Center for Tumor Diagnostics and Therapy

Osnabrück, Germany

Center for Tumor Diagnostics and Therapy

Osnabrück, Germany
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Fruehauf S.,Center for Tumor Diagnostics and Therapy | Fruehauf S.,University of Heidelberg | Ehninger G.,Carl Gustav Carus Institute | Hubel K.,University of Cologne | And 8 more authors.
Bone Marrow Transplantation | Year: 2010

This report describes the first investigational use of plerixafor in Europe and the determination of tumor cell mobilization by polymerase chain-reaction after plerixafor treatment in a subset of patients with multiple myeloma (MM). Thirty-five patients (31 MM and 4 NHL) received granulocyte colony-stimulating factor (G-CSF) (10 g/kg) each morning for 4 days. Starting the evening of Day 4, patients recieved plerixafor 0.24 mg/kg. Apheresis was initiated 10-11 h later, in the morning of Day 5. This regimen of G-CSF treatment each morning before apheresis and plerixafor treatment in the evening was repeated for up to 5 consecutive days. Mobilization with plerixafor and G-CSF resulted in a median 2.6-fold increase in peripheral blood (PB) CD34 cell count compared with before plerixafor treatment. All patients collected >2 × 10 6 CD34 cells/kg and 32 of 35 patients collected >5 × 10 6 CD34 cells/kg. After plerixafor treatment, 3 of 7 patients had a small increase and 4 of 7 patients had a small decrease in PB tumor cells. No G-CSF was given post transplant. The median number of days to polymorphonuclear leukocyte and platelet engraftment was 14.0 and 11.0, respectively. There were no reports of graft failure. Plerixafor was generally well tolerated. Mobilization of PB CD34+ cells was consistent with previous clinical trials. The addition of plerixafor did not significantly increase the relative number of PB MM tumor cells. © 2010 Macmillan Publishers Limited All rights reserved.

Heckmann D.,German Cancer Research Center | Heckmann D.,University of Heidelberg | Maier P.,University of Mannheim | Laufs S.,German Cancer Research Center | And 10 more authors.
Clinical Cancer Research | Year: 2014

Purpose: In colorectal cancer, increased expression of the CXC chemokine receptor 4 (CXCR4) has been shown to provoke metastatic disease due to the interaction with its ligand stromal cell-derived factor-1 (SDF-1). Recently, a second SDF-1 receptor, CXCR7, was found to enhance tumor growth in solid tumors. Albeit signaling cascades via SDF-1/CXCR4 have been intensively studied, the significance of the SDF-1/CXCR7-induced intracellular communication triggering malignancy is still only marginally understood. Experimental Design: In tumor tissue of 52 patients with colorectal cancer, we observed that expression of CXCR7 and CXCR4 increased with tumor stage and tumor size. Asking whether activation of CXCR4 or CXCR7 might result in a similar expression pattern, we performed microarray expression analyses using lentivirally CXCR4- and/or CXCR7-overexpressing SW480 colon cancer cell lines with and without stimulation by SDF-1α. Results: Gene regulation via SDF-1α/CXCR4 and SDF-1α/CXCR7 was completely different and partly antidromic. Differentially regulated genes were assigned by gene ontology to migration, proliferation, and lipid metabolic processes. Expressions of AKR1C3, AXL, C5, IGFBP7, IL24, RRAS, and TNNC1 were confirmed by quantitative real-time PCR. Using the in silico gene set enrichment analysis, we showed that expressions of miR-217 and miR-218 were increased in CXCR4 and reduced in CXCR7 cells after stimulation with SDF-1α. Functionally, exposure to SDF-1α increased invasiveness of CXCR4 and CXCR7 cells, AXL knockdown hampered invasion. Compared with controls, CXCR4 cells showed increased sensitivity against 5-FU, whereas CXCR7 cells were more chemoresistant. Conclusions: These opposing results for CXCR4- or CXCR7-overexpressing colon carcinoma cells demand an unexpected attention in the clinical application of chemokine receptor antagonists such as plerixafor. © 2014 American Association for Cancer Research.

DiPersio J.F.,Washington University in St. Louis | Ho A.D.,University of Heidelberg | Hanrahan J.,Genzyme | Hsu F.J.,Genzyme | Fruehauf S.,Center for Tumor Diagnostics and Therapy
Biology of Blood and Marrow Transplantation | Year: 2011

Autologous transplantation of peripheral blood (PB) hematopoietic stem cells (HSCs) is a widely used strategy for reconstitution of blood cells following high-dose chemotherapy for hematologic malignancies such as multiple myeloma (MM), non-Hodgkin lymphoma (NHL), and acute myeloid leukemia (AML), among others. Stem cells for transplantation are usually obtained from PB after treatment with chemotherapy with or without cytokine, usually granulocyte-colony stimulating factor (G-CSF), or after treatment with cytokine alone. The use of autologous peripheral blood stem cells (PBSCs) for transplantation is associated with the risk of contamination of the graft with tumor cells; whether this impacts response rates, progression-free survival (PFS), and overall survival (OS) is still debatable. This review summarizes the controversy surrounding tumor cell mobilization (TCM), the complexity of detection of minimal residual diseases, the available diagnostic tools, differences in TCM with available mobilization regimens, and the potential effect of TCM on clinical outcome. Collectively, these data suggest that new treatment paradigms to manage hematologic malignancies, such as MM, NHL, and AML, are needed and should focus on increasing the chemosensitivity of the tumor and eliminating residual disease. © 2011.

Heckmann D.,University of Heidelberg | Laufs S.,University of Heidelberg | Maier P.,University of Heidelberg | Zucknick M.,German Cancer Research Center | And 8 more authors.
Onkologie | Year: 2011

Background: The development of distant metastasis is associated with poor outcome in patients with colorectal cancer (CRC). The stromal cell-derived factor-1 (SDF-1) and its receptor CXC chemokine receptor 4 (CXCR4) have pivotal roles in the chemotaxis of migrating tumor cells during metastasis. Thus, hampering the SDF-1/CXCR4 cross-talk is a promising strategy to suppress metastasis. Methods: We investigated the invasive behavior of the lentivirally CXCR4overexpressing CRC cell lines SW480, SW620 and RKO in chemotaxis and invasion assays toward an SDF-1α gradient. Low endogenous CXCR4 expression levels were determined by quantitative realtime polymerase chain reaction (PCR) and fluorescence-activated cell sorting (FACS) analyses. Results: A lentiviral CXCR4 overexpression and knockdown model was established in these CRC cells. In transwell migration assays, CXCR4 overexpression favored chemotaxis and invasion of cells in all 3 lines depending on an SDF-1α gradient (p < 0.001 vs. untransduced cells). Functional CXCR4 knockdown using lentiviral short hairpin RNA (shRNA) vectors significantly decreased the migration behavior in CRC cell lines (p < 0.001), confirming a CXCR4-specific effect. Pharmacologic inhibition of the SDF-1α/CXCR4 interaction by the bicyclam Plerixafor TM at 100 μM significantly abrogated CXCR4-dependent migration and invasion through Matrigel TM (SW480, SW620, RKO; p < 0.05). Conclusion: Our results indicate that a CXCR4-antagonistic therapy might prevent tumor cell dissemination and metastasis in CRC patients, consequently improving survival. © 2011 S. Karger AG, Basel.

Jeltsch K.S.,German Cancer Research Center | Radke T.F.,Cell Therapeutics | Laufs S.,German Cancer Research Center | Giordano F.A.,German Cancer Research Center | And 8 more authors.
Cytotherapy | Year: 2011

Background aims: Transplantation of allogeneic hematopoietic stem cells (HSC) within the framework of hematologic oncology or inherited diseases may be associated with complications such as engraftment failure and long-term pancytopenia. HSC engraftment can be improved, for example by co-transplantation with mesenchymal stem cells (MSC). Recently, a new multipotent MSC line from umbilical cord blood, unrestricted somatic stem cells (USSC), has been described. It was demonstrated that USSC significantly support proliferation of HSC in an in vitro feeder layer assay. Methods. A NOD/SCID mouse model was used to assess the effect of USSC on co-transplanted CD34+ cells and look for the fate of transplanted USSC. The migration potential of USSC was studied in a Boyden chamber migration assay and in vivo. Quantitative real-time polymerase chain reaction (qRT-PCR) for CXCR4, CD44, LFA1, CD62L, VLA4, RAC2, VLA5A and RAC1 were performed. NMR1 nu/nu mice were used for a tumorigenicity test. Results. After 4 weeks, homing of human cells (CD45+) to the bone marrow of NOD/SCID mice was significantly increased in mice co-transplanted with CD34+ cells and USSC (median 30.9%, range 7-50%) compared with the CD34+ cell-only control group (median 5.9%, range 3-10%; P = 0.004). Homing of USSC could not be shown in the bone marrow. A cell-cell contact was not required for the graft enhancing effect of USSC. An in vivo tumorigenicity assay showed no tumorigenic potential of USSC. Conclusions. This pre-clinical study clearly shows that USSC have an enhancing effect on engraftment of human CD34+ cells. USSC are a safe graft adjunct. © 2011 Informa Healthcare.

Maier P.,German Cancer Research Center | Maier P.,University of Heidelberg | Spier I.,German Cancer Research Center | Laufs S.,German Cancer Research Center | And 5 more authors.
Gene Therapy | Year: 2010

Myelotoxicity is a dose-limiting effect of many chemotherapeutic regimens. Thus, there is great interest in protecting human hematopoietic stem cells by the transfer of drug resistance genes. The main focus of this study was the simultaneous overexpression of multidrug resistance 1 (MDR1) and the O 6-benzylguanine (O 6-BG)-resistant mutant MGMT P140K (O 6-methylguanine-DNA methyltransferase) with a bicistronic lentiviral vector (HR′SIN-MDR1-IRES- MGMT P140K), with regard to the capability to convey chemoprotection in the leukemia cell line, HL60, and human hematopoietic stem cells (CD34). Combination therapy with O 6-BG/1-(2-chloroethyl)-3-(4-amino-2-methylpyrimidine-5-yl) methyl-1-nitrosourea) (ACNU) plus paclitaxel showed a significant survival advantage of HL60 cells transduced with this combination vector. In CD34 cells, monotherapy with O 6-BG/temozolomide (TMZ) resulted in an increased percentage of MGMT-positive cells (vs untreated cells) after transduction with HR′SIN-MDR1-IRES-MGMT P140K (28.3%). For combination therapy with O 6-BG/temozolomide plus paclitaxel the increase was higher with the combination vector (52.8%) than with a vector expressing MGMT P140K solely (29.1%). With regard to MDR1-positive cells the protective effect of the combination vector (88.5%) was comparable to the single vector HR′SIN-MDR1 (90.0%) for monotherapy with paclitaxel and superior for combination therapy with O 6-BG/temozolomide plus paclitaxel (84.6 vs 69.7%). In conclusion, the combination vector presents simultaneous protective effects of two drug-resistance genes, offering an opportunity to increase the cancer therapeutic index. © 2010 Macmillan Publishers Limited All rights reserved.

Heckmann D.,German Cancer Research Center | Heckmann D.,University of Heidelberg | Maier P.,University of Heidelberg | Laufs S.,German Cancer Research Center | And 5 more authors.
Translational Oncology | Year: 2013

BACKGROUND: Signaling through stromal cell-derived factor-1α (SDF-1α), strongly secreted by bone marrow stromal cells and the CXC chemokine receptor 4 (CXCR4) exposed on tumor cells has pivotal roles in proliferation, metastasis, and tumor cell "dormancy." Dormancy is associated with cytostatic drug resistance and is probably a property of tumor stem cells andminimal residual disease. Thus, hampering the SDF-1α/CXCR4 cross talk by a CXCR4 antagonist like Plerixafor (AMD3100) should overcome tumor cell dormancy bymobilization of tumor cells from "sanctuary" niches.Our aimwas to elucidate the direct effects exerted by SDF-1α and Plerixafor on proliferation, chemosensitivity, and apoptosis of CXCR4- expressing tumor cells. METHODS: The ability of SDF-1α and Plerixafor to regulate intracellular signaling, proliferation, and invasion was investigated using two colon cancer cell lines (HT-29 and SW480) with either high endogenous or lentiviral expression of CXCR4 compared to their respective low CXCR4-expressing counterparts as a model system. Efficacy of Plerixafor on sensitivity of these cell lines against 5-fluorouracil, irinotecan, or oxaliplatin was determined in a cell viability assay as well as stroma-dependent cytotoxicity and apoptosis assays. RESULTS: SDF-1α increased proliferation, invasion, and ERK signaling of endogenously and lentivirally CXCR4-expressing cells. Exposure to Plerixafor reduced proliferation, invasion, and extracellular signal-regulated kinase 1/2 (ERK1/2) signaling. Combination of chemotherapy with Plerixafor showed an additive effect on chemosensitivity and apoptosis in CXCR4-overexpressing cells. An SDF-1-secreting feeder layer provideda"protective niche" forCXCR4-overexpressing cells resulting in decreased chemosensitivity.CONCLUSION: CXCR4-antagonistic therapy mobilizes and additionally sensitizes tumor cells toward cytoreductive chemotherapy. © 2013 Neoplasia Press, Inc. All rights reserved.

Fruehauf S.,Center for Tumor Diagnostics and Therapy | Tricot G.,University of Utah
Biology of Blood and Marrow Transplantation | Year: 2010

Autologous and allogeneic hematopoietic stem cell transplantation (HSCT) are considered the standard of care for many malignancies, including lymphoma, myeloma, and some leukemias. In many cases, mobilized peripheral blood has become the preferred source of hematopoietic stem cells. The efficacy of different mobilization regimens and transplantation outcomes based on cell doses has been well studied; however, the characteristics of the stem cell graft may be of equal importance with respect to patient outcomes following autologous or allogeneic HSCT. This review summarizes available preclinical and clinical data for bone marrow and mobilized peripheral blood HSCT characteristics, defined as the cell types found in the graft as well as their gene expression profiles. It also explores how graft characteristics can affect bone marrow homing, engraftment, immune reconstitution, and other posttransplantation outcomes in both the allogeneic and autologous HSCT settings. © 2010 American Society for Blood and Marrow Transplantation.

Jantunen E.,Kuopio University Hospital | Fruehauf S.,Center for Tumor Diagnostics and Therapy
Bone Marrow Transplantation | Year: 2011

Although blood stem cells have been widely used to support high-dose therapy in the autologous setting, limited data are available on the effects of graft characteristics in patient outcomes other than haematopoietic engraftment. Retrospective studies suggest that patients who mobilize more CD34+ cells have better outcomes than do patients who mobilize less well. Furthermore, immunological reconstitution may be important in terms of post-transplant outcome and is apparently affected by graft composition. There is accumulating evidence that the mobilization regimen used may be an important determinant of graft content. Plerixafor has been recently introduced combined with G-CSF in patients who mobilize poorly. In addition to enhancing mobilization of CD34 + cells, there are indications that plerixafor may also affect other graft components. A combination of chemotherapy plus G-CSF with plerixafor has been shown to be very effective in stem-cell mobilization, but more data are needed in regard to other graft characteristics in this setting. Prospective studies are needed to evaluate whether higher CD34+ doses or other modifications to graft composition translate into better long-term outcomes in the autologous setting. These studies are not only important in regard to defining the optimal stem-cell graft in the autologous setting, but also in identifying the optimal mobilization regimen. © 2011 Macmillan Publishers Limited All rights reserved.

Kamioner D.,AFSOS and Hopital Prive de lOuest Parisien | Fruehauf S.,Center for Tumor Diagnostics and Therapy | Maloisel F.,Clinique Saint Anne | Cals L.,CHRU de Besancon | Lepretre S.,Center Henri Becquerel
BMC Cancer | Year: 2013

Background: Nivestim™ (filgrastim) is a follow-on biologic agent licensed in the EU for the treatment of neutropenia and febrile neutropenia induced by myelosuppressive chemotherapy. Nivestim™ has been studied in phase 2 and 3 clinical trials where its efficacy and safety was found to be similar to its reference product, Neupogen®. Follow-on biologics continue to be scrutinised for safety. We present a design for two observational phase IV studies that are evaluating the safety profile of Nivestim™ for the prevention and treatment of febrile neutropenia (FN) in patients treated with cytotoxic chemotherapy in general clinical practice.Methods/Design: The NEXT (Tolérance de Nivestim chez les patiEnts traités par une chimiothérapie anticancéreuse cytotoXique en praTique courante) and VENICE (VErträglichkeit von NIvestim unter zytotoxischer Chemotherapie in der Behandlung malinger Erkrankungen) trials are multicentre, prospective, longitudinal, observational studies evaluating the safety profile of Nivestim™ in 'real-world' clinical practice. Inclusion criteria include patients undergoing cytotoxic chemotherapy for malignancy and receiving Nivestim as primary or secondary prophylaxis (NEXT and VENICE), or as treatment for ongoing FN (NEXT only). In accordance with European Union pharmacovigilance guidelines, the primary objective is to evaluate the safety of Nivestim™ by gathering data on adverse events in all system organ classes. Secondary objectives include obtaining information on patient characteristics, efficacy of Nivestim™ therapy (including chemotherapy dose intensity), patterns of use of Nivestim™, and physician knowledge regarding filgrastim prescription and the reasons for choosing Nivestim™. Data will be gathered at three visits: 1. At the initial inclusion visit, 2. At a 1-month follow-up visit, and 3. At the end of chemotherapy. Recruitment for VENICE commenced in July 2011 and in November 2011 for NEXT. VENICE completed recruitment in July 2013 with 407 patients, and NEXT in September 2013 with 2123 patients. Last patient, last visit for each study will be December 2013 and March 2014 respectively.Discussion: The NEXT and VENICE studies will provide long-term safety, efficacy and practice pattern data in patients receiving Nivestim™ to support myelosuppressive chemotherapy in real world clinical practice. These data will improve our understanding of the performance of Nivestim™ in patients encountered in the general patient population. Trial registration: NEXT NCT01574235, VENICE NCT01627990. © 2013 Kamioner et al.; licensee BioMed Central Ltd.

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