David versus Goliath

Article Outline

• The Lundbeck Foundation
• CBN—The Lundbeck Foundation Center for Biomembranes in Nanomedicine
• LUNA—The Lundbeck Foundation Nanomedicine Centre for Individualized Management of Tissue Damage and Regeneration
• NanoCAN—The Lundbeck Foundation Nanomedicine Research Center for Cancer Stem Cell Targeting Therapeutics
• Quo vadis Danish Nanomedicine: NanoMED-Alliance—The Nanomedicine Research Center Strategic Alliance
• Copyright

In 2010 three new Nanomedicine Research Centers have been launched in Denmark. The three Centers of Excellence emerged via an open call for proposals posted by the private funding organization The Lundbeck Foundation. The foundation will provide a total financing of €13 million (US $17.7 million) for an initial period of 5 years to the three Centers. For a country like Denmark, this represents a massive investment into frontline research. The real dimension can best be visualized by relating it to the gross domestic product of Denmark and scaling it to the gross domestic products of Germany or the United States. Doing so, the financial support would correspond to investments of €139 million (US $189 million) and €540 million (US $735 million) for Germany and the United States, respectively, which is well in the scale of larger national programs. The three Centers will investigate novel strategies for exploiting nanoscaled biostructures in combating major human diseases, including cancer as well as neurological, infectious, cardiovascular, and musculoskeletal disorders.

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The Lundbeck Foundation 

The Lundbeck Foundation (http://www.lundbeckfonden.dk) is a private commercial organization established in 1954 by Grete Lundbeck, the widow of Hans Lundbeck, who founded the pharmaceutical company H. Lundbeck A/S. This Danish company has its main business area in the field of drug development for central nervous system diseases. The Lundbeck Foundation holds the majority of the stocks of H. Lundbeck A/S, as well as shares of further companies, summing up to an impressive equity of €3.7 billion (US $5 billion) on the basis of the 2008 market values.

The mission of the Lundbeck Foundation is to maintain and expand the activities of the Lundbeck Group, and to provide financial support for research of the highest quality in biomedical and natural sciences. Commercial operations are carried out through the wholly owned subsidiary daughter company LFI A/S, and thus are separated from research funding activities. In 2009, the Lundbeck Foundation provided research grants of a total of €45.7 million (US $63 million), which is well beyond the sums distributed by individual Danish Research Councils and comparable national funding programs in the fields of biomedical and natural sciences (Figure 1). Remarkably, program development, calls, grant review, decision making, grant distribution, and follow-up are accomplished via a relatively small staff, pointing to a quite effective management. Research funding via the foundation addresses various relevant levels, including awards, 5-year professorships, junior group leader fellowships, and individual research projects. Since 2005 the Lundbeck Foundation has established 15 Centers of Excellence in Denmark with a total financing of €64.5 million (US $88.9 million). The calls are developed through a bottom-up approach, wherein the Foundation discusses suggestions from the scientific community so as to identify relevant topics. Applications are selected via a transparent two-stage process, each with international review by an external scientific expert committee. For the 2009 Center of Excellence call, nanomedicine was identified as the topic, resulting in the establishment of the three new Nanomedicine Research Centers in Denmark (Figure 2) that start their operation in 2010.

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• Figure 1. 

  Funding volumes provided by major Danish funding organizations in 2009. Values are presented both in U.S. dollars and Euros. LF, the Lundbeck Foundation. The Danish Council for Strategic Research (DCSR) had three programs in 2009, which related to health sciences or nano/biotechnology and are relevant for comparison: the programs for Health, Food and Welfare (HFW), for Individual, Disease and Society (IDS), and for Strategic Growth Technologies (SGT). Also displayed are the volumes of the Danish Council for Independent Research (DCIR), Natural Sciences (NS) and Medical Sciences (MS) as well as the total volume of funds distributed by the Danish Cancer Society (DCS).

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• Figure 2. 

  Distribution of the three Nanomedicine Research Centers and associated Nanobiotechnology Centers throughout Denmark. Aarhus University hosts LUNA (the Lundbeck Foundation Nanomedicine Centre for Individualized Management of Tissue Damage and Regeneration) and the Interdisciplinary Nanoscience Center (iNANO), whereas the CBN (the Lundbeck Foundation Center for Biomembranes in Nanomedicine) and the NSC (Nano-Science Center) are located at the University of Copenhagen. NanoCAN (the Lundbeck Foundation Nanomedicine Research Center for Cancer Stem Cell Targeting Therapeutics), the NAC (Nucleic Acid Center), and MEMPHYS (Center for Biomembrane Physics) are at the University of Southern Denmark in Odense.

The three Centers use complementary techniques and approaches adapted to their individual goals but also share certain core approaches and provide or develop platform technologies that may create strong synergies in the future. The Centers are highly interdisciplinary, combining know-how and experts in basic and clinical research as well as from the areas of nanotechnology, biotechnology, molecular biology, biomedicine, chemistry, and physics.

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CBN—The Lundbeck Foundation Center for Biomembranes in Nanomedicine 

The CBN (http://www.nanomedicine.ku.dk/english) is located at the University of Copenhagen and jointly led by U.G. (gether@sund.ku.dk) of the Institute for Neuroscience and Pharmacology, and D.S. (stamou@nano.ku.dk) of the Institute for Neuroscience and Pharmacology and Nano-Science Center, as directors. Lipid biomembranes, which are the focus of research within this Center, not only represent barriers for compartmentalization of the cell versus the environment and of cell organelles versus the cytoplasm but actively participate in a variety of fundamental biological processes (Figure 3). Membrane lipids mediate, together with a complex composition of embedded membrane proteins, processes such as transport of nutrients and ions into and out of the cell, cell-cell contacts, and transmission of chemical signals into, from, and between cells. Because of the particular importance of these processes, it is not surprising that the underlying principles and mechanisms are evolutionarily conserved and thus very similar when comparing human and bacterial cell-cell communication.

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• Figure 3. 

  Concepts and strategies of the CBN. Biomembranes possess nanoscaled domains that differ in composition (1) and/or architecture (2), which modulates their functional properties. Via type VI secretion systems, bacteria secrete vesicles that contain quorum sensing factors and toxins for cell-cell communication within the species and for delivery of factors to host cells (3). The reciprocal processes of exocytosis and endocytosis via lipid vesicles take place during neurotransmitter-mediated communication between neurons, which employs exocytotic release from intracellular liposomal particles (4), inside-outside orientation (inverted). Lipid domain composition and architecture (1, 2) exert critical influences on these processes. The CBN will employ state-of-the-art tools to image and manipulate communication vesicles with nanoparticles (5), to create advanced nanobiosensors (6) and to utilize this knowledge for creating a new generation of drug delivery systems.

The neurotransmitter dopamine is one of the central interests of the CBN. Dopamine is essential for the human brain's ability to regulate motivation, reward, learning, and voluntary movements. Aberrations in dopamine function play a key role in several psychiatric and neurological disorders including schizophrenia, drug addiction, attention deficit hyperactivity disorder, and Parkinson's disease. Upon excitation, the neurotransmitter is released from intracellular lipid vesicles to the synaptic cleft between neurons. After dopamine has bound to its target receptors on the postsynaptic neuron, its action is shut down again by endocytosis via lipid vesicles. Thus, these naturally occurring nanoscaled biomembrane devices regulate fundamental processes in the central nervous system. It is the goal of the CBN to develop ultrasensitive assays for the detection of individual molecules of receptors and transporters in the cell membrane as well as novel nanobiosensors for dopamine and related transmitters. This will improve the understanding of these communication processes and in the long-term contribute to improved diagnosis and therapies for the associated diseases.

A second part of the CBN's project focuses on the fission and fusion of membranes during endo-and exocytosis, respectively. In addition to being involved in the release of neurotransmitters by nerve cells, fission and fusion also function in cell-cell signaling in bacteria, such as quorum sensing. This is a communication process by which the bacteria inform each other about their actual population density. Depending on the density, bacteria might organize into biofilms, which alters their pathogenicity and reduces their sensitivity against antibiotics. Thus, understanding the principles of this communication is of interest for identifying more effective means to combat infectious diseases and nosocomial infections with multidrug-resistant bacteria.

The CBN's activities could turn out to be more than simply the sum of its parts. The mechanisms underlying exo- and endocytosis could aid in constructing more effective or selective liposomal nanodrug delivery systems in the future.

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LUNA—The Lundbeck Foundation Nanomedicine Centre for Individualized Management of Tissue Damage and Regeneration 

Two additional epidemiologically important disease areas are covered by LUNA (http://www.nanomedicine.dk), which is located at Aarhus University and is led by A.F. (allan.flyvbjerg@dadlnet.dk) of Aarhus University Hospital and Health Sciences Faculty, and J.K. (jk@mb.au.dk) of the Interdisciplinary Nanoscience Center (iNANO), as directors. LUNA's activities are dedicated to cardiovascular and musculoskeletal diseases, which—conceptually—can be regarded as having in common as their origin a destructive imbalance between tissue damage and tissue regeneration (Figure 4). Again, naturally occurring nanostructures may have a critical role. So-called pattern recognition molecules (PRMs) are a class of human proteins that are known for their ability to specifically recognize nanostructured patterns and to consecutively trigger corresponding responses. Pattern recognition (PR) is of importance as the first frontline of defense against invading bacteria and viruses, to discern friend (host cells) from foe (pathogens), and for the removal of cell debris, as well as for cells to read out information from the surrounding extracellular matrix for regulation of migration, proliferation, differentiation, and regeneration. Essentially, PR also traces back to ancient evolutionary roots, defined by necessities emerging with the organization of the first multicellular organisms about 1 billion years ago. In this critical evolutionary step, recognition of self and non-self (i.e., of potential pathogens), and the capacity to develop specialized cell types and organ structures became relevant for survival via specialization and adaptation. The enormous regenerative potential of sponges, for example, is based on PR: After dissociation of a sponge into single cells, PRMs allow for reassembly, followed by consecutive regeneration of an intact sponge. PR is also the principle on which the first primitive immune system was based. For example, sea urchins, which lack antibodies, rely on PRMs for defending themselves against invading pathogens. Research within LUNA aims at gaining clues for conceiving novel nanopatterned surfaces and three-dimensional matrices for the guided differentiation of human stem cells into tissues and organs. Furthermore, a targeted intervention via nanodevices (e.g., RNA aptamers and devices that confer RNA interference) will be explored to inhibit PR for the purpose of restoring the balance in chronic disease away from dominance of tissue destruction back to the advantage of regeneration. Thus, the development and optimization of nanoparticles for targeted short interfering RNA (siRNA) delivery and for packaging of RNA aptamers will compose one of the central activities within the Centre. LUNA builds on the excellent framework created by iNANO (http://www.inano.au.dk) at Aarhus University, which is directed by F.B. Since its establishment in 2002, iNANO has grown to an internationally visible nanotechnology center with more than 200 scientific employees at the end of 2009.

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• Figure 4. 

  Research strategies in LUNA. The Center's research strategies comprise four major components. Nanoparticles and nanostructured surfaces modulating or mimicking pattern recognition processes are employed for cell-based therapies and three-dimensional scaffold-based tissue engineering. The approaches are supported by state-of-the-art bioimaging at the nanoscale and the development of nanodrug-delivery devices. Information gained from these efforts is funneled into drug design and validation, which finally uses animal models for proof of principle at the preclinical stage.

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NanoCAN—The Lundbeck Foundation Nanomedicine Research Center for Cancer Stem Cell Targeting Therapeutics 

NanoCAN (http://www.nanocan.dk) is located at The University of Southern Denmark, Odense, and is led by J.M. (jmollenhauer@health.sdu.dk) of the Institute for Molecular Medicine. The central goal of NanoCAN is to develop novel nanodevices that would allow for a selective targeting and consecutive eradication of cancer stem cells (Figure 5). However, the general strategies employed in the Center's research might likewise be applicable to pharmacologically address normal human stem cells for the purpose of regenerative medicine. The main cancer type within the focus of NanoCAN is breast cancer, which is still the major cause for cancer mortality among women and is one of the best investigated tumors with regard to cancer stem cells and molecular profiles.

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• Figure 5. 

  Schematic presentation of NanoCAN's research strategies. Core activities within the Center comprise the construction of cancer cells with cancer stem cell–specific molecular fingerprints, which consecutively are processed via high-throughput genome-wide siRNA screens and analyses via an advanced biochip format. In parallel, aptamers for targeting are generated and packaging of the nucleic acid–based drugs via self-assembly, liposomes, and solid nanospheres is tested. The optimal configurations are selected for integration of selected siRNA hits from high-throughput screening. The goal is to yield nanodrugs that would selectively kill cancer stem cells (red cells), thereby cutting off supply for the cancer cells (orange cells), leading to tumor eradication with only minimal adverse side effects on normal cells (blue cells) and normal adult stem cells (green cells).

The term cancer stem cells circumscribes a small population of cells within a tumor that is particularly potent in cancer formation. A few cells of this type, or even only a single cell, can be sufficient to create a full-blown tumor, which contrasts with the bulk of tumor cells that have no or low capacity to re-create a tumor. Based on the analogy to the regenerative capacity of normal human stem cells, this population is often referred to as cancer stem cells. There are a number of further similarities between these two cell types. They share specific protective mechanisms, which—in the case of the cancer stem cells—are thought to be associated with resistance against radiation therapy and chemotherapy. Cancer stem cells have also been shown to have an extraordinary high metastatic potential. Based on these properties, they are suspected to be responsible for failure of cancer treatment and to substantially contribute to cancer mortality. One of the major challenges is to design new drugs that kill the cancer stem cells but do not exert deleterious side effects on the very similar normal stem cells within the human organism.

NanoCAN aims at constructing nucleic acid–based nanodrugs consisting of aptamers for targeting and siRNAs for selective killing of the cancer stem cells. For optimization of the pharmacological properties, the nucleic acid devices will be equipped with self-assembly structures or incorporated into liposomal and solid nanospheres. These approaches profit from previous work in the Center for Biomembrane Physics (MEMPHYS; http://www.memphys.sdu.dk) and the Nucleic Acid Center (NAC; http://www.nac.sdu.dk), funded by the Danish National Research Foundation and directed by Ole Mouritzen and J.W., respectively. Both Centers are located at The University of Southern Denmark. NanoCAN will employ genome-wide siRNA screens with genetically engineered cancer cells that differ with respect to molecular fingerprints of cancer stem cells so as to recover siRNAs with selective killing activity (Figure 5). The activities also include the development of novel biochip formats to increase the information contents of the screens and approaches to construct synthetic cancer stem cells as novel research tools.

A superimposed goal of NanoCAN is to create a toolkit within which components can be permutated to create other, different targeting selectivities, pharmaceutical effects, or pharmacological properties. This would allow for rapid projection of the strategies to cancer types other than breast cancer and for targeted modulation of normal stem cells for the purpose of regenerative medicine. Furthermore, such a flexible toolkit could contribute to fulfilling the future demands posed by individualized medicine, because component permutation theoretically enables the rapid assembly of drugs with improved match to the molecular profiles of individual cancer patients or individual groups of cancer patients.

Similar to the CBN and LUNA, NanoCAN also independently selected approaches that, in a certain sense, employ mimics of molecular nanodevices that have developed during evolution. Aptamers resemble configurations in early evolution, wherein RNA-based molecules with three-dimensional folds are thought to have exerted critical effects, whereas siRNAs mimic the gene-silencing effects of endogenous cellular micro RNAs, and liposomal drug delivery is reminiscent of natural cellular uptake processes.

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Quo vadis Danish Nanomedicine: NanoMED-Alliance—The Nanomedicine Research Center Strategic Alliance 

How to build up sustainable and effective nanomedicine research infrastructures in Denmark within the national and international context? As initially may have become evident, there exist a fair number of sites of intersection between the three new Centers. Basically, all three Centers independently selected approaches that employ “nanobionics” strategies—in other words, aim at imitating and utilizing the properties of evolutionarily developed nanoscaled biomolecules. One of the predominant reasons is probably that biomolecules allow for creating the high diversity commonly needed as a starting point for drugs that would permit a targeted modulation of processes within a complex organism. Basic principles of biomembrane-nanoparticle interactions deciphered by the CBN obviously might provide useful information also for LUNA and NanoCAN for drug assembly and encapsulation. LUNA's activities to unravel processes underlying PR can be imagined to recover also novel starting points for NanoCAN, because driving cancer stem cells into a targeted differentiation via drugs that mimic nanostructured signaling matrices represents an additional option for achieving positive therapeutic effects. Tools and screening strategies developed in NanoCAN might become relevant for systematic research in the CBN and LUNA as well. Finally, NanoCAN and LUNA not only share an interest in general processes associated with stem cell–mediated regeneration but also have independently selected aptamers and siRNAs as the building blocks for therapeutic nanodevices. Based on these convergent interests, the three Centers have decided to configure themselves in The Nanomedicine Research Center Strategic Alliance (NanoMED-Alliance; http://www.nanomed-alliance.com). In the first step, this organization serves to create the necessary framework for a free exchange of scientific results, know-how, technologies, students, and scientific employees so as to establish synergies. In parallel, the alliance plans to organize workshops and targeted symposia at regular intervals. It is certain, however, that sustainability in this new area will require the development of public funding programs in Denmark, dedicated to strengthening and expanding national activities in nanomedicine.

As a highly interdisciplinary field in and of itself, nanomedicine might further profit from interaction with other emerging fields. From the description of the activities of the three new Nanomedicine Research Centers, experts might recognize that several approaches essentially include top-down and bottom-up synthetic biology strategies. This includes, for example, conception of liposomal nanodrug delivery devices, which in conjunction with nucleic acid–based targeting devices and drugs, reach the complexity of a (nonreplication-capable) virus. Imitation of natural nanopatterned surfaces and construction of synthetic cancer stem cells likewise entail strong synthetic biology elements. Recognizing this momentum, the Nano-Science Center Copenhagen, led by T.B. (http://www.nano.ku.dk; contact: tb@nano.ku.dk), joined the NanoMED-Alliance as the first external partner with the goal to develop a joint roadmap for synthetic biology approaches in nanomedicine. The NanoMED-Alliance has an open architecture and invites other centers in the area of nanomedicine and related fields to participate (interim contact: J.M.; jmollenhauer@health.sdu.dk).