Sub theme 3.12
Clinical and experimental immuno (-gene) therapy


Goals of research: general outline
Scientific achievements
Future plans: special goals and approach
Running projects
Associated staff


Goals of research: general outline

Goal

Therapeutic use of T lymphocytes genetically endowed with specificity to cancer in general and renal cell cancer and melanoma in particular.

Solid tumors such as renal cell carcinoma and melanoma are amenable to immune therapy. In particular, adoptive transfer of antigen-specific T cells has demonstrated significant clinical successes. We and others have demonstrated that transfer of antibody or TCRab genes into T cells (i.e. genetic T cell retargeting) represents a feasible and attractive rationale to provide tumor-specific immunity to patients. TCR gene transfer introduces TCRab genes, generally derived from T cell clones with known antigen specificities, into a predetermined T cell population and preserves both avidity and peptide fine specificity of parental T cell clones. Currently, gene transduction and T cell expansion protocols have been optimized, meet good clinical practice (GCP) criteria, and are implemented in a Rotterdam phase I study to treat metasta­tic renal cell cancer with autologous gene modified T cells.

Current research efforts are directed towards further improvement of efficacy and safety of TCR gene therapy. To this end, genetic T cell engineering is further optimized, and molecular strategies are designed to enhance the immune control of tumors.

Core research elements include:

  • Target antigens of choice, such as MAGE antigens, show an expression that is restricted to malignant tissue and are therefore considered safe. Pre-clinical in vivo models used to test adoptive T cell therapy deal primarily with melanoma.
  • Design of alternative receptor formats that show high surface expression levels and are less prone to result in possible dangerous T cell reactivities when compared to ‘natural’ TCR.
  • (Genetic) strategies to generate T cells with enhanced anti-tumor activity as well as an improved ability to counteract the inflammatory and often immunosuppressive milieu of tumors.

The Department of Medical Oncology has 20-year track record in clinical trials of immunotherapy of solid tumors and extensive experience in monitoring immunotherapeutic interventions, and as such fulfils the role of reference and expertise center at national level (Stichting Kwaliteitsbewaking Medische Laboratorium Diagnostiek) and in various European consortia (such as EU FP6 ATTACK).

Available technologies:

  • Design of antigen-specific immune receptors and their gene transfer.
  • Selection and validation of antibodies from an in-house phage-display library (repertoire 8x109).
  • Non-genetic technologies to redirect T cells.
  • GMP T cell processing and conduct of clinical immuno(gene) therapy.
  • CCKL-accredited immunological monitoring of therapeutic interventions including up to 6-color flow cytometry, in vitro assays and real-time PCR to assess absolute numbers, phenotype and function of patient T lymphocyte subsets.

Available facilities for adoptive cellular (gene) therapy:

  • Experimental Animal Center providing DM1 and 2 rooms for (gene) therapy research in rodents and nonhuman primates.
  • Hospital Pharmacy having > 10 years of experience in supporting clinical gene therapy trials.
  • GMP-facility for T cell procedures is combined with the GMP facility for clinical hematopoietic stem cell processing and operational since 1999.
  • Clinical isolation rooms and an outpatient facility suitable for patient treatment.

Scientific achievements

Achievements:

  • Contributed significantly to the field of human T cell retargeting to tumors or viruses via gene transfer of antibody-based and T cell receptors, and participated in various EU consortia on this topic.
  • Molecularly engineered TCR to enhance efficacy and safety of TCR gene therapy.
  • Translated gene transfer technology into a GMP-level, clinically applicable protocol.
  • Currently performing a phase I/II study to treat metasta­tic renal cell cancer with autologous gene modified T lymphocytes, the first clinical study of its nature in Europe (P00.0040C, v14).
  • Clinical lessons learnt:
    1. Maintenance of chimeric receptor mediated functions of gene modified T cells upon adoptive transfer to patients.
    2. On-target toxicities are mediated by chimeric receptor-positive T lymphocytes (in case receptor is directed against self-antigen).
    3. Immunogenicity of chimeric receptor gene modified T cells: limiting long-term or repeated treatment.

Future plans: special goals and approach

·         Implementation of clinical TCR gene therapy directed against metastatic melanoma patients. Clinical lessons learnt from our renal cell carcinoma trial (see above) are currently addressed pre-clinically in an effort to uniquely optimize trial design (including choice of antigen; receptor format; and cytokine treatment of T lymphocytes prior to adoptive transfer).

·         Immune monitoring of patients with emphasis on detection of rare events (in collaboration with subtheme 3.10: John Foekens, Els Berns, Stefan Sleijfer and Arzu Umar).

·         Development and testing of strategies that reverse the pro-inflammatory and immune evading milieu within tumors, which are expected to facilitate immune-mediated tumor eradication and support adoptive T cell therapy.

·         Development and testing of nanomedicines targeting the tumor milieu again to support adoptive T cell therapy (in collaboration with subtheme 3.7: Alexander Eggermont and Timo ten Hagen).


Most recent publications

Clinical (immuno-)gene therapy:

  1. Lamers C, Gratama J, Pouw N, Langeveld S, van Krimpen B, Kraan J, Stoter G, Debets R. Parallel Detection of Transduced T Lymphocytes After Immunogene Therapy of Renal Cell Cancer by Flow Cytometry and Real-Time Polymerase Chain Reaction: Implications for Loss of Transgene Expression. Hum Gene Ther, 2005 16:1. IF 4.3
  2. Lamers CHJ, Sleijfer S, Vulto AG, Kruit WHJ, Kliffen M, Oosterwijk E, Debets R, Gratama JW, Stoter G. Treatment of metastatic renal cell carcinoma with autologous T-lymphocytes genetically retargeted against carbonic anhydrase IX: first clinical experience. J Clin Oncol, 2006 24:e20. IF 15.5
  3. Lamers C, Willemsen R, van Elzakker P, van Krimpen B, Gratama J, Debets R. Phoenix-ampho outperforms PG13 as retroviral packaging cells to transduce human T cells with tumor-specific receptors: implications for clinical immunogene therapy of cancer. Cancer Gene Ther, 2006 13:503. IF 3.0
  4. Lamers CHJ, Langeveld SCL, Groot-van Ruijven CM, Debets R, Sleijfer S, Gratama JW. Gene-modified T cells for adoptive immunotherapy of renal cell cancer maintain transgene-specific immune functions in vivo. Cancer Immunol Immunother, 2007 56:1875. IF 3.8
  5. Lamers CHJ, Willemsen RA, van Elzakker PMML, Gratama JW, Debets R. Gibbon Ape Leukemia Virus poorly replicates in primary human T lymphocytes: implications for safety testing of primary human T lymphocytes transduced with GALV-pseudotyped vectors. J Immunother, 2009 32:272. IF 4.8

Experimental (immuno-)gene therapy:

  1. Willemsen RA, Weijtens ME, Ronteltap C, Eshhar Z, Gratama JW, Chames P, and Bolhuis RL. 2000. Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR. Gene Ther. 2000 7:1369. IF 4.5
  2. Schaft N, Willemsen RA, de Vries J, Lankiewicz B, Essers BWL, Gratama JW, Figdor CG, Bolhuis RLH, Debets R, Adema G. Peptide fine-specificty of anti-gp100 CTL is preserved following transfer of engineered TCRa/b genes into primary human T lymphocytes. J Immunol, 2003 170:2186. IF 6.1
  3. Schaft N, Lankiewicz B, Drexhage J, Berrevoets C, Moss DJ, Levitsky V, Bonneville M, Lee SP, McMichael AJ, Gratama JW, Bolhuis RLH, Willemsen R, Debets R. T cell retargeting to Epstein-Barr virus antigens following T cell receptor gene transfer: CD28-containing receptors mediate enhanced antigen-specific interferon-g production. Int Immunol, 2006 18:591. IF 3.3
  4. Willemsen R, Sebestyen Z, Ronteltap C, Berrevoets C, Drexhage J, Debets R. CD8a co-receptor to improve TCR gene transfer to treat melanoma: down-regulation of tumor-specific production of IL-4, IL-5 and IL-10. J Immunol, 2006 177:991. IF 6.1
  5. Sebestyén Z, Schooten E, Sals T, Zaldivar I, San José E, Alarcón B, Bobisse S, Rosato A, Szöllősi J, Gratama J, Willemsen R, Debets R. Human TCR incorporating CD3z induce highly preferred pairing of TCRa and b chains following gene transfer. J Immunol, 2008 180:7736. IF 6.1