Hematopoietic stem cell transplantation (HSCT) is a groundbreaking therapy that has transformed the treatment of severe hematological and immune disorders. It involves infusing healthy or corrected hematopoietic stem cells (HSCs) into a patient to restore their blood cell production. Despite its remarkable success, HSCT is currently limited to the most severe cases due to several key challenges. These bottlenecks include inefficient HSC mobilization, limited engraftment leading to insufficient clonal diversity, and the toxicity of chemotherapy-based conditioning regimens. This comprehensive proposal outlines a transformative approach to address these limitations head-on, enhancing both the efficacy and safety of HSCT. The project aims to: 1. Uncover the key factors governing HSC engraftment and egress: Using optimized genome-wide gain- and loss-of-function in vivo screening in HSCs, I will identify the pivotal players responsible for HSC engraftment and egress. These insights will inform the development of innovative mobilization strategies and the engineering of HSCs to confer a transient engraftment advantage. 2. Establish a mobilization-based conditioning regimen: I will exploit the temporary niche depletion induced by mobilizers to optimize a genotoxic-free regimen. By harnessing cell surface profiling data and potentially using mobilizer-resistant variants generated through directed evolution, I will tilt the competitive balance between freshly mobilized and infused HSCs, favoring the engraftment of infused cells. This project seeks to pioneer avant-garde methods that enhance mobilization and engraftment while avoiding the use of toxic conditioning regimens, thereby addressing major HSCT limitations. These advancements hold the potential to transform HSCT into a safer, more broadly applicable therapy, extending its benefits to a wider demographic and diverse healthcare settings. This aligns with the ultimate objective of achieving global health equity.

Guillain-Barré syndrome (GBS) is a rare heterogenous inflammatory peripheral neuropathy usually triggered by a preceding infection and causing a potentially life-threatening progressive muscle weakness. While the disease is generally considered to have an autoimmune basis, the underpinning immune-mediated mechanisms remain mostly elusive. This aspect poses a significant medical challenge in terms of accurate diagnosis, prognosis, and treatment. Building on our recent findings revealing that autoreactive T cell immunity play a key role in a subset of GBS patients, the project proposed herein aims at systematically elucidating its relative contribution to distinct GBS subtypes and disease stages. To this end, we will use cutting-edge technical approaches that enable the study of potentially rare autoreactive T cells in human biological samples (i.e. blood, cerebrospinal fluid and tissue biopsies) through three specific aims: Aim 1 is to perform in-depth immunophenotyping of blood-circulating immune cells for identification of potential variations in their frequency and distribution across GBS subtypes and stages; Aim 2 is to provide a systematic characterization of self-reactive T cells in patients’ biological samples by combining in vitro T cell screenings and ex vivo tetramers staining with high-throughput single cell analysis, such as single T cell clone generation, single cell RNA sequencing and TCR sequencing; Aim 3 is to dissect the cellular and molecular bases of cross-reactive T cell immunity in GBS by integrating in vitro sequential stimulation, single T cell clone generation and TCR sequencing with mass spectrometry-based immunopeptidomics or lipidomics. The outcomes of this project will significantly expand our understanding of the immunopathology of inflammatory peripheral neuropathies and advance our basic knowledge of human autoreactive T cell biology, with substantial implications for biomedical applications.

Due to an aging population and the spiralling cost of brain disease in Europe and beyond, EDEN2020 aims to develop the gold standard for one-stop diagnosis and minimally invasive treatment in neurosurgery. Supported by a clear business case, it will exploit the unique track record of leading research institutions and key industrial players in the field of surgical robotics to overcome the current technological barriers that stand in the way of real clinical impact. EDEN2020 will provide a step change in the modelling, planning and delivery of diagnostic sensors and therapies to the brain via flexible surgical access, with an initial focus on cancer therapy. It will engineer a family of steerable catheters for chronic disease management that can be robotically deployed and kept in situ for extended periods. The system will feature enhanced autonomy, surgeon cooperation, targeting proficiency and fault tolerance with a suite of technologies that are commensurate to the unique challenges of neurosurgery. Amongst these, the system will be able to sense and perceive intraoperative, continuously deforming, brain anatomy at unmatched accuracy, precision and update rates, and deploy a range of diagnostic optical sensors with the potential to revolutionise today’s approach to brain disease management. By modelling and predicting drug diffusion within the brain with unprecedented fidelity, EDEN2020 will contribute to the wider clinical challenge of extending and enhancing the quality of life of cancer patients – with the ability to plan therapies around delicate tissue structures and with unparalleled delivery accuracy. EDEN2020 is strengthened by a significant industrial presence, which is embedded within the entire R&D process to enforce best practices and maximise translation and the exploitation of project outputs. As it aspires to impact the state of the art and consolidate the position of European industrial robotics, it will directly support the Europe 2020 Strategy.

Liver metastases commonly develop in up to 50% of patients with various cancer types. The most common cancer that metastasizes to the liver is colorectal cancer (CRC). At least 25% of CRC patients develop colorectal liver metastases (CRLM) during their illness. CRLM represent the major unmet clinical need for this malignancy, as the 5-year survival rate of patients with unresectable disease does not exceed 2%. New therapies that promote antitumor immunity have been recently developed, mostly focusing on enhancing T cell responses. Although these therapies have led to unprecedented successes, only a minority of patients benefit from these treatments, highlighting the need to identify new cells and molecules that could be exploited in next generation immunotherapies. Given the crucial role of innate immune responses in immunity, targeting these responses opens up new possibilities for tumour control. We hypothesize that the immunotherapy of liver metastases can be significantly improved through harnessing the biology of innate lymphoid cells (ILC), such as Natural Killer (NK) cells and ILC1s, and myeloid cells such as macrophages and DCs. Our team brings together experts in the biology of tissue-resident myeloid (Ginhoux, PI4) and lymphoid (Gasteiger, cPI) cells, in liver immunology (Fumagalli, PI3), and in the development of novel immunotherapeutic strategies that modulate immune cells in the fight against cancer (Vivier, PI2). By combining cutting-edge single cell and spatial transcriptomics of human patient samples with cross-species analyses in advanced genetic mouse models, we aim (1) to identify cellular interactions defining the metastatic tumor microenvironment across murine and human tissue-specimens, (2) to investigate immune cell functions regulating metastatic disease using a unique combination of advanced genetic mouse and human tissue models, and (3) to harness the anti-tumoral functions of innate immune cells via next generation cell engagers.

With falling birth rates and increasing life expectancy we are an ageing society. 20% of boys and 25% of girls born in 2019 are expected to reach their 100th birthday. This would be good news if it were not for the fact that healthy life span has not kept pace with increasing longevity and now on average adults spend the last 15-20 years of life in ill health. Frailty is a major component of ill health in old age and refers to an enhanced vulnerability to stressors, such as falls, surgery or infections, which was demonstrated clearly in the mortality data for the COVID19 pandemic. The transition from robust health to frailty is a critical factor in the loss of independence and places increased pressure on health and social care. All of this has led governments to prioritise the enhancement of healthspan. The Doctorate Network on UnderstandiNg fraIlty tOwards a future of healthy ageiNg (UNION) is a multi-partner joint doctoral research training network with the overall aims of educating 13 Early Stage Researchers (DCs), advancing the current understanding of frailty, and providing innovative solutions on how healthy ageing can be achieved. UNION brings together world leaders in a range of relevant disciplines (frailty, ageing biology and ageing medicine, inflammation, immunosenescence, immunometabolism, stem cell biology) with state-of-the art technologies including mass spectrometry metabolomics, advanced imaging, artificial intelligence, CRISPR-CAS9 libraries, SPECTRA 35-colour flow cytometry, global and conditional knockout mice. This technological excellence is applied to the clinical situation through unique longitudinal human ageing and frailty cohorts, longitudinal assessment of age-related multimorbidity in animal models and innovative multidimensional frailty indices in humans. Together this integrated activity will provide the highest quality training and research environment in our rapidly ageing society.