Dendritic Cell Immunotherapy

Dendritic cells (DC) are the most potent antigen-presenting cells of the immune system specialized in initiating primary T cell responses against pathogens and tumors. In vitro and murine in vivo data suggest that immature DC are primarily involved in antigen uptake, while the stimulation of T cell-mediated immunity is regulated by mature DC. These findings have stimulated several investigators to exploit tumor antigen-loaded DC as an anti-cancer vaccine. The studies so far appeared to be safe and effective. Sanquin collaborates with the group of Prof Carl Figdor and Prof Jolanda de Vries of the department of Tumor Immunology of the UMC St Radboud on the implementation of a  protocol for clinical grade blood derived DCs for the treatment of stage IIIb and IIIc melanoma. This trial will start at the end of 2016 and will include 178 patients, 119 in the DC group and 59 in the control cohort.

Adoptive T cell therapy

The LCT actively participates in a clinical trial initiated at the AvL/NKI by Prof dr T Schumacher and Prof dr J. Haanen. This trial on adoptive therapy consists of culturing melanoma-reactive T cells from resected metatastasis ex vivo and administering them to patients that receive high dose chemotherapy and IL-2 bolus. During the second half of 2013 and beginning of 2014 the protocol was implemented and improved at the LCT, initially the tumor infiltrating lymphocytes were expanded on an autologous feeder layer harvested by leukapheresis. During the startup phase of the trial it appeared that often not enough cells could be harvested by leukapherisis. Therefore the protocol was adjusted including the use of pooled buffycoats of healthy donors as feeder layer during the expansion phase.  In December 2014 the first TIL product was generated within the LCT and successfully administered to the patient, and 10 more products are expected for 2016/2017.  In addition the LCT is involved in the clinical trial of Novartis for the application of CAR-T cells to patients in the AMC. In this trial the LCT participates in the cryopreservation of autologous T-cell products and the shipping to the gene-therapy facility of Novartis in the US. After generation of the CAR-T products the ATMPS is stored at the AMC  and bed-side administered in the AMC.

Expansion of bone-marrow derived MSC s to treat acute graft versus host disease
Mesenchymal stromal cells (MSC) are a heterogenous population of multi-potent cells that have the ability to migrate to sites of inflammation and injury and display immunomodulatory capacities. Clinical applications of MSC include treatment of therapy resistant acute graft versus host disease (GvHD), rejection after hematopoietic stem cell or solid organ transplantation and refractory Crohn’s disease. A clinical grade MSC expansion protocol has been developed to generate a standardized bone marrow-derived MSC product from third-party donors. Recently the LCT became the second production facility for the European Hovon 113 trial (coordinated by the LUMC in Leiden) in which patients with acute GVHD grade 2-4 and refractory on steroid treatment will receive 2 portions of MSC (1.5-2x10*6 MSC/kg) or placebo and standard therapy (75 patients per arm). In December 2014 the first MSC for clinical application were generated in the LCT. Currently this trial is experiencing a delay in its inclusion, so at the moment no MSC products are generated. 

Novel Therapies

Novel therapies that are being developed in close collaboration with the LCT. The LCT promotes collaboration in an early phase to avoid large adaptations to GMP conditions in a late stage that may alter the characteristics of the cellular products. Pre-clinical research is performed by several groups within Sanquin: the group of Emile van den Akker/Marieke von Lindern within the department of Hematopoiesis that focuses on the ex vivo generation of red blood cells for transfusion purposes (ZONMW TAS grant) as well as by the Wolkers group that mainly studies the translational regulation of effector function of T cells. In analogy of the application of TIL for melanoma patients, research in collaboration with the AvL/NKI is aimed to investigate whether TIL generation for the treatment of prostate and lung cancer is feasible. 

Basic research underlying translational approaches

The Laboratory for Cell therapy became part of the department of Hematopoiesis (HEP) (headed by Dr M. von Lindern) which focuses on molecular and cellular aspects of hematopoiesis. In recent years her group published numerous articles in well-cited peer-reviewed international journals and her lab has extensive expertise in the development of new blood-derived cellular products. Her research group consists of 1 postdoc, 4PhD students, 2 technicians and 1 vacancy for a postdoc (interviews pending)

MSC-related research

Since MSC are a potential cell source for cellular therapies, this is also one of the main topics within the group of Dr C. Voermans. The therapeutic use of MSC is hampered by the lack of knowledge on MSC identity and function. Based on expression of the new MSC markers CD271 and CD146, we identified three new subpopulations within the classical MSC population in the bone marrow (BM). (Maijenburg et al., Haematologica 2012). Therefore we study the immunomodulatory properties and hematopoietic support of conventional MSC and our newly defined MSC subsets and our results indicate that the CD271+ subset display a higher and long term hematopoietic support compared to the conventional culture expanded MSC (manuscript submitted). Furthermore, To optimize expansion and function of MSC in general, we explored the differences between Adult BMSC and Fetal BMSC. Gene expression profiling implicated differential expression of genes encoding proteins in the Wnt signaling pathway, including excreted inhibitors of Wnt signaling, particularly by ABMSC. While FBMSC produced factors, including Wnt signals, that enhanced MSC proliferation, ABMSC produced Wnt factors in a setting that enhanced hematopoietic support.(paciejewska et al 2016) Thus, further unraveling the molecular basis of this phenomenon may lead to improvement of clinical expansion protocols of MSCs. Other aspects currently being studied are the roles of MSC-derived vesicles on the hematopoietic support and to unravel the MSC compartment of neuroblastoma patients after chemo- and radiotherapy as they display a delayed hematopoietic recovery to other patient groups after stem cell transplantation.

HSPC function in regenerative bone marrow

A long lasting interest of Dr. Voermans is bone marrow regeneration and improved HSC function following stem cell transplantation. Cell adhesion and migration are important determinants of homing and development of hematopoietic stem and progenitor cells (HSPCs) in bone marrow niches (BM). The extracellular matrix protein Transforming growth factor-beta (TGF-beta) inducible gene H3 (BIGH3) is involved in adhesion and migration, although the effect of BIGH3 is highly cell type dependent. BIGH3 is abundantly expressed by mesenchymal stromal cells, while its expression in HSPCs is relatively low unless induced by certain BM stressors. We established that BIGH3 overexpression in HSPCs decreased adhesion and migration and downstream RAC1 and ERK activation The upregulation of BIGH3 may help loosen the adhesive contacts of the hematopoietic cells with their originating niche and may make them susceptible for polarization and subsequent egress. Therefore, upregulation of BIGH3 in regenerative BM might contribute to HSPC homeostasis, through decreased adhesion and an altered GTPase balance of HSPCs, which may regulate homing and mobilization of HSPCs.(Klamer et al 2013, 2014 and 2016). This is now further studied together with the groups of M Nolte,  and JA van Buul within Sanquin and S. Huveneers from AMC within the program Joined forces: understanding and rebuilding the vascular bone marrow niche.

Graft versus host disease

Together with Sacha Zeerleder, MD, PhD haematologist in AMC we initiated a research line on Graft versus host disease as it is one of the main limitations of allogeneic HSCT which is associated with high morbidity and mortality. Up to now it is still not possible to predict the onset and the severity of GvHD in the individual patients undergoing allogeneic HSCT. The therapy-induced tissue damage may lead to the release of damage-associated molecular patterns (DAMPs) which can then lead to alloreactive immune responses by activating T and B cell responses that in turn lead to GvHD.. In this project are developing tools to identify patients at high risk to develop severe GvHD at time points before and early after allogeneic SCT. The main goals of this project are to demonstrate that the release of DAMPs induced by the conditioning therapy in humans and using mouse models and to evaluate therapeutic strategies to attenuate systemic inflammation induced by the conditioning procedure by blocking the release or the signalling of DAMPs.

Key publications

• Klamer SE, Nota B, Moorhouse M, Voermans C, Schoot CE. Gene-expression-based monocyte-specific clustering of acute myeloid leukemias reveals novel associations. Leuk Lymphoma. 2017 Jul;58(7):1721-1725
• van der Valk FM, Kuijk C, Verweij SL, Stiekema LC, Kaiser Y, Zeerleder S, Nahrendorf M, Voermans C, Stroes ES. Increased haematopoietic activity in patients with atherosclerosis.  Eur Heart J. 2017 Feb 7;38(6):425-432
• Munneke JM, Spruit MJ, Cornelissen AS, van Hoeven V, Voermans C, Hazenberg MD. The Potential of Mesenchymal Stromal Cells as Treatment for Severe Steroid-Refractory Acute Graft-Versus-Host Disease: A Critical Review of the Literature. Transplantation. 2015 Dec 28.
• Cornelissen AS, Maijenburg MW, Nolte MA, Voermans C. Organ-specific migration of mesenchymal stromal cells: Who, when, where and why? Immunol Lett. 2015 Dec;168(2):159-69. doi: 10.1016/j.imlet.2015.06.019.