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Bonde,Schwendener, Mete, Kumar

MODULATION OF THE TUMOR MICROENVIRONMENT BY BISPHOSPHONATES: BASIC BIOLOGICAL AND IMMUNOLOGICAL PROPERTIES AND EVALUATION AS A NOVEL APPROACH TO CANCER THERAPY

Background

Modulation of solid tumors with bisphosphonates and bisphosphonate-liposomes leads to growth inhibition and re-polarization to a growth-suppressing tumor microenvironment. We study the potential of this method by two ways:1) as a tool to study basic biological and immunological effects in the tumor microenvironment and 2) as cancer therapy approach in mouse tumor models. Our aim is to contribute to a better understanding of the complex processes of tumor development, growth and metastatic dissemination.

Introduction

Solid tumors are not only composed of malignant cells; they are complex organ-like structures comprising many cell types, including a wide variety of migratory hematopoietic (macrophages, neutrophils, myeloid-derived suppressor cells, immune cells) and resident stromal cells. Migration of these cell types into tumors has been interpreted as evidence for an immunological response of the host against a growing tumor. It is acknowledged that tumors are largely recognized as "self" and lack strong antigens. Instead, they have the property to manipulate the host immune system to prevent rejection and to facilitate their own growth and spread. This led to the proposal that hematopoietic cell infiltrates have a causal role in carcinogenesis. Clinical data collected from a wide range of solid tumors underscore these results, given that those high densities of leukocytic infiltrations, most notably tumor associated macrophages (TAM) and neutrophils (TAN), correlate with poor prognosis of the diseases.

TAMs are derived from circulating monocytes and are activated macrophages of the polarized type II (M2 macrophages), mainly induced by IL-4, IL-10, IL-13 and corticosteorids. Differential cytokine and chemokine production and coordinated temporal and spatial activities of these cells in the tumor stroma are key features of polarized macrophages that promote tumor angiogenesis and growth. Due to their tumorigenic role, M2-TAMs have been proposed as potential therapeutic targets. Similarly, TANs have been shown assume comparable polarized phenotypes that suppress anti-tumorigenic immune cells in the tumor microenvironment. To study the role of TAMs and TANs we use bisphosphonates and bisphosphonate-liposomes in various subcutaneous mouse tumor models. Tumor growth is monitored by immunohistochemistry, flow cytometry, microscopy and by genomic and proteomic approaches. Additionally, we use in vitro conditioned-medium co-culture models to identify emerging proteins translated in cancer and endothelial cells in response to macrophages.

Most conventional tumor therapies are flawed due to the genetic instability of cancer cells, which leads to drug resistance. Since macrophages and neutrophils are shown to be involved in assisting tumor properties e.g., polarization, invasion and angiogenesis, it is important to reveal the molecular mechanisms responsible for these events. Moreover, it is essential to identify new drug targets in signaling pathways in these cells and apply their inhibitors in liposomal formulations to specifically target myeloid cell types. To achieve this, we have established co-culture models of tumor cells and macrophages, which are used to screen inhibitors of inflammatory signaling pathways. We also exploit our co-culture models to identify nascent proteins translated in cancer and endothelial cells in response to macrophages. This project will shed light on the understanding of the role of the tumor microenvironment and for the development of new therapies.

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Figure 1. Recruitment of bone marrow-derived cells such as monocytes/ macrophages and neutrophils to the tumor microenvironment. Tumor and stromal cells mobilize tumor promoting myeloid cells to the peripheral blood through secretion of cytokines and chemokines followed by the mobilization and infiltration of these cells to the tumor microenvironment, where they assume tumorigenic properties. Bisphosphonate-liposomes inhibit tumor growth, either by depletion of M2-polarized TAMs or by causing neutrophil infiltration and skewing of N2-neutrophils to anti-tumorigenic N1-neutrophils, thereby creating an anti-tumorigenic microenvironment (Modified from Schmid & Varner, J. Oncol. 2010).

Tumor-associated macrophages regulate tumor cell malignancy by induction of epithelial to mesenchymal transition

Macrophages are important components of the tumor microenvironment and their cancer-promoting properties are widely acknowledged. Besides regulating the “angiogenic switch” and remodeling the extracellular matrix, a number of studies have suggested that macrophages orchestrate the migration and invasion of epithelial tumor cells. Epithelial-mesenchymal transition (EMT) is a well-characterized cellular process, through which cells down-regulate epithelial adherence molecules and acquire motile and invasive properties. In this project we are addressing the potential involvement of TAMs in the regulation of an EMT-associated phenotypic shift in tumor cells. We have used liposome-encapsulated clodronate to deplete macrophages in a murine F9-teratocarcinoma model. Gene expression analysis indicated a reduction in mesenchymal gene expression in macrophage depleted tumors. Our data suggest that macrophages can contribute to the regulation of an EMT-associated phenotypic shift in tumor cells. Using conditioned medium culturing we identified macrophage-derived TGF-β as the main regulator of the mesenchymal phenotype in F9-cells and mammary gland NMuMG-cells. Moreover, macrophage conditioned medium, as well as recombinant TGF-β, stimulated the invasive properties of the cells. The clinical relevance of our findings was addressed in a cohort of 491 non-small cell lung cancer patients by an immunohistochemical analysis. This study confirmed a significant correlation between CD68+ macrophage density, a pronounced mesenchymal tumor cell profile and tumor grade. In conclusion, this project has identified a regulatory role for TAMs in EMT-associated phenotypic shift of tumor cells.

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Figure 2. Immunohistochemical analysis identifies correlations between tumor cell expression of EMT markers and intra-tumoral CD68+ macrophage density. Whereas E-cadherin (red, left panel) and β-catenin (red, middle panel) localize to the plasma membrane of tumor cells in areas with low CD68+ macrophage (green) densities (upper row), expression of both proteins is compromised and partially lost in areas with high CD68+ densities (lower row). Conversely, fibronectin (red, right panel) expression is increased in areas with high CD68+ densities. Thus, intra-tumoral TAM density correlates with a mesenchymal tumor cell phenotype in F9-tumors. The nuclei are stained with DAPI (blue). Scale bar=0.02 mm.

Zoledronate-mediated modulation of the tumor microenvironment leads to impaired tumor growth

Zoledronate, an inhibitor of osteoclastic bone resorption, is commonly used to prevent and treat osteoporosis. There is emerging interest in the use of zoledronate as an anticancer agent based on preclinical evidence of its anti-tumor properties. Due to its high affinity for bone matrix, most models addressed the ability of zoledronate to reduce skeletal tumor burden and prevent bone metastases. However, whether zoledronate prevents tumor progression in soft tissue tumors and the mechanism of its antitumor effects is still under investigation.

To address these issues, we treated mice bearing syngeneic subcutaneous tumors with zoledronate and zoledronate-liposomes. A significant reduction in growth of Lewis lung (LLC) and colon carcinoma (MC38), but not B16 melanoma, tumors in mice was observed. We examined the effect of the drug on the tumor microenvironment focusing on tumor infiltrating myeloid cells. We saw an increase of CD11b+ myeloid cells in the tumor microenvironment as well as in spleen, blood and peritoneum of treated animals. It is known that solid tumors actively recruit myeloid cells and divert their functions toward an immune-suppressive and pro-tumorigenic M2-like phenotype. The inverse correlation between myeloid cell density and tumor growth in zoledronate-treated animals points to a  reprogramming of these cells: myeloid cells from treated tumors were found to acquire an M1 anti-tumorigenic phenotype, as shown by increased expression of pro-inflammatory and immunostimulatory and reduced expression of the immunosuppressive factors. Furthermore, these cells displayed an enhanced ability to stimulate proliferation of naive CD8+ T cells. Further characterization of these cells identified the neutrophils as increasingly accumulating myeloid cell types in tumors of zoledronate-treated animals. Accordingly, zoledronate was found to increase the production of neutrophil-attracting chemokines by cancer cells as well as tumor infiltrating myeloid cells. Further analysis of CD11b+Ly6G+ neutrophils and CD11b+Ly6G- monocytes/macrophages revealed that zoledronate exerts inflammatory and immunogenic transcriptional changes specifically in neutrophils, but not in macrophages. Another key finding was that recombinant TGF-β administration reduced therapeutic efficacy of zoledronate by reducing neutrophil infiltration. To improve the antitumor efficacy of zoledronate, we encapsulated the drug into liposomes, which significantly improved the antitumor efficacy of zoledronate by altering its pharmacokinetics and biodistribution profiles. Collectively, our findings reveal novel anti-tumorigenic properties of zoledronate that may assist in the design of more effective immunotherapeutic approaches for cancer.

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Figure 3. Model of neutrophil re-polarization induced by zoledronate. Upon in vivo administration, zoledronate preferentially accumulates in bone where it inhibits tumor-induced osteoclast activity and bone resorption. Thereby, zoledronate prevents the release of bone matrix-embedded growth factors (e.g. TGF-β, IGF-1, BMPs) that, induce enhanced expression of neutrophil-attracting chemokines, and thus cause increased accumulation of neutrophils in tumors. Zoledronate skews neutrophil polarization away from the pro-tumorigenic and immunosuppressive N2-like phenotype that is known to be regulated by TGF-β. Accordingly, TGF-β administration reverses the anti-tumor effects. Free zoledronate primarily accumulates in bones while liposomal encapsulation of the drug improves its bioavailability in extraskeletal tumor sites that leads to stronger inhibition of tumor growth.

Liposomes as drug carriers

Our laboratory has a continuous interest in liposome technology and development of liposomal formulations with antitumor and antiviral drugs, peptide vaccines, macrophage depletion and lipophilic contrast agents. For a comprehensive overview of our activities including recent publications we recommend a visit to the website of the Laboratory of Liposome Research.

Publications

da Costa, Maria Helena Bueno; Sant'anna, Osvaldo A; Quintilio, Wagner; Schwendener, Reto Albert; de Araujo, Pedro Soares (2012). A Rational Design for the Nanoencapsulation of Poisonous Animal Venoms in Liposomes Prepared with Natural Phospholipids. Current Drug Delivery, 9(6):637-644.

Warchol, Mark E; Schwendener, Reto A; Hirose, Keiko (2012). Depletion of resident macrophages does not alter sensory regeneration in the avian cochlea. PLoS ONE, 7(12):e51574.

Linkermann, Andreas; Bräsen, Jan H; De Zen, Federica; Weinlich, Ricardo; Schwendener, Reto A; Green, Douglas R; Kunzendorf, Ulrich; Krautwald, Stefan (2012). Dichotomy between RIP1- and RIP3-mediated necroptosis in tumor necrosis factor-α-induced shock. Molecular Medicine, 18(1):577-586.

Panigrahy, D; Edin, M L; Lee, C R; Huang, S; Bielenberg, D R; Butterfield, C E; Barnés, C M; Mammoto, A; Mammoto, T; Luria, A; Benny, O; Chaponis, D M; Dudley, A C; Greene, E R; Vergilio, J A; Pietramaggiori, G; Scherer-Pietramaggiori, S S; Short, S M; Seth, M; Lih, F B; Tomer, K B; Yang, J; Schwendener, R A; Hammock, BD; Falck, J R; Manthati, V L; Ingber, D E; Kaipainen, A; D'Amore, P A; Kieran, M W; Zeldin, D C (2012). Epoxyeicosanoids stimulate multiorgan metastasis and tumor dormancy escape in mice. Journal of Clinical Investigation, 122(1):178-191.

Zhang, Yi; Zhang, Ruihua; Zhang, Huafeng; Liu, Jing; Yang, Zhuoshun; Xu, Pingwei; Cai, Wenqian; Lu, Geming; Cui, Miao; Schwendener, Reto A; Shi, Huang-Zhong; Xiong, Huabao; Huang, Bo (2012). Microparticles released by Listeria monocytogenes-infected macrophages are required for dendritic cell-elicited protective immunity. Cellular & Molecular Immunology, 9(6):489-496.

Nasser, M W; Qamri, Z; Deol, Y S; Ravi, J; Powell, C A; Trikha, P; Schwendener, R A; Bai, X F; Shilo, K; Zou, X; Leone, G; Wolf, R; Yuspa, S H; Ganju, R K (2012). S100A7 Enhances Mammary Tumorigenesis through Upregulation of Inflammatory Pathways. Cancer Research, 72(3):604-615.

Vuarchey, C; Kumar, S; Schwendener, R A (2011). Albumin coated liposomes: a novel platform for macrophage specific drug delivery. Nanotechnology Development, 1(1):e2.

Zattoni, M; Mura, M L; Deprez, F; Schwendener, R A; Engelhardt, B; Frei, K; Fritschy, J M (2011). Brain infiltration of leukocytes contributes to the pathophysiology of temporal lobe epilepsy. Journal of Neuroscience, 31(11):4037-4050.

Bijnsdorp, I V; Schwendener, R A; Schott, H; Fichtner, I; Smid, K; Laan, A C; Schott, S; Losekoot, N; Honeywell, R J; Peters, G J (2011). Cellular pharmacology of multi- and duplex drugsconsisting of ethynylcytidine and 5-fluoro-2'-deoxyuridine. Investigational New Drugs, 29(2):248-257.

Schott, H; Goltz, D; Schott, T C; Jauch, C; Schwendener, R A (2011). N(4)-[Alkyl-(hydroxyphosphono)phosphonate]-cytidine-new drugs covalently linking antimetabolites (5-FdU, araU or AZT) with bone-targeting bisphosphonates (alendronate or pamidronate). Bioorganic & Medicinal Chemistry, 19(11):3520-3526.

Egilmez, N K; Harden, J L; Virtuoso, L P; Schwendener, R A; Kilinc, M O (2011). Nitric oxide short-circuits interleukin-12-mediated tumor regression. Cancer Immunology, Immunotherapy, 60(6):839-845.

Yang, H; Kim, C; Kim, M J; Schwendener, R A; Alitalo, K; Heston, W; Kim, I; Kim, W J; Koh, G Y (2011). Soluble vascular endothelial growth factor receptor-3 suppresses lymphangiogenesis and lymphatic metastasis in bladder cancer. Molecular Cancer, 10:36.

Treiger Borema, S E; Schwendener, R A; Osso, J A; de Andrade , H F; Nascimento, N (2011). Uptake and antileishmanial activity of meglumine antimoniate-containing liposomes in Leishmania (Leishmania) major-infected macrophages. International Journal of Antimicrobial Agents, 38(4):341-347.

Schwendener, R A; Schott, H (2010). Liposome formulations of hydrophobic drugs. In: Weissig, V. Liposomes, Methods and Protocols, Vol. 1: Pharmaceutical Nanocarriers. New York, NY, USA, 129-138. ISBN 978-1-60327-359-6.

Schwendener, R A; Ludewig, B; Cerny, A; Engler, O (2010). Liposome-based vaccines. In: Weissig, V. Liposomes, Methods and Protocols, Vol. 1: Pharmaceutical Nanocarriers. New York, NY, USA, 163-175. ISBN 978-1-60327-359-6.

Westwood, J A; Haynes, N M; Sharkey, J; McLaughlin, N; Pegram, H J; Schwendener, R A; Smyth, M J; Darcy, P K; Kershaw, M H (2009). Toll-like receptor triggering and T-cell costimulation induce potent antitumor immunity in mice. Clinical Cancer Research, 15(24):7624-7633.

Tsushima, Yukio; Jang, Jae-Hwi; Yamada, Yoshito; Schwendener, Reto; Suzuki, Kenji; Weder, Walter; Jungraithmayr, Wolfgang (2014). The depletion of donor macrophages reduces ischaemia-reperfusion injury after mouse lung transplantation. European Journal of Cardio-Thoracic Surgery, 45(4):703-709.

König, Simone; Regen, Tommy; Dittmann, Kai; Engelke, Michael; Wienands, Jürgen; Schwendener, Reto; Hanisch, Uwe-Karsten; Pukrop, Tobias; Hahn, Heidi (2013). Empty liposomes induce antitumoral effects associated with macrophage responses distinct from those of the TLR1/2 agonist Pam3CSK 4 (BLP). Cancer Immunology, Immunotherapy, 62(10):1587-1597.

Jurgeit, Andreas; McDowell, Robert; Moese, Stefan; Meldrum, Eric; Schwendener, Reto; Greber, Urs F (2012). Niclosamide is a proton carrier and targets acidic endosomes with broad antiviral effects. PLoS Pathogens, 8(10):e1002976.

Kuonen, François; Laurent, Julien; Secondini, Chiara; Lorusso, Girieca; Stehle, Jean-Christophe; Rausch, Thierry; Faes-Van't Hull, Eveline; Bieler, Grégory; Alghisi, Gian-Carlo; Schwendener, Reto; Andrejevic-Blant, Snezana; Mirimanoff, René-Olivier; Rüegg, Curzio (2012). Inhibition of the Kit Ligand/c-Kit Axis Attenuates Metastasis in a Mouse Model Mimicking Local Breast Cancer Relapse after Radiotherapy. Clinical Cancer Research, 18(16):4365-4374.

Bonde, Anne-Katrine; Tischler, Verena; Kumar, Sushil; Soltermann, Alex; Schwendener, Reto (2012). Intratumoral macrophages contribute to epithelial-mesenchymal transition in solid tumors. BMC Cancer, 12(1):35.

Schilling, Joel D; Machkovech, Heather M; Kim, Alfred H J; Schwendener, Reto; Schwedwener, Reto; Schaffer, Jean E (2012). Macrophages modulate cardiac function in lipotoxic cardiomyopathy. American Journal of Physiology - Heart and Circulatory Physiology, 303(11):H1366-H1373.

Lee, Y S; Li, P; Huh, J Y; Hwang, I J; Lu, M; Kim, J I; Ham, M; Talukdar, S; Chen, A; Lu, W J; Bandyopadhyay, G K; Schwendener, R; Olefsky, J; Kim, J B (2011). Inflammation is necessary for long-term but not short-term high-fat diet-induced insulin resistance. Diabetes, 60(10):2474-2483.

Rizzo, A; Monteleone, I; Fina, D; Stolfi, C; Caruso, R; Fantini, M C; Franzè, E; Schwendener, R; Pallone, F; Monteleone, G (2011). Inhibition of colitis by IL-25 associates with induction of alternatively activated macrophages. Inflammatory Bowel Diseases, 18(3):449-459.

Haynes, N M; Hawkins, E D; Li, M; McLaughlin, N M; Hämmerling, G J; Schwendener, R; Winoto, A; Wensky, A; Yagita, H; Takeda, K; Kershaw, M H; Darcy, P K; Smyth, M J (2010). CD11c(+) dendritic cells and B cells contribute to the tumoricidal activity of anti-DR5 antibody therapy in established tumors. Journal of Immunology, 185(1):532-541.

Hofer, U; Schlaepfer, E; Baenziger, S; Nischang, M; Regenass, S; Schwendener, R; Kempf, W; Nadal, D; Speck, R F (2010). Inadequate clearance of translocated bacterial products in HIV-infected humanized mice. PLoS Pathogens, 6(4):e1000867.

Ibrahim-Granet, O; Jouvion, G; Hohl, T M; Droin-Bergère, S; Philippart, F; Kim, O Y; Adib-Conquy, M; Schwendener, R; Cavaillon, J-M; Brock, M (2010). In vivo bioluminescence imaging and histopathopathologic analysis reveal distinct roles for resident and recruited immune effector cells in defense against invasive aspergillosis. BMC Microbiology, 10:105.

Andrés, G; Leali, D; Mitola, S; Coltrini, D; Camozzi, M; Corsini, M; Belleri, M; Hirsch, E; Schwendener, R; Christofori, G; Alcamí, A; Presta, M (2009). A pro-inflammatory signature mediates FGF2-induced angiogenesis. Journal of Cellular and Molecular Medicine, 13(8b):2083-2108.

Torroella-Kouri, M; Silvera, R; Rodriguez, D; Caso, R; Shatry, A; Opiela, S; Ilkovitch, D; Schwendener, R; Iragavarapu-Charyulu, V; Cardentey, Y; Strbo, N; Lopez, D M (2009). Identification of a subpopulation of macrophages in mammary tumor-bearing mice that are neither M1 nor M2 and are less differentiated. Cancer Research, 69(11):4800-4809.

Gunsten, S; Mikols, C L; Grayson, M H; Schwendener, R; Agapov, E; Tidwell, R M; Cannon, C L; Brody, S L; Walter, M J (2009). IL-12 p80-dependent macrophage recruitment primes the host for increased survival following a lethal respiratory viral infection. Immunology, 126(4):500-513.

Schott, H; Hamprecht, K; Schott, S; Schott, T C; Schwendener, R (2009). Synthesis and in vitro activities of a new antiviral duplex drug linking Zidovudine (AZT) and Foscarnet (PFA) via an octadecylglycerol residue. Bioorganic & Medicinal Chemistry, 17(1):303-310.

Kataru, R P; Jung, K; Jang, C; Yang, H; Schwendener, R; Baik, J E; Han, S H; Alitalo, K; Koh, G Y (2009). Critical role of CD11b+ macrophages and VEGF in inflammatory lymphangiogenesis, antigen clearance, and inflammation resolution. Blood, 113(22):5650-5659.

Zumsteg, A; Baeriswyl, V; Imaizumi, N; Schwendener, R; Rüegg, C; Christofori, G (2009). Myeloid cells contribute to tumor lymphangiogenesis. PLoS ONE, 4(9):e7067.

Kim, K E; Koh, Y J; Jeon, B H; Jang, C; Han, J; Kataru, R P; Schwendener, R; Kim, J M; Koh, G Y (2009). Role of CD11b+ macrophages in intraperitoneal lipopolysaccharide-induced aberrant lymphangiogenesis and lymphatic function in the diaphragm. American Journal of Pathology, 175(4):1733-1745.

Schott, H; Schott, S; Schwendener, R (2009). Synthesis and in vitro activities of new anticancer duplex drugs linking 2'-deoxy-5-fluorouridine (5-FdU) with 3'-C-ethynylcytidine (ECyd) via a phosphodiester bonding. Bioorganic & Medicinal Chemistry, 17(19):6824-6831.

Sayi, A; Kohler, E; Hitzler, I; Arnold, I; Schwendener, R; Rehrauer, H; Müller, A (2009). The CD4+ T cell-mediated IFN-gamma response to Helicobacter infection is essential for clearance and determines gastric cancer risk. Journal of Immunology, 182(11):7085-7101.

Kim, D D; Miwa, T; Kimura, Y; Schwendener, R; van Lookeren Campagne, M; Song, W C (2008). Deficiency of decay-accelerating factor and complement receptor 1-related gene/protein y on murine platelets leads to complement-dependent clearance by the macrophage phagocytic receptor CRIg. Blood, 112(4):1109-1119.

Hippert, C; Dubois, G; Morin, C; Disson, O; Ibanes, S; Jacquet, C; Schwendener, R; Antignac, C; Kremer, E J; Kalatzis, V (2008). Gene transfer may be preventive but not curative for a lysosomal transport disorder. Molecular Therapy, 16(8):1372-81.

Bijnsdorp, I V; Schwendener, R; Schott, H; Schott, S; Fichtner, I; Honeywell, R J; Losekoot, N; Laan, A C; Peters, G J (2008). In vitro activity and mechanism of action of a duplex and multidrug of ethynylcytidine and 5-fluorodeoxyuridine. Nucleic Acids Symposium Series (2004), (52):651.

Shen, W; Liu, Y; Zhu, J; Schwendener, R; Huard, J (2008). Interaction between macrophages, TGF-beta1, and the COX-2 pathway during the inflammatory phase of skeletal muscle healing after injury. Journal of Cellular Physiology, 214(2):405-412.

Ren, W; Markel, D C; Schwendener, R; Ding, Y; Wu, B; Wooley, P H (2008). Macrophage depletion diminishes implant-wear-induced inflammatory osteolysis in a mouse model. Journal of Biomedical Materials Research Part A, 85(4):1043-1051.

Wang, J; Barke, R A; Charboneau, R; Schwendener, R; Roy, S (2008). Morphine induces defects in early response of alveolar macrophages to Streptococcus pneumoniae by modulating TLR9-NF-kappaB signaling. Journal of Immunology, 180(5):3594-3600.

Hauk, T G; Müller, A; Lee-Thorp, J; Schwendener, R; Fischer, D (2008). Neuroprotective and axon growth promoting effects of intraocular inflammation do not depend on oncomodulin or the presence of large numbers of activated macrophages. Experimental Neurology, 209(2):469-482.

Kim, E Y; Battaile, J T; Patel, A C; You, Y; Agapov, E; Grayson, M H; Benoit, L A; Byers, D E; Alevy, Y; Tucker, J; Swanson, S; Tidwell, R; Tyner, J W; Morton, J D; Castro, M; Polineni, D; Patterson, G A; Schwendener, R; Allard, J D; Peltz, G; Holtzman, M J (2008). Persistent activation of an innate immune response translates respiratory viral infection into chronic lung disease. Nature Medicine, 14(6):633-640.

Jeon, B H; Jang, C; Han, J; Kataru, R P; Piao, L; Jung, K; Cha, H J; Schwendener, R; Jang, K Y; Kim, K S; Alitalo, K; Koh, G Y (2008). Profound but dysfunctional lymphangiogenesis via vascular endothelial growth factor ligands from CD11b+ macrophages in advanced ovarian cancer. Cancer Research, 68(4):1100-9.

Zehnder-Fjällman, A H M; Marty, C; Halin, C; Hohn, A; Schibli, R; Ballmer-Hofer, K; Schwendener, R (2007). Evaluation of anti-VEGFR-3 specific scFv antibodies as potential therapeutic and diagnostic tools for tumor lymph-angiogenesis. Oncology Reports, 18(4):933-941.

Cho, C H; Koh, Y J; Han, J; Sung, H K; Jong Lee, H; Morisada, T; Schwendener, R; Brekken, R A; Kang, G; Oike, Y; Choi, T S; Suda, T; Yoo, O J; Koh, G Y (2007). Angiogenic role of LYVE-1-positive macrophages in adipose tissue. Circulation Research, 100(4):e47-e57.

Schwendener, R (2007). Delivery Systems for Peptides/Oligonucleotides and Lipophilic Nucleoside Analogs. In: Gad, S C. Handbook of Pharmaceutical Biotechnology. Hoboken, New Jersey, USA, 1149-1172. ISBN 978-0-471-21386-4.

Bijnsdorp, I V; Schwendener, R; Schott, H; Fichtner, I; Smid, K; Schott, S; Laan, A C; Peters, G J (2007). In vivo and in vitro activity and mechanism of action of the multidrug cytarabine-L-glycerylyl-fluorodeoxyuridine. Nucleosides, Nucleotides and Nucleic Acids, 26(10-12):1619-1624.

Schwendener, R (2007). Liposomes in biology and medicine. In: Chan, W C W. Bio-Applications of Nanoparticles. New York, 117-128. ISBN 978-0-387-76712-3.

Zeisberger, S M; Odermatt, B; Marty, C; Zehnder-Fjällman, A H M; Ballmer-Hofer, K; Schwendener, R (2006). Clodronate-liposome-mediated depletion of tumour-associated macrophages: a new and highly effective antiangiogenic therapy approach. British Journal of Cancer, 95(3):272-281.

Marty, C; Langer-Machova, Z; Sigrist, S; Schott, H; Schwendener, R; Ballmer-Hofer, K (2006). Isolation and characterization of a scFv antibody specific for tumor endothelial marker 1 (TEM1), a new reagent for targeted tumor therapy. Cancer Letters, 235(2):298-308.

Lasbury, M E; Durant, P J; Ray, C A; Tschang, D; Schwendener, R; Lee, C H (2006). Suppression of alveolar macrophage apoptosis prolongs survival of rats and mice with pneumocystis pneumonia. Journal of Immunology, 176(11):6443-6453.

Beck-Schimmer, B; Schwendener, R; Pasch, T; Reyes, L; Booy, C; Schimmer, R C (2005). Alveolar macrophages regulate neutrophil recruitment in endotoxin-induced lung injury. Respiratory Research, 6:61.

Tyner, J W; Uchida, O; Kajiwara, N; Kim, E Y; Patel, A C; O'Sullivan, M P; Walter , M J; Schwendener, R; Cook, D N; Danoff, T M; Holtzman, M J (2005). CCL5-CCR5 interaction provides antiapoptotic signals for macrophage survival during viral infection. Nature Medicine, 11(11):1180-1187.

Marty, C; Schwendener, R (2005). Cytotoxic tumor targeting with scFv antibody-modified liposomes. In: Ludewig, B; Hoffmann, M W. Adoptive Immunotherapy: Methods and Protocols. Springer, 389-401. ISBN 978-1-58829-406-7 (P) 978-1-59259-862-5 (E).

Probst, H C; Tschannen, K; Odermatt, B; Schwendener, R; Zinkernagel, R M; Van Den Broek, M (2005). Histological analysis of CD11c-DTR/GFP mice after in vivo depletion of dendritic cells. Clinical and Experimental Immunology, 141(3):398-404.

Schwendener, R; Schott, H (2005). Lipophilic arabinofuranosyl cytosine derivatives in liposomes. Methods in Enzymology, 391:58-70.

Buchholz, Julia; Kaser-Hotz, Barbara; Khan, Tania; Rohrer Bley, Carla; Melzer, Katja; Schwendener, Reto; Roos, Malgorzata; Walter, Heinrich (2005). Optimizing photodynamic therapy: in vivo pharmacokinetics of liposomal meta-(tetrahydroxyphenyl)chlorin in feline squamous cell carcinoma. Clinical Cancer Research, 11(20):7538-7544.

Robinson, J A; Shankaramma, S C; Jetter, P; Kienzl, U; Schwendener, R; Vrijbloed, J W; Obrecht, D (2005). Properties and structure-activity studies of cyclic beta-hairpin peptidomimetics based on the cationic antimicrobial peptide protegrin I. Bioorganic & Medicinal Chemistry, 13(6):2055-2064.

Beck-Schimmer, B; Rosenberger, D S; Neff, S B; Jamnicki, M; Suter, D; Fuhrer, T; Schwendener, R; Booy, C; Reyes, L; Pasch, T; Schimmer, R C (2005). Pulmonary aspiration: new therapeutic approaches in the experimental model. Anesthesiology, 103(3):556-566.

Ludwig, P S; Schwendener, R; Schott, H (2005). Synthesis and anticancer activities of amphiphilic 5-fluoro-2'-deoxyuridylic acid prodrugs. European Journal of Medicinal Chemistry, 40(5):494-504.

Rubio Demirovic, A; Marty, C; Console, S; Zeisberger, S M; Ruch, C; Jaussi, R; Schwendener, R; Ballmer-Hofer, K (2005). Targeting human cancer cells with VEGF receptor-2-directed liposomes. Oncology Reports, 13(2):319-324.

Engler, O B; Schwendener, R; Dai, W J; Wölk, B; Pichler, W; Moradpour, D; Brunner, T; Cerny, A (2004). A liposomal peptide vaccine inducing CD8+ T cells in HLA-A2.1 transgenic mice, which recognise human cells encoding hepatitis C virus (HCV) proteins. Vaccine, 23(1):58-68.

Marty, C; Meylan, C; Schott, H; Ballmer-Hofer, K; Schwendener, R (2004). Enhanced heparan sulfate proteoglycan-mediated uptake of cell-penetrating peptide-modified liposomes. Cellular and Molecular Life Sciences, 61(14):1785-1794.

Storni, T; Ruedl, C; Schwarz, K; Schwendener, R; Renner, W A; Bachmann, M F (2004). Nonmethylated CG motifs packaged into virus-like particles induce protective cytotoxic T cell responses in the absence of systemic side effects. Journal of Immunology, 172(3):1777-1785.

Console, S; Marty, C; García-Echeverría, C; Schwendener, R; Ballmer-Hofer, K (2003). Antennapedia and HIV transactivator of transcription (TAT) "protein transduction domains" promote endocytosis of high molecular weight cargo upon binding to cell surface glycosaminoglycans. Journal of Biological Chemistry, 278(37):35109-35114.

Madjdpour, C; Jewell, U R; Kneller, S; Ziegler, U; Schwendener, R; Booy, C; Kläusli, L; Pasch, T; Schimmer, R C; Beck-Schimmer, B (2003). Decreased alveolar oxygen induces lung inflammation. American Journal of Physiology: Lung Cellular and Molecular Physiology, 284(2):L360-L367.

Aichele, P; Zinke, J; Grode, L; Schwendener, R; Kaufmann, S H E; Seiler, P (2003). Macrophages of the splenic marginal zone are essential for trapping of blood-borne particulate antigen but dispensable for induction of specific T cell responses. Journal of Immunology, 171(3):1148-1155.

Marty, C; Odermatt, B; Schott, H; Neri, D; Ballmer-Hofer, K; Klemenz, R; Schwendener, R (2002). Cytotoxic targeting of F9 teratocarcinoma tumours with anti-ED-B fibronectin scFv antibody modified liposomes. British Journal of Cancer, 87(1):106-112.

Schwendener, R; Friedl, K; Depenbrock, H; Schott, H; Hanauske, A R (2001). In vitro activity of liposomal N4octadecyl-1-beta-D-arabinofuranosylcytosine (NOAC), a new lipophilic derivative of 1-beta-D-arabinofuranocylcytosine on biopsized clonogenic human tumor cells and hematopoietic precursor cells. Investigational New Drugs, 19(3):203-210.

Seiler, P; Schwendener, R; Bandermann, S; Brinkmann, V; Grode, L; Kaufmann, S H E; Aichele, P (2001). Limited mycobacterial infection of the liver as a consequence of its microanatomical structure causing restriction of mycobacterial growth to professional phagocytes. Infection and Immunity, 69(12):7922-7926.

Marty, C; Scheidegger, P; Ballmer-Hofer, K; Klemenz, R; Schwendener, R (2001). Production of functionalized single-chain Fv antibody fragments binding to the ED-B domain of the B-isoform of fibronectin in Pichia pastoris. Protein Expression and Purification, 21(1):156-164.

Roscic-Mrkic, B; Schwendener, R; Odermatt, B; Zuniga, A; Pavlovic, J; Billeter, M A; Cattaneo, R (2001). Roles of macrophages in measles virus infection of genetically modified mice. Journal of Virology, 75(7):3343-3351.

Cattaneo-Pangrazzi, R M; Schott, H; Wunderli-Allenspach, H; Rothen-Rutishauser, B; Guenthert, M; Schwendener, R (2000). Cell-cycle arrest and p53-independent induction of apoptosis in vitro by the new anticancer drugs 5-FdUrd-P-FdCydOct and dCydPam-P-FdUrd in DU-145 human prostate cancer cells. Journal of Cancer Research and Clinical Oncology, 126(5):247-256.

Horber, D H; Cattaneo-Pangrazzi, R M; von Ballmoos, P; Schott, H; Ludwig, P S; Eriksson, S; Fichtner, I; Schwendener, R (2000). Cytotoxicity, cell-cycle perturbations and apoptosis in human tumor cells by lipophilic N4-alkyl-1-beta-D-arabinofuranosylcytosine derivatives and the new heteronucleoside phosphate dimer arabinocytidylyl-(5'-->5')-N4-octadecyl-1-beta-D-arabinofuranosylcytosi ne. Journal of Cancer Research and Clinical Oncology, 126(6):311-319.

Ludewig, B; Barchiesi, F; Pericin, M; Zinkernagel, R M; Hengartner, H; Schwendener, R (2000). In vivo antigen loading and activation of dendritic cells via a liposomal peptide vaccine mediates protective antiviral and anti-tumour immunity. Vaccine, 19(1):23-32.

Cattaneo-Pangrazzi, R M; Schott, H; Wunderli-Allenspach, H; Derighetti, M; Schwendener, R (2000). Induction of cell cycle-dependent cytotoxicity and apoptosis by new heterodinucleoside phosphate dimers of 5-fluorodeoxyuridine in PC-3 human prostate cancer cells. Biochemical Pharmacology, 60(12):1887-96.

Cattaneo-Pangrazzi, R M; Schott, H; Schwendener, R (2000). The novel heterodinucleoside dimer 5-FdU-NOAC is a potent cytotoxic drug and a p53-independent inducer of apoptosis in the androgen-independent human prostate cancer cell lines PC-3 and DU-145. Prostate, 45(1):8-18.

Koller-Lucae, S K; Schott, H; Schwendener, R (1999). Low density lipoprotein and liposome mediated uptake and cytotoxic effect of N4-octadecyl-1-beta-D-arabinofuranosylcytosine in Daudi lymphoma cells. British Journal of Cancer, 80(10):1542-9.

Koller-Lucae, S K; Suter, M J; Rentsch, K M; Schott, H; Schwendener, R (1999). Metabolism of the new liposomal anticancer drug N4-octadecyl-1-beta-D-arabinofuranosylcytosine in mice. Drug Metabolism and Disposition, 27(3):342-350.

Ludewig, B; Oehen, S; Barchiesi, F; Schwendener, R; Hengartner, H; Zinkernagel, R M (1999). Protective antiviral cytotoxic T cell memory is most efficiently maintained by restimulation via dendritic cells. Journal of Immunology, 163(4):1839-1844.

Schwendener, R; Peghini, P A; Ludwing, P S; Schott, H (1999). Synthesis, in vitro anti-HIV and anti-hepatitis B activities and pharmacokinetic properties of amphiphilic heterodinucleoside phosphates containing ddC and AZT. Nucleosides and Nucleotides, 18(4-5):949-950.

Peghini, P A; Zahner, R; Kuster, H; Schott, H; Schwendener, R (1998). In vitro anti-human immunodeficiency virus and anti-hepatitis B virus activities and pharmacokinetic properties of heterodinucleoside phosphates containing AZT or ddC. Antiviral Chemistry & Chemotherapy, 9(2):117-126.

Rentsch, K M; Schwendener, R; Pestalozzi, B C; Sauter, C; Wunderli-Allenspach, H; Hänseler, E (1998). Pharmacokinetic studies of mitoxantrone and one of its metabolites in serum and urine in patients with advanced breast cancer. European Journal of Clinical Pharmacology, 54(1):83-89.

van Borssum Waalkes, M; Goris, H; Dontje, B H; Schwendener, R; Scherphof, G; Nijhof, W (1998). Toxicity of liposomal 3'-5'-O-dipalmitoyl-5-fluoro-2'-deoxyuridine in mice. Anti-Cancer Drug Design, 13(4):291-305.

Rentsch, K M; Horber, D H; Schwendener, R; Wunderli-Allenspach, H; Hänseler, E (1997). Comparative pharmacokinetic and cytotoxic analysis of three different formulations of mitoxantrone in mice. British Journal of Cancer, 75(7):986-992.

Seiler, P; Aichele, P; Odermatt, B; Hengartner, H; Zinkernagel, R M; Schwendener, R (1997). Crucial role of marginal zone macrophages and marginal zone metallophils in the clearance of lymphocytic choriomeningitis virus infection. European Journal of Immunology, 27(10):2626-2633.

Koller-Lucae, S K; Schott, H; Schwendener, R (1997). Interactions with human blood in vitro and pharmacokinetic properties in mice of liposomal N4-octadecyl-1-beta-D-arabinofuranosylcytosine, a new anticancer drug. Journal of Pharmacology and Experimental Therapeutics, 282(3):1572-1580.

Frankenberger, M; Hofmann, B; Emmerich, B; Nerl, C; Schwendener, R; Ziegler-Heitbrock, H W (1997). Liposomal 1,25 (OH)2 vitamin D3 compounds block proliferation and induce differentiation in myelomonocytic leukaemia cells. British Journal of Haematology, 98(1):186-194.

Rentsch, K M; Schwendener, R; Schott, H; Hänseler, E (1997). Pharmacokinetics of N4-octadecyl-1-beta-D-arabinofuranosylcytosine in plasma and whole blood after intravenous and oral administration to mice. Journal of Pharmacy and Pharmacology, 49(11):1076-1081.

Rentsch, K M; Schwendener, R; Hänseler, E (1996). Determination of mitoxantrone in mouse whole blood and different tissues by high-performance liquid chromatography. Journal of Chromatography. B, Biomedical Applications, 679(1-2):185-192.

Schwendener, R; Schott, H (1996). Lipophilic 1-beta-D-arabinofuranosyl cytosine derivatives in liposomal formulations for oral and parenteral antileukemic therapy in the murine L1210 leukemia model. Journal of Cancer Research and Clinical Oncology, 122(12):723-726.

Schwendener, R; Horber, D H; Odermatt, B; Schott, H (1996). Oral antitumour activity in murine L1210 leukaemia and pharmacological properties of liposome formulations of N4-alkyl derivatives of 1-beta-D-arabinofuranosylcytosine. Journal of Cancer Research and Clinical Oncology, 122(2):102-108.

Schott, H; Schwendener, R (1996). Synthesis and structure-activity studies in vivo of liposomal phospholipid-N4-palmitoyl- and N4-hexadecyl-1-beta-D-arabinofuranosylcytosine conjugates. Anti-Cancer Drug Design, 11(6):451-62.

Horber, D H; von Ballmoos, P; Schott, H; Schwendener, R (1995). Cell cycle-dependent cytotoxicity and induction of apoptosis by liposomal N4-hexadecyl-1-beta-D-arabinofuranosylcytosine. British Journal of Cancer, 72(5):1067-1073.

Horber, D H; Schott, H; Schwendener, R (1995). Cellular pharmacology of N4-hexadecyl-1-beta-D-arabinofuranosylcytosine in the human leukemic cell lines K-562 and U-937. Cancer Chemotherapy and Pharmacology, 36(6):483-492.

Horber, D H; Schott, H; Schwendener, R (1995). Cellular pharmacology of a liposomal preparation of N4-hexadecyl-1-beta-D-arabinofuranosylcytosine, a lipophilic derivative of 1-beta-D-arabinofuranosylcytosine. British Journal of Cancer, 71(5):957-962.

Horber, D H; Ottiger, C; Schott, H; Schwendener, R (1995). Pharmacokinetic properties and interactions with blood components of N4-hexadecyl-1-beta-D-arabinofuranosylcytosine (NHAC) incorporated into liposomes. Journal of Pharmacy and Pharmacology, 47(4):282-288.

Pestalozzi, B C; Vass, A; Adam, H; Horber, D H; Schwendener, R; Sauter, C (1995). Phase II study of liposome-complexed mitoxantrone in patients with advanced breast cancer. European Journal of Cancer, 31A(6):1024.

Rentsch, K M; Schwendener, R; Schott, H; Hänseler, E (1995). Sensitive high-performance liquid chromatographic method for the determination of N4-hexadecyl- and N4-octadecyl-1-beta-D-arabinofuranosylcytosine in plasma and erythrocytes. Journal of Chromatography. B, Biomedical Applications, 673(2):259-266.

Schwendener, R; Gowland, P; Horber, D H; Zahner, R; Schertler, A; Schott, H (1994). New lipophilic alkyl/acyl dinucleoside phosphates as derivatives of 3'-azido-3'-deoxythymidine: inhibition of HIV-1 replication in vitro and antiviral activity against Rauscher leukemia virus infected mice with delayed treatment regimens. Antiviral Research, 24(1):79-93.

Condrau, M A; Schwendener, R; Niederer, P; Anliker, M (1994). Time-resolved flow cytometry for the measurement of lanthanide chelate fluorescence: I. Concept and theoretical evaluation. Cytometry Part A, 16(3):187-194.

Condrau, M A; Schwendener, R; Zimmermann, M; Muser, M H; Graf, U; Niederer, P; Anliker, M (1994). Time-resolved flow cytometry for the measurement of lanthanide chelate fluorescence: II. Instrument design and experimental results. Cytometry Part A, 16(3):195-205.

Pestalozzi, B; Schwendener, R; Sauter, C (1992). Phase I/II study of liposome-complexed mitoxantrone in patients with advanced breast cancer. Annals of Oncology, 3(6):445-449.

Schwendener, R; Schott, H (1992). Treatment of L1210 murine leukemia with liposome-incorporated N4-hexadecyl-1-beta-D-arabinofuranosyl cytosine. International Journal of Cancer, 51(3):466-469.

Schwendener, R; Fiebig, H H; Berger, M R; Berger, D P (1991). Evaluation of incorporation characteristics of mitoxantrone into unilamellar liposomes and analysis of their pharmacokinetic properties, acute toxicity, and antitumor efficacy. Cancer Chemotherapy and Pharmacology, 27(6):429-439.

Schwendener, R; Wüthrich, R; Duewell, S; Wehrli, E; von Schulthess, G K (1990). A pharmacokinetic and MRI study of unilamellar gadolinium-, manganese-, and iron-DTPA-stearate liposomes as organ-specific contrast agents. Investigative Radiology, 25(8):922-932.