Grant Projects (ongoing in the year 2026)
View ongoing grants in the year: 2009, 2010, 2011, 2012, 2013, 2014, 2015, 2016, 2017, 2018, 2019, 2020, 2021, 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030.
Advanced genomic characterization of secondary myeloid malignancies: implications for diagnostics and prognostication
AZV NU23-03-00401 [2023 – 2026]
RNDr. Monika Beličková, Ph.D. MHA
Mgr. Jana Kotašková, Ph.D., Fakultní nemocnice Brno
Therapy-related myeloid neoplasms (t-MNs) are secondary malignancies that occur after chemotherapy and/or radiation therapy of a previous cancer. The t-MN category accounts for approximately 10-20% of newly diagnosed patients with myeloid neoplasms (MNs) and the latency between treatment and development of t-MN ranges from a few months to years. The prognosis of these patients is generally poor, and unlike primary MNs, there are no specific prognostic scoring systems for this group nor a defined complex genomic background. We hypothesized that the combined effect of acquired somatic mutations and numerous germline variants, including common and rare variants, which underlie the multistep molecular pathogenesis, contributes to t-MN development. Thus, we propose an advanced analysis of the t-MN genome, which will include the examination of somatic and germline mutations using whole-genome and targeted sequencing. Furthermore, transcriptome analysis of protein-coding and noncoding RNAs at the level of bone marrow CD34+ cells will be performed to determine the signaling pathways involved in t-MN development. Genomic data along with cytogenetic findings will be correlated with clinical parameters. Relevant molecular markers will be incorporated into the diagnostic and prognostic evaluation of t-MN patients. Knowledge of the molecular background of secondary malignancies will lead to a more detailed subclassification of patients, determination of a more accurate prognosis and selection of an appropriate treatment, ultimately strengthening personalized approaches for patients.
Next generation CAR-T cells for the therapy of hematological malignancies
AZV NU23-03-00188 [2023 – 2026]
doc.MUDr. Pavel Otáhal, Ph.D.
Prof. MUDr. Pavel Klener, Ph.D., 1. LF UK Praha
The proposed project deals with new methods of cell therapy of hematological malignancies such as AML, B-ALL and B-NHL using CAR-T T lymphocytes. The aim of the project is to build on the ongoing clinical study with CD19-specific CAR-T lymphocytes, which are produced in IHBT for therapeutic purposes (UHKT CAR19) and are tested in a clinical trial in patients with refractory/relapsing B-ALL/B-NHL. Treatment with commercial CD19 CAR-T products cures about 20-30% of patients indicated for this treatment, the remaining patients either do not respond to treatment or relapse even when complete remission is achieved. The causes of CAR-T treatment failure are being intensively studied. One of the ways to prevent lies in the development of so-called next-generation CAR-T cells that have enhanced antitumor activity, e.g. by co-expression of cytokines (IL-12, IL-15, IL-18, IL-21) or—by secreted bi-specific antibodies. The first goal of this project is to develop such modified CD19-specific CAR-T lymphocytes. The second goal is the development of CD123-specific CAR-T lymphocytes, for the treatment of acute myeloid leukemia. These new types of CAR-T products will be analogically to the UHKT CAR19 T cells that are now being produced at UHKT. During the solution of the project, we will establish GMP-certified production of both CAR-T products in the Clean Rooms of the IHBT, prepare pharmaceutical documentation and submit an application for two clinical trials with these products - one for AML and the other for refractory/relapsing B-ALL/B-NHL.
Changes in expression of adhesion molecules: a tool for prediction of extramedullary relapse of B cell acute lymphoblastic leukemia following blinatumomab treatment
AZV NU23J-03-00033 [2023 – 2026]
Blinatumomab (Blincyto ®) is a bi-specific therapeutic antibody mediating the connection of the CD19 receptor on malignant B-cells to CD3 on effector T-lymphocytes. This enables the formation of a cytolytic synapse and effective elimination of the tumor. According to current knowledge, however, the relapse into extramedullary disease (EMD) is unusually common in blinatumomab-treated patients with B-cell acute lymphoblast leukemia (B-ALL). The main hypothesis of the project assumes that blinatumomab affects leukemia cell adhesion properties and thereby homing of B-ALL cells to tissues where blinatumomab is not effective. This may eventually lead to EMD. The sole interaction of blinatumomab with CD19 can affect the signaling of several adhesion-related molecules (integrins, Lyn). Furthermore, immune synapse formation can lead to changes in the expression of surface cell adhesion molecules (CAM) on both involved cells. Additionally, cytoskeleton changes induced by B-cell receptor (BCR), a vital component of the immune synapse, are dependent on Lyn kinase signaling. The project aims to analyze the effect of blinatumomab on B-cell adhesion processes. Mechanisms of blinatumomab-induced changes in adhesion signaling will be studied on B-cell leukemia (B-ALL) cell lines, both in the absence and in the presence of model T-lymphocytes. We will assess changes in the cell affinity to proteins of the extracellular matrix and to model stromal cells and, on a molecular level, changes in the activation status of selected proteins to find a clinically useful marker of the changes induced by blinatumomab. In primary samples (B-ALL, ~60 patients and up to 250 samples during the project course), changes in expression and activation of relevant CAMs (integrins, CD44, CD62L), as well as selected intracellular markers (Lyn) after blinatumomab treatment, will be evaluated, and correlated with the treatment results.
VEXAS syndrome; characterization and clinical consequences of individual UBA1 genetic variants among patients with rheumatic and hematologic disorders
AZV NU23-10-00160 [2023 – 2026]
MUDr. Heřman Mann, Ph.D., Revmatologický ústav, Praha
RNDr. Monika Beličková, Ph.D. MHA
VEXAS (vacuoles, E1 activating enzyme, X-linked, auto-inflammatory, somatic) syndrome is a clinically serious and potentially fatal adult-onset disease caused by somatic mutations in the UBA1 (Ubiquitin-like modifier activating enzyme 1) gene in hematopoietic progenitor cells. It was first described in 2020 therefore data regarding its clinical manifestations, genetic associations, prognosis and treatment efficacy are limited. UBA1 missense mutations in VEXAS syndrome patients cause activation of inflammatory pathways, which clinically present in older patients with systemic inflammation and a potential to progress into myelodysplastic, myeloproliferative, or lymphoproliferative disease. Patients may meet diagnostic criteria for several rheumatic and/or hematological disorders. Given its heterogeneous clinical manifestations a high degree of clinical suspicion is required for pursuing the diagnosis of VEXAS syndrome. Demonstration of known pathogenic variants in hot spot sites of p.Met41 in exon 3 in UBA1 sequencing analysis of hematopoietic cells is currently required to establish the diagnosis of VEXAS syndrome. However, this approach may not be sufficient since an extended analysis of all coding regions of the UBA1 gene may uncover other mutations with putative functional consequences as we have shown. VEXAS syndrome is associated with high morbidity and mortality. There is currently no information regarding the impact of VEXAS syndrome on patients’ quality of life. Many patients experience partial response to treatment with glucocorticoids and/or immunosuppressive therapy. Limited evidence suggests that hematopoietic stem cell transplantation may be curative in some cases. More data from careful clinical observations is clearly needed to provide much evidence regarding the optimal diagnostic approach, impact of the disease on patients’ quality of life, prognosis and value of individual treatments.
IHBT: Successful Commercialization of Hematology, Hematooncology and Immunotherapy Products
TQ11000057 [2024 – 2028]
The primary objective of the project is to enhance the existing infrastructure at the Institute of Hematology and Blood Transfusion (IHBT) for proof-of-concept activities aimed at testing the potential of new gene therapy drugs and modern diagnostic methods and increasing their market value in the period of project implementation. The secondary objective is to improve the economic performance of the Institute in the field of applied research and experimental development and to establish a self-sufficient system for financing technology transfer, including proof-of-concept projects in the future. The tertiary objective is the benefit of the patients of IHBT with the availability of innovative medicines and diagnostic methods in the Czech Republic. The technical parameters of these objectives are as follows: primary objective - to achieve the planned project outputs, including at least 3 modern therapeutics in clinical trials (TRL4 to TRL6), at least 4 innovative diagnostic methods (TRL4 to TRL7), at least 6 intermediate products or services supporting the innovation ecosystem in the Czech Republic in the field of modern treatments by the end of the sustainability period (target higher than TRL7). Secondary objective - to find funding mechanisms for at least 1/3 of the outputs of the sub-projects by the end of the sustainability period. Based on our experience with technology transfer, we consider these objectives to be achievable.
Single-cell analysis for deep characterization of leukemic cell subtypes responsible for aggressiveness and relapse of Ph+ leukemias
AZV NW25J-03-00098 [2025 – 2028]
Tyrosine kinase inhibitors (TKIs) targeting the oncoprotein BCR::ABL1 play a main role in the therapy of Ph+ leukemias, which significantly improved the outcome of chronic myeloid leukemia (CML) and Ph+ acute lymphoblastic leukemia (Ph+ ALL) and the overall survival of patients. However, the challenge how to prevent the progression of both diseases still remains. In Ph+ ALL, the problem is an aggressive relapse, in CML it is a progression to blast crisis, which in both cases require innovative therapeutic approaches combining TKIs with other drugs. In the proposed project, we suppose that next-generation sequencing at the single-cell level will identify leukemic cells responsible for the progression of Ph+ leukemias and that we will be able to characterize their molecular background and therapeutic targets. The aim of this project is also to distinguish lymphoblastic relapsed CML, Ph+ ALL, and Ph+ MPAL at the time of diagnosis. Methodologically, we will focus on single-cell RNA-seq with simultaneous analysis of surface markers. We will analyze cells at the time of disease relapse (for CML it will be a myeloid and/or lymphoid blast crisis) and cells from a paired sample from the time of diagnosis. To distinguish the diagnosis of Ph+ leukemias, we will analyze primary patient cells.
Molecular base for therapy with BH3 mimetics to overcome refractory/relapsed Ph+ leukemias
AZV NU24-03-00056 [2024 – 2027]
doc. Mgr. Kateřina Machová Poláková, Ph.D.
doc. MUDr. Mgr. Cyril Šálek, Ph.D.
The project is focused on experimental therapy of preclinical models of refractory/relapsed Ph+ leukemias based on the measurement of protein expression of the "apoptotic panel" by mass cytometry. Failure of several tyrosine kinase inhibitor (TKI) treatment lines, including ponatinib, and relapse after eventual hematopoietic stem cell transplantation in Ph+ leukemias, exhausts therapeutic options in most patients, often with a fatal outcome. BH3 mimetics represent a promising group of drugs that inhibit the function of antiapoptotic proteins and induce apoptosis of malignant cells. In previous works led by a member of the proposed research team Prof. Pavel Klener, it was found that diffuse large B cell lymphoma can be divided into subgroups based on dependence on the antiapoptotic proteins BCL2 and MCL1. The efficacy of combination therapy with BCL2 and MCL1 inhibitors has been demonstrated in patient-derived models (PDX models) in vivo. The effect of BH3 mimetics has not been studied in Ph+ malignancies in detail, especially in combination with promising 3rd generation TKIs (ponatinib; STAMP inhibitor asciminib). Mass cytometry is a technology that enables multiparametric analyzes of phenotypes and cellular processes at the single-cell level. We suppose that an “apoptotic panel” analysed via mass cytometry in combination with NGS-based analysis of mutational background and gene expression changes will find its spot in diagnostic procedures in haematological malignancies with conventional therapy failure. We believe that the results of this project may lead to the design of clinical trials using effective combination therapy with BH3 mimetics to overcome the progression of Ph+ leukemias after failure of standard therapeutic options.
Dissecting the role of bone marrow microenvironment behind chemoresistance development in acute lymphoblastic leukemia
AZV NU24-03-00376 [2024 – 2027]
RNDr. Júlia Starková, PhD., 2. LF UK, Praha
MUDr. Robert Pytlík, Ph.D., Meritxell Alberich Jorda, Ph.D., ÚMG AV ČR, Praha
In addition to the intrinsic factors of tumor cells, the effect of chemotherapy, which is currently the gold-standard treatment for patients with acute lymphoblastic leukemia, is influenced to a significant extent by the external environment. It has been previously shown that the bone marrow microenvironment contributes to chemoresistance. Our data show that metabolic rewiring plays an inevitable role in this process and will be tightly orchestrated by the crosstalk of leukemic cells and tumor environment. Moreover, the basal metabolic activity of leukemic cells already at diagnosis varies and is associated with the therapy response of patients. We plan to study the interaction of bone marrow cells with leukemic cells from patients with different metabolic profiles. Identifying the critical processes involved in leukemic bone marrow niche interactions will contribute to target optimization and combinatorial drug treatment strategies in order to overcome acquired drug resistance and prevent relapse.
Role of DNA damage response in suppressing malignant progression in preleukemia conditions of some myeloproliferative and myelodysplastic neoplasms
GAČR 24-11730S [2024 – 2026]
Doc. RNDr. Vladimír Divoký, Ph.D., LF UP Olomouc
RNDr. Monika Beličková, Ph.D. MHA
DNA damage response (DDR) regulation and inflammatory signaling cooperate in checkpoint control of myeloproliferative and myelodysplastic neoplasms (MPN and MDS). We have elaborated on a concept of protection mechanisms that guard progenitors of polycythemia vera (PV), a JAK2 V617F+ MPN, and lower-risk (LR)-MDS from rapid malignant transformation. Using induced pluripotent stem cells and organoids from patients’ CD34+ cells, mouse models and patients’ samples we will test our hypothesis that adaptations of PV progenitors to cell autonomous and microenvironment-dependent inflammatory stress involve multiple JAK2 V617F-dependent factors that mitigate the potential genotoxic impact of overall oncogenic program, allowing for chronic long-term proliferation with stable genome. In LR-MDS, intrinsic and extrinsic stresses cooperate in building up a threshold of DNA damage and DDR, resulting in DDR-mediated anti-tumor senescence barrier, which in LR-MDS could be eliminated through RUNX1 loss-of-function mutation. The results should be useful for novel therapeutic approaches to PV and LR-MDS.
Safety and efficacy of anti-CD123 chimeric antigen receptor-modified autologous T cells (CART123) in patients with relapsed/refractory CD123+ hematologic malignancies.
NW26-03-00535 [2026 – 2029]
Treatment with genetically modified T lymphocytes expressing a chimeric antigen receptor (CAR) is a highly innovative approach to cancer therapy. Currently, this treatment is available for CD19 and BCMAA positive malignancies. At IHBT, previous projects successfully developed CAR targeting the CD123 antigen, which is expressed in most cases of acute myeloid leukemia (AML), blastic plasmacytoid dendritic cell neoplasm (BPDCN), and some cases of advanced myelodysplastic syndrome (MDS) and ALL. An innovative method for manufacturing autologous CAR-T cells containing anti-CD123 CAR (CART123) under GMP conditions was established and validated, utilizing non-viral transduction of linear DNA. Preclinical trials were successfully conducted, and clinical trial was approved and initiated. The subject of the proposed project is a Phase I clinical trial (CT) of CART123 in patients with refractory or relapsed CD123-positive AML, MDS, ALL, or BPDCN. This is an open-label, dose-escalation trial in three cohorts, using a Bayesian Optimal Interval (BOIN) approach to optimize the process of determining the maximum tolerated dose (MTD). The primary objectives of the study are to assess the safety of CART123, evaluate hematopoietic recovery, and determine the appropriate dose for further research based on MTD assessment. The secondary objectives include evaluating efficacy and the feasibility of subsequent allogeneic hematopoietic transplantation. In addition to clinical outcomes, correlative analyses of CART123 phenotype and persistence will be conducted using flow cytometry, histology, immunohistochemistry, and molecular genetic methods. Up to 18 evaluable patients will be enrolled in the clinical trial, depending on the occurrence of dose-limiting adverse events assessed by an independent DSMB committee. The proposal builds on the results of the preclinical projects AZV NU23-03-00188 and AZV NU22-05-00374, whose principal investigator is part of the project team.