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NCI experience using engineered T cells for the treatment of cancer (Implication for RCR testing). Richard A. Morgan, Ph.D. Surgery Branch National Cancer Institute. 20 Years of Clinical Gene Therapy at NCI. Since this initial report, >200 patients have been treated

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nci experience using engineered t cells for the treatment of cancer implication for rcr testing

NCI experience using engineered T cells for the treatment of cancer(Implication for RCR testing)

Richard A. Morgan, Ph.D.

Surgery Branch

National Cancer Institute

20 years of clinical gene therapy at nci
20 Years of Clinical Gene Therapy at NCI

Since this initial report, >200 patients have been treated

at NCI with transduced lymphocytes

slide3

Development of gene therapy as

an alternative to TIL therapy

production process mcb
Production Process - MCB

MSGV1 backbone:

Combination of MSCV+SFG

Hughes et al., Hum Gene Ther 2005; 16:457--‐472.

Clone Selection

(RNA Dot Blot)

Transduction

Transient Transfection

PG13

Phoenix-Eco

  • Select 6 best clones
  • Functional evaluation
  • Select final clone
  • Produce master cell bank
production process retroviral vector
Production Process - Retroviral Vector

SBVPF

Preclinical

Vector Development

25 x 1700cm2

Roller bottles

Generate PG13 MCB

Cell Thaw

120/240 ml

Upstream

Cell Expansion

Seed Roller Bottles

Medium Exchange/Harvest

Clarification

Downstream

Fill

Modified from: Reeves and Cornetta, Gene Ther. 23:1993-8, 2000

slide10

Procedure for TCR Gene Therapy Protocols

Carry out patient pheresis to obtain PBMC

Activate T cells using anti-CD3 antibody for two days

Transduce with gammaretroviral vectors using RetroNectin-coated plates

Pre-condition patients with cyclophosphamide and fludarabine

Administered T cells with IL-2 to tolerance

+/- Vaccine

Post-treatment----Test/archive samples for further RCR testing

RCR-PCR

slide11

NY-ESO-1 TCR gene therapy:

Tumor regression in melanoma and synovial cell sarcoma patients

* No on or off target toxicities have been observed

that could be attributed to TCR engineered cells.

Paul F. Robbins, et al., In press, Journal of Clinical Oncology

summary clinical findings
Summary: clinical findings:
  • Both TCR and CAR gene therapy targeting multiple tumor antigens can lead to cancer regression in humans (melanoma, colorectal cancer, synovial cell sarcoma, and lymphoma).
  • Targeting self-antigens also leads to on-target recognition of antigen expressing normal cells and mild to severe toxicity.
rcr testing implications i
RCR testing implications(I)
  • A total of 144 patients have been treated on active INDs. The total number of all current and former patient treatments is 231.
  • All 231 patient treatments have been tested by PCR-based lot release assays for RCR and all 231 patient treatment products were negative. 22 patient treatments were tested by cell-based RCR assays and all were negative. All remaining patient samples have been archived for the purposes of future testing if necessary
rcr testing implications ii
RCR testing implications(II)
  • Issues with cell-based RCR testing:
    • It is difficult and unwise to perform a cell culture based RCR “in house” because it requires an infectious retrovirus control, which could lead to contamination.  Therefore, the assay must be contracted to an outside vendor. 
    • The test is extremely expensive (>$3000/sample), and contractors capable of performing the assay are scarce.    The government procurement procedures for outside contractors are complex, involved, and protracted.
    • Achieving samples is also an onerous prospect in particular when long-term storage is required. 
rcr testing implications iii
RCR testing implications(III)
  • Why RCR testing at all?
  • Current gamma-retroviral vector backbones in combination with split-gene packaging cell lines do not generate RCR. Furthermore, each and every retroviral product and master cell bank is tested for RCR using cell based assays before it can be released for patient use. It is highly unlikely that RCR (which is a theoretical risk to begin with) would escape detection in this extremely thorough pre-transduction testing. So why test for something that is not there to begin with????
rcr testing implications iv
RCR testing implications(IV)
  • Recommendations:
  • The simplest solution is to eliminate patient-specific cell-based RCR testing.  The previously certified vectors and processes are theoretically extremely unlikely to generate RCR (or RCL) and other patient safety issues vastly supersede the effort and expenses spent on this issue. 
  •  An alternative would be to archive all samples, and test later only if a problem were suspected for clinical or other reasons.  The EU model currently does not require cell-based RCR testing of transduced patient.
acknowledgments
Acknowledgments:

Morgan Lab

Steven A. Rosenberg, M.D., Ph.D.

Chief, Surgery Branch NCI

Surgery Branch Colleagues:

Mark Dudley

Steve Feldman

Carolyn Laurencot

John Wunderlich

Paul Robbins

James Yang

Nick Restifo

Jim Kochenderfer

Maria Parkhurst

TIL LAB

FACS LAB

Clinical Staff

Vector production:

Kenneth Cornetta

Indiana University

Takara Bio for supplying

clinical grade RetroNectin

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