Beneficiary: Alex Rojas Bie Thomsen, Columbia University Medical Center

Grant: 3.600.308 DKK

Melanoma is the deadliest form of skin cancer with few treatment options to patients with advanced metastatic disease.

Melanoma metastasis to lymph nodes is associated with expression of the chemokine receptor CCR7, a member of the G protein-coupled receptors (GPCRs) superfamily that promote cell migration of immune cells. Classically, upon agonist stimulation, GPCRs at the cell membrane activate heterotrimeric G proteins, causing downstream signaling throughout the cell. In order to terminate G protein signaling, cells have devised a specialized desensitization mechanism that includes receptor phosphorylation by GPCR kinases and subsequent recruitment of β-arrestins (βarrs) to the phosphorylated receptors. The GPCR–βarrs interaction both blocks the G proteinbinding site and promotes receptor endocytosis.

However, we recently discovered that some GPCRs interact with G proteins and βarrs simultaneously to form GPCR–G protein–βarr ‘megaplexes’, which allows the receptor to continue to stimulate G protein signaling while being internalized into endosomes by βarrs. Our preliminary results suggest that CCR7 forms megaplexes and promotes G protein signaling from internalized compartments. Interestingly, endosomal signaling, in general, is highly involved in cell migration, and different proteins are trafficked between plasma membrane and endosomes during this process. Thus, the proposed project aims to investigate the involvement of endosomal CCR7 signaling in melanoma cell migration. Furthermore, using a combination of highly advanced cryo-electron tomography and APEX2 proteomics, we will visualize the mechanism of CCR7-mediated melanoma cell migration protein-by-protein. Such detailed mechanistic knowledge will assist in designing innovative therapeutics to treat metastatic malignant melanoma.

 

Beneficiary: Jan J. Enghild, Aarhus University

Grant: 3.045.231 DKK

Severe eczema, also known as atopic dermatitis (AD) is the most common inflammatory skin disease resulting in itchy, inflamed, and swollen skin that is very susceptible to infection. It is estimated that 15-20% of all children and 2-10% of adults are affected, without effective treatment.

Because of this, significant public health burden and the lack of safe and effective treatments, there is a need for novel targeted therapeutics that can help manage symptoms and improve the quality of life for the patients.

The protein periostin is expressed in the skin and is implicated in AD. Significantly, studies have shown that the elimination of periostin in an AD mouse model reduces or completely removes the symptoms making periostin an apparent therapeutic target. However, the physiological functions of periostin remains unclear and a reduction or elimination of the protein in the skin could have severe side effects.

Therefore, a deeper understanding of the physiological role in healthy and diseased skin must be established. The interdisciplinary research team behind this project propose to address these issues and establish the function of periostin using in vitro and in vivo experimental setups including primary cell cultures, zebrafish, mouse models and human specimens combined with advanced biochemical methods. Novel therapeutics are urgently needed, and this project aim is to establish a strategy for the development of new treatment paradigms for AD, leading towards novel, innovative therapeutic strategies.

Beneficiary: Samir Mitragotri, Harvard John A. Paulson School of Engineering and Applied Sciences

Grant: 2.000.043 DKK

We will develop a novel ionic-liquid formulation for topical delivery of nucleic acids into the skin.

Our ionic liquid enhances lipophilicity of nucleic acids by ~100,000,000 fold and enhances their penetration into the skin. We will demonstrate the feasibility of the platform through delivery and efficacy of two siRNAs. The resulting platform is expected to have broad applications to other nucleic acid drugs including mRNA for the treatment of a wide range of dermatological conditions.

Our ionic liquid platform will open new opportunities for the treatment of dermatological conditions.

Beneficiary: David M. Sabatini, Whitehead Institute of Biomedical Research

Grant: 1.278.270 DKK

Studies in human populations have identified dozens of pigmentation genes, many of which encode proteins with well-understood functions, such as in melanocyte development, melanin biosynthesis, and the biogenesis and trafficking of specialized melanin-containing organelles called melanosomes.

Yet, there are other pigmentation genes that we know much less about, such as those that encode putative transporter proteins on the melanosome surface. These putative melanosomal transporters have been reported to import precursor metabolites for melanin synthesis or regulate melanosomal pH; however, many of these findings have been controversial or speculative. Deciphering the molecular function of these putative transporters and their physiological substrates is crucial to our understanding of pigmentation.

To address this problem, we propose to determine the metabolite composition of melanosomes and define the role of individual transporters in melanosomal function. We will develop a purification method to rapidly isolate intact melanosomes and analyse them by liquid chromatography and mass spectrometry to compile the first catalog of melanosomal metabolites. We will subsequently characterize SLC45A2, a putative melanosomal transporter that modulates human pigmentation in response to sunlight. By comparing the metabolite profile of wild-type versus SLC45A2-deficient melanosomes, we will identify candidate SLC45A2 substrates and validate them using biochemical assays, a workflow that will be applied to other putative melanosomal transporters.

This study will present the first detailed analysis of melanosome metabolites, as well as identify key metabolites and their transporters essential for melanogenesis. This work could inform new ways to modulate pigmentation and treat pigmentation pathologies.

Beneficiary: Merete Hædersdal, Bispebjerg Hospital

Grant: 2.358.825 DKK

Squamous cell carcinoma (SCC) together with basal cell carcinoma comprises the absolute majority of non-melanoma skin cancers, affecting 150,000 persons in Denmark, equivalent to 3% of the population.

SCC’s cost is consequently substantial, reflected by notable patient morbidity, heavy socioeconomic burdens and significant mortality in immunosuppressed populations.

In oncology, systemic immunotherapies with PD1 and CTLA4 antibodies have had revolutionizing impact on clinical cancer treatment. Recognizing the immense potential of these strategies also for SCC, our vision is to pioneer a new local treatment approach by harnessing the immune system to combat SCC, while at the same time avoiding side effects associated with systemic treatment.

In a three-tiered translational project, we thus aim to deliver PD1 and CTLA4 antibodies through the skin using ablative fractional laser (AFL), effectively opening the door to implementation of topical SCC immunotherapy. The project is executed in collaboration with the Wellman Center at Harvard Medical School and Center for Cancer Immune Therapy at Herlev Hospital. The 3-year research plan comprises preclinical studies on biodistribution and pharmacokinetics in healthy skin, a proof-of-concept study in a well-established murine model for human SCC, and an explorative clinical study in SCC patients from the skin cancer clinic at Bispebjerg Hospital. For patients, topical immunotherapy may constitute a safe treatment with decreased morbidity and the prospect of potentially reduced risk of future SCC occurrence. This in turn will lower the socioeconomic burden of repeated treatments for a large cancer patient group, including high-risk immunosuppressed patients such as organ transplant recipients.

Beneficiary: Kai Kisand, University of Tartu, Institute of Biomedicine and Translational Medicine; University of Tartu; Ravila Str. 19; Tartu 50411; Estonia

Grant: DKK 4.908.566

Melanoma is a very aggressive type of cancer that affects people at their most productive period of the life. As most of the diagnosed patients should have a long life still ahead a cure of the disease is highly desired.

Cancer immunotherapy with checkpoint inhibitors and T cell adoptive therapy have established the crucial role of T cell responses in melanoma as well as in many other cancers. Successful immunotherapy of melanoma is often associated with vitiligo as a side effect indicating the importance of targeting the antigenic epitopes that are shared between melanocytes and melanoma cells.

However, melanocyte antigens are “self” and T cell receptor (TCR) specificities that recognise such epitopes with high affinity are deleted during their maturation in the thymus. To find high-affinity TCRs specific for melanocyte/melanoma antigens we will interrogate the TCR repertoire of a patient population that is defective in their central (thymic) tolerance induction mechanisms due to mutations in autoimmune regulator gene, and who develop vitiligo as one of their disease manifestations.

We expect to identify several TCR specificities that recognise melanocyte/melanoma antigenic epitopes. This information can be used for designing genetically modified T cells for adoptive treatment of melanoma patients, and to advance the knowledge about vitiligo pathogenesis and mechanisms of central tolerance induction.

Beneficiary: Kathryn Browning, LEO Foundation Centre for Cutaneous Drug Delivery (LFCCDD), Department of Pharmacy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen Ø, Denmark

Grant: DKK 2.234.415

Atopic dermatitis (AD) is a chronic disorder caused by the improper function of the skins barrier layer, the stratum corneum (SC). It is thought to affect between 15 and 30% of children and up to 10% of adults.

The need to develop drugs and drug delivery vehicles which effectively, and possibly specifically, interact with the compromised skin of AD patients is of great importance.

However, to date most pre-clinical trials utilise healthy skin, excised from surgical procedures, to investigate the penetration and interactions of drugs targeted to skin disease. This approach does not accurately represent lesional or diseased stratum corneum.

This project aims to develop models of the stratum corneum to compare the lipid multilayer structure and interactions of healthy and atopic dermatitis (AD) lesional stratum corneum. Key to the success of these models is access to a wide variety of skin lipids not commercially available and crucial to the realistic self-assembly of the lipid multilayers observed in SC.

An example of a currently unavailable lipid is the long chain esterified ceramides, Ceramide[EOS], which has been linked to the formation of long periodicity phases and lower permeability in SC and is often deficient in AD patients.2-4 These lipids will be obtained by extraction, separation and purification of ceramides from pig skin SC. Lipid mixtures of ratios found in healthy and AD lesional skin will then be self-assembled on a solid support and investigated for interactions with drugs and drug delivery vehicles using neutron reflection, which offers unique opportunities for angstrom level structural resolution and, through selective deuteration, the ability to highlight specific components of the system to improve contrast.

Beneficiary: Mariena van der Plas, University of Copenhagen, Faculty of Health and Medical Sciences, Department of Pharmacy, Universitetsparken 2, Copenhagen 2100, Denmark

Grant: DKK 2.745.375

Proper wound healing is a fundamental survival mechanism and dysfunctions cause significant disease, such as seen in infections after burns, trauma and surgery, as well as in non-healing ulcers.

Currently, the prevalence of non-healing wounds is estimated to be over 40 million worldwide, a number projected to rise with 6-9% annually, due to aging of the population and the increasing incidence of diseases that contribute to nonhealing ulcer development, such as obesity and diabetes.

There is a great and unmet need for novel treatments for improved healing, and thus better predictors for wound healing outcomes are essential. Given the importance of innate immunity and microbial interactions for development of impaired wound healing, the aim of this project is to define novel prognostic and diagnostic biomarkers for assessment of wound healing and infection risk.

For this purpose, we will use state-of-the-art techniques for peptidomics mass spectrometry. This unique approach, without the classical trypsin digestion of the samples, will give actual insight in processes occurring in the wound bed, e.g. enzymatic activity, infection, inflammation, and angiogenesis, instead of just reporting the presence of a protein, independent of the state it is in.

Furthermore, we will set up biological models for validation of biomarkers, as well as novel treatments. Together, the outcomes of these studies have the potential to improve diagnostic evaluations of wounds, and will enable us to develop novel treatment concepts for early prevention of infection, leading to improved healing results for large and significant patient groups.

Beneficiary: Dr. Rudolf Jaenisch, Member, Whitehead Institute and Professor, Department of Biology, Massachusetts Institute of Technology, MA, USA

Grant: DKK 2.476.836

Two major challenges when using mouse models to model human cancers such as melanoma are that the human tumor cells transplanted to mice 1) represent the end-stage of the disease and 2) that the host animals are usually immunocompromised.

Thus, these models fail to actually show development of the disease and they fail to display the ongoing interaction between melanoma cells and the immune system as the disease progresses.

To curb these two shortcomings, the team led by Rudolf Jaenisch of Massachusetts Institute of Technology, has set out to create an experimental model system that will make it possible to study initiation, progression, and manifestation of human melanoma in immune competent host animals.

Their basis is generation of human-mouse neural crest chimeras – where mice embryos are introduced with human neural crest cells carrying the genetic dispositions alleged to lead to development of the particular cancer – and their goal is a model that has the potential to show how melanoma cells evade the immune system.

Given a positive outcome, this innovative project can help devise strategies to improve the effectiveness of current immunotherapies, to test novel immunotherapies, and to identify novel targets in melanoma treatment.

 

Project group

Malkiel Cohen, Postdoctoral researcher

Kristin Andrykovich, Graduate Assistant

Beneficiary: Prof. Dr. Andrew G. Myers, Amory Houghton Professor of Chemistry Harvard University, Cambridge, MA, USA

Grant: DKK 3.108.110

Many common skin infections are caused by the Gram-positive bacterial species Staphylococcus aureus and Streptococcus pyogenes. The infections lead to conditions ranging in severity from minor folliculitis to life threatening skin reactions. If not managed successfully, they may escalate into lethal systemic infections.

Commonly, these diseases are treated with clindamycin, a prototypical member of the wide-ranging so-called lincosamide antibiotic class. Its clinical importance is underlined by the World Health Organization’s listing of it as an essential medicine. In the past decades, however, prevalence of in particular lincosamide resistant Staphylococci and Streptococci has risen sharply. The rise threatens to diminish clindamycin’s usability in the future, even render it obsolete.

In the course of this project, the team led by Andrew G. Myers of Havard University, will seek to address this growing unmet medical need by synthetic discovery efforts focused on the lincosamide class.

The team’s preliminary results indicate that new lincosamides uncovered in this fashion are able to address the contemporary resistance threats: Many of the compounds designed, synthesised, and evaluated by the team to date have shown themselves active against multidrug-resistant clinical isolates of Staphylococci and Streptococci, and at the same time they demonstrate favourable pharmacokinetic and safety profiles.

The team expects that it can uncover new candidates displaying expanded spectra of action against MDR and Gram-negative bacteria. The expected results can then be used to advance refined lead candidates capable of demonstrating efficacy in in vivo murine models of skin infection, and thus yield substantial promise for further clinical development of actual treatments.

 

Project group

Dr. Amarnath Pisipati, Postdoctoral microbiologist

Matthew J. Mitcheltree, PhD student, chemistry

Ioana Moga, PhD student, chemistry

Katherine J. Silvestre, PhD student, chemistry

 

International affiliation

The Institut Pasteur, Annecy, France – International Course on Antibiotics and Resistance (ICARe), Organizing Committee, Core Faculty

Beneficiaries: Professor Gregor B. Jemec, Department of Dermatology, Zealand University Hospital, Roskilde, Denmark, and Assoc. Professor Ole B. V. Pedersen, Department of Clinical Immunology, Næstved Hospital, Denmark

Grant: DKK 5.770.000

In this study, the group led by Professor Gregor Jemec of Roskilde Hospital has set out to identify new genes for the development of a long line of common dermatological conditions, including deep skin infections, warts, fungal infections, and eczema.

Many of these common skin diseases are still poorly understood and the treatments often insufficient. A study of the genetics of these disorders will help increase the understanding of the pathogenic mechanisms. The study will have its origin in Denmark and be based on unique national biobanks, national registries, and with extensive genetic analyses done in collaboration with deCODE Genetics, Iceland.

This is possible due to the growing number of Danish large-scale biobanks as well as biobank based scientific studies suited for further genetic studies. The largest genetic study in Denmark is the Danish Blood Donor Study (DBDS) in which the genome wide association (GWA) arrays have been analysed on 110,000 research participants.

In addition to this cohort, Jemec’s group is currently pursuing genetic testing on the Copenhagen Hospital Biobank (CHB) that includes samples from around 350,000 patients. Both of these biobanks have established a collaboration with deCODE Genetics, Iceland – one of the leading genetic research centers in the world.

 

Project Group

Henrik Ullum, Professor, Department of Clinical Immunology, Rigshospitalet

Søren Brunak, Professor, Center for Protein Research (CPR), Copenhagen University

Simon Francis Thomsen, Professor, Department of Dermatology, Bispebjerg Hospital

Claus Zachariae, Professor, Department of Dermatology, Gentofte Hospital

 

International affiliations

Ingileif Jonsdottir, Professor, deCODE Genetics, Iceland

Errol Prens, Professor, Department of Dermatology, Erasmus University, Rotterdam, Netherlands

Christos Zouboulis, Professor, Department of Dermatology, Brandenburg Medical School Theodor Fontane, Dessau, Germany

Beneficiaries: Dr Simon G. Danby, Independent Research Fellow, The University of Sheffield Medical School, United Kingdom

Grant: DKK 390.506

With this grant, the group led by Simon G. Danby seeks a potentially important technological addition to the ongoing A longitudinal investigation of skin barrier development from birth and the validation of early predictors of Atopic dermatitis (AD) risk: the skin testing for atopic dermatitis risk (STAR) trial (see Grants 2017).

This addition may prove valuable to the group’s envisioned paradigm shift – from management of established AD to primary prevention of the condition.

More specifically, the group will include enhanced ATR-FTIR spectroscopy to quantify biomarkers of skin barrier condition and AD severity in newborns. While existing spectroscopy works in adults and children, its sensitivity has been proven unsatisfactory when measuring newborns.

Working with the equipment manufacturer, the group has developed a solution that increases sensitivity 6-fold. This increase can help better prediction of the risk of AD in the newborn and thus enable targeted emollient intervention right from birth – potentially leading to a reduction of the incidence of the condition as increasing evidence suggests that topical emollient therapy can prevent the initial onset of AD by 50%.

AD is a very common chronic inflammatory skin condition affecting around 20% of children worldwide. The disease often heralds development of allergic diseases such as food allergy, asthma, and allergic rhinitis.

Project Group

Prof. Michael J. Cork and Mr J. Chittock, The University of Sheffield, United Kingdom

Dame Prof. Tina Lavender and Dr Alison Cooke, The University of Manchester, United Kingdom