Patrik Ernfors

Pain in rheumatoid arthritis (RA) is highly debilitating, impacts quality of life and lacks adequate treatment options. Although biologics have proven benefits on inflammatory disease activity, up to 1 in 5 of the treated patients still report continuous significant problems with pain and fatigue. Recent extensive experimental work from co-applicant Patrik Ernfors’ research group reveals strong evidence for a causative pain pathway involving the Type 1 interferon (IFN) system (IFN1). Our proposal includes a translational approach with analysis of patient reported outcomes including remaining pain in an early RA cohort, effects of biologics and potential associations with Type 1 IFN signaling and continous experimental mapping of the background mechanisms of this novel pain signaling pathway. We will also plan for a RCT with current available IFN-blockade, anifrolumab (Saphnelo) in RA with remaining pain.The proposal has very good potential to answer important questions on effects of biologics on pain and other patient reported outcomes. The recently identified causative pathway through IFN1 constitutes a potential breakthrough in general pain research, with vast clinical implications. The increased knowledge will also lay the ground for development of new and specific targeted treatment strategies for remaining pain in RA and other pain conditions. Thereby the results may have vital importance on future treatment strategies both nationally and in an international perspective.

Pain is a result of the activation of molecularly unique sensory cell types that form assemblies of different neuronal and glial cells, located in the dorsal root ganglion (DRG). Under normal circumstances, pain is beneficial as a protective mechanism. However, under pathological conditions, painful stimuli cause stronger pain, and even non-painful stimuli can cause unpleasantness. To understand mechanisms of chronic pain we need to identify the causative molecular perturbations in the pain causing cell types. In this proposal, I will employ multiomics sequencing on the single cell level in the mouse DRG, namely single cell RNA sequencing to determine molecularly defined sensory cell types combined with single cell ATAC sequencing to identify open chromatin regions, which allows to identify enhancer gene regulatory networks in the respective sensory cell types. I will compare the gene expression and chromatin topology in the naïve mouse to chronic pain mouse models to find enhancer elements responsible for chronic pain. I aim at utilizing the discovered cell-type and disease specific DNA regulatory elements to silence the activity in the pain causing cell types in vivo. Therefore, I will designing pain specific enhancer constructs which will be delivered to the mouse sensory cell types in vivo using adeno-associated viruses and lipid nanoparticles. The constructs will drive the expression of a optogenetic tool fused to a flourescent protein to allow validation of the delivery using immunohistochemical methods and functional silencing of the neurons by light stimulation and sensory behavior analysis in the mice. This application is timely, since technologies for identifying accessible chromatin in single cells, deep learning methods to design optimal cell type-specific enhancers and lipid nanoparticle technologies are just emerging, opening for discoveries previously unreachable. I anticipate that deep insights into the cellular and molecular basis of chronic pain will be enabling for highly needed new therapeutic options to treat pain.

Pain is a problem affecting almost 20% of the population and despite an increase in the prevalence of pain-related disorders there has been a general failure in the development of pain relieving drugs. A major reason is that the cellular basis for pain transduction is not understood. Due to technical limitations, an unbiased system-wide approach to resolve the complexity of cell types and their involvement in the development of pain has yet not been tried.The proposed environment will make use of the recent discovery and molecular classification of pain transducing neurons using single-cell RNA sequencing (scRNAseq) by the applicant. Based on this, we will determine the cellular basis for pain by establishing the role of individual cell types in various chronic pain disorders and by developing enabling technologies allowing an activity-based Cre-dependent permanent labeling and identification by scRNAseq the exact cell types transducing particular types of pain. The role of these cells and cell assemblies will be functionally studied by genetic silencing, ablation or artificial activation in mouse models. Thus, this work will for the first time reveal the full complexity of different cell types engaged in particular types of pain, and unravel by mouse genetics the role of that these play in chronic pain disorders. The multidisciplinary nature requires recruitment of new expertize, building knowledge, expertize and experimental platforms with support functions.

Each year, about 400 people are affected by glioblastoma in Sweden. It is the most aggressive type of brain cancer. Despite surgical, radiation, and chemotherapy combination therapies, average survival is only 15 months. A tumor does not consist of a type of cancer cells, but many different types with different properties. Drugs that have an effect on a tumor cell type leave the others untouched. It is therefore a great challenge to find better treatment methods for this type of malignant brain tumor. In the project, we identify exactly which cell types are present in a glioblastoma (GBM) tumor and how they differ between patients. We identify cell types by so-called single cell RNA sequencing. Advanced analysis of such large-scale experiments answers the molecular properties of different types of cells and allows us to identify common traits that could be useful for finding new strategies for treating the disease. We will also test in mouse models of human GBM whether the identified tumor-specific properties that could be used in a possible therapy also have an effect on tumor development and survival. In recent years, various strategies for treating GBM have been tried. These have to a large extent been focused on "correcting" biological processes in the cells that contribute to the cancer. But many different changes occur in the cells and in addition various changes in different cells in the same tumor. The goal of our project is instead to find common changes that can be used to find new possible treatment strategies. We also hope to explain from which cells in the brain that GBM occurs, which could partly explain why different GBMs differ so much.

Each year, about 400 people are affected by glioblastoma in Sweden. It is the most aggressive type of brain cancer. Despite surgical, radiation, and chemotherapy combination therapies, average survival is only 15 months. It is therefore a great challenge to find better treatment methods against this type of malignant brain tumor. We have discovered that the substance Vacquinol-1 can burst the cancer cells by causing the cells to bud off their cytoplasmic membrane into the cell so that the membrane that previously encircled it forms small bubbles inside the cell (called vacuolation). When we expose cancer cells to Vacquinol-1, it eventually becomes so many bubbles that the cell does not have enough membrane left and then it bursts and dies. The mechanism is unique to cancer cells as it appears that when the glial cells in the brain turn into cancer cells, they get an acquired hypersensitivity to vacuolation. The origin cells in the brain do not have that hypersensitivity and therefore they are not affected by Vacquinol-1. A possible drug based on this principle would thus attack glioblastoma in a whole new way, and it might also be able to function with other cancer disease. The proposed research aims to increase understanding of exactly how Vacquinol-1 works in the cells and how it can best be used to eliminate brain cancer in animals bearing human tumors. The hope is that we can generate enough results to be able to go ahead and test the molecule in clinical trials on patients affected by glioblastoma. We also hope to be able to clarify whether the hypersensitivity to vacuolation exists in other types of cancer and in such cases try in animal models if a treatment with Vacquinol-1 can inhibit / cure the cancer.

Each year, about 400 people are affected by glioblastoma in Sweden. It is the most aggressive type of brain cancer. Despite surgical, radiation, and chemotherapy combination therapies, average survival is only 15 months. It is therefore a great challenge to find better treatment methods against this type of malignant brain tumor. We have discovered that the substance Vacquinol-1 can burst the cancer cells by causing the cells to bud off their cytoplasmic membrane into the cell so that the membrane that previously encircled it forms small bubbles inside the cell (called vacuolation). When we expose cancer cells to Vacquinol-1, it eventually becomes so many bubbles that the cell does not have enough membrane left and then it bursts and dies. The mechanism is unique to cancer cells as it appears that when the glial cells in the brain turn into cancer cells, they get an acquired hypersensitivity to vacuolation. The origin cells in the brain do not have that hypersensitivity and therefore they are not affected by Vacquinol-1. A possible drug based on this principle would thus attack glioblastoma in a whole new way, and it might also be able to function with other cancer disease. The proposed research aims to increase understanding of exactly how Vacquinol-1 works in the cells and how it can best be used to eliminate brain cancer in animals bearing human tumors. The hope is that we can generate enough results to be able to go ahead and test the molecule in clinical trials on patients affected by glioblastoma. We also hope to be able to clarify whether the hypersensitivity to vacuolation exists in other types of cancer and in such cases try in animal models if a treatment with Vacquinol-1 can inhibit / cure the cancer.