ERC FUNDED PROJECTS

Pain is one of the most problematic symptoms of rheumatic disease such as rheumatoid arthritis (RA) and fibromyalgia (FM). We have earlier discovered that antibodies (immunoglobulin, IgG) purified from blood of seropositive rheumatoid arthritis (RA) patients induce pain-like behavior when transferred to mice, independent of inflammatory reactions. Even though FM is not considered an autoimmune disease, it has been suggested that neuroimmune dysregulation contribute to the pathogenesis. Therefore, we purified IgG from FM patients and found that also IgG from FM patients, but not healthy controls, have pronociceptive properties in mice, and surprisingly, bind to satellite glial cells in dorsal root ganglia. Our findings highlights the importance of expanding our view on which chronic pain conditions that could have an underlying autoimmunity as part of the pain pathology. Thus, the overall objective of this project is to investigate both general, and disease specific, pain-inducing mechanisms mediated by RA and FM IgG. Objective 1. Investigate how IgG from RA and FM patients induce pain-like behavior after transfer to mice Objective 2. Search for RA and FM IgG induced maladaptive changes in sensory neurons that mediate hyperexcitability and long-term pain-like behavior Using patient and healthy control samples, in vivo mouse behavioral assays, primary neuronal and non-neuronal cell cultures together with stat-of-the-art methodology, we will investigate how RA and FM-associated autoantibodies alter sensory neuronal excitability. If successful our project will not only challenge the view of how antibodies can contribute to pain but also pin-point specific mechanisms by which disease-relevant antibodies induce and maintain pain independent of previously described inflammatory mechanisms. Such findings promise to resolve currently unanswered questions concerning symptoms of pain in RA and FM, and to pave the way for the development of new pain-relieving therapies.

1 993 763 €

Start date: 2020-10-01, End date: 2025-09-30

The human hematopoietic system is a paradigmatic, stem cell-maintained organ with enormous cell turnover. Hundreds of billions of new blood cells are produced each day. The process is tightly regulated, and susceptible to perturbation due to genetic variation. In this project, we will explore an innovative, population-genetic approach to find regulators of blood cell formation. Unlike traditional studies on hematopoiesis in vitro or in animal models, we will exploit natural genetic variation to identify DNA sequence variants and genes that influence blood cell formation in vivo in humans. Instead of inserting artificial mutations in mice, we will read out ripples from the experiments that nature has performed during evolution. Building on our previous work, unique population-based materials, mathematical modeling, and the latest genomics and genome editing techniques, we will: 1. Develop high-resolution association data and analysis methods to find DNA sequence variants influencing human hematopoiesis, including stem- and progenitor stages. 2. Identify sequence variants and genes influencing specific stages of adult and fetal/perinatal hematopoiesis. 3. Define the function, and disease associations, of identified variants and genes. Led by the applicant, the project will involve researchers at Lund University, Royal Institute of Technology and deCODE Genetics, and will be carried out in strong environments. It has been preceded by significant preparatory work. It will provide a first detailed analysis of how genetic variation influences human hematopoiesis, potentially increasing our understanding, and abilities to control, diseases marked by abnormal blood cell formation (e.g., leukemia).

2 000 000 €

Start date: 2018-10-01, End date: 2023-09-30

The research I propose here will provide an enabling technology; spatially resolved transcriptomics, to address important problems in cell- and developmental-biology, in particular: How are stem cells in the skin and gut proliferating without turning into cancers? How are differentiated cells related, in their transcriptome and spatial positions, to their progenitors? To investigate these problems on a molecular level and open up paths to find completely new spatiotemporal interdependencies in complex biological systems, I propose to use our newly developed DNA-origami strategy (Benson et al, Nature, 523 p. 441 (2015) ), combined with a combinatorial cloning technique, to build a new method for deep mRNA sequencing of tissue with single-cell resolution. These new types of origami are stable in physiological salt conditions and opens up their use in in-vivo applications. In DNA-origami we can control the exact spatial position of all nucleotides. By folding the scaffold to display sequences for hybridization of fluorophores conjugated to DNA, we can create optical nano-barcodes. By using structures made out of DNA, the patterns of the optical barcodes will be readable both by imaging and by sequencing, thus enabling the creation of a mapping between cell locations in an organ and the mRNA expression of those cells. We will use the method to perform spatially resolved transcriptomics in small organs: the mouse hair follicle, and small intestine crypt, and also perform the procedure for multiple samples collected at different time points. This will enable a high-dimensional data analysis that most likely will expose previously unknown dependencies that would provide completely new knowledge about how these biological systems work. By studying these systems, we will uncover much more information on how stem cells contribute to regeneration, the issue of de-differentiation that is a common theme in these organs and the effect this might have on the origin of cancer.

1 923 263 €

Start date: 2017-08-01, End date: 2022-07-31

Mitochondria are required to convert food into usable energy forms and every cell contains thousands of them. Unlike most other cellular compartments, mitochondria have their own genomes (mtDNA) that encode for 13 of the about 90 proteins present in the respiratory chain. All proteins necessary for mtDNA replication, as well as transcription and translation of mtDNA-encoded genes, are encoded in the nucleus. Mutations in nuclear-encoded proteins required for mtDNA maintenance is an important cause of neurodegeneration and muscle diseases. The common result of these defects is either mtDNA depletion or accumulation of multiple deletions of mtDNA in postmitotic tissues. The long-term goal (or vision) of research in my laboratory is to understand in molecular detail how mtDNA is replicated and how this process is regulated in mammalian cells. To this end we use a protein biochemistry approach, which we combine with in vivo verification in cell lines. My group was in 2004 the first to reconstitute mtDNA replication in vitro and we have continued to develop even more elaborate system ever since. In the current application, the major focus is studies of the mitochondrial D-loop region, a triple-stranded structure in the mitochondrial genome. The D-loop functions as a regulatory hub and we will determine how initiation and termination of mtDNA replication is controlled from this region. We will also determine the physical organization of the mtDNA replication machinery at the replication fork and establish how mtDNA deletions, a classical hallmark of human ageing, are formed.

1 999 985 €

Start date: 2016-11-01, End date: 2021-10-31