Sub theme 3.7
Innovative tumor model development/ tumor vasculature/ tumor targeting/ locoregional control

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

Goals of research: general outline

Combination therapy, mechanistic studies and clinical and translation studies

The major fields of interest of Surgical Oncology are the treatment of solid tumors, in particular melanoma, pancreatic cancer, breast cancer, colorectal and hepatic tumors and soft tissues sarcoma. The laboratory of Experimental Surgical Oncology aims at improvement of existing (local regional or targeted) therapies, development of new treatment modalities, defining new targets with emphasis on the tumor vascular network, and to provide insight in mechanism of therapy. The laboratory has a strong background in the development of innovative in vitro and in vivo tumor models, such as tumor cell and endothelial cell motility assays, tumor window models (including hyperthermic tumor chambers), intravital (confocal) microscopy, locoregional treatment models and nano-particle targeted treatment models. Fundamental research lines regard investigation of the mechanisms of tumor-vascular development and the formation of metastasis, with emphasis on cell-motility dynamics and cell survival.

There are 3 Main Areas of Research:

a) Tumor Manipulation and Targeting working group headed by Dr. Gerben Koning. The main subjects are: Loco-regional solid tumor therapy (TLM ten Hagen, AMM Eggermont). Manipulation of the tumorpathophysiology with vaso-active agents in solid tumor therapy (ALB Seynhaeve). Targeted Nanomedicine; development and application of targeted nanocarriers for drug delivery, molecular imaging and image-guided drug delivery. Development of nanotools for cell specific targeting and controlled release of contents from nanocarriers for instance using hyperthermia.

b) Mechanisms in Angiogenesis and Metastasis group headed by Dr. Timo ten Hagen. The main subjects are: Angiogenesis and metastasis as primary target for therapy in Malignant Melanoma. Analysis, prediction and therapeutic targeting of the metastatic potential of breast cancer. Mechanisms of early tumor vascular development (ALB Seynhaeve).

c) Translational Research linked to various clinical trials is performed mainly involving tissues from melanoma and pancreatic cancer patients. Interferon adjuvant therapy, prognositic and predictive biomarkers in melanoma (AMM Eggermont). Prognostics markers in pancreatic cancer (CHJ van Eijck).

A         Tumor Manipulation and Targeting

a) Loco-regional therapy of solid tumors. Treatment modalities are developed which improve local drug delivery by either surgical methodology, such as isolated perfusion or infusion approaches.

b) Manipulation of the tumorpathophysiology with vaso-active agents and /or hyperthermia in solid tumor therapy. With the aid of vaso-active agents drug delivery and activity of concomitant chemotherapy is improved both in the regional and systemic setting. Vaso-active agents are TNF, IL-2, Histamine and Cilengitide. The mechanism of TNF on endothelial cell function, tumor vascular permeability and vascular function is studied. Currently the interaction between endothelial cells and stroma cells, with respect to response to vaso-active agents, is under investigation.

Cellular processing of liposomal compounds (DXR and alike) is studied in vitro and in vivo. The fate of liposomal compounds in the tumor microenvironment is unclear. Here we study the fate op liposomes in the tumor, cellular uptake and intracellular processing of liposomes and their contents.

Flow kinetics in solid tumor. We have shown that manipulation of the vascular leakage with vaso-active agents and hyperthermia influences drug distribution in tumors. The fluid flow in tumors and transport of active agents (e.g. chemotherapeutics) is studied.

c) Nano-particle Drug Delivery Tools: Targeting, Nanotools and Hyperthermia (GA Koning). Receptor targeted radiotherapy (GA Koning).

Targeted nanomedicine research aims at optimizing solid tumor therapy by developing and applying novel nanocarriers for the delivery of chemotherapy, radionuclides and molecular imaging probes and/or combinations thereof. To achieve this the following strategies are being followed:

        increased drug targeting using lipid-based nanocarriers.

        triggered drug release from thermosensitive lipid nanocarriers.

        enhanced intracellular delivery of cytostatic drugs by modulating tumor cell membranes.

        receptor-specific targeting of radionuclides and drug-containing nanocarriers.

        molecular imaging probes for imageable drug delivery using nanocarriers.

B         Mechanisms in Angiogenesis and Metastasis

Angiogenesis and metastasis as primary target for therapy in malignant melanoma. In vitro as well as in vivo the effect of melanoma cells on endothelial cell survival, growth and migratory behavior is studied. The migratory capacity of melanoma cells is studied in relation to occurrence of metastasis.

Analysis, prediction and therapeutic targeting of the metastatic potential of breast cancer. In vitro studies are performed on breast cancer metastasis with emphasis on biological characteristics such as survival, interaction with stromal cells and migratory behavior.

Mechanisms of early tumor vascular development. Initiation of angiogenesis and early sprout formation is studied. Main focus is the behavior of the Tip cell in tumor vascular development.

C         Translational Research

Interferon adjuvant therapy in melanoma. In large trials subgroup analysis is performed on predictive markers for Interferon–based therapy of malignant melanoma. Studies are performed on immunologic markers, Interferon pharmacokinetics and SNPs in melanoma patients.

Prognostics markers in pancreatic cancer. In resected pancreatic cancer specimens biomarker analysis by immunohistochemistry, Q-PCR and Western Blotting is performed.

Advanced animal model systems for solid tumor therapy research.

An important area of technological development is the realization of an intravital mouse and rat model, which enables real-time studies on tumor vascular events, drug delivery and tumor response. The in-house established model allows imaging with a confocal microscope at the subcellular level. The model is based on the skin fold window chamber and is used for most studies mentioned above. Currently, transgenic mice, with specific fluorescent cells are acquired to study endothelial and stromal cell interaction. In addition a heating device has been developed to study behavior of tumor cells and tumor vasculature during and after hyperthermia and to study heat-triggered drug delivery approaches (coll. with Gerard van Rhoon, subtheme 3.4 Hyperthermia: a treatment for cancer).

Targeted therapy

Nanotools are deployed to develop tumor targeted devices which will be used in the adoptive T cell therapy (in collaboration with subtheme 3.12: Reno Debets)'. Specific antibodies to melanoma markers of T Cell receptor constructs are coupled to liposomal nanoparticles to device a targeted delivery tool for chemotherapy.

Scientific achievements
  • Combination chemo-immunotherapy in loco-regional therapy: (isolated perfusion).
    Addition of Histamine to an Isolated Limb Perfusion (ILP) with Melphalan results in profoundly improved tumor response as is also observed with IL-2. Histamine had comparable effect in treatment of liver cancer in an Isolated Hepatic Perfusion. Strikingly, combination of Histamine and IL-2 in and melphalan-based ILP abrogates each other’s effect.
    Histamine strongly affects the tumor endothelial lining resulting in massive apoptosis and endothelial detachment, which is also seen with TNF. TNF seems to work not directly on tumor endothelial cells but leukocyte produced factors such as IL-1 are necessary to inflict the observed tumor vascular effects. The early vascular effects inflicted by TNF and locally expressed cytokines are crucial for improved tumor response to concomitant chemotherapy.
    Studies on hypoxic perfusion of liver, abdomen and pelvis were performed in patients. While application of these techniques appeared feasible in pigs, with favorable pharmacokinetics, usefulness in patients was shown to be rather limited.
  • Combination chemo-immunotherapy in systemic setting.
    Previously it was shown that TNF improved systemic chemotherapy with doxorubicin-loaded liposomes. The activity of TNF was independent of tumor type or liposomal formulation used. An elegant study by Ann Seynhaeve showed that addition of TNF to systemic therapy with doxorubicin liposomes (Doxil) resulted in a more homogeneous distribution of the liposomes. A novel observation, only possible because of the in-house developed intravital window chamber, is the intracellular accumulation of the liposomes followed by doxorubicin release with consequent nuclear localization. This is contrary to current believe that the liposomes release the doxorubicin in the interstitial space after which the drug is taken up by the cells.
  • Real-time evaluation of angiogenesis, tumor vascular functionality by intravital microscopy.
    The developed intravital models are employed to study early vascular formation. With the aid of the eNOS-Tag mouse endothelial cells can be visualized and their behavior studied.
  • Endothelial cell survival under hypoxia is induced by a melanoma-derived factor.
    When cultured under hypoxia endothelial cells (EC) undergo swift apoptosis. Addition of melanoma-conditioned medium (MCM) protects the EC from apoptosis. Detailed studies reveal that the factor involved is relatively small, not heat, trypsin, nor pH sensitive. This capacity to protect EC appears to be specific for melanoma.
  • Melanoma cell migration and metastatic capacity.
    Migratory capacity of malignant melanoma cells correlate directly with their metastatic potential.
  • NICE
    Due to their ample expertise and major research activities in this area, the establishment of a nanotools facility, and the fast growing number of local, national and international collaborations LECO has taken the lead in setting up the Nanomedicine Innovation Center Erasmus MC (NICE). NICE aims to provide a platform for innovation, development, production and application of nanotools for basic, translational and clinical research. NICE will focus on novel developments and innovation in nanomedicine and serve as an open facility where various disciplines within Erasmus MC meet, collaborate and innovate. The idea for such a platform has been welcomed by many leading laboratories and clinical disciplines in Erasmus MC.

Future plans: special goals and approach

3.1 Tumor manipulation and targeting

  • Our studies with TNF to improve drug accumulation in solid tumors, both in the loco-regional perfusion systems as well as in the systemic setting initiated the exploration of other vaso-active drugs, which could mimic the activity of TNF. Next to Histamine and IL-2 also Cilengitide appears effective. This compound has no shown toxicity and is therefore tested in organ perfusion and systemic setting.
  • In addition to the use of vaso-active drugs, also hyperthermia will be used to manipulate the tumor vasculature with the aim to enhance drug delivery to solid tumors. In this a two-step approach is followed in which hyperthermia firstly enhances the extravasation of lipid-based drug carriers and secondly can trigger the release of drugs from the nanocarriers. For this detailed studies towards hyperthermia-induced extravasation of nanocarriers will be performed. Thermosensitive liposomes will be developed and studied for temperature dependent drug release in solid tumors. This new line of research has already lead to important national and international collaborations with TU Delft, TU Eindhoven, Philips, University of Munich and Duke University. 
  • Short chain sphingolipids have been discovered as new tools to enhance delivery of amphiphilic drugs across cell membranes. Current research aims to combine these newly identified lipids with liposomal drug delivery systems to further optimize drug delivery to solid tumors. This program is financed by the Dutch Cancer Society and involves a close collaboration with the Netherlands Cancer Institute.
  • Targeted receptor specific delivery of radionuclides (collaboration with Nuclear Medicine) and nanocarriers is being studied to further enhance specificity of tumor treatment. In addition these nanocarriers can and will be used for molecular imaging.

3.2 Mechanisms in Angiogenesis and Metastasis

  • Melanoma-conditioned medium protects endothelial cells from hypoxia-induced apoptosis. Effort will be made to define the factor(s) involved using biological test systems as well as genomics and proteomics.
  • The onset of angiogenesis is considered as an important moment in cancer development. With the aid of transgenic mouse models, intravital microscopy and in vitro models early angiogenesis is studied. In particular the activity of the Tip cell, leading cell in vascular sprout, is studied.
  • Metastasis is the main cause of cancer related deaths. We have found that the cells capacity to migrate strongly correlates with the metastatic potential of the tumor. At present we study intrinsic factors of melanoma cells, which affect its migratory potential.
  • During metastasis the characteristics of the tumor cells may change. We will compare biological parameters, survival, growth rate, angiogenic potential, migratory behavior of breast cancer cells isolated from the primary tumor, local metastasis and distant growth.

3.3 Translational Research

Biomarker discovery in the context of adjuvant therapy melanoma trials (Interferon, Ipilimumab, vaccines). Serum and plasma samples from these trials will be assessed in collaboration with proteomics facilities and novel antibody discovery platforms. Moreover, new melanoma candidate genes and identification of immunotherapy-response driving genes will be part of collaborations with the University of Leeds and the University of Essen and the Deutsche Krebs Forschungs Zentrum.

Most recent publications
  1. Bouwhuis MG, Suciu S, Collette S, Aamdal S, Kruit WH, Bastholt L, Stierner U, Salès F, Patel P, Punt CJ, Hernberg M, Spatz A, ten Hagen TL, Hansson J, Eggermont AM; EORTC Melanoma Group and the Nordic Melanoma Group. Autoimmune antibodies and recurrence-free interval in melanoma patients treated with adjuvant interferon. J Natl Cancer Inst. 2009 Jun 16;101(12):869-77. IF 14.9
  2. 2. ten Hagen TL, Seynhaeve AL, Eggermont AM. Tumor necrosis factor-mediated interactions between inflammatory response and tumor vascular bed. Immunol Rev. 2008 Apr;222:299-315. IF 11.8
  3. Seynhaeve AL, Hoving S, Schipper D, Vermeulen CE, de Wiel-Ambagtsheer G, van Tiel ST, Eggermont AM, Ten Hagen TL. Tumor necrosis factor alpha mediates homogeneous distribution of liposomes in murine melanoma that contributes to a better tumor response. Cancer Res. 2007 Oct 1;67(19):9455-62. IF 7.5
  4. Drabek K, van Ham M, Stepanova T, Draegestein K, van Horssen R, Sayas CL, Akhmanova A, Ten Hagen T, Smits R, Fodde R, Grosveld F, Galjart N. Role of CLASP2 in microtubule stabilization and the regulation of persistent motility. Curr Biol. 2006 Nov 21;16(22):2259-64. IF 10.8
  5. van Horssen R, Eggermont AM, ten Hagen TL. Endothelial monocyte-activating polypeptide-II and its functions in (patho)physiological processes. Cytokine Growth Factor Rev. 2006 Oct;17(5):339-48. IF 7.0
  6. Koning GA, Schiffelers RM, Wauben MH, Kok RJ, Mastrobattista E, Molema G, ten Hagen TL, Storm G. Targeting of angiogenic endothelial cells at sites of inflammation by dexamethasone phosphate-containing RGD peptide liposomes inhibits experimental arthritis. Arthritis Rheum. 2006 Apr;54(4):1198-208. IF 6.8
  7. Grünhagen DJ, de Wilt JH, ten Hagen TL, Eggermont AM. Technology insight: Utility of TNF-alpha-based isolated limb perfusion to avoid amputation of irresectable tumors of the extremities. Nat Clin Pract Oncol. 2006 Feb;3(2):94-103. IF 9.1
  8. Hoving S, Brunstein F, aan de Wiel-Ambagtsheer G, van Tiel ST, de Boeck G, de Bruijn EA, Eggermont AM, ten Hagen TL. Synergistic antitumor response of interleukin 2 with melphalan in isolated limb perfusion in soft tissue sarcoma-bearing rats. Cancer Res. 2005 May 15;65(10):4300-8. IF 7.5
  9. Brunstein F, Hoving S, Seynhaeve AL, van Tiel ST, Guetens G, de Bruijn EA, Eggermont AM, ten Hagen TL. Synergistic antitumor activity of histamine plus melphalan in isolated limb perfusion: preclinical studies. J Natl Cancer Inst. 2004 Nov 3;96(21):1603-10. IF 14.9
  10. Brouckaert P, Takahashi N, van Tiel ST, Hostens J, Eggermont AM, Seynhaeve AL, Fiers W, ten Hagen TL. Tumor necrosis factor-alpha augmented tumor response in B16BL6 melanoma-bearing mice treated with stealth liposomal doxorubicin (Doxil) correlates with altered Doxil pharmacokinetics. Int J Cancer. 2004 Apr 10;109(3):442-8. IF 4.7