20 innovative ideas receive Off Road grant

Multipronged treatment of untreatable bacteria: hampering the membrane, efflux, and internal targets
Dr. A. Melcrová, University Medical Center Groningen
Resistant bacterial infections were responsible for almost 5 million deaths worldwide in a single year of 2019. Without any effective treatment, as is the case today, this number will only grow. Recently, I discovered a new antibiotic mechanism, in which the antibiotics disrupt the organization of the bacterial membrane. The full effects of these new antibiotics on bacteria are unknown. This research will test the hypothesis that the new antibiotics deactivate the proteins responsible for the efflux of intracellular antibiotics out of the bacteria, hence deleting the resistance of bacteria to intracellular antibiotics. If successful, this research will open a way for repurposing intracellular antibiotics that are approved for clinical use worldwide but cannot be effectively used due to the high resistance occurrence in bacteria. In the long term, we foresee the development of new treatments to treat the currently untreatable and deadly bacterial infections.

Measuring the unmeasurable: laser spectroscopy for non-invasive measurement of mitochondrial oxygen
Dr. A.L. Peters, Stichting Amsterdam UMC
Oxygen is essential for energy production in the human body. If too little oxygen reaches the tissues, the organs become damaged. This ultimately leads to death. Patients who experience insufficient oxygen supply to their organs are treated in the intensive care. The amount of oxygen is measured in the blood using a fingertip clip. However, it is not possible to measure the oxygen level directly in the organs. As a result, doctors cannot effectively monitor whether the organs are receiving enough oxygen. This study investigates whether the amount of oxygen in the organs can be measured with laser. For this purpose, a prototype spectrometer will be assembled and laser-based oxygen measurements are performed on the skin of healthy volunteers and compared with the oxygen concentration in the blood. This method could ultimately enable doctors to measure whether the organs are receiving enough oxygen. This can prevent organ damage and assist in optimizing treatments.

Intravital 3-/4-photon microscopy for deep-tissue detection of kidney tubule remodeling
Dr. P.A. Leermakers, Radboudumc
The kidney has a crucial function in regulating the salt balance in the body. High blood pressure is caused by a disturbed salt balance, and is associated with an increased risk of both cardiovascular diseases and kidney damage. Recent studies have shown that structural changes in the kidney can underlie a disturbed salt balance, but these changes are extremely difficult to map with current technology and are therefore not used in drug development. Here I describe the development of a new innovative microscopy technique which implements the latest developments in the field of pulsed lasers. This technique is able to measure at currently unreachable locations in the kidney. By using this out-of-the-box technique to simultaneously measure both structural changes and the regulation of the salt balance in living kidney, I am able to investigate how structural changes influence the salt balance in the kidney for the first time ever.

A molecular snapshot of the substantia nigra of living Parkinson’s Disease patients
Dr. G. van Mierlo, Radboudumc
Parkinson’s disease (PD) is a progressive neurodegenerative disorder and leads to symptoms such as tremors and impaired movement. While treatments like dopamine agonists provide temporary relief, they lose effectiveness over time. Deep Brain Stimulation (DBS) offers a promising alternative by delivering electrical signals to the substantia nigra (SN) via implanted electrodes. However, not all patients respond to DBS, and no reliable biomarkers exist to predict success.This project aims to molecularly profile SN cells from living PD patients, collected during DBS surgery. By leveraging cutting-edge techniques like single-cell RNA sequencing, we will define cell types, assess molecular signatures, and explore mechanisms driving neuron vulnerability in PD. These insights could revolutionize PD care by identifying biomarkers for DBS and advance our understanding of SN dysfunction, ultimately paving the way for more effective, personalized PD treatment.

VR-STEM: Virtual Reality Schema Therapy Exercises with parent and child Modes
Dr. E.C.D. van der Stouwe, University Medical Center Groningen
Experiential psychological therapies, such as EMDR or schema therapy, in which imagination exercises are used have become increasingly popular. While many patients benefit from these kinds of therapies, effects in a substantial group of patients with less mental imagery abilities stay behind. Therefore, in this project we will develop dialogues with virtual avatars that visually represent schema modi such as the 'punitive parent' and 'vulnerable child' and compare this with regular imaginary dialogues. Patients will be invited to the Virtual Reality lab one time to perform one of both exercises based on randomization. We expect that VR exercises are more effective, especially for patients with less mental imagery abilities. If the developed VR exercises are feasibile and usefull, than this potentially provides new treatment options for patients that currently can profit from schema therapy. Next steps would be to develop and investigate a therapy protocol existing of such VR exercises.

A novel angle: manipulating knee position to understand muscle deterioration during immobilization
Dr. ir. M.L. Dirks, Wageningen University
When people are sick, injured, or in hospital, even short periods of inactivity can cause rapid muscle loss and weakness which require a long recovery period. Despite decades of research, we still do not fully understand why this muscle deterioration occurs or how to prevent it. This project will test the novel concept of whether keeping muscles stretched while a leg is immobilized can help maintain muscle mass and function. In a randomized controlled study, healthy volunteers will have one leg immobilized for five days, either in a straight position or bent at a 60-degree angle. We will use advanced techniques like MRI scans, muscle biopsies, and stable isotope methods to track changes in muscle mass, strength, and metabolism. By studying how muscle stretch affects inactive muscles, we hope to find better ways to prevent muscle weakening during periods of physical inactivity. These insights could improve recovery strategies for patients, benefiting families and healthcare systems.

Small vesicles, great changes in treatment strategy: enriched milk EVs preventing gut barrier loss.
Dr. I.H. de Lange, Maastricht University
Baby’s with a suboptimal start, such as preterm birth, are at risk for a broad range of diseases later in life. On the short term, these infants suffer from gut barrier loss, predisposing to the severe gut disease necrotizing enterocolitis (NEC). Breastmilk promotes gut barrier function, but is not available to all neonates. Nutritional interventions using single breastmilk components work insufficiently, probably due to the complex nature of NEC requiring a multi-component intervention and the fact that the effect of digestion is omitted. In the current project, we aim to overcome both challenges with milk extracellular vesicles (EVs) as natural delivery system. Milk EVs are present in breastmilk, highly resistant to gastrointestinal digestion, and already contain components beneficial for gut health. By enriching milk EVs with mixtures of other beneficial breastmilk components, we aim to develop new interventions for optimal gut barrier preservation and long term health protection.

Fighting superbugs: repurposing an existing vaccine to provide cross-pathogen protection
Dr. A. Hendriks, Stichting Amsterdam UMC
Bacterial infections remain a major cause of death, and this will only increase within the next years because of emerging antibiotic-resistant bacteria (so called superbugs). New treatment options need to be developed to prevent future health care burden. Boosting the immune system through vaccination has been highly successful in reducing the global burden of infectious diseases. Unfortunately, there still are no effective vaccines for a number of major human pathogens, including antibiotic-resistant Staphylococcus aureus (MRSA). The current dogma is that vaccines have a narrow specificity, as it only provides protection for specific types of bacteria. The researcher challenges this dogma and will investigate in this project, at the molecular and functional level, how an existing, seemingly unrelated childhood vaccine may be employed to provide cross-pathogen protection against MRSA.

Induced Cancer Stem Cells as a Basis for Pancreatic Cancer Organoids
Dr. N.G. Kooreman, Erasmus Medisch Centrum
Pancreatic cancer is aggressive and difficult to treat. Laboratory-grown tumor clusters (organoids) provide a powerful platform for modeling drug responses to a patient's tumor, enabling personalized testing and identification of effective treatments. However, current organoid models rely on growing patient-specific tumor tissue, which is often influenced by prior treatments and takes significant time to generate, delaying drug response modeling at diagnosis. This project uses advanced stem cell technologies to create cancer stem cells with different genetic profiles from which 3D organoid models can be derived. By matching the genetic profile of a patient’s cancer to a biobank of organoids, this innovative approach addresses current limitations of organoids and allow for a genetically similar organoid to be available at time of diagnosis. In turn, this will allow for rapid and personalized drug testing and would help develop predictive models for drug responses in patients.

From HIV to Hope: pioneering super-antibodies for increased immune defense in vulnerable populations
Dr. L.M. Verhagen,  Radboudumc
Respiratory tract infections can be life-threatening, especially for the millions of people with weakened immune systems. For them, common infections can become severe, yet current treatments do not protect against newly emerging and harmful pathogens.This project takes an unconventional approach, exploring an unexpected source for a solution: people living with HIV. Due to their chronic infection, their immune systems produce unique, broadly protective antibodies. I aim to determine whether these antibodies could help safeguard vulnerable patients from respiratory infections in ways that existing therapies cannot. To investigate their potential, I will replicate these special antibodies in the lab, and test their ability to fight respiratory pathogens. If successful, this could pave the way for a groundbreaking new treatment, offering long-term broad protection to people at risk by preventing lung damage and improving the lives of those with weakened immunity.

Modeling stem cell transplantation in a dish
Dr. A.A. Soto Gamez, University Medical Center Groningen
Transplantation of stem/progenitor cells is a promising strategy to rescue organ function. However, its effectivity in the clinic is largely variable depending on the organ treated. Novel strategies to enhance stem cell transplantation are necessary but typically reach a bottleneck due to resource intensive animal models. Notably, such models often have limited translatability because of histological and immunological differences between humans and animal models. In this project, we will develop a New Approach Methodology where stem cell engraftment can be tested ex vivo using patient-derived biopsies and advanced culture models. We will use salivary gland stem/progenitor cell transplantation, now under investigation in a first-in-man clinical trial, to provide proof of concept of this methodology, and answer clinical questions that remain unanswered.

Innovative Models to Uncover Brain Vulnerabilities to Harmful Compounds During Fetal Development
Dr. N. Antón Bolaños, Universitair Medisch Centrum Utrecht
This project aims to create a pioneering platform to test the safety of drugs prescribed to pregnant women with medical conditions requiring ongoing treatment. As a proof of concept, we focus on lithium, a standard treatment for bipolar disorder, to evaluate its effects on fetal brain development. By innovating 3D stem-cell-based models that mimic vulnerable brain regions, such as the thalamocortical axis, we will investigate how these regions develop and interact under drug exposure. This system provides a controlled, animal-free approach to assess vulnerabilities, guide safer drug use during pregnancy, and generate critical insights into human brain development. The outcomes will support targeted therapeutic strategies, improve public health, and set a foundation for future drug safety evaluations.

Changes in Brain DNA: Elucidating the Human Brain-Specific Response to DNA Breaks
Dr. J.A. Kamp, Erasmus Medisch Centrum
By examining DNA across distinct cells within the same human brain, differences are found in the DNA among these cells. Having an increased amount of these DNA changes is linked to psychiatric brain disorders. DNA changes are caused by DNA damage and faulty repair, but mechanisms and consequences of DNA repair in brain remain poorly understood. I will make brain cells from human stem cells to investigate this. I will make DNA breaks in the human brain cells and then identify active repair mechanisms. I will also develop a technique to correct disease-causing DNA variants. This would form the basis for potential treatment for patients with a hereditary brain disorder. The results of this project will clarify how DNA changes occur in brain cells and may lead to novel insights for treating brain diseases.

(Phase) transforming the rigid perspective on Lewy bodies in Parkinson's disease
Dr. T.E. Moors, Stichting Amsterdam UMC
Parkinson’s disease is a common neurological disease with currently no cure available. In this disease, brain cells form inclusions (‘Lewy bodies’) containing the protein alpha-synuclein. An influential hypothesis has been that alpha-synuclein forms rigid aggregates that are packaged into Lewy bodies. However, the current models do not recapitulate spontaneous Lewy body formation by cells, hampering the development of effective therapies. Therefore, I will explore an alternative hypothesis by scrutinizing the relation between cellular transport and phase separation of proteins such as alpha-synuclein, inspired by recent insights in ALS and Huntington’s disease. Using highly advanced microscopy techniques, I will analyze these pathways in cultured cells and the postmortem brain tissue of donors Parkinson's disease. I hope my project will yield scalable cell models for Lewy body formation that contribute to developing highly-required new strategies to halt Parkinson's disease.

Exploiting aberrant proteoglycan expression profiles for targeted therapy of cancer
Dr. J.D. Campeiro, Erasmus Medisch Centrum
Triple-negative breast cancer (TNBC) is the most aggressive form of breast cancer. This subtype has a very poor prognosis. A successful strategy for the treatment of cancer is image-guided surgery and internal radiation for primary and advanced disease, respectively. These treatment methods have great potential for TNBC but they require a cancer cell-specific property against which the treatment can be targeted. However, this is a challenge for TNBC, as there is a lack of cancer cell-specific molecules. The proposal of this study is to use Crotamine, a peptide, to develop molecules for image-guided surgery and internal radiation of TNBC. Crotamine has a positive charge and has been shown to bind to proteoglycans (PG) that have a negative charge. Studies have shown that PG with a high negative charge are abundant on TNBC cells. This study will therefore use the properties of Crotamine and the abnormal pattern of PG in TNBC to develop effective treatment methods for the disease.

Disrupting the core: the effect of lymph node stromal cells on immunity in Lyme disease
Dr. M.M.J. van Gool, Stichting Amsterdam UMC
Lyme disease is a growing health concern caused by the Borrelia burgdorferi bacteria, transmitted through tick bites. Some patients struggle to develop long-term immunity against the bacteria, leading to persistent symptoms. This project aims to understand how Borrelia burgdorferi disrupts the structure of lymph nodes, which are essential for a strong immune response.Using innovative precision-cut human lymph node slices, we will study how the bacteria affect immune responses in a laboratory setting. Our goal is to uncover why some people do not build lasting protection and identify key mechanisms involved in this process. This research can help improve Lyme disease treatment and prevention strategies, ultimately benefiting patients by reducing long-term symptoms and improving recovery outcomes.

Immune regulation by molecular exclusion at the cell-matrix interface
Dr. M.V.F. Farrell, AMOLF
Balance is crucial in the immune system as over-activation leads to autoimmunity and under-activation can result in pathogens and cancerous cells going unnoticed. To maintain this balance immune cells are equipped with inhibitory receptors that recognise changes in their environment and send 'stop' signals to prevent immune overreaction and damage. How inhibitory receptor recognition of the environment leads a to signal in the cell is not understood. In this project, I will research the mechanism behind this interaction by seeing immune cells in action using powerful microscopes. I will combine two microscopy approaches to visualise immune cells in their 3D environment, and then zoom right down to the nanoscale to see the inhibitory receptors themselves. In doing this, fundamental questions of how inhibitory immune receptors activate will be resolved and this knowledge will support the development of drugs that help to direct and regulate the immune system in cancer and inflammation.

Molecular unravelling the FGR placenta
Dr. J. Kooiman, Erasmus Medisch Centrum
Fetal growth restriction (FGR) complicates 5:1000 pregnancies and is an important cause of preterm birth, neonatal morbidity and mortality. The molecular mechanisms behind the placental insufficiency causing FGR are poorly understood and therapeutic options are non-existent. I will use a state-of-the-art molecular microscope able to classify cellular sub-types using artificial intelligence and highlight active metabolic pathways to study the FGR placenta. Gaining understanding of the functional pathways and tissue homeostasis of the placenta has major scientific impact as it is of vital importance to prevent serious obstetric complications such as FGR, but also preterm birth and intra-uterine fetal death.

Studying amyloid disassembly by protein design
Dr. N. J. Rinzema, Hubrecht Institute
Proteins have essential functions in the body. When they are not folded properly they may aggregate and thereby inflict damage. Alzheimer’s and Parkinson’s disease are examples of disorders associated with such protein clumps or “fibrils”. I will try to tackle these fibrils with molecular machines that will disturb their structure. By studying how they do that I will try to understand how fibrils grow or shrink in the body. Maybe I will even find new strategies for the design of therapeutics for Alzheimer’s and Parkinson’s disease. The design of these kinds of machines has only recently been made possible by breakthroughs in algorithms for the prediction of protein structures by means of artificial intelligence.

RabSwab — using wound swabs of dog-bite-victims as a novel rabies virus surveillance tool
Dr. C.W.E. Embregts, Erasmus Medisch Centrum
Rabies is a neglected disease caused by the rabies virus and related viruses, transmitted through the saliva of infected animals. Infection leads to encephalitis, which is 100% lethal due to the lack of effective treatments. Eradicating rabies is challenging, as the virus circulates among dog and wildlife populations. Accurate surveillance is crucial for informing policy-making, including mass dog and wildlife vaccination efforts and awareness campaigns. However, current rabies surveillance programs severely underestimate the problem, leading to delays in vaccination programs, rising human cases and fatalities, and setbacks in disease eradication. This underscores the urgent need for novel and accurate surveillance tools. In this proposed study, I will evaluate the performance of wound swabs from dog-bite victims for detecting viral RNA. This approach is sensitive, easy to implement, and has the potential to serve as a real-time surveillance tool with minimal infrastructure.