Host Institution: Assistance Publique Hopitaux De Paris (France)
Supervisor: Paul Coppo, Agnès Veyradier
Duration: 36 months

Introduction: Immune-mediated TTP has become a disease of favorable prognosis at the acute phase. However, relapses and comorbidities are prevalent after the acute phase. Comorbidities, especially cardiovascular risk factors, combined with relapses in patients unresponsive to usual immunosuppressive strategies, shorten life expectancy in these patients. These unmet needs will be addressed here.

Aims
Science: 1: To identify risk factors associated with morbidity and shortened life expectancy following an acute iTTP episode; Science: 2: To address whether ADAMTS13 activity levels are predictive of morbidity and shortened life expectancy; Science: 3: To gain clinical experience in the use of immunomodulators for the prevention of clinical relapses in patients refractory to rituximab; Science: 4: To address whether newest therapeutic regimens at the acute phase and preemptive treatments during follow-up can reduce the incidence of comorbidities and prevent premature death. Training: 1: Experience in clinical diagnosis, treatment and follow-up of patients with iTTP; Training: 2: Learn how to approach a patient suffering of iTTP; Training: 3: Management of clinical databases and statistical analyses; design and execution of research based on registry data.

Expected Results: To identify risk factors linked to morbidity and shortened life expectancy; adapt treatment to improve life expectancy and quality of life. To provide definitive evidence about whether abnormal ADAMTS13 levels during follow-up exposes patients to more comorbidities, such as ischemic stroke, and if ADAMTS13 activity improvement affects the prevalence of comorbidities.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) SMW, 6 months: To understand the immune response in iTTP during long-term follow-up; (3) Werfen(Biokit Research & Development S.L.U.), 1 month: To get acquainted with commercialization of commercial assays.

Enrolment in Doctoral degree(s): DC will be enrolled at Sorbonne Université

Host Institution: Katholieke Universiteit Leuven (Belgium)
Supervisor: Karen Vanhoorelbeke
Duration: 48 months

Introduction: Universal chimeric antigen receptor (UniCAR)-T cell therapy was originally developed to treat cancer by HZDR. Here UniCAR-T cell therapy will be introduced for the first time to treat autoimmune diseases. The disease model used will be the autoimmune disease iTTP. The efficacy of the UniCAR-T cell therapy will be studied in vitro and in vivo in a mouse model of iTTP.

Aims
Science: 1: To generate UniCAR-T cells and the CS- and CUB-targeting modules; Science: 2: To study in vitro efficacy as well as the in vivo efficacy of UniCAR-T cell therapy in a mouse model of iTTP; Science: 3: To compare UniCAR-T cell therapy with other innovative immune therapies (WP2 and WP3). Training: 1: To learn to generate UniCAR-T cells via lentiviral transduction and transduce T cells, to isolate T cells via FACS sorting to learn to perform cytotoxicity assays and how to use the preclinical mouse model for iTTP (trigger, sign and symptoms).

Expected Results: Having designed human and murine UniCAR-T cells and CS-and CUB-targeting modules; Having realized in vitro cytotoxicity using the UniCAR-T cell therapy; Having demonstrated the UniCAR-T cell therapy reduces signs and symptoms in the mouse model of iTTP.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) HZDR, 3 months: Being trained UniCAR-T cell therapy; (3) Immudex, 3 months: Trained in Klickmer technology to follow depletion of antigen specific B cells.

Enrolment in Doctoral degree(s): DC will be enrolled at Katholieke Universiteit Leuven

Host Institution: Helmholtz-Zentrum Dresden-Rossendorf e.V. (Germany)
Supervisor: Michael Bachmann
Email address: Interested candidates are invited to apply for this position via the HZDR career website
Duration: 36 months

Introduction: Reverse chimeric antigen receptor (RevCAR)-T cell therapy was originally developed to treat cancer by HZDR. Here RevCAR-T cell therapy will be introduced for the first time to treat autoimmune diseases. The disease model used will be the autoimmune disease iTTP. The efficacy of the RevCAR-T cell therapy will be studied in vitro and in vivo in a mouse model of iTTP.

Aims
Science: 1: To generate RevCAR-T cells and CS- and CUB-targeting modules; Science: 2: To study in vitro efficacy and in vivo efficacy of RevCAR-T cell therapy in a mouse model of iTTP; Science: 3: To compare RevCAR-T cell therapy with other innovative immune therapies (WP2 and WP3). Training: 1: Learn to generate RevCAR-T cells via lentiviral transduction and to isolate T cells via MACS/FACS sorting; Training: 2: Learn to perform cytotoxicity assays; Training: 3: Learn how to use the preclinical mouse model for iTTP (trigger, sign and symptoms).

Expected Results: Having designed human and murine RevCAR-T cells and CS-and CUB-targeting modules. Having proven in vitro cytotoxicity using the RevCAR-T cell therapy. Having demonstrated the RevCAR-T cell therapy reduces signs and symptoms of iTTP in the mouse model.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) KUL, 3 months: Test RevCAR-T cell therapy in the mouse model for iTTP; (3) Immudex, 1 month: Trained in Klickmer technology to follow depletion of antigen specific B cells.

Enrolment in Doctoral degree(s): DC will be enrolled at Technische Universitaet Dresden

Host Institution: ANICELLS (Belgium)
Supervisor: Nathalie Cools
Duration: 48 months

Introduction: To date, cell therapies are also being evaluated to treat autoimmune diseases, such as type I diabetes, rheumatoid arthritis and multiple sclerosis. In this perspective, the essential role of regulatory T cells (Tregs) in preventing autoimmunity and controlling responses to self-antigens is well established. In current proposal, we aim to develop "designer" Tregs that are engineered to express a disease-specific TCR to enforce cell interactions that may play a key role in resolving the disease pathogenesis. The disease model used will be the autoimmune disease iTTP. The efficacy of the TCR-engineered Tregs will be studied in vitro and in vivo in a mouse model of iTTP.

Aims
Science: 1: Having generated TCR transgenic Tregs and tested their stability; Science: 2: Having tested the functionality and suppressive potential of TCR transgenic Tregs towards key immune cells. Training: 1: Having learned to isolate Tregs via FACs, expand them and study their stability; Training: 2: Having learned how to generate TCR transgenic Tregs via T cell electroporation with mRNA; Training: 3: Having learned how to study the suppressive effect of the TCR transgenic Tregs on selected immune cells.

Expected Results: Having generated TRC transgenic Tregs that are suppressive towards key immune cells.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) SQN, 4 months: To understand and study T cell responses in iTTP patients.

Enrolment in Doctoral degree(s): DC will be enrolled at Universiteit Antwerpen

Host Institution: Stichting Sanquin Bloedvoorziening (The Netherlands)
Supervisor: Jan Voorberg
Duration: 48 months

Introduction: DC5 will develop a novel strategy to induce antigen-specific tolerance in iTTP based on targeting of surface receptors on highly tolerogenic liver sinusoidal endothelial cells (LSEC). In addition, CRISPR/Cas9 mediated genome engineering of red blood cell precursors will be employed to hijack the tolerogenic properties of ageing red blood cells.

Aims
Science: 1: To design novel protein-based tolerogenic vaccines based on our current knowledge of B cell and T cell epitopes involved in iTTP; Science: 2: Design peptide-based tolerogenic vaccines that hijack the tolerogenic properties of ageing red blood cells; Science: 3: Provide in vivo proof of the efficacy of “designer” tolerogenic vaccines for the treatment of iTTP in a mouse model. Training: 1: Rational design of tolerogenic vaccines to treat autoimmune disorders; Training: 2: Characterize the tolerogenic properties of LSEC and different macrophage populations; Training: 3: To learn how to use the preclinical mouse model for iTTP (trigger, sign and symptoms).

Expected Results: Having designed tolerogenic vaccines that can potentially be used for treatment of autoimmune disorders. Having designed peptide-based vaccines that have the potential to re-direct CD4+ T cells towards a regulatory phenotype thereby restoring tolerance in autoimmune disorders like iTTP. Having tested the tolerogenic vaccines in the mouse model of iTTP.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) AHT, 3 months: To test whether incorporation of tolerogenic vaccines in nanoparticles will provide a novel treatment option for autoimmune disorders; (3) KUL, 4 months: Test tolerogenic vaccines in the mouse model for iTTP.

Enrolment in Doctoral degree(s): DC will be enrolled at Academisch Medisch Centrum bij de Universiteit van Amsterdam

Host Institution: AHEAD THERAPEUTICS SL (Spain)
Supervisor: Marta Vives-Pi
Duration: 36 months

Introduction: Ahead’s PS-liposomes induce antigen specific immune tolerance by biomimicry, stopping the autoimmune reaction. The immunotherapy was preclinically developed for type 1 diabetes and has been successfully validated in other autoimmune diseases. Considering the potential of PS-liposomes therapy, it is worth exploring the use of this technology platform to solve the autoimmune attack that undergoes in iTTP. Liposomes will be generated and tested both in vitro and in vivo in iTTPs patients’ lymphocytes and in a mouse model of iTTP respectively.

Aims
Science: 1: To design, synthesize, characterize (composition, charge, size) and optimize iTTP PS-Liposomes; Science: 2: To validate PS-Liposomes potential for iTTP treatment; Science: 3: To validate tolerogenicity using mouse model of iTTP and ex-vivo using human dendritic cells from 10 patients with iTTP. Training: 1: Acquisition of full competence in laboratory work, cytometer sample processing, nanoparticle preparation, animal manipulation, data analysis as well as acquisition of industrial skills.

Expected Results: Having designed and manufactured iTTP PS-liposomes. Having selected the peptide to boost a tolerance signal. Both laboratory scale and scalable production process. Having produced 2 or 3 iTTP PS-liposome candidates. Having validated iTTP PS-Liposomes potential for treatment in the mouse model for iTTP, and in vitro via autologous proliferation assays with iTTPs patients’ lymphocytes.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) KUL, 4 months: Test PS-liposomes in the mouse model for iTTP; (3) RCSI, 1 month: To get insight into how ADAMTS13 can be retargeted to antigen presenting cells.

Enrolment in Doctoral degree(s): DC will be enrolled at Universidad Autonoma de Barcelona

Host Institution: Royal College of Surgeons in Ireland (Ireland)
Supervisor: James O’Donnell
Duration: 36 months

Introduction: The metalloprotease ADAMTS13 plays a key role in preventing the development of pathological thrombosis. ADAMTS13 deficiency is associated with life-threatening microvascular thrombosis in patients with TTP. In addition, variation in plasma ADAMTS13 levels has been implicated in the pathogenesis of other important human diseases including myocardial infarction, stroke, cerebral malaria and sickle cell disease. This project will investigate the biological clearance mechanisms involved in regulating ADAMTS13 half-life.

Aims
Science: 1: Characterize the biological mechanisms underlying ADAMTS13 clearance in vivo; Science: 2: Based on clearance biology, retarget ADAMTS13 to specific clearance pathways to promote tolerance. Training: 1: Having learned how to study in vivo regulation of ADAMTS13 clearance; Training: 2: Having learned how to retarget ADAMTS13 to specific clearance pathways to promote tolerance.

Expected Results: Having defined the role of macrophages and specific macrophage receptors in modulating ADAMTS13 clearance; Having investigate the importance of specific ADAMTS13 domains and glycan determinants in triggering clearance. Having identified how to retarget ADAMTS13 to specific clearance pathways to promote tolerance.

Planned secondment(s): (1) AP-HP, 6 weeks: To gain understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP; (2) KUL, 2 months: To learn mouse model for iTTP; (3) AHT, 3 months: study targeting liposomes to dendritic cells.

Enrolment in Doctoral degree(s): DC will be enrolled at Royal College of Surgeons in Ireland

Host Institution: Semmelweis Egyetem (Hungary)
Supervisor: Zoltán Prohászka
Duration: 48 months

Introduction: Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is mediated by anti-ADAMTS13 antibodies, however, the mechanism and course of the development of these antibodies are not yet elucidated. Here novel immunoassays will be developed and performed in sets of longitudinal iTTP samples and the role of antibody-, antigen- and host related factors will be studied in the anti-ADAMTS13 immune response.

Aims
Science: 1: To develop a novel immunoassay for patient-derived “opening” antibodies that modulate ADAMTS13 conformation; Science: 2: To measure opening, inhibitory and total anti-ADAMTS13 IgG antibodies in longitudinal samples; Science: 3: To study the role of antigen-related and host-related factors in the anti-ADAMTS13 immune response; Science: 4: To study the relationship between changes in anti-ADAMTS13 antibody concentrations, complement and cytokine parameters and the development of clinical endpoints. Training: 1: Learn how to develop immunoassays to determine the concentration of opening ADAMTS13-antibodies and antibodies against citrullinated or differently glycosylated ADAMTS13; Training: 1: Learn how to study host factors that modulate autoimmunity.

Expected Results: Having identified host- and antigen-related factors that could drive autoimmunity against ADAMTS13 in iTTP, and having explored how opening, inhibiting or binding autoantibodies are related to the development of an acute iTTP episode.

Planned secondment(s): (1) AP-HP, 6 weeks: Understanding of the medical needs of patients with iTTP and challenges linked to diagnosis and treatment of patients with iTTP. To gain skills in the standardisation of clinical databases and ADAMTS13 measurements; (2) Werfen (Biokit Research & Development S.L.U.), 3 months: Skills on regulatory/quality requirements to develop/market diagnostic assays; (3) RCSI, 1 month: To learn how to measure glycoforms of ADAMTS13.

Enrolment in Doctoral degree(s): DC will be enrolled at Semmelweis Egyetem