BrainMap

Mrs. Marioara Chiritoiu-Butnaru

Dr.

Senior Researcher III - INSTITUTUL DE BIOCHIMIE

Researcher | Scientific reviewer

Publons/ResearcherID: Q-3983-2017

Curriculum Vitae (21/01/2022)

Expertise & keywords

Molecular mechanisms of protein aggregation and unconventional secretion in the context of inflammation and neurodegenerative disorders Call name: P 1 - SP 1.1 - Proiecte de cercetare pentru stimularea tinerelor echipe independente
PN-III-P1-1.1-TE-2019-1705 2021 - 2023

Role in this project: Project coordinator

Coordinating institution: INSTITUTUL DE BIOCHIMIE

Project partners: INSTITUTUL DE BIOCHIMIE (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: https://marichiritoiu4.wixsite.com/te156

Abstract:

Background: Inflammation is a hallmark of many pathologies such as autoimmune and neurodegenerative disorders, diabetes and atherosclerosis. Cytokines of the IL-1 family are primary mediators of inflammation. These are cytoplasmic proteins exported from macrophages and monocytes via the unconventional secretory pathway (UPS). I have previously shown that interleukin-1β (IL-1β) secretion form activated primary macrophages is regulated by the GRASP55-IRE1α axis and disrupting this pathway leads to intracellular aggregation of IL-1β, hampers its secretion and causes general proteome instability.
Hypothesis: Intracellular aggregation of cytoplasmic proteins (such IL-1β, α-synuclein and other) might be correlated with cargo availability for unconventional secretion, therefore modulating this process could provide potential methods to control pathological outcomes for inflammation and neurodegeneration.
Aim: Dissect the molecular mechanism and identify intracellular factors modulating aggregation and secretion of UPS cargoes. For this purpose we will use as model cargo IL-1β, produced by cells of the immune system and further corroborate the functional conservation of the identified factors for α-synuclein physiology in neurons and C. elegans model for Parkinson’s disease.
Expected results: This project should facilitate the understanding of signaling pathways and identify the intermediates transducing the information for the stress sensing machinery to induce cytoplasmic proteome instability. Also we estimate to indentify key proteins involved in UPS cargo aggregation, which could be used as targets to control the secretion of cytoplasmic proteins in the context of inflammation and neurodegeneration.

Validation of Iba1 protein as a new therapeutic target in human microglia Call name: P 2 - SP 2.1 - Proiect experimental - demonstrativ
PN-III-P2-2.1-PED-2019-3103 2020 - 2022

Role in this project: Key expert

Coordinating institution: UNIVERSITATEA BUCURESTI

Project partners: UNIVERSITATEA BUCURESTI (RO); INSTITUTUL DE BIOCHIMIE (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: http://neurobiologie.ro/ro/47-romana/contracte-nationale/209-validarea-proteinei-iba1-ca-o-noua-tinta-terapeutica-in-microgliile-umane

Abstract:

Microglia are the resident immune cells of the central nervous system (CNS). In the healthy CNS microglia are not dormant, but survey the environment by extending and retracting their ramified processes without overall cell displacement. After an injury to the CNS or peripheral nervous system, microglia rapidly activates: the cell body increases in size, proximal processes become thicker, distal branches are less ramified, specific membrane ruffles develop and the cells move to the damaged site where they show increased phagocytic activity and release of pro-inflammatory mediators aimed to restore CNS homeostasis. Iba1 is a cytoskeleton protein specifically located only in microglia and macrophages where it acts as an actin-cross linking protein. In a previous study we showed that by silencing Iba1 expression in BV2 microglia cell line using specific anti-Iba1 siRNA, the migration, proliferation and cell adhesion were significantly reduced, while other activities that involve cytoskeleton and potentially Iba1, like phagocytosis and the functioning of P2x7 receptors, were significantly increased. In the present study we aim to investigate if our custom-made anti-Iba1 siRNA is equally efficient in silencing Iba1 protein in human microglia. All the human microglia experiments will be performed on the HMC3 human microglia cell line and will be doubled by similar experiments on rat microglia primary cultures which will serve as a reference. Silencing Iba1 in human microglia might be relevant for CNS diseases associated with increased microglia activation.

High-throughput screening platform for small-molecules with anti-inflammatory potential Call name: P 2 - SP 2.1 - Proiect experimental - demonstrativ
PN-III-P2-2.1-PED-2019-3297 2020 - 2022

Role in this project: Project coordinator

Coordinating institution: INSTITUTUL DE BIOCHIMIE

Project partners: INSTITUTUL DE BIOCHIMIE (RO); INSTITUTUL NATIONAL DE CERCETARE-DEZVOLTARE IN DOMENIUL PATOLOGIEI SI STIINTELOR BIOMEDICALE "VICTOR BABES" (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: https://marichiritoiu4.wixsite.com/ped337

Abstract:

Background: The immune system is the main pillar of the human body’s health. Accurate understanding of the immune modulators will enable us to precisely unveil the abilities of the immune system to fight virtually all human diseases. Chronic inflammation is the hallmark of many human pathologies including cancer and currently, anti-inflammatory therapy is focused on inhibiting production of eicosanoid mediators of inflammation (prostaglandings, thromboxanes, prostacyclins, leukotrienes) by inhibiting their producing enzymes (COX-1/2). Although efficient, recent discoveries emphasized potentially dangerous side effects of these drugs. Therefore, identification and validation of new molecules with anti-inflammatory potential and reduced side effects, becomes of major importance. In the recent years the development of molecules targeting signaling pathways involved in production and release of pro-inflammatory mediators, such as cytokines and chemokines, has gained field and forwards the concept of new-age therapies.
Hypothesis: Our hypothesis is that identifying small molecules, which target the production or secretion of pro-inflammatory cytokines from macrophages and monocytes, such as interleukin-1β (IL-1β), will provide an improved alternative for treating chronic inflammation associated with numerous pathologies.
Aim: This project aims to develop a sensitive high-throughput screening platform by generating an endogenously tagged interleukin-1β reporter cell line by CRISPR-Cas9 technology, able to monitor stimulated IL-1β secretion with the purpose to identify new chemical compounds with anti-inflammatory activity that will be validated in primary macrophages and a mouse model for sepsis.
Expected results: Implementation of this project will deliver a versatile screening platform for anti-inflammatory compounds and a list of validated small molecules with anti-inflammatory properties, which can be easily translated to pre-clinical studies.

Integrated development project for advanced medical treatment technologies Call name: P 1 - SP 1.2 - Proiecte complexe realizate in consorții CDI
PN-III-P1-1.2-PCCDI-2017-0728 2018 - 2021

Role in this project: Key expert

Coordinating institution: INSTITUTUL NATIONAL DE CERCETARE DEZVOLTARE PENTRU FIZICA LASERILOR, PLASMEI SI RADIATIEI - INFLPR RA

Project partners: INSTITUTUL NATIONAL DE CERCETARE DEZVOLTARE PENTRU FIZICA LASERILOR, PLASMEI SI RADIATIEI - INFLPR RA (RO); INSTITUTUL NATIONAL DE CERCETARE-DEZVOLTARE MEDICO-MILITARA „CANTACUZINO” (RO); INSTITUTUL DE BIOCHIMIE (RO); UNIVERSITATEA BUCURESTI (RO); UNIVERSITATEA PITESTI (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: http://teramed.inflpr.ro/

Abstract:

In order to increase community’s quality of life, the aim of the project proposal entitled “Integrated development project for advanced medical treatment technologies” (TERAMED) is to develop novel technologies with respect to the treatment of osseous and cutaneous conditions and oncological disorders. Given our experience in healthcare research and the current requirements of multidisciplinary and interinstitutional collaboration towards the personalized treatment purpose, the TERAMED project aims genuine synthesis and processing of biomaterials, but also functional and therapeutic evaluation relevant for clinical trials. The main objectives of the “Medical devices functionalized by laser technologies and alternatives for enhanced osseous integration and regeneration” subproject are to design and produce inorganic, composite or hybrid coatings for superior osteoconductive and osteoinductive performances of titanium-based implants. Smart wound patches and polymeric gels functionalized with antimicrobial and wound healing biomolecules incorporated within micro- and nanoparticles constitute the purpose of the “Medical devices (patches and gels) based on composite biomaterials obtained by laser, plasma and radiation technologies and alternatives for enhanced healing of cutaneous injuries” subproject. The “Technologies based on magnetic triggered nanostructures for oncological therapy: early diagnosis and targeted treatment” subproject aims the development of multifunctional medical devices for specific and selective diagnosis and treatment of breast cancer and melanoma. The general impact of the TERAMED project ensues from the beneficial conjunction of the clinical potential of the proposed medical devices, the feasible technological transfer and the economic advantages of interinstitutional collaboration.

Free energy prediction of biomolecular processes using high speed robotics algorithms and its use in in-silico driven experimental research. Call name: P 1 - SP 1.1 - Proiecte de cercetare pentru stimularea tinerelor echipe independente
PN-III-P1-1.1-TE-2016-1852 2018 - 2020

Role in this project: Key expert

Coordinating institution: INSTITUTUL DE BIOCHIMIE

Project partners: INSTITUTUL DE BIOCHIMIE (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: http://old.biochim.ro/ib/projects/robofep/robofep.php

Abstract:

Critical quantities in experimental biomolecular research such as affinity constants, IC50s or rate constants are a direct reflection of the free energy in their associated processes. Therefore, in silico prediction of free energy can greatly enhance research effectiveness both in terms of costs and system detailed understanding. However, to predict in silico the free energy of a bio-molecular process is at the time being overwhelmingly difficult, especially when large biomolecules such as proteins or nucleic acids are involved - as a huge number of their configurations needs to be drawn from their practically infinite statistical thermodynamic ensemble.
To address these difficulties, we aim in this project to improve, adapt and use high speed robotics algorithms for molecular sampling that can easily escape the spatial and temporal scales in which classical methods are usually trapped. This project builds further upon a Hamiltonian Monte Carlo algorithm that we have recently developed to reproduce the Boltzmann distribution necessary for correct free energy predictions. Herein we plan to develop two new multiscaling methods, namely Generalized Adaptive Mixing (GAM) and Dynamic Adaptive Constraing (DAC), that can hike between scales without losing any level of force field detail or statistical mechanics rigor. These new methods can be a major leap in molecular simulation strategies.
We also aim by this project to implement these in the already existing Application Programming Interfaces (APIs) Simbody and Molmodel – for which we collaborate with groups from Stanford and Upsalla universities.
Last but not least we aim herein to use our methods to assist experimental work performed either by members of our team or in cooperation with various partners in the following hot topics in molecular biosciences: plant-pathogen interaction, HIV and HCV drug design and immunotherapy.

Validation of the combined use of RNAi and mass spectrometry for the identification of microglial therapeutic targets important for neuropathic pain and Alzheimer disease Call name: Joint Applied Research Projects - PCCA 2013 - call
PN-II-PT-PCCA-2013-4-0294 2014 - 2017

Role in this project: Key expert

Coordinating institution: UNIVERSITATEA BUCURESTI

Project partners: UNIVERSITATEA BUCURESTI (RO); INSTITUTUL DE BIOCHIMIE (RO); PHARMA SERV INTERNATIONAL SRL (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: http://neurobiologie.ro/Contracte-nationale/validarea-folosirii-combinate-a-tehnicii-de-rnai-si-spectrometrie-de-masa-pentru-identificarea-unor-tinte-terapeutice-din-microglii-importante-pentru-durerea-neuropatica-si-maladia.html

Abstract:

Neuropathic pain is prevalent among older patients, mainly because many diseases that cause neuropathic pain increase in incidence with age. Furthermore, older patients frequently suffer of neurodegenerative diseases like Alzheimer, which complicates drug regimens and increase the burden in economic and therapeutic terms. Therefore, is very important to better understand the pathologic mechanism of these two ailments, in order to find a better treatment with less side effects and drug-drug interactions. Iba-1, a specific marker for microglia, is an actin-cross linking protein involved in membrane ruffling and phagocytosis in microglia and macrophages which seems to be crucial for their survival and pro-inflammatory activity. Its silencing could represent a new therapeutic target for neuropathic pain and potentially for Alzheimer disease, given its strong up-regulation in microglia activated in these two conditions. In this project we want to show that by using specially designed siRNA molecules we can silence the Iba-1 gene which can have consequences on microglia activation, cytokinesis and phagocytosis potentially through P2X7 inactivation. The end-product of this project will be an intellectual property (IP) patent for the discovery of a new drug targeting a specific protein in microglia/macrophages and new mass spectrometry protocols to measure the silencing efficiency. We firmly believe that the outcome of this project will lead to a better understanding of the Iba-1 role in microglia, of the P2X7 functioning at cellular and molecular level and will open future clinical development pathways for other treatments for neuropathic pain, and potentially Alzheimer disease.

Fabrication of osteoinductive orthopedic implants with gradual 3D hierarchical structure Call name: Joint Applied Research Projects - PCCA-2011 call, Type 2
PN-II-PT-PCCA-2011-3.2-0898 2012 - 2016

Role in this project: Key expert

Coordinating institution: INSTITUTUL NATIONAL DE CERCETARE DEZVOLTARE PENTRU FIZICA LASERILOR, PLASMEI SI RADIATIEI - INFLPR RA

Project partners: INSTITUTUL NATIONAL DE CERCETARE DEZVOLTARE PENTRU FIZICA LASERILOR, PLASMEI SI RADIATIEI - INFLPR RA (RO); INSTITUTUL DE BIOCHIMIE (RO); INSTITUTUL DE BIOLOGIE SI PATOLOGIE CELULARA ,,NICOLAE SIMIONESCU'' (RO); INSTITUTUL NATIONAL DE CERCETARE - DEZVOLTARE PENTRU FIZICA MATERIALELOR BUCURESTI RA (RO); TEHNOMED IMPEX CO S.A. (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: http://lspi.inflpr.ro/Contracts/Contracts.html

Abstract:

The project “Fabrication of osteoinductive orthopedic implants with gradual 3D hierarchical structure” proposes a novel approach for the ossteointegration of permanent metallic prostheses. The originality of the project consists in the development of implants with 3D hierarchical porous structure characterized by increased surface roughness and wettability, covered with soluble thin multilayer of biocatalytic composites. The resulting structures have the advantage of combining the benefits of deposited biomaterials and the mechanical strength of the metallic collector. The aim of our research is the manufacturing of permanent implants with biofunctional surface which are able to trigger certain healing phases typical for injured bone tissues. The implant fabricated as result of our research will induce complex chronological biological reactions which in the first stage will initiate a vasogenic and angiogenic response succeeded then by the migration and differentiation of osteoblasts and mesenchymal cells and accomplished with the formation of new bone tissue. Thus, the surface structure of the implant with appropriate topology and roughness will serve as framework for osteoconductivity, while further coating with biocomposites acts, in addition to osteoconductivity, as delivery vehicles for cytokines, such as bone morphogenetic proteins, insulin-like growth factors and transforming growth factors, vascular endothelial growth factor, etc. The proposed 3D structures for osteoinduction will mimic bone morphology, structure and function in order to improve the integration into surrounding tissue.

Selection of protein degradative pathways in the pathogenesis of human diseases Call name: Exploratory Research Projects - PCE-2012 call
PN-II-ID-PCE-2012-4-0350 2013 - 2016

Role in this project: Key expert

Coordinating institution: INSTITUTUL DE BIOCHIMIE

Project partners: INSTITUTUL DE BIOCHIMIE (RO)

Affiliation: INSTITUTUL DE BIOCHIMIE (RO)

Project website: http://www.biochim.ro/ib/projects/prodegrad/prodegrad.php

Abstract:

A considerable fraction of all newly synthesized secretory polypeptides fail to attain their native conformation due to mutations, transcriptional and translational errors, folding defects or endoplasmic reticulum (ER) stress conditions. Besides the lack of function, the accumulation of aggregated proteins jeopardize the ER homeostasis and the cell functioning. Terminally misfolded polypeptides are retained and eventually removed by the ER associated degradation pathway(ERAD).We have recently found that besides the mannosidase-like domain, EDEM1 protein, one of the important players of the ERAD has an intrinsically disordered domain with high prediction for favoring protein-protein interactions. Indeed, we showed that this domain is responsible for the interaction with a misfolded tyrosinase mutant. In sum, we suggest that the ER associated degradation of glycoproteins depends less on the glycan recognition signals while rather relying on the direct recognition of the misfolded region of the polypeptide. The aim of this project is to validate this working hypothesis using ERAD substrates involved in tumor antigen degradation/peptide presentation and viral disassembly. Finally, we will use transgenic C.elegans as models for Huntington, Alzheimer and Parkinson diseases, to decipher the pathology and toxicity of protein aggregation. Overall, our approach will reveal new insights into the mechanistic details of the ERAD pathway and help identify new therapeutic targets.

Modeling molecular complexes and assemblies with experimental and bioinformatic constraints Call name: Exploratory Research Projects - PCE-2011 call
PN-II-ID-PCE-2011-3-0342 2011 - 2016

Role in this project: Key expert

Coordinating institution: Institutul de Biochimie al Academiei Romane

Project partners: Institutul de Biochimie al Academiei Romane (RO)

Affiliation:

Project website: http://www.biochim.ro/ib/projects/molint/molint_e.php

Abstract:

Protein-protein and protein-DNA assembiles in three types of systems will be investigated by constrained molecular modeling and simulation, aiming to assist and drive experimental research within the particular fields explored. The first field relates to the domain architecture of resistance gene products in plants and their interaction with effector proteins, an interaction that is instrumental in plant pathogen defense mechanisms. The second explores the structural basis of the differential functioning of human topoisomerases and the effects of some point mutations proven to induce differential malignant effects in topo II alpha and beta. Finally the third line of research tackles the structural biology of RAG-DNA synaptic complexes, involved in the assembly of the gene segments coding for the variable portions of lymphocyte antigen receptors. The three directions reflect the diversity of macromolecular assemblage in biological systems but also unveil common roots residing in the molecular geometry, interactions and dynamics at atomic level, and hence they can be tackled by common approach in which we intend to bring some contributions. On this line we intend to continue the endeavor started in a previous PN2-ID project to develop and implement in-silico techniques and use them as a motive force in molecular life sciences, when intertwined with the experimental approach.

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