Vladislav Sergeev - Academia.edu (original) (raw)

Papers by Vladislav Sergeev

Research paper thumbnail of Xenotransplantation of Human Hematopoietic Stem Cells into NBSGW Mice: A Basic Model for Preclinical Development of Gene Therapy Approaches

Kliničeskaâ onkogematologiâ, Apr 1, 2024

Актуальность. Генная терапия на основе транспланта ции кроветворных клеток становится мощной и ун... more Актуальность. Генная терапия на основе транспланта ции кроветворных клеток становится мощной и универ сальной стратегией лечения все более расширяющегося спектра различных заболеваний человека. Одна из акту альных проблем при внедрении технологий модифика ции генома в гемопоэтических стволовых клетках (ГСК) в клиническую практику -обеспечение качества и безо пасности продуктов генной и клеточной терапии при их использовании у человека. Это достигается путем тести рования на животных моделях на этапе доклинических исследований. В этом отношении линия мышей NBSGW представляется уникальной и перспективной моделью, которая позволяет обеспечивать приживление ГСК че ловека без предварительного кондиционирования. Цель. Апробация модели мышей NBSGW для прижив ления ГСК человека, оптимизация методов оценки со стояния животных и мониторинга уровня химеризма для трансляционной доклинической разработки про дуктов генной и клеточной терапии на основе ГСК. Материалы и методы. Для создания моделей ксе нотрансплантатов мышей NBSGW использовали образ цы отобранных CD34+ ГСК периферической крови здо рового донора. Серийную трансплантацию проводили методом внутривенной инъекции клеток костного мозга от первичных реципиентов с высоким уровнем химериз ма. Эффективность приживления оценивали методами проточной цитофлюориметрии (ПЦФ) и цифровой ка пельной полимеразной цепной реакции (цкПЦР). Субпо

Research paper thumbnail of Triple-responsive inorganic–organic hybrid microcapsules as a biocompatible smart platform for the delivery of small molecules

Journal of Materials Chemistry B, 2016

We designed novel hybrid inorganic/organic capsules with unique physicochemical features enabling... more We designed novel hybrid inorganic/organic capsules with unique physicochemical features enabling multimodal triggering by physical (UV light, ultrasound) and chemical (enzymatic treatment) stimuli. Notably, the UV-and ultrasound response was achieved by a synergetic combination of TiO 2 and SiO 2 nanostructures which were in situ deposited into the polymer shell of microcapsules during sol-gel synthesis. This results in the formation of a composite hybrid shell with enhanced mechanical stability. Such sol-gel modification reduces the permeability of the capsule shell to allow for small molecule encapsulation. At the same time, these hybrid capsules consist of degradable polypeptides and polysaccharides and can be decomposed in response to enzymatic reaction. Upon employing different modes of treatment (UV-light, ultrasound or enzymatic degradation) we can stimulate different mechanisms of cargo release at desired times. Importantly, such capsules have been shown to be non-cytotoxic and can be internalized into human mesenchymal stem cells (MSCs) and cervical cancer cell lines (HeLa) revealing intracellular degradation. This work demonstrates that our hybrid capsules possess a triple stimuli-responsive effect, which is of capital importance for the future design and application of multimodal responsive platforms to improve externally stimulated release of bioactive compounds and their healthcare performance.

Research paper thumbnail of Safe and Effective Delivery of Antitumor Drug Using Mesenchymal Stem Cells Impregnated with Submicron Carriers

ACS Applied Materials & Interfaces, Mar 18, 2019

An important area in modern malignant tumor therapy is the optimization of the antitumor drugs ph... more An important area in modern malignant tumor therapy is the optimization of the antitumor drugs pharmacokinetics. The use of some antitumor drugs is limited in clinical practice due to their high toxicity. Therefore, the strategy for optimizing the drug pharmacokinetics is focusing on the generation of high local concentrations of these drugs in the tumor area with minimal systemic and tissue-specific toxicity. This can be achieved by encapsulation of highly toxic antitumor drug (vincristine/VCR that is 20-50 times more toxic than widely used in antitumor therapy doxorubicin) into nano-and microcarriers with their further association into therapeutically relevant cells that possess ability to migrate to sites of tumor. Here we fundamentally examine the effect of drug carrier size on behavior of human mesenchymal stem cells (hMSCs), incl. internalization efficiency, cytotoxicity, cell movement, to optimize the conditions for the development of carrier-hMSCs drug delivery platform. Using the malignant tumors derived from patients, we evaluated the capability of hMSCs associated with VCR loaded carriers to target tumors using a 3D spheroid model in collagen gel. Compared to free VCR, developed hMSCsbased drug delivery platform showed enhanced antitumor activity regarding to those tumors that express CXCL12 (SDF-1) gene, inducing directed migration of hMSCs via CXCL12 (SDF-1)/CXCR4 pathway. These results show that the combination of encapsulated antitumor drugs and hMSCs, which possess the properties of active migration into tumors, is therapeutically beneficial and demonstrated high efficiency and low systematic toxicity, revealing novel strategies for chemotherapy in the future.

Research paper thumbnail of Efficient gene editing via non-viral delivery of CRISPR–Cas9 system using polymeric and hybrid microcarriers

Nanomedicine: Nanotechnology, Biology and Medicine, 2018

CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the trea... more CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarriers, made of degradable polymers such as polypeptides and polysaccharides and modified by silica shell, for delivery of all CRISPR-Cas9 components. We found that these microcarriers mediate more efficient transfection than a commercially available liposome-based transfection reagent (>70% vs. <50% for mRNA, >40% vs. 20% for plasmid DNA). For proofof-concept, we delivered CRISPR-Cas9 components using our capsules to dTomato-expressing HEK293T cellsa model, in which loss of red fluorescence indicates successful gene editing. Notably, transfection of indicator cells translated in high-level dTomato knockout in app. 70% of transfected cells. In conclusion, we have provided proof-of-principle that our micro-sized containers represent promising non-viral platforms for efficient and safe gene editing.

Research paper thumbnail of Клеточная терапия несовершенного остеогенеза

Research paper thumbnail of Генная терапия на основе трансплантации гемопоэтических стволовых клеток с использованием сайт-специфического редактирования генома

Research paper thumbnail of High prevalence of CD3, NK, and NKT cells in the graft predicts adverse outcome after matched-related and unrelated transplantations with post transplantation cyclophosphamide

Bone Marrow Transplantation, Sep 20, 2019

The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT... more The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT is well known for conventional GVHD prophylaxis based on calcineurin inhibitors with or without antithymocyte globulin. Currently, there is limited data whether these results could be translated to post transplantation cyclophosphamide (PTCy). The prospective extension cohort of NCT02294552 trial enrolled 79 adult patients with acute leukemia in CR. Twenty-six received matchedrelated bone marrow (BM) grafts with single-agent PTCy and 53 received unrelated peripheral blood stem cell graft (PBSC) with PTCy, tacrolimus, and MMF. The grafts were studied by the flow cytometry, and plasma samples were analyzed by ELISA. In the cluster and major component analysis, we determined that transplantation from donors with high content of CD3, NKT, and CD16-CD56 + subpopulations in the PBSC grafts was associated with poor immunological recovery and compromised event-free survival (50% vs. 80%, HR 2.93, p = 0.015) both due to increased relapse incidence and nonrelapse mortality. The significant independent predictor of moderate and severe chronic GVHD was the high prevalence of and iNKT, Vβ11, and double-positive cells in the PBSC grafts from young donors (HR 2.75, p = 0.0483). No patterns could be identified for BM grafts and for plasma biomarkers.

Research paper thumbnail of Острый миелобластный лейкоз и миелодиспластический синдром: применение азацитидина с профилактической и превентивной целью после аллогенной трансплантации гемопоэтических стволовых клеток

Клиническая онкогематология. Фундаментальные исследования и клиническая практика, 2017

Research paper thumbnail of Mobilization of hematopoietic stem cells (HSCs) by using Pleriksafor (Mozobail) in combinationwith colony-stimulating factor in patients with previous failure transplant procurement for autotransplantation of HSCs

Modern Oncology, 2014

AutoHSCT is an effective treatment for patients with relapsed and refractory lymphomas, multiple ... more AutoHSCT is an effective treatment for patients with relapsed and refractory lymphomas, multiple myeloma, some solid tumors. For a successful transplantation it is necessary to obtain an adequate quality and quantity transplant. Due to the heavy pretreatment this category of patients receiving a transplant can be a serious problem, which can be solved with the use of the new mobilizing agent inhibitor CXCR4 - plerixafor. In our study, 10 patients, of which 7 were confirmed as «poor mobilizers» were stimulated with colony stimulating factor, together with a plerixafor dose 0,24 mg/kg body weight on day 5 and 6 of stimulation. In 9 of the 10 patients we had obtained an adequate transplant. In all patients before the administration of plerixafor low or intermediate number of CD34+ cells per microliter of peripheral blood was obtained. The patient failed to mobilize with the use of plerixafor mentioned the smallest number of CD34+ cells per microliter in peripheral blood both before and...

Research paper thumbnail of Functionalization of bone marrow mesenchymal stromal cells by multifunctional capsules for targeted drug delivery

Cellular therapy and transplantation, 2018

Research paper thumbnail of Production of autologous T-lymphocytes with anti-CD19 chimeric antigen receptor using an automated, completely closed and serum-free technological process

Cellular therapy and transplantation, Dec 29, 2023

Autologous anti-CD19 CART cells can be produced by means of closed automated systems, which seems... more Autologous anti-CD19 CART cells can be produced by means of closed automated systems, which seems to be the most convenient platform for the point-of-care production. However, to fully comply with good practices and to utilize relatively low-grade of clean air environments, the production processes should be carried out in a completely closed format, without usage of pooled human blood products. The purpose of the present study was to test a production process that fully meets the necessary requirements.

Research paper thumbnail of Gene-Cell Therapy of Hiv and Hematological Malignances Based on Hematopoietic Stem Cell Transplantation and Site-Specific Genome Editing

Journal Infectology, 2017

Резюме По данным ежегодных докладов UNAIDS, c 1983 г. количество ВИЧ-инфицированных пациентов неу... more Резюме По данным ежегодных докладов UNAIDS, c 1983 г. количество ВИЧ-инфицированных пациентов неуклонно растет. Антиретровирусная терапия (АРВТ) позволяет увеличить продолжительность жизни, но проблемы качества жизни и долгосрочной выживаемости не решены. Одной из основных причин летальности у пациентов с ВИЧ в эру АРВТ являются онкологические заболевания, среди которых значительную часть составляют злокачественные лимфомы. Лечение лимфом включает этапную противоопухолевую химиотерапию (ПХТ) с потенциалом полного излечения пациента. Высокодоз ная ПХТ с аутологичной трансплантацией гемопоэтических стволовых клеток (ТГСК)-основное средство лечения рецидивов и рефрактерных форм агрессивных злокачественных лимфом. В последние годы с появлением технологии редактирования генома аутологичная ТГСК становится также одним из самых перспективных методов лечения ВИЧ-инфекции. История излечения от ВИЧ «Берлинского пациента» благодаря аллогенной ТГСК от донора с мутацией гена CCR5 продемонстрировала эффективность подхода генной терапии на основе трансплантации. Высокодозная химиотерапия с трансплантацией аутологичных, подвергнутых редактированию (инактивации гена CCR5, с помощью генно-инженерных нуклеаз), гемопоэтических стволовых клеток потенциально представляет собой способ комплексного лечения этих пациентов. С одной стороны, высокодозная ПХТ с ауто-ТГСК обеспечивает излечение от злокачественной опухоли, с другой стороны, она является способом доставки отредактированных клеток и создания условий для реализации лечебного эффекта против ВИЧ.

Research paper thumbnail of Screening for predictive biomarkers after related and unrelated grafts with posttransplantation cyclophosphamide in adult patients

Research paper thumbnail of Functionalization of bone marrow mesenchymal stromal cells by multifunctional capsules for targeted drug delivery

Research paper thumbnail of A highly efficient and safe gene delivery platform based on polyelectrolyte core–shell nanoparticles for hard-to-transfect clinically relevant cell types

Journal of Materials Chemistry B

The polyelectrolyte nanocarriers’ based on nanosized vaterite particles as a novel tool for genet... more The polyelectrolyte nanocarriers’ based on nanosized vaterite particles as a novel tool for genetic material delivery into the clinically relevant cell types and potential application of described technology in gene therapy approaches.

Research paper thumbnail of High prevalence of CD3, NK, and NKT cells in the graft predicts adverse outcome after matched-related and unrelated transplantations with post transplantation cyclophosphamide

Bone Marrow Transplantation

The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT... more The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT is well known for conventional GVHD prophylaxis based on calcineurin inhibitors with or without antithymocyte globulin. Currently, there is limited data whether these results could be translated to post transplantation cyclophosphamide (PTCy). The prospective extension cohort of NCT02294552 trial enrolled 79 adult patients with acute leukemia in CR. Twenty-six received matched-related bone marrow (BM) grafts with single-agent PTCy and 53 received unrelated peripheral blood stem cell graft (PBSC) with PTCy, tacrolimus, and MMF. The grafts were studied by the flow cytometry, and plasma samples were analyzed by ELISA. In the cluster and major component analysis, we determined that transplantation from donors with high content of CD3, NKT, and CD16-CD56 + subpopulations in the PBSC grafts was associated with poor immunological recovery and compromised event-free survival (50% vs. 80%, HR 2.93, p = 0.015) both due to increased relapse incidence and non-relapse mortality. The significant independent predictor of moderate and severe chronic GVHD was the high prevalence of and iNKT, Vβ11, and double-positive cells in the PBSC grafts from young donors (HR 2.75, p = 0.0483). No patterns could be identified for BM grafts and for plasma biomarkers.

Research paper thumbnail of Safe and Effective Delivery of Antitumor Drug Using Mesenchymal Stem Cells Impregnated with Submicron Carriers

ACS Applied Materials & Interfaces

An important area in modern malignant tumor therapy is the optimization of antitumor drugs pharma... more An important area in modern malignant tumor therapy is the optimization of antitumor drugs pharmacokinetics. The use of some antitumor drugs is limited in clinical practice due to their high toxicity. Therefore, the strategy for optimizing the drug pharmacokinetics focuses on the generation of high local concentrations of these drugs in the tumor area with minimal systemic and tissue-specific toxicity. This can be achieved by encapsulation of highly toxic antitumor drug (vincristine (VCR) that is 20-50 times more toxic than widely used the antitumor drug doxorubicin) into nano- and microcarriers with their further association into therapeutically relevant cells that possess the ability to migrate to sites of tumor. Here, we fundamentally examine the effect of drug carrier size on the behavior of human mesenchymal stem cells (hMSCs), including internalization efficiency, cytotoxicity, cell movement, to optimize the conditions for the development of carrier-hMSCs drug delivery platform. Using the malignant tumors derived from patients, we evaluated the capability of hMSCs associated with VCR-loaded carriers to target tumors using a three-dimensional spheroid model in collagen gel. Compared to free VCR, the developed hMSC-based drug delivery platform showed enhanced antitumor activity regarding those tumors that express CXCL12 (stromal cell-derived factor-1 (SDF-1)) gene, inducing directed migration of hMSCs via CXCL12 (SDF-1)/CXCR4 pathway. These results show that the combination of encapsulated antitumor drugs and hMSCs, which possess the properties of active migration into tumors, is therapeutically beneficial and demonstrated high efficiency and low systematic toxicity, revealing novel strategies for chemotherapy in the future.

Research paper thumbnail of Acute Myeloblastic Leukemia and Myelodysplastic Syndrome: Azacitidine for Prophylactic and Preventive Purposes after Allogeneic Hematopoietic Stem Cell Transplantation

Clinical oncohematology

Aim. To evaluate the effectiveness of preventive and prophylactic post-transplantation therapy us... more Aim. To evaluate the effectiveness of preventive and prophylactic post-transplantation therapy using azacitidine (5-AZA) in patients at high risk of post-transplantation relapse. Methods. 136 patients were included in the study performed by the pairwise comparison: 68 of them received 5-AZA after allo-HSCT and 68 patients were included in the historical control group. 5-AZA was prescribed for prophylactic or preventive purposes. The results were assessed according to the OS, RR, EFS, DUM, and relapse-free and GVHR-free survival. Results. 1-year OS was 76 % in the 5-AZA group (95% CI 60-84 %) and 44 % in the reference group (95% CI 33-55 %) (р = 0.001); 2-year OS was 63 % (95% CI 39-67 %) and 37 % (95% CI 26-48 %) (р = 0.007), respectively. The relapse rate (RR) in the 5-AZA group was 34 % (95% CI 22-46 %) during 1 year and 51 % (95% CI 38-64 %) in the reference group (р = 0.02). 1- and 2-year disease unrelated mortality (DUM) was similar: 5 % in the 5-AZA group (95% CI 0.1-14.0 %) a...

Research paper thumbnail of Potential clinical applications of bone marrow-derived mesenchymal stem cells in bone regeneration

Cellular Therapy and Transplantation

Bone marrow (BM) contains a small resident cell population referred to as 'multipotent mesenchyma... more Bone marrow (BM) contains a small resident cell population referred to as 'multipotent mesenchymal stem cells' (MSC). These adherent cells could be isolated and expanded in simple culture media and may differentiate in adipogenic or osteogenic pathway. So far an opportunity of MSC differentiation to hepatocytes, brain, or renal cells is not proven yet. Meanwhile, two potential clinical applications are considered for MSC: (1) as a tool for immune modulation in graft-versus-host disease (GVHD) and autoimmune diseases, or, (2) as a potential source of growth-promoting factors in specialized tissues. This heterogenous population may support hematopoiesis by secreting growth factors, cytokines and other biologically active substances. Upon injection, MSCs are able to migrate into damaged tissues, thus promoting their repair. However, only small MSC fraction may reach bone marrow niches following intravenous infusion. Multiple experiments with MSCs in different injury models show their ability to suppress apoptosis initiated by hypoxia, chemical agents/acidity and other deteriorating factors. This protective effect is mediated by a number of secreted growth factors, e.g., granulocyte-macrophage colony-stimulating growth factor (GM-CSF). A big number of clinical trials show high level of safety for the MSC therapy. Both clinical and experimental studies demonstrated only weak immunogenic effects of allogeneic MSC upon injection into immunocompetent recipients. At the present time, injections of in vitro expanded MSCs were performed in the patients developing acute GVHD after hematopoietic stem cell transplantation (HSCT), and in some autoimmune disorders. Over last decade, several studies concerned potentially curative effects of MSCs injected into affected bone areas in the patients with osteogenesis imperfecta (OI), a severe inherited disease with altered collagen structure resulting into increased bone fragility. Here we present a synopsis of clinical protocol aimed for assessing safety, immunogenicity, and clinical effects of MSC injected to the OI patients during corrective osteotomy. One may suggest that a minor MSC subpopulation may migrate to the damaged areas differentiating to chondrocytes and osteoblasts, and, hence, contributing to the bone repair.

Research paper thumbnail of Efficient gene editing via non-viral delivery of CRISPR–Cas9 system using polymeric and hybrid microcarriers

Nanomedicine: Nanotechnology, Biology and Medicine

CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the trea... more CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarriers, made of degradable polymers such as polypeptides and polysaccharides and modified by silica shell, for delivery of all CRISPR-Cas9 components. We found that these microcarriers mediate more efficient transfection than a commercially available liposome-based transfection reagent (&gt;70% vs. &lt;50% for mRNA, &gt;40% vs. 20% for plasmid DNA). For proof-of-concept, we delivered CRISPR-Cas9 components using our capsules to dTomato-expressing HEK293T cells-a model, in which loss of red fluorescence indicates successful gene editing. Notably, transfection of indicator cells translated in high-level dTomato knockout in approx. 70% of transfected cells. In conclusion, we have provided proof-of-principle that our micro-sized containers represent promising non-viral platforms for efficient and safe gene editing.

Research paper thumbnail of Xenotransplantation of Human Hematopoietic Stem Cells into NBSGW Mice: A Basic Model for Preclinical Development of Gene Therapy Approaches

Kliničeskaâ onkogematologiâ, Apr 1, 2024

Актуальность. Генная терапия на основе транспланта ции кроветворных клеток становится мощной и ун... more Актуальность. Генная терапия на основе транспланта ции кроветворных клеток становится мощной и универ сальной стратегией лечения все более расширяющегося спектра различных заболеваний человека. Одна из акту альных проблем при внедрении технологий модифика ции генома в гемопоэтических стволовых клетках (ГСК) в клиническую практику -обеспечение качества и безо пасности продуктов генной и клеточной терапии при их использовании у человека. Это достигается путем тести рования на животных моделях на этапе доклинических исследований. В этом отношении линия мышей NBSGW представляется уникальной и перспективной моделью, которая позволяет обеспечивать приживление ГСК че ловека без предварительного кондиционирования. Цель. Апробация модели мышей NBSGW для прижив ления ГСК человека, оптимизация методов оценки со стояния животных и мониторинга уровня химеризма для трансляционной доклинической разработки про дуктов генной и клеточной терапии на основе ГСК. Материалы и методы. Для создания моделей ксе нотрансплантатов мышей NBSGW использовали образ цы отобранных CD34+ ГСК периферической крови здо рового донора. Серийную трансплантацию проводили методом внутривенной инъекции клеток костного мозга от первичных реципиентов с высоким уровнем химериз ма. Эффективность приживления оценивали методами проточной цитофлюориметрии (ПЦФ) и цифровой ка пельной полимеразной цепной реакции (цкПЦР). Субпо

Research paper thumbnail of Triple-responsive inorganic–organic hybrid microcapsules as a biocompatible smart platform for the delivery of small molecules

Journal of Materials Chemistry B, 2016

We designed novel hybrid inorganic/organic capsules with unique physicochemical features enabling... more We designed novel hybrid inorganic/organic capsules with unique physicochemical features enabling multimodal triggering by physical (UV light, ultrasound) and chemical (enzymatic treatment) stimuli. Notably, the UV-and ultrasound response was achieved by a synergetic combination of TiO 2 and SiO 2 nanostructures which were in situ deposited into the polymer shell of microcapsules during sol-gel synthesis. This results in the formation of a composite hybrid shell with enhanced mechanical stability. Such sol-gel modification reduces the permeability of the capsule shell to allow for small molecule encapsulation. At the same time, these hybrid capsules consist of degradable polypeptides and polysaccharides and can be decomposed in response to enzymatic reaction. Upon employing different modes of treatment (UV-light, ultrasound or enzymatic degradation) we can stimulate different mechanisms of cargo release at desired times. Importantly, such capsules have been shown to be non-cytotoxic and can be internalized into human mesenchymal stem cells (MSCs) and cervical cancer cell lines (HeLa) revealing intracellular degradation. This work demonstrates that our hybrid capsules possess a triple stimuli-responsive effect, which is of capital importance for the future design and application of multimodal responsive platforms to improve externally stimulated release of bioactive compounds and their healthcare performance.

Research paper thumbnail of Safe and Effective Delivery of Antitumor Drug Using Mesenchymal Stem Cells Impregnated with Submicron Carriers

ACS Applied Materials & Interfaces, Mar 18, 2019

An important area in modern malignant tumor therapy is the optimization of the antitumor drugs ph... more An important area in modern malignant tumor therapy is the optimization of the antitumor drugs pharmacokinetics. The use of some antitumor drugs is limited in clinical practice due to their high toxicity. Therefore, the strategy for optimizing the drug pharmacokinetics is focusing on the generation of high local concentrations of these drugs in the tumor area with minimal systemic and tissue-specific toxicity. This can be achieved by encapsulation of highly toxic antitumor drug (vincristine/VCR that is 20-50 times more toxic than widely used in antitumor therapy doxorubicin) into nano-and microcarriers with their further association into therapeutically relevant cells that possess ability to migrate to sites of tumor. Here we fundamentally examine the effect of drug carrier size on behavior of human mesenchymal stem cells (hMSCs), incl. internalization efficiency, cytotoxicity, cell movement, to optimize the conditions for the development of carrier-hMSCs drug delivery platform. Using the malignant tumors derived from patients, we evaluated the capability of hMSCs associated with VCR loaded carriers to target tumors using a 3D spheroid model in collagen gel. Compared to free VCR, developed hMSCsbased drug delivery platform showed enhanced antitumor activity regarding to those tumors that express CXCL12 (SDF-1) gene, inducing directed migration of hMSCs via CXCL12 (SDF-1)/CXCR4 pathway. These results show that the combination of encapsulated antitumor drugs and hMSCs, which possess the properties of active migration into tumors, is therapeutically beneficial and demonstrated high efficiency and low systematic toxicity, revealing novel strategies for chemotherapy in the future.

Research paper thumbnail of Efficient gene editing via non-viral delivery of CRISPR–Cas9 system using polymeric and hybrid microcarriers

Nanomedicine: Nanotechnology, Biology and Medicine, 2018

CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the trea... more CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarriers, made of degradable polymers such as polypeptides and polysaccharides and modified by silica shell, for delivery of all CRISPR-Cas9 components. We found that these microcarriers mediate more efficient transfection than a commercially available liposome-based transfection reagent (>70% vs. <50% for mRNA, >40% vs. 20% for plasmid DNA). For proofof-concept, we delivered CRISPR-Cas9 components using our capsules to dTomato-expressing HEK293T cellsa model, in which loss of red fluorescence indicates successful gene editing. Notably, transfection of indicator cells translated in high-level dTomato knockout in app. 70% of transfected cells. In conclusion, we have provided proof-of-principle that our micro-sized containers represent promising non-viral platforms for efficient and safe gene editing.

Research paper thumbnail of Клеточная терапия несовершенного остеогенеза

Research paper thumbnail of Генная терапия на основе трансплантации гемопоэтических стволовых клеток с использованием сайт-специфического редактирования генома

Research paper thumbnail of High prevalence of CD3, NK, and NKT cells in the graft predicts adverse outcome after matched-related and unrelated transplantations with post transplantation cyclophosphamide

Bone Marrow Transplantation, Sep 20, 2019

The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT... more The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT is well known for conventional GVHD prophylaxis based on calcineurin inhibitors with or without antithymocyte globulin. Currently, there is limited data whether these results could be translated to post transplantation cyclophosphamide (PTCy). The prospective extension cohort of NCT02294552 trial enrolled 79 adult patients with acute leukemia in CR. Twenty-six received matchedrelated bone marrow (BM) grafts with single-agent PTCy and 53 received unrelated peripheral blood stem cell graft (PBSC) with PTCy, tacrolimus, and MMF. The grafts were studied by the flow cytometry, and plasma samples were analyzed by ELISA. In the cluster and major component analysis, we determined that transplantation from donors with high content of CD3, NKT, and CD16-CD56 + subpopulations in the PBSC grafts was associated with poor immunological recovery and compromised event-free survival (50% vs. 80%, HR 2.93, p = 0.015) both due to increased relapse incidence and nonrelapse mortality. The significant independent predictor of moderate and severe chronic GVHD was the high prevalence of and iNKT, Vβ11, and double-positive cells in the PBSC grafts from young donors (HR 2.75, p = 0.0483). No patterns could be identified for BM grafts and for plasma biomarkers.

Research paper thumbnail of Острый миелобластный лейкоз и миелодиспластический синдром: применение азацитидина с профилактической и превентивной целью после аллогенной трансплантации гемопоэтических стволовых клеток

Клиническая онкогематология. Фундаментальные исследования и клиническая практика, 2017

Research paper thumbnail of Mobilization of hematopoietic stem cells (HSCs) by using Pleriksafor (Mozobail) in combinationwith colony-stimulating factor in patients with previous failure transplant procurement for autotransplantation of HSCs

Modern Oncology, 2014

AutoHSCT is an effective treatment for patients with relapsed and refractory lymphomas, multiple ... more AutoHSCT is an effective treatment for patients with relapsed and refractory lymphomas, multiple myeloma, some solid tumors. For a successful transplantation it is necessary to obtain an adequate quality and quantity transplant. Due to the heavy pretreatment this category of patients receiving a transplant can be a serious problem, which can be solved with the use of the new mobilizing agent inhibitor CXCR4 - plerixafor. In our study, 10 patients, of which 7 were confirmed as «poor mobilizers» were stimulated with colony stimulating factor, together with a plerixafor dose 0,24 mg/kg body weight on day 5 and 6 of stimulation. In 9 of the 10 patients we had obtained an adequate transplant. In all patients before the administration of plerixafor low or intermediate number of CD34+ cells per microliter of peripheral blood was obtained. The patient failed to mobilize with the use of plerixafor mentioned the smallest number of CD34+ cells per microliter in peripheral blood both before and...

Research paper thumbnail of Functionalization of bone marrow mesenchymal stromal cells by multifunctional capsules for targeted drug delivery

Cellular therapy and transplantation, 2018

Research paper thumbnail of Production of autologous T-lymphocytes with anti-CD19 chimeric antigen receptor using an automated, completely closed and serum-free technological process

Cellular therapy and transplantation, Dec 29, 2023

Autologous anti-CD19 CART cells can be produced by means of closed automated systems, which seems... more Autologous anti-CD19 CART cells can be produced by means of closed automated systems, which seems to be the most convenient platform for the point-of-care production. However, to fully comply with good practices and to utilize relatively low-grade of clean air environments, the production processes should be carried out in a completely closed format, without usage of pooled human blood products. The purpose of the present study was to test a production process that fully meets the necessary requirements.

Research paper thumbnail of Gene-Cell Therapy of Hiv and Hematological Malignances Based on Hematopoietic Stem Cell Transplantation and Site-Specific Genome Editing

Journal Infectology, 2017

Резюме По данным ежегодных докладов UNAIDS, c 1983 г. количество ВИЧ-инфицированных пациентов неу... more Резюме По данным ежегодных докладов UNAIDS, c 1983 г. количество ВИЧ-инфицированных пациентов неуклонно растет. Антиретровирусная терапия (АРВТ) позволяет увеличить продолжительность жизни, но проблемы качества жизни и долгосрочной выживаемости не решены. Одной из основных причин летальности у пациентов с ВИЧ в эру АРВТ являются онкологические заболевания, среди которых значительную часть составляют злокачественные лимфомы. Лечение лимфом включает этапную противоопухолевую химиотерапию (ПХТ) с потенциалом полного излечения пациента. Высокодоз ная ПХТ с аутологичной трансплантацией гемопоэтических стволовых клеток (ТГСК)-основное средство лечения рецидивов и рефрактерных форм агрессивных злокачественных лимфом. В последние годы с появлением технологии редактирования генома аутологичная ТГСК становится также одним из самых перспективных методов лечения ВИЧ-инфекции. История излечения от ВИЧ «Берлинского пациента» благодаря аллогенной ТГСК от донора с мутацией гена CCR5 продемонстрировала эффективность подхода генной терапии на основе трансплантации. Высокодозная химиотерапия с трансплантацией аутологичных, подвергнутых редактированию (инактивации гена CCR5, с помощью генно-инженерных нуклеаз), гемопоэтических стволовых клеток потенциально представляет собой способ комплексного лечения этих пациентов. С одной стороны, высокодозная ПХТ с ауто-ТГСК обеспечивает излечение от злокачественной опухоли, с другой стороны, она является способом доставки отредактированных клеток и создания условий для реализации лечебного эффекта против ВИЧ.

Research paper thumbnail of Screening for predictive biomarkers after related and unrelated grafts with posttransplantation cyclophosphamide in adult patients

Research paper thumbnail of Functionalization of bone marrow mesenchymal stromal cells by multifunctional capsules for targeted drug delivery

Research paper thumbnail of A highly efficient and safe gene delivery platform based on polyelectrolyte core–shell nanoparticles for hard-to-transfect clinically relevant cell types

Journal of Materials Chemistry B

The polyelectrolyte nanocarriers’ based on nanosized vaterite particles as a novel tool for genet... more The polyelectrolyte nanocarriers’ based on nanosized vaterite particles as a novel tool for genetic material delivery into the clinically relevant cell types and potential application of described technology in gene therapy approaches.

Research paper thumbnail of High prevalence of CD3, NK, and NKT cells in the graft predicts adverse outcome after matched-related and unrelated transplantations with post transplantation cyclophosphamide

Bone Marrow Transplantation

The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT... more The predictive value of graft composition and plasma biomarkers on the outcome of allogeneic HSCT is well known for conventional GVHD prophylaxis based on calcineurin inhibitors with or without antithymocyte globulin. Currently, there is limited data whether these results could be translated to post transplantation cyclophosphamide (PTCy). The prospective extension cohort of NCT02294552 trial enrolled 79 adult patients with acute leukemia in CR. Twenty-six received matched-related bone marrow (BM) grafts with single-agent PTCy and 53 received unrelated peripheral blood stem cell graft (PBSC) with PTCy, tacrolimus, and MMF. The grafts were studied by the flow cytometry, and plasma samples were analyzed by ELISA. In the cluster and major component analysis, we determined that transplantation from donors with high content of CD3, NKT, and CD16-CD56 + subpopulations in the PBSC grafts was associated with poor immunological recovery and compromised event-free survival (50% vs. 80%, HR 2.93, p = 0.015) both due to increased relapse incidence and non-relapse mortality. The significant independent predictor of moderate and severe chronic GVHD was the high prevalence of and iNKT, Vβ11, and double-positive cells in the PBSC grafts from young donors (HR 2.75, p = 0.0483). No patterns could be identified for BM grafts and for plasma biomarkers.

Research paper thumbnail of Safe and Effective Delivery of Antitumor Drug Using Mesenchymal Stem Cells Impregnated with Submicron Carriers

ACS Applied Materials & Interfaces

An important area in modern malignant tumor therapy is the optimization of antitumor drugs pharma... more An important area in modern malignant tumor therapy is the optimization of antitumor drugs pharmacokinetics. The use of some antitumor drugs is limited in clinical practice due to their high toxicity. Therefore, the strategy for optimizing the drug pharmacokinetics focuses on the generation of high local concentrations of these drugs in the tumor area with minimal systemic and tissue-specific toxicity. This can be achieved by encapsulation of highly toxic antitumor drug (vincristine (VCR) that is 20-50 times more toxic than widely used the antitumor drug doxorubicin) into nano- and microcarriers with their further association into therapeutically relevant cells that possess the ability to migrate to sites of tumor. Here, we fundamentally examine the effect of drug carrier size on the behavior of human mesenchymal stem cells (hMSCs), including internalization efficiency, cytotoxicity, cell movement, to optimize the conditions for the development of carrier-hMSCs drug delivery platform. Using the malignant tumors derived from patients, we evaluated the capability of hMSCs associated with VCR-loaded carriers to target tumors using a three-dimensional spheroid model in collagen gel. Compared to free VCR, the developed hMSC-based drug delivery platform showed enhanced antitumor activity regarding those tumors that express CXCL12 (stromal cell-derived factor-1 (SDF-1)) gene, inducing directed migration of hMSCs via CXCL12 (SDF-1)/CXCR4 pathway. These results show that the combination of encapsulated antitumor drugs and hMSCs, which possess the properties of active migration into tumors, is therapeutically beneficial and demonstrated high efficiency and low systematic toxicity, revealing novel strategies for chemotherapy in the future.

Research paper thumbnail of Acute Myeloblastic Leukemia and Myelodysplastic Syndrome: Azacitidine for Prophylactic and Preventive Purposes after Allogeneic Hematopoietic Stem Cell Transplantation

Clinical oncohematology

Aim. To evaluate the effectiveness of preventive and prophylactic post-transplantation therapy us... more Aim. To evaluate the effectiveness of preventive and prophylactic post-transplantation therapy using azacitidine (5-AZA) in patients at high risk of post-transplantation relapse. Methods. 136 patients were included in the study performed by the pairwise comparison: 68 of them received 5-AZA after allo-HSCT and 68 patients were included in the historical control group. 5-AZA was prescribed for prophylactic or preventive purposes. The results were assessed according to the OS, RR, EFS, DUM, and relapse-free and GVHR-free survival. Results. 1-year OS was 76 % in the 5-AZA group (95% CI 60-84 %) and 44 % in the reference group (95% CI 33-55 %) (р = 0.001); 2-year OS was 63 % (95% CI 39-67 %) and 37 % (95% CI 26-48 %) (р = 0.007), respectively. The relapse rate (RR) in the 5-AZA group was 34 % (95% CI 22-46 %) during 1 year and 51 % (95% CI 38-64 %) in the reference group (р = 0.02). 1- and 2-year disease unrelated mortality (DUM) was similar: 5 % in the 5-AZA group (95% CI 0.1-14.0 %) a...

Research paper thumbnail of Potential clinical applications of bone marrow-derived mesenchymal stem cells in bone regeneration

Cellular Therapy and Transplantation

Bone marrow (BM) contains a small resident cell population referred to as 'multipotent mesenchyma... more Bone marrow (BM) contains a small resident cell population referred to as 'multipotent mesenchymal stem cells' (MSC). These adherent cells could be isolated and expanded in simple culture media and may differentiate in adipogenic or osteogenic pathway. So far an opportunity of MSC differentiation to hepatocytes, brain, or renal cells is not proven yet. Meanwhile, two potential clinical applications are considered for MSC: (1) as a tool for immune modulation in graft-versus-host disease (GVHD) and autoimmune diseases, or, (2) as a potential source of growth-promoting factors in specialized tissues. This heterogenous population may support hematopoiesis by secreting growth factors, cytokines and other biologically active substances. Upon injection, MSCs are able to migrate into damaged tissues, thus promoting their repair. However, only small MSC fraction may reach bone marrow niches following intravenous infusion. Multiple experiments with MSCs in different injury models show their ability to suppress apoptosis initiated by hypoxia, chemical agents/acidity and other deteriorating factors. This protective effect is mediated by a number of secreted growth factors, e.g., granulocyte-macrophage colony-stimulating growth factor (GM-CSF). A big number of clinical trials show high level of safety for the MSC therapy. Both clinical and experimental studies demonstrated only weak immunogenic effects of allogeneic MSC upon injection into immunocompetent recipients. At the present time, injections of in vitro expanded MSCs were performed in the patients developing acute GVHD after hematopoietic stem cell transplantation (HSCT), and in some autoimmune disorders. Over last decade, several studies concerned potentially curative effects of MSCs injected into affected bone areas in the patients with osteogenesis imperfecta (OI), a severe inherited disease with altered collagen structure resulting into increased bone fragility. Here we present a synopsis of clinical protocol aimed for assessing safety, immunogenicity, and clinical effects of MSC injected to the OI patients during corrective osteotomy. One may suggest that a minor MSC subpopulation may migrate to the damaged areas differentiating to chondrocytes and osteoblasts, and, hence, contributing to the bone repair.

Research paper thumbnail of Efficient gene editing via non-viral delivery of CRISPR–Cas9 system using polymeric and hybrid microcarriers

Nanomedicine: Nanotechnology, Biology and Medicine

CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the trea... more CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarriers, made of degradable polymers such as polypeptides and polysaccharides and modified by silica shell, for delivery of all CRISPR-Cas9 components. We found that these microcarriers mediate more efficient transfection than a commercially available liposome-based transfection reagent (&gt;70% vs. &lt;50% for mRNA, &gt;40% vs. 20% for plasmid DNA). For proof-of-concept, we delivered CRISPR-Cas9 components using our capsules to dTomato-expressing HEK293T cells-a model, in which loss of red fluorescence indicates successful gene editing. Notably, transfection of indicator cells translated in high-level dTomato knockout in approx. 70% of transfected cells. In conclusion, we have provided proof-of-principle that our micro-sized containers represent promising non-viral platforms for efficient and safe gene editing.