Project goal
The main goal of the project is to study the regulation of adipose tissue inflammation in aging by mesenchymal stem cells after the inhibition of Cdc42.

Project tasks:

1. To study the activity of Cdc42 in murine mesenchymal stem cells from adipose tissue (ADMSCs) of aged obese mice
2. To study the effects of inhibition of Cdc42 on the secretome of aged murine adipose-derived mesenchymal stem cells
3. To evaluate the therapeutic potential of ADMSCs after Cdc42 inhibition on adipose tissue inflammaging in vivo

Abstract: It is known that, with aging, there is a change in fat distribution, including a relative increase in intra-abdominal fat and ectopic fat deposition, which is often accompanied by obesity. It is important to note that obesity with aging accelerates the aging process and the development of such chronic diseases as ischemic heart disease, hypertension, type 2 diabetes mellitus (DM2), osteoarthritis, cataracts, etc. In the last decade, it has been shown that slowly progressive inflammation of adipose tissue is one of the key pathogenetic factors in obesity and aging. One of the important endogenous regulators of inflammation in tissues is mesenchymal stem cells (MSC), due to their powerful secretory activity. However, with aging, the secretory activity of MSCs changes and becomes pro-inflammatory. One of the methods for normalizing the activity of MSC functioning is the transplantation of MSCs that secrete anti-inflammatory cytokines. The most effective operation can be performed by introducing autologous MSCs. In our previous studies, it was shown that the Cdc42 protein is an important regulator of the aging of MSCs and the activity of Cdc42 had increased the level of senescent MSCs in tissues isolated from adipose tissue of aging animals. In several works, including this previous study, it was shown that inhibition of Cdc42 activity improves the secretion of MSCs and partially rejuvenates cells. We suggest that the transplantation of MSCs, which undergoes inhibition of Cdc42 activity in animals with inflammaging of adipose tissue, may be a method of therapy for senile obesity.

Project goal
The project goal is to study the expression levels of pro-inflammatory and anti-inflammatory cytokines in blood serum in association with APOE genotype and inflammation-related gene polymorphisms in DNA samples isolated from AD patients and age- and sex-matched healthy individuals.

Project tasks:

1. Study of APOE genotypes in DNA samples isolated from the peripheral blood of patients with dementia and the control group
2. Study of serum pro-inflammatory and anti-inflammatory cytokine levels, as well as blood inflammatory profiles in the patients diagnosed with AD and the control group
3. Evaluation of inflammation-related genes polymorphisms in DNA samples isolated from the peripheral blood of patients with dementia and the control group
4. Study the associations between APOE genotype, inflammation-related genes polymorphisms, and pro-inflammatory and anti-inflammatory cytokines and chemokines levels in patients with dementia and the control group. Our special focus of interest is a comparative analysis of the inflammation-related gene polymorphisms in APOE4 carriers with and without dementia.

Abstract: Over the last decade, it become evident that chronic peripheral inflammation is one of the important factors contributing significantly to the development and progression of Alzheimer’s disease (AD), and that chronic inflammation in AD involves innate immune cells and inflammatory mediators, such as cytokines and chemokines. However, despite increasing evidence supporting the association between abnormal inflammation and AD, no well-established inflammatory biomarkers are currently available for AD and there has been a sparse expansion of inflammation-based biomarkers and preventive strategies. In turn, Apolipoprotein E (APOE) polymorphism is the primary genetic risk factor for sporadic AD. The human form of the APOE gene possesses three polymorphic alleles: E2, E3, and E4, and the E4 allele occurs in 40% of AD patients. ApoE is a key regulator of lipid homeostasis and data is indicating that ApoE is associated with increased inflammatory responses before and after the onset of AD pathogenesis. In this regard, we propose to study the interplays between APOE genotype, blood inflammatory biomarkers, and inflammation-related gene polymorphisms in AD patients and cognitively healthy aged people. Our special focus of interest is studying the inflammation-related gene polymorphisms in APOE4 carriers with dementia in comparison with APOE4 carriers who are not demented. We hypothesize that APOE4 and some inflammation-related gene polymorphisms may exert synergistic effects significantly increasing the risks of AD. The proposed research will provide novel insights into how APOE and pro-inflammatory gene isoforms impact the risks of AD. This study can potentially contribute to the discovery of new genetic and blood-based biomarkers for earlier diagnosis of AD and give impetus to examine new therapeutic measures (e.g. CRISPR/Cas9 therapy) for the management/alleviation of AD-associated CNS outcomes.

Project goal
To study the relationships between the human gut and oral microbiome diversity and composition, peripheral inflammation markers, biogeography, ethnicity, lifestyle (e.g., diet), and AD.

Project tasks:

1. To study the gut metagenome in patients with AD and age and gender-matched healthy seniors.
2. To study the oral metagenome in patients with AD and age and gender-matched healthy seniors.
3. To perform a comparative analysis of gut and oral microbiome diversity and composition in association with food consumption patterns in patients with dementia and healthy controls from different ethnic groups residing in two different geographic locations of Kazakhstan (North vs. South).
4. To study the blood expression levels of pro-inflammatory and anti-inflammatory cytokine and chemokine biomarkers in AD patients and a group of controls in association with gut and oral microbiomes diversity and composition

Abstract: Recent studies strongly suggest that the gut microbiome can influence brain functions and contribute to the development of Alzheimer's disease (AD). However, there are no studies comparing gut microbiome alterations associated with AD in different human populations, although evidence exists that the changes in the gut microbiome in AD patients from different countries are not similar. These differences might be attributed to such facts as biogeography, ethnicity, lifestyle, and eating habits. There is also evidence aside from the gut microbiome that oral microflora can influence brain functions; however, studies of the oral microbiome in AD are very limited. There is also an assumption that the development of sporadic AD might be driven by microbiome-associated peripheral inflammation. Therefore, more research is needed to reveal the relationships between human microbiome alterations, inflammation, and lifestyle in different ethnic populations and their influence on cognitive functions and risks of AD. Yet, no studies have been done on the associations between the gut and oral microbiomes and AD risks in Central Asia. Based on the foregoing, the main goal of the proposed research is to study the relationships between the human gut and oral microbiome diversity and composition, peripheral inflammation markers, biogeography, ethnicity, lifestyle (e.g., diet), and AD.

Project goal:
The goal of the project is to develop a new drug delivery system based on PEG-PLGA nanoparticles containing Cdc42 inhibitor (CASIN) loaded into a thermosensitive liquid rectal suppository for targeted therapy of colorectal cancer.

Project tasks:

1. Synthesis and characterization of PEG-PLGA nanoparticles containing a Cdc42 inhibitor
2. Evaluation of the effect of PLGA-PEG-CASIN nanoparticles on the viability, migratory and invasive potential of colorectal cancer cells.
3. Development and characterization of thermosensitive enemas loaded with PLGA-PEG-CASIN nanoparticles.
4. Evaluation of the effect of PLGA-PEG-CASIN nanoparticles loaded into thermosensitive enemas on the viability, migratory and invasive potential of colorectal cancer cells.
5. In vivo evaluation of distribution and retention of PLGA-PEG nanoparticles loaded into thermosensitive enemas

Abstract: Cdc42 is abnormally expressed in colorectal cancer and promotes tumor growth and metastasis. Therefore, Cdc42 is a promising target for targeted cancer therapy. However, commercially available Cdc42 inhibitors have low bioavailability and are rapidly eliminated from the body. In this regard, it is necessary to develop a drug delivery system for small molecules of Cdc42 inhibitors. In this project, we propose to develop a rectal system based on PEG-PLGA nanoparticles loaded with a Cdc42 inhibitor, which can effectively pass through the mucus and deliver the drug to the cells, as a temperature-sensitive gel. It will facilitate the effective distribution and retention of nanoparticles in the intestine.

Project goal
This project aims to study the functional dependence of Aβ phagocytosis in blood monocytes, isolated from AD patients, on the phenotype (M1/M2) and IDE concentration in these cells, and to study the possibility of pharmacological correction of Aβ proteolysis disturbance.

Project tasks:

1. Study of monocytes’ phenotype (M1/M2)
2. Study of monocytes’ phagocytic activity in association with monocyte’s polarization phenotype (M1/M2)
3. Study of somatostatin and tuftsin effects on IDE activity in blood monocytes in association with their phagocytic activity in vitro.

Abstract: According to the amyloid hypothesis the progression of Alzheimer’s disease (AD) is driven by the disturbance of amyloid-β peptide (Аβ) metabolism and its accumulation in the brain. Clearance of Аβ from the brain and its further utilization in the blood is regulated via several pathways. The main mechanism of Аβ utilization in the central nervous system (CNS) is phagocytosis performed by brain microglia and blood-derived monocytes. According to the literature, 40%-60% of Аβ is utilized by peripheral blood monocytes and disturbance of this process can be a reason for Аβ accumulation in the brain. Monocytes represent a heterogeneous population of mononuclear cells with functionally different subpopulations of different phenotypes (M1 and M2). They are characterized by a certain degree of plasticity, and depending on age or pathological conditions, they can change their phenotypes. It is believed that the M2 phenotype has higher phagocytic activity, however, both phenotypes are involved in Аβ phagocytosis in AD patients. Nevertheless, the effectiveness of Aβ phagocytosis and its dependence on the monocytes’ phenotype in AD is still unknown. Several enzymes are responsible for the proteolysis of Аβ, however, therapeutically, the most interesting is the insulin-degrading enzyme (IDE). IDE is capable of cleaving Aβ oligomers into monomers and submolecular fragments. Dysfunction of IDE enzyme is observed in AD patients; however, mechanisms of pathogenesis are still poorly understood. Assessment of phagocytosis disturbance and its induction if AD patients are offered in the frame of this project.

Project goal
The goal of the project is to develop cellular delivery methods for CRISPR/Cas9 complexes using polymeric conjugates that enable cellular penetration of the complex and gene editing.

Project tasks:

1. Development, synthesis, and characterization of Cas9-polymer conjugates.
2. Development of cellular delivery of CRISPR/Cas9-polymer complexes
3. Editing of cancer marker genes in cancer cell lines using CRISPR/Cas9-polymer complexes

Abstract: CRISPR/Cas9 delivery into cells has been extensively studied and several methods have been developed. However, all these methods have their own advantages and limitations. Delivery of the Cas9 in a protein form allows for the fastest onset of gene editing, while the delivery of the protein in a complex with RNA and DNA is considered the most difficult pathway. Herein, we propose to develop a Cas9 enzyme conjugated to polymers to enable cell membrane penetration and efficient gene editing. The data obtained may serve as a basis for the development of new therapeutic strategies for genome editing.

Project goal
The main aim of the project is pre-clinical testing of the new composites for fracture bone regeneration based on the modifications of the tricalcium phosphate (TCP) ceramic porous granules with Strontium (Sr) and Copper (Cu).

Project tasks:

1. In vitro assessments of the effects of TCP ceramic porous granules modified with Sr and Cu on osteogenic differentiation and migration properties of bone marrow-derived mesenchymal stem cells (BM-MSCs) and evaluation of its anti-osteoclastic activity
2. In vitro evaluation of the angiogenic potential of TCP ceramic porous granules in the HUVEC model and assessment of its biocompatibility and antibacterial properties
3. In vivo evaluation of the therapeutic potential of the TCP ceramic porous granules modified with Sr and Cu in a rat model of the ulna fracture

Abstract: Bioceramics and, in particular, calcium-phosphate-based materials are widely recognized for their excellent biocompatibility, osteoconductivity, and their ability to induce biomineralization. In the present project, tricalcium phosphate (TCP) ceramic porous granules modified with Strontium (Sr) and Copper (Cu) will be developed. Our choice of the metals is dictated by the fact that Sr has been reported to possess osteogenic and bone regeneration properties while Cu is capable of increasing the antibacterial and angiogenic potential of the TCP granules. Sr is present in the mammalian bones’ mineral phase. It is hypothesized that Sr plays a role in the early bone formation and induction of the positive effects on osteogenesis, osteoclastogenesis, and angiogenesis. Furthermore, Sr has positive therapeutic effects on osteoporosis and osteoarthritis. Sr has been also shown to increase osteoblast proliferation, differentiation, and mineral formation, and to slow down the osteoclastic resorption of the bone. Sr-doped hydroxyapatite has a positive effect on osseointegration and bone regeneration, according to our experience and literature reports. The use of Cu in the biomedical field is desirable because it is an essential cofactor of several enzymes and possesses proangiogenic and antibacterial properties. In this regard, in this project, we propose to assess the efficiency of the deposition of TCP granules modified with Sr and Cu in vitro and in vivo. We suggest that implantation of the TCP ceramic porous granules modified with Sr and Cu in the zone of bone fracture will be an effective method for stimulating reparative osteogenesis and may improve the outcomes of pathological bone fractures.

Project goal
The research aims to develop new innovative biocompatible coatings and related technological processes for the effective manufacture of affordable medical adaptive orthopedics implants with improved biocompatibility and osseointegration properties.

Project tasks:

1. Comprehensive analysis of titanium alloys for medical implants and different techniques of coatings with bioactive materials
2. Development of advanced technologies for the effective production of small-scale samples of titanium medical implants with bioactive coatings
3. In vitro analysis of small-scale samples of titanium medical implants with bioactive coatings for biocompatibility and antibacterial activity
4. Development of 2 types of medical implants prototypes and their quality characterization

Abstract: Currently, the priority direction of solving problems related to public health protection is the development of high-tech competitive technologies to produce affordable medical products that meet international quality standards, particularly metal implants for dentistry and orthopedics, and are patient-specific or adaptive. Such implants are needed to speed up the patient’s recovery, reduce the risk of post-surgery complications associated with implant rejection, and ensure long service life. Nowadays, more than 5% of embedded implants are rejected by the patient’s body due to the development of bone necrosis, which occurs when the implant surface is destroyed, deformed, and metal particles penetrate the surrounding tissues of the body; or due to the development of periprosthetic bacterial infections. Therefore, the development of new technologies for surface treatment of medical implants to increase their biocompatibility, ensure their non-toxic and antibacterial properties, and provide osteoconductivity osseointegration to shorten the patient’s recovery time and prolong the life of the implant is of great importance. In this project, we propose additive manufacturing of metal products by selective laser melting, and development of innovative coatings from biocompatible materials with controlled structure and properties, and the automation of related technological processes to produce affordable medical implants for dentistry and orthopedics.

Project goal: The aim of the project is to explore the potential preventive and therapeutic effects of recently proposed antioxidants of natural and synthetic origin using molecular, histological and cognitive behavioral research methods in vitro and in vivo in the APP/PS1 mouse model of Alzheimer's disease.

Project description: Increasing evidence suggests that oxidative stress and neuro inflammation significantly contribute to the mechanisms of AD. However, therapeutic and preventive options with these targets are currently insufficiently investigated and largely underdeveloped. At the same time, several antioxidants of various origins, from synthetic nanoparticles to natural plant extracts and food compounds, were shown to interfere with key molecular and cellular processes underlying the AD. For example, sialic-acid binding lectins ameliorate AD pathology in the CNS [1]. Rich in phenolic acids and flavonols plant extract of Limonium gmelinii suppressed oxidative stress and pro-inflammatory responses induced by NMDA and TNF-α in human primary neurons and astrocytes [2]. Several new nanoparticles based on the fusion of fullerene or fullerenol with dihydroquercetine, an extract obtained from the Dahurian larch, were shown to exert powerful anti-oxidant effect and inhibit amyloid fibril formation [3]. However, most of these studies have been carried out in vitro and need to be expanded to the established in vivo models. One of the most commonly used AD model is APP/PS1, in which transgenic modification leads to the expression of amyloid precursor protein and presinelin 1 in a mouse brain. This mutation results in the development of hallmarks of AD in mutant mice APP/PS1: the formation of pathological amyloid plaques, cognitive and emotional disturbances [4]. Based on the foregoing, we propose to study potential preventive and therapeutic effects of recently proposed antioxidants of natural and synthetic origins, using molecular, histological, and cognitive readouts in vitro and in the above-mentioned AD mouse model. We will utilize previously established protocols of in vitro assays and of cognitive tests, MDA assay, expression of cytokines and cardiovascular markers (platelet activation) in the brain and periphery as described elsewhere

PI: Sholpan Askarova
Co-PI: Andrey Tsoy, Aliya Kassenova

Project partners: First Moscow State Medical University named after Sechenov; Al-Farabi Kazakh National University

Realisation period: 2024-2026

Expected results: The efficacy of new anti-oxidants and flavonoid-containing extract to treat or prevent AD-like pathology will be studied that can lead to the development of new remedies of AD; new data linking neuroinflammation, oxidative stress, microbiome will be obtained in the APP/PS1 mouse model that can be extrapolated to the clinical situation in patients with AD and result in the identificantion of new mechansims of this disorder.