PROJECTS IN THE LAB

Webpage

Project Code: PNRR-III-C9-2022-I8-186

Project Title: Targeting Cardiac Fibrosis in Heart Failure; Challenges and Potential Solutions Based on ncRNA Therapeutics

Contract number: 760062/23.05.2023

Coordinator: Dr. Fabio Martelli

Labs involved in project implementation:

• Laboratory of Stem Cell Biology

• Laboratory of Inflammation Research

Budget: 1.400.000 Euro

Implementation period: 18.07.2023 – 31.06.2026

Abstract: FibroThera aims to fight against myocardial infarction (MI)-induced heart failure, a major human health problem with high socio-economic implications. In this project, we will abandon the traditional focus on protein-coding genes and explore the noncoding RNA-mediated regulatory pathways. The FibroThera concept is based on the assumption that identifying the ncRNA specifically involved in MI-induced heart failure will increase the probability of finding new druggable therapeutic targets with clinical applicability, which can improve the quality of life in ischemic heart disease patients. FibroThera will characterize the dynamics of cells contributing to fibrosis during post-MI cardiac repair and will propose solutions for limiting the adverse remodelling after MI.

The specific objectives of FibroThera are to:

(i) Define the pro-fibrotic cell types sequentially involved in the post-MI repair and identify ncRNA candidates for modulation of cardiac fibrosis;

(ii) Identify cell-specific changes induced by Mesenchymal Stromal Cell therapy in fibrosis signalling cascades activated after MI;

(iii) Explore the function of activated fibroblasts and provide insights into the molecular mechanisms of cardiac fibrosis;

(iv) Explore miR-210-controlled pathways that interfere with MI-induced cardiac fibrosis in human and mouse samples.

In reaching these objectives, the team will work under the direct coordination of a highly specialized researcher in the field of ncRNA and will greatly benefit from his extensive network of collaborators. We will use innovative in vitro and in vivo models, high-throughput RNA sequencing methods, and relevant human samples for data validation.

Project's code: PN-III-P4-ID-PCE-2020-1340

Project's title: Fundamental mechanisms of post-infarction remodeling in old heart at fibroblast population level

Contract number: PCE122/12.02.2021

Coordinator: Alexandrina Burlacu

Budget: 1.197.912 Lei

Period: 04.01.2021 – 31.12.2023

Abstract:  Cardiovascular diseases (CVD) are the major cause of mortality in the world, with myocardial infarction (MI)-based injury and subsequent heart failure as major sequela of this disorder. Despite the importance of fibrosis in CVD, the contribution of cardiac fibroblasts (cFb) to disease progression remains poorly understood, and interventions effectively targeting this cell have only recently emerged in the spotlight. In-fOLD project focuses on the active role of cFb in the ventricular remodeling of old animals and aims to unveil cFb-specific signaling pathways in natural ageing process upon interference with MI and cell therapy. The objectives of the project are: (i) generate a reference transcriptome of cFb derived from old mice with MI; (ii) identify changes in cFb-specific signaling cascades induced by cell therapy post-MI; (iii) provide insights into the role of cFb in the modulation of the reparatory process of MI in old mice. The first two objectives are expected to provide data about the molecular changes over the MI time course that produce the impaired responsiveness of cFb in old individuals. By the third objective, the cardiac repair in old animals will be addressed as a T cell-mediated phenomenon, by focusing on the crosstalk between cFb and T cells in vitro, which has not been carried out before. These studies will provide a more integrated view of cardiac biology by illustrating the role of different cell types in the cardiac repair process after MI in old organisms.

Project's code: PN-III-P1-1.1-TE-2019-1893

Project's title: MIR-210 genomic locus at the overlap between hypoxia signalling and inflammatory networks

Contract number: TE186/07.01.2021

Coordinator: Dr. Mihai Bogdan Preda

Budget: 431.900 Lei

Period: 07.01.2021 – 31.12.2022

Abstract:  Accumulating evidence suggests that hypoxia and inflammation are intimately linked on many levels, with a significant crosstalk between the transcription factors that are classically understood to respond to each one of them. The molecular machinery that regulates the activity of genes in response to hypoxia is subject to many regulatory feedback loops, mediated by transcription factors, metabolites, and the more recently appreciated, yet insufficiently understood, non-coding RNAs. To date, the best characterised hypoxia-responsive non-coding RNA is miR210, which has diverse functions that cover almost every aspect of cellular hypoxic response.

An integrated inspection of mouse and human genomic loci shows multiple long-noncoding RNA transcripts and a promoter/enhancer regulatory element, suggesting that the miR210 locus functions as a coordinated unit with the neighbouring genetic elements. Furthermore, the consequences of genetically disrupting this locus are not equivalent to a conventional inactivation of the mature miR210.   

The goal of this project is to determine cell-specific cis- and trans- effects of the miR210 locus during hypoxia and inflammation. The specific objectives of this project are: (i) Dissection of miR210 genomic locus in immune cells (T cells, B cells, and monocytes/ macrophages) in two transgenic mouse models for miR210; (ii) Evaluate the impact of hypoxia and inflammation on the inactivation of miR210 genomic locus after CRISPR-mediated experimental dissection.

Dissecting RNA biology of non-coding RNAs is generally a difficult and laborious process, however to get these objectives, we will work under the direct supervision of recognized experts in the hypoxia signalling field, and will use powerful and modern tools for deciphering genome organization (CRISPR-mediated genome editing, RNA-seq, 3C assay), with the goal of shedding the light into the biology of a genomic region long considered important for the immune and inflammatory response.

Project's code: PN-III-P4-ID-PCCF-2016-0172

Project's title: Targeting innate immune mechanisms to improve risk stratification and to identify future therapeutic options in myocardial infarction

Contract number: 05/18.07.2018

Coordinator: Acad.Maya Simionescu

Research Partners:

CO: Institute of Cellular Biology and Pathology "Nicolae Simionescu", BUCHAREST

P1: University of Medicine, Pharmacy, Science and Technology, TARGU MURES

P2: University of Medicine and Pharmacy "Carol Davila" BUCHAREST

Budget: 8.500.000 Lei

Period: 18.07.2018 – 31.06.2022

Abstract: Myocardial infarction (MI) is a major cause of morbidity and mortality. At present, clinicians lack specific biomarkers for accurate post-MI risk stratification and therapeutic tools to modulate myocardial inflammation and to promote efficient recovery. Innate immune processes mediated by polymorphonuclear neutrophils (PMN) and macrophages (MAC) in the immediate post-MI period determine the extent of myocardial damage but also induce repair. Our major goal is to identify central molecules that mediate the crosstalk between sub-populations of PMN and MAC, and determine their involvement in MI. Additionally, we will test the ability of specific therapies to regulate myocardial inflammation and to improve cardiac function in vivo. 

Project objectives are: 

(1) To identify key mediators that determine post-MI myocardial remodeling and prognosis. We will investigate the relationships between the soluble PMN/MAC mediators, post-MI cardiac function, and prognosis in MI patients. 

(2) Investigation of the crosstalk between PMN and MAC in MI. We will use in vitro studies, genomics, and proteomics to identify mediators that govern the PMN-induced MAC polarization into sub-populations that promote repair. The role of the identified molecules will be tested in vivo. 

(3) Development of a mesenchymal stem cell (MSC)-based therapy in MI. We will investigate the ability of MSC treatment to shift PMN and MAC polarization towards reparatory phenotypes and to improve cardiac recovery. 

(4) To investigate S100A8/A9 blockade as a potential therapy in MI – S100A8/A9 is a potent pro-inflammatory molecule secreted by PMN and MAC. 

We will assess whether S100A8/A9 blockade inhibits inflammation and improves post-MI cardiac function. The expected outcome is to identify biomarkers that can be used to accurately identify patients at high risk to suffer new events. MSC therapy or S100A8/A9 blockade might improve the recovery of MI patients and reduce morbidity and mortality in this large patient group.

Project's code: PN-III-P1-1.1-PD-2016-1903 

Project's title: Interference with hypoxia-signaling pathways in mesenchymal stem cells prior to transplantation as a strategy to enhance myocardial recovery post infarction 

Contract number: 133/02/05/2018 

Coordinator: Dr. Mihai Bogdan Preda

Budget: 250.000 Lei

Period: 02.05.2018 – 31.10.2020 

Abstract:  The aim of OXI-SCENARIO project is to improve the effector properties of human mesenchymal stem cells (MSC) by in vitro targeting the oxygen-sensing mechanisms and evaluating the therapeutic potential of the hypoxia-modulated cells in a pre-clinical mouse model of myocardial infarction (MI). Although MSC have a native potential to promote a regenerative microenvironment in an injured tissue and are the focus of intensive efforts for developing cell-based therapies for a wide range of diseases, increasing evidence points towards a rather abnormal behaviour of cultured MSC. One possible explanation is the collection of these cells from a low oxygen environment, where they normally reside in vivo, into an atmospheric oxygen environment for in vitro expansion before therapeutic use. In this context, we hypothesized that modulating the culture conditions so as to better mimic the in vivo environment would produce MSC with superior functionality and therapeutic potential. To assess our hypothesis, three specific objectives are envisioned: (i) Assessing the effector functions (pro-angiogenic, anti-inflammatory and immunomodulatory) of MSC cultured for up to three passages in an hypoxic environment; (ii) Functional dissection of MSC to uncover the way by which miR-210 host gene locus (a master regulator of hypoxia) interfere with the effector properties; (iii) Evaluate the migration efficacy and regenerative capacity of modified human MSCs in a clinically-relevant animal model of MI. The importance of this project resides in its potential to yield new advances with regard to the molecular mechanisms of hypoxia-responsive pathways in MSC, with direct implications for human health. We anticipate that, at the conclusion of this project, the collected data might impact not only the treatment of MI, but also many other biomedical fields, given that hypoxia and its intricate regulation are at the epicenter of the cardiovascular, organ transplantation or cancer research.

Project's code: POC-A.1-A.1.1.4-E-2015, ID: P_37_668

​Project co-funded by The European Regional Development Fund through the Competitiveness Operational Programme 2014-2020.

Priority Axis 1 - "Research, technological development and innovation (RDI) in support of economic competitiveness and business development".

Action 1.1.4: Attracting personnel with advanced skills from abroad to strengthen the research-development capacity

​Project's title: The improvement of the institutional competitiveness in the treatment of type 1 diabetes by developing an innovative concept of immunotherapy based on mesenchymal stromal cells

​Contract number: 118/16.09.2016

​Coordinators:

Dr. Alexandrina Burlacu (LSCB, ICBP, Romanian Academy, Romania)

Dr. Nadir Askenasy (Frankel Laboratory, Centre for Stem Cell Research, Schneider Children’s Medical Centre of Israel)

​Budget: 8,630,843.2 Lei

​Period: 04.01.2016 – 31.12.2020

Abstract: Type 1 diabetes is an inflammatory reaction against pancreatic islets triggered by extrinsic precipitating factors in subjects with particular genetic configurations favorable to autoimmunity. Despite intensive interest in etiology, pathogenesis, and mechanisms of this inflammatory reaction, reliable curative therapies are yet to be developed. Therefore, the scientific objective of DIABETER project is to design, refine, and consolidate a clinically relevant cell therapeutic approach to cure diabetic autoimmunity by targeted delivery of apoptotic signals using mesenchymal stromal cells (MSC) as vehicles.

The specific aims designed to accomplish the scientific objective are:

Specific aim 1: Arrest of inflammatory insulitis by immunomodulation with killer MSC;

Specific aim 2: Abrogation of autoimmunity using repeated killer MSC infusions;

Specific aim 3: Indefinite restoration of self-tolerance through hematopoietic chimerism.

Versatility of MSC function offers, in principle, the possibility to provide support on all three levels mentioned above: arrest insulitis, abrogate autoimmunity and restore self-tolerance; however, sustained effects and robust restoration of endogenous insulin production have not yet been achieved with naïve MSC. Therefore, a systematic approach will be undertaken to consolidate a pragmatic pre-clinical therapeutic strategy that integrates the three mechanistic steps based on unique principle: use of killer MSC in conjunction with maximal immunomodulation and minimal lymphoreduction. The significance of this new therapy lies in its innovative nature, the use of already approved therapeutic tools, and the potential transfer into the clinic for further development. Moreover, the proof of concept of this therapy may be translated to treat a wide variety of immune and autoimmune disorders.

Transnational Research Projects on Cardiovascular Diseases

Project's title: Exploring new pathways in age-related heart diseases 

Coordinator: Dr. Thomas THUM

Budget: ~ 1.450.000 €

Period: 2017 - 2020  

Participants:

(i)  Dr. Thomas THUM, Hannover Medical School, GERMANY;

(ii) Dr. Leon DE WINDT, Maastricht University, NETHERLANDS;

(iii) Dr. Gianluigi CONDORELLI, Humanitas Research Hospital, Milan, ITALY;

(iv) Dr. Valentin FUSTER and Dr. Vicente ANDRES, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, SPAIN;

(v) Dr. Faiez ZANNAD, University of Lorraine, Inserm-CHU of Nancy, FRANCE;

(vi) Dr. Alexandrina BURLACU, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, Bucharest, ROMANIA. 

Abstract:

EXPERT is an interdisciplinary 3-year program involving 6 European nations exploiting the power of RNA biology to fight age-related heart failure (HF). EXPERT will identify pathways characteristic of age-related HF and will abandon the traditional focus on protein-coding genes, instead exploring the non-coding RNA (ncRNA)-mediated regulatory pathways of the human genome. EXPERT uses innovative in vitro/in vivo genetic models, ncRNA gender-specific diagnostic and therapeutic approaches, and clinical/population-based cohorts to explore age-related HF. The project will lead to novel diagnostic and therapeutic strategies for the treatment of HF in the elderly.

EXPERT will a) validate pathways of pathologic cardiac ageing to develop therapeutic ncRNA-based strategies, b) identify an age-specific pattern of circulating ncRNA to predict future cardiac events/death.

The specific objectives were to:

• Generate a cell-type specific ncRNA blueprint of the ageing heart.

• Study ncRNA regulation in cardiac ageing upon interference with cardiac stress leading to HF.

• Validate important age-related cardiovascular pathways by testing ncRNA interventions in small and large animal models.

• Define the predictive value of circulating age-related ncRNA patterns in clinical and population-based cohorts.

• Project's code: PN-III-P2-2.1-PED-2016-1881  

• Project's title: Consolidating the subcutaneous transplantation of MSC as a warranted therapy for myocardial infarction

• Contract number: 251PED/01/09/2017 

• Coordinator: Dr. Mihai Bogdan Preda

• Budget: 475.000 Lei

• Period: 01.09.2017 – 11.12.2018

• Abstract:  Taking advantage of the trophic effects of mesenchymal stem cells (MSCs) and the possibility of evoking myocardial protection through extra-cardiac approaches, we hypothesized that remote transplantation of MSCs might confer protection of the heart against ischemic injury.  As the number of patients with infarction who develop heart failure, a disease with poor prognosis and few treatment options, is increasing, there is a substantial unmet need for novel therapies.

• By this project, we aim to expand our previous studies beyond the proof-of-concept by refining and consolidating the approach of subcutaneous (s.c.) transplantation of MSCs as a warranted therapy for myocardial infarction (MI). This approach offers several advantages compared to other routes of stem cell delivery. Firstly, it is minimally invasive, does not require general anaesthesia, and does not imply any blood loss. Secondly, this approach can be repeated periodically without any risk for the patients. Thirdly, this approach may represent an adjuvant therapy in combination with conventional therapies to add further benefits, or it may be applied as a stand-alone therapy for patients with increased surgical risk.

• The objectives of the project are: (i) To determine the number of human MSCs in a transplantation dose that produces maximum benefits to the infarcted heart; (ii) To assess the extra-benefits produced by repetitive doses of s.c. transplanted MSCs on the cardiac function after MI; (iii) To evaluate the effectiveness of allogenic MSCs in conferring cardioprotection.

• We are aware that consolidation of this stem cell-based therapy requires a lot of work to precisely define the protocol and mechanisms standing behind it and to carefully define the clinical circumstances where this strategy can be utilized. Yet, the great advantages this therapy will bring in comparison to current approaches give us hope that the progress we will achieve can substantially contribute to life-saving clinical approaches.