Investigating the role of human periostin in healthy skin and severe eczema

Grantee: Jan J. Enghild, Aarhus University

Amount: DKK 3,045,231

Grant category: Research Grants in open competition

Year: 2018

Geography: Denmark

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.

Endosomal Chemokine Receptor Signaling as Basis for Metastasis in Malignant Melanoma

Grantee: Alex Rojas Bie Thomsen, Columbia University Medical Center

Amount: DKK 3,600,308

Grant category: Research Grants in open competition

Year: 2018

Geography: USA

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.

Developing a Cell-Based Therapy for Alopecia

Grantee: George Cotsarelis, Perelman School of Medicine, University of Pennsylvania, Philadelphia

Amount: DKK 3,793,808

Grant category: Research Grants in open competition

Year: 2018

Geography: USA

Androgenetic alopecia (AGA, Male or Female Pattern Baldness) is the most common type of hair loss, affecting approximately 50% of men and 30% of women by the age of 50.

Current therapies, including pharmaceutical and surgical interventions, are either marginally effective or expensive with significant limitations. Over the last decade, breakthroughs made in the field of adult stem cells have laid the foundation for a cell-based approach to tissue and organ regeneration. Cell-based therapies will comprise a new wave of medical breakthroughs.

In this study we propose to produce human hair follicles from induced pluripotent stem (iPS) cells by directing these cells to form the two types of cells that are needed for human hair formation, namely the human hair follicle epidermal cells and the hair inductive dermal fibroblasts.

We will combine our hair biology and tissue-engineering expertise to generate early stage human hair follicles in culture that can be implanted into an animal model to grow into a mature hair. The long-term goals are to develop an innovative cell-based treatment for hair loss and an in vitro platform for testing hair growth compounds.

Psoriasis: a microbiome-driven disease?

Grantee: Patrick Zeeuwen, Radboud University Medical Center, Nijmegen

Amount: DKK 2,545,944

Grant category: Research Grants in open competition

Year: 2018

Geography: Netherlands

Psoriasis is highly prevalent and has a significant medical and socio-economic impact.

The prevailing dogma has been that abnormalities of the adaptive immune system were primary, but genetic studies have highlighted the importance of local skin-specific factors. We and others have identified epidermis-specific innate immunity genes, like beta-defensins and Late Cornified Envelope (LCE) genes, to be associated with disease development.

We recently made two exciting observations. First, deletion of LCE3B and LCE3C does not merely imply the loss of two genes but has a genomic effect that leads to a strong induction of the flanking LCE3A gene. Secondly, we found that LCE proteins, and LCE3A in particular, have broad-spectrum antimicrobial activity. We hypothesize that the LCE3B/C-del status affects the cutaneous host defense repertoire thereby shaping the skin microbiome. We aim to investigate the biology of LCE genes and to translate these findings to our understanding of psoriasis pathogenesis.

Key objectives are to:

  1. assess the antimicrobial activity and specificity of all LCEs and their synergy with other antimicrobial proteins. This will be investigated by metagenomic approaches and classical in vitro microbiological assays, using recombinant and synthetic LCE proteins and peptides derived thereof
  2. investigate LCE3B/C-del in isogenic 3D-skin equivalents in vitro generated from the immortalized human keratinocyte N/TERT cell line. Deletions of other LCE genes or their regulatory sequences will be made using CRISPR/Cas9 technology. Effects on epidermal biology relevant to psoriasis will be studied and include antimicrobial host defense, innate immune response and skin barrier function

Investigating the tumor suppressive functions of Notch signaling during skin cancer initiation and progression

Grantee: Sunny Y. Wong, Assistant Professor, University of Michigan, Ann Arbor

Amount: DKK 2,486,354

Grant category: Research Grants in open competition

Year: 2018

Geography: USA

Basal cell carcinoma (BCC) is the world’s most common cancer and is defined by uncontrolled activation of the Hedgehog (Hh) signaling pathway.

Although previous studies have suggested that elevated Hh may be sufficient for BCC formation, mutations in the Notch pathway are also commonly observed. Furthermore, Notch-deficient mice are susceptible to forming BCCs, and our recent studies have shown that Notch can modulate tumor-drug response.

These studies seek to understand whether Notch affects multiple aspects of BCC tumorigenesis. Using a combination of animal studies and human BCC specimens, we will investigate how Notch modulates tumor progression and stem cell origin.

We will also model in mice a recent clinical trial, where Alzheimer’s patients treated with a Notch inhibitor reported increased incidence of BCC. We hypothesize that Notch may suppress tumorigenesis at multiple levels by controlling cell differentiation, apoptosis and turnover, similar to its function in normal skin and hair follicles.

These studies will ultimately build on the novel premise that BCCs may originate from a precursor lesion. Given that Notch mutations are the most commonly observed genetic aberrations in human skin, a deeper understanding of the tumor suppressive properties of this pathway is urgently needed.

Characterizing the disease memory in atopic dermatitis

Grantee: Patrick M. Brunner, Medical University of Vienna

Amount: DKK 2,920,541

Grant category: Research Grants in open competition

Year: 2018

Geography: Austria

Atopic dermatitis (AD), the most common chronic inflammatory skin disease, typically starts very early in life.

While many patients outgrow their disease, some develop chronic disease for the rest of their lives. Mechanisms responsible, however, are completely unknown, and no biomarker exists that can predict the course of the disease.

Thus, we want to compare skin from young adults that have outgrown their AD, with skin from patients with active disease (namely normal appearing AD under topical glucocorticoid treatment, which can be expected to flare up again after cessation of treatment, thus harbouring a “disease memory”).

Skin from healthy control subjects will serve as baseline comparators. Due to low immune cell numbers in this type of tissue, we want to use in vivo suction blistering of AD patients to obtain (i) skin resident immune cells and (ii) skin proteins. Suction blister fluid will be analysed with low cytometry and single cell RNAseq (for cells) as well as a proteomic multiplex assays (OLINK) for soluble proteins. The blister roof (i.e. the epidermis) will also be harvested, and keratinocytes will be stored in liquid nitrogen for functional experiments.

Results obtained from flow cytometry, single cell RNAseq and proteomic approaches will then be used for such functional in vitro experiments (e.g. co-culturing, skin equivalents, stimulation experiments) in future research projects.

Overall, we hope that the identification of cellular and/or molecular factors influencing the natural course of AD could possibly identify targets for novel therapeutic approaches in AD, that could induce long term remission – or even lead to a cure – of AD.

Compartmentalized and Systemic Interactions of the Skin Microbiome in Cancer Immunotherapy Response

Grantee: Julia Oh, Jackson Laboratory, Farmington, Connecticut

Amount: DKK 2,107,529

Grant category: Research Grants in open competition

Year: 2018

Geography: USA

My vision is to use metagenomics to better predict patient responses to immunotherapy and rationally design microbial adjuvant cocktails and engineered microbes to improve therapeutic outcomes.

However, a central question is the role of the local microbiota vs. systemic effects in potentiating these immunotherapeutics. In skin cancer, we have been studying how the skin microbiome affects predisposition and progression. Specific gut microbes have been implicated in the outcomes for immunotherapy response in melanoma skin cancer, supporting a role of systemic immune interactions via the gut in potentiating immunotherapy response.

However, because many aspects of cutaneous immunity are compartmentalized from systemic immune effects, we hypothesize that the skin microbiome could uniquely impact skin cancer outcomes during immunotherapy by modulating the cutaneous immune milieu.