignant Gliomaand Glioblastoma, sarcoma and brain tumor



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MULTICENTER CLINCAL TRIALSCardiovascular Cell Therapy Research Network (CCTRN)

Endomyocardial BiopsyAutologousAutologous

C-Kit Cardiac Stem Cells (CSCs)

Digestion ExpansionMagnetic Selection

cGMP Compliant ManufacturingcGMP Compliant Manufacturing

CC--Kit CSCsKit CSCs

Safe Safe PotentPotent

CharacterizedCharacterizedFull DoseFull Dose

• GLP Manufacturing


• 1phase I/II INDs at ISCI

• Manufactures for 7 center for phase II trial

Examples of iPSC studies at ISCI

Basic Research (Human and mouse iPSCs)

Preclinical Research(Human and swine iPSCs)

• Modeling molecular and cellular • Development of clinical grade• Modeling molecular and cellular mechanisms of myocardial lineage development

• Development of clinical-grade, autologous and allogeneic iPSC-derived cardiomyoblasts for cell-based therapy in response to heart damage

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Clinical-grade manufacturing of iPSC-derived CPCs expressing the novel receptor

Human and porcine iPSC MCBs from cGMP-grade

MSCs Establishment of xeno-free cGMP

compliant monolayer differentiation

protocols

Scale-up differentiation

and purificationof CPCs

Quantum System (Terumo)

SOP and quality control Porcine model of MISOP and quality control documents

Safety, identity, purity, viability

Assay Qualification

Xeno-free, Virus-freeFeeder-free, Transgene

Free MCB

SOP and quality control documents

Safety, identity, purity, viability

Assay qualification & Validation

Xeno-free, Virus-freeFeeder-free, Transgene

Free product

Development Capabilities

Lab Capabilities

IncuCyte® applications for cell monitoring & workflows

Cells are monitored non-invasively, within the incubator and images are automatically analyzed so investigator can always see what happened when and where.

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Quantum® Cell Expansion

System

QiIACube

This instrument allows for high-throughput extraction of DNA and RNA.

Automated cell culture platform that can help simplify the open, labor-intensive manual tasks associated with flask-based culture.

MoFlow Cell Sorter High-speed sorter, compatible with array of laser options. High viability, yield and purity with an analysis rate of 100,000 events per second and sort rates of up-to 70,000 events per second are standard.

Thank you

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The Center for Biomedicine and Genetics at COH

Joseph Gold, Ph.D.Director, manufacturing

March 14, 2017

Center for Applied TechnologyBeckman Research Institute, City of Hope

GMP Capacity at COHCenter for Biomedicine and Genetics (CBG)

•Designated NGVL Sept, 2001

•Designated ICRC, Sept, 2001•Designated PACT May 2010; Aug 2016•CIRM TC partner facility October 2017

2000

C ll h d i C (C C)

20,000 ft2

GMP Manufacturing Core at COH

2012

•Small molecules

•Reference standards

•Oligo therapeutics

3,000 ft2

2010

Cell Therapy Production Center (CTPC)

•Cadaveric islet cells

•CAR‐T work6,000ft2

Chemical GMP Synthesis Facility (CGSF)

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CBG production experience

• 17 years in operation

• >400 released product lots

Lentivirus

Neural stem cells

hiPSC

hESC

Islet cells

MVA

AdenovirushESC/hiPSC ‐cardiomyocytes

hESC‐RPE/NSC/DA neurons

Small molecule fills

Biologics fills

MSC

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Notable product expertise and capabilities at CBG

• Lentivirus production 100L scale High transduction efficiency

• Neural stem cell modification and banking

• hPSC culture and banking

• hPSC differentiated products: production and dose prepsp p p p

Scalable suspension adapted cardiomyocytes NSC, NPC, DA neuron progenitors Retinal pigment epithelia

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hESC on CELLstart(or Matrigel, Synthemax)

single cells

Accutase

125-1000 mL spinner flask

ROCK inhibitor

StemPro SFM or E8

(40ng/mL bFGF)70 rpm

passage cells every 3 days

Scalable hPSC suspension culture

MCB/WCB

2.5x105

cells/ml

(Accutase)

200 mm

cell aggregates

Vincent C. Chen et al., Stem Cell Research, May, 2012

(Accutase)

differentiation process

•Defined reagents

•No substrates

•No beads, matrices, gels

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d5

cell harvesting

d0 d1 d3 d18-21d2 d4 d6 d7d-2

hPSC expansion

StemPro hESC SFM

cardiac differentiation

RPMI + B27-insulin RPMI + B27

IWP4 CHIR 99021

Wave bag cardiac differentiation

d20(4X)d0 (4X)

Vessel cells/L input cells/L output cell viability cTnT hPSC‐CM/LCardiac/hESC

2L bag 2.5E+08 1.78E+09 96.1% 92.9% 1.65E+09 6.6

cTnT TE7 OCT4

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Target: Macular degeneration (CPCB)Strategy: hESC‐RPE monolayer on synthetic substrate

hESC‐RPE

Parylene substrate

Day 9 D103

hESCMCB

ExpandEnrichDifferentiate ICB

Cryopreserve

hESC‐

RPE

hESC‐

RPE

hESC‐

RPE

Thaw/culture on Parylene

7 Months 1 Month

Full testing

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Wolfram ZimmermannDeepak Srivastava

Academic Collaborators and Clients

Kevin Healy

Tim Nelson

Joseph WuCharles Murry

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Mark Humayun

Production Assistance for Cell Therapy CIRM Translating Center

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Moffitt Cancer CenterCell Processing Facility

Linda L. Kelley, PhDy

Director

Moffitt CPF Staff and Facility

Project ManagementMoffitt Cell Therapies

Cheryl Cox, BSManager

Emily Hopewell, PhDAssistant Technical Director

Nermin Gerges, PhDDevelopment Specialist

Renee Smilee, BSFacility Operations Manager

Martha Hackett, MSQuality Assurance Director

Virginia Garcia-Gobaira, MATraining Specialist

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T Regulatory (Treg) Cells for GVHD

Dendritic Cell (DC) Vaccines

Tumor Infiltrating Lymphocytes (TIL)

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Chimeric Antigen Receptor (CAR) T Cells

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Center for Cell & Gene Therapy cGMP Activities

Adrian GeeBaylor College of Medicine

Houston, Texas

Location

Feigin Center, TCH

Support of Blood & Marrow Transplant Programs

30 beds expanding to 60

• Bone Marrow

• PBPC

• Cord BloodCord Blood

• DLI

• CD34 selected

Special Protocols

Bellicum

Novartis etc.Jeannette Bloom et al.

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Virus‐specific T cells• Used to prevent or treat viral infections inimmune‐suppressed patients

Cliona Rooney

Specialized Products & Services

patients• EBV, followed by CMV,Adeno, HHV‐6, BK & HIVvirus

• Used across HLA barriers

• Manufacturing time reduced from 3 months to 14 days

HelenHeslop

Ann Leen

Chimeric Antigen Receptor (CAR) T Cells

• Directed towards solidtumors

StephenGottschalk


tumors

• Overcoming tumor resistance mechanism

• Combining tumor and viral specificity to prolong persistence &provide means to reactivate

Maksim Mamomkin

NabilAhmed

Mesenchymal Stromal Cells ‐MSC

• Regenerative and Immunosuppressive

Zhuyong Mei


Immunosuppressive actions in

• Cardiac disease

• Stroke

• Parkinson’s Disease

• Kidney Transplantation

Adrian Gee

Mariola Klis

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Cell Culture Systems• Simpler and automated systems formanufacturing of

JuanVera


manufacturing of therapeutic cells

• G‐Rex bioreactor

• Gather

Suicide Genes• Designed to destroy cells in the case of anadverse reaction

MalcolmBrenner


adverse reaction

• Activated by administration of a small molecule drug

• Used to switch off tumor‐directed T cellsin cases where they may cause GvHD

Therapeutic Cell Sorting• Sorting of Aldehyde‐bright progenitor cells April Durett


• Used by the CCTRN to treat peripheral arterydisease

• PACE protocol – some promising results

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Quality Control • Endotoxin

• MycoplasmaDeborah Lyon


• Bioburden

• Replication‐competent virus detection

• Viral titers

• Vector identity

• Stability testing

• EBV assays


Quality Assurance Services

• Generation of SOPs and worksheets for

Sara Richman

specialized protocols

• Review of records

• Generation ofCertificates of Analysis

• Assistance with CMCsection of IND

NatashaLapteva

MygiaoLe

Contact us at: [email protected]

through the PACT Program

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APPLICATION PROCESSAPPLICATION PROCESS

Debbie WoodEmmes Corporation Rockville, Maryland

PACT Application ProcessPACT Application Process

Web‐based Application• Scope Review by NHLBI

• Full Application invited if scope is approved

Scientific Review Board Review & NHLBI Vote

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TO‐RFP issued; competitive bids

TO awarded (1 CPF)

Contract, Timeline & Budget

Scope Review Application ContentScope Review Application Content

Investigator information (Biosketch, CV, etc.)

Scope

• Relevance to NHLBI scientific interests and PACT Scope

Proposal Overview

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Proposal Overview

• Synopsis of proposed study

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Full Application ContentFull Application Content

Investigator information (Biosketch, CV, etc.)

Scope ‐ Scientific interest and programmatic scope

Proposal Overview

Product information

Current funding ‐ product manufacturing and other activities/tasks not covered

Scope Review

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g p f g /by PACT.

Manufacturing information

Production methodology

Product development

Regulatory information

Timeline

Estimated budget

Institutional/Organizational business office information

Application Review/CriteriaApplication Review/Criteria

NHLBI and an external, independent Scientific Review Board (SRB) comprised of cell therapy experts review:

• Relevance to the scientific mission and programmatic scope

• Applicant and the collaborative team qualifications

• Evaluation of the strengths and weaknesses of preliminary data,

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manufacturing feasibility, significance to the field, degree of innovation,and overall scientific merit of the application

• Detailed translational development plan summary

• Product specifications (proposed release criteria and potency assay)

• Supplemental manufacturing funding

• Demonstration of an adequate plan for eventual use of the cell therapyproduct in a clinical trial

• Proposed work supports regulatory pathway for this product

Next StepsNext Steps

Contract

• Contract negotiations with designated facility

• Intellectual property and/or indemnification

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Timeline

• Development of production task order

• Scheduling of manufacturing tasks

• Scheduling of production

Budget

• CPF assistance

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Website Website

www.pactgroup.net

• Online application process and information

• Education Initiatives

/ d

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• Events/Updates

• Links to PACT facilities, regulatory references & other resources

Contact InformationContact Information

Debbie Wood or Lani IbenanaPhone: 301‐251‐1161Email: [email protected]

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