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Gene and Cell Therapy in Regenerative Medicine

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A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Cell and Gene Therapy".

Deadline for manuscript submissions: closed (15 July 2023) | Viewed by 37239

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Prof. Dr. Albert Rizvanov

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Guest Editor

Institute of Fundamental Medicine and Biology, Department of Genetics, Kazan Federal University, Kazan, Russia
Interests: gene therapy; stem cell therapy; multiomics; regenerative medicine; translational medicine
Special Issues, Collections and Topics in MDPI journals

Dr. Ayşegül Doğan

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Guest Editor

Department of Genetics and Bioengineering, T.C. Yeditepe Universitesi, Istanbul, Turkey
Interests: pluripotent stem cells; organoids; gene editing; neuromesodermal progenitors; apelin signalling
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Despite recent advances in biomedical and clinical research, many diseases and conditions still lack effective treatments, resulting in inadequate recovery and poor patient quality of life. Gene and cell therapy represent a promising approach to overcoming the natural restrictions of the body relating to regeneration. This approach can be used to eliminate the cause of the disease or condition, such as the correction of genetic mutations and hereditary factors. Another approach is based on cell replacement strategies designed to compensate for the lack of, or increase in, the pool of certain cells. Yet another approach is based on the paracrine theory behind regeneration, where gene therapy or transplanted cells result in the secretion of different growth factors; other biological molecules such as proteins, metabolites, RNA, and others; and microvesicles, which are able to stimulate angiogenesis, neuroprotection/neuroregeneration, osteo- and/or chondrogenesis as well as other regenerative processes. Finally, to further increase the efficiency of regenerative treatments, and to overcome problems related to the targeted delivery of gene therapy in vivo, combination gene–cell therapy can be used to correct genetic mutations ex vivo, achieve transgene expression or use cells as delivery systems for gene therapy targeted delivery.

Potential topics include, but are not limited to, the following:

Isolation, expansion and analysis of stem cells from various tissues and organs for biomedical and clinical applications;
Pre-clinical and clinical applications of autologous and allogeneic natural and genetically modified cells;
Stem cells in tissue engineering (3D printing, organoids, decellularized tissues and organs, natural and artificial matrixes etc.);
Immunomodulatory effects of stem cells;
Gene therapy (vector development, therapeutic gene editing, transgene delivery etc.);
Using genetic modification and chemical treatments for modulating biological properties and therapeutic efficiency of stem and differentiated;
Characterization and application of natural and artificial membrane vesicles from various natural and genetically modified cells for regenerative medicine.

Prof. Dr. Albert A. Rizvanov
Dr. Ayşegül Doğan
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

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Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

gene therapy
cell therapy
regenerative medicine
growth factors
stem cells

Published Papers (11 papers)

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Research

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25 pages, 17946 KiB

Open AccessArticle

TGFβ Inhibitor A83-01 Enhances Murine HSPC Expansion for Gene Therapy

by Jenni Fleischauer, Antonella Lucia Bastone, Anton Selich, Philipp John-Neek, Luisa Weisskoeppel, Dirk Schaudien, Axel Schambach and Michael Rothe

Cells 2023, 12(15), 1978; https://doi.org/10.3390/cells12151978 - 31 Jul 2023

Viewed by 1343

Abstract

Murine hematopoietic stem and progenitor cells (HSPCs) are commonly used as model systems during gene therapeutic retroviral vector development and preclinical biosafety assessment. Here, we developed cell culture conditions to maintain stemness and prevent differentiation during HSPC culture. We used the small compounds A83-01, pomalidomide, and UM171 (APU). Highly purified LSK SLAM cells expanded in medium containing SCF, IL-3, FLT3-L, and IL-11 but rapidly differentiated to myeloid progenitors and mast cells. The supplementation of APU attenuated the differentiation and preserved the stemness of HSPCs. The TGF

β

inhibitor A83-01 was identified as the major effector. It significantly inhibited the mast-cell-associated expression of Fc

ε

α

and the transcription of genes regulating the formation of granules and promoted a 3800-fold expansion of LSK cells. As a functional readout, we used expanded HSPCs in state-of-the-art genotoxicity assays. Like fresh cells, APU-expanded HSPCs transduced with a mutagenic retroviral vector developed a myeloid differentiation block with clonal restriction and dysregulated oncogenic transcriptomic signatures due to vector integration near the high-risk locus Mecom. Thus, expanded HSPCs might serve as a novel cell source for retroviral vector testing and genotoxicity studies. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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18 pages, 8057 KiB

Open AccessArticle

Adenovirus-Based Gene Therapy for Bone Regeneration: A Comparative Analysis of In Vivo and Ex Vivo BMP2 Gene Delivery

by Tatiana Borisovna Bukharova, Irina Alekseevna Nedorubova, Viktoria Olegovna Mokrousova, Anastasiia Yurevna Meglei, Viktoriia Pavlovna Basina, Andrey Anatolevich Nedorubov, Andrey Vyacheslavovich Vasilyev, Timofei Evgenevich Grigoriev, Yuriy Dmitrievich Zagoskin, Sergei Nicolaevich Chvalun, Sergey Ivanovich Kutsev and Dmitry Vadimovich Goldshtein

Cells 2023, 12(13), 1762; https://doi.org/10.3390/cells12131762 - 01 Jul 2023

Cited by 4 | Viewed by 1356

Abstract

Adenovirus-mediated gene therapy is a promising tool in bone regenerative medicine. In this work, gene-activated matrices (GAMs) composed of (1) polylactide granules (PLA), which serve as a depot for genetic constructs or matrices for cell attachment, (2) a PRP-based fibrin clot, which is a source of growth factors and a binding gel, and (3) a BMP2 gene providing osteoinductive properties were studied. The study aims to compare the effectiveness of in vivo and ex vivo gene therapy based on adenoviral constructs with the BMP2 gene, PLA particles, and a fibrin clot for bone defect healing. GAMs with Ad-BMP2 and MSC(Ad-BMP2) show osteoinductive properties both in vitro and in vivo. However, MSCs incubated with GAMs containing transduced cells showed a more significant increase in osteopontin gene expression, protein production, Alpl activity, and matrix mineralization. Implantation of the studied matrices into critical-size calvarial defects after 56 days promotes the formation of young bone. The efficiency of neoosteogenesis and the volume fraction of newly formed bone tissue are higher with PLA/PRP-MSC(Ad-BMP2) implantation (33%) than PLA/PRP-Ad-BMP2 (28%). Thus, ex vivo adenoviral gene therapy with the BMP2 gene has proven to be a more effective approach than the in vivo delivery of gene constructs for bone regeneration. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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13 pages, 2857 KiB

Open AccessArticle

Olfactory Regeneration with Nasally Administered Murine Adipose-Derived Stem Cells in Olfactory Epithelium Damaged Mice

by Tomoko Ishikura, Hideaki Shiga, Yuka Nakamura, Takako Kanitani, Yasuhito Ishigaki and Takaki Miwa

Cells 2023, 12(5), 765; https://doi.org/10.3390/cells12050765 - 27 Feb 2023

Cited by 1 | Viewed by 2015

Abstract

In this study, we aimed to determine whether nasally administered murine adipose-derived stem cells (ADSCs) could support olfactory regeneration in vivo. Olfactory epithelium damage was induced in 8-week-old C57BL/6J male mice by intraperitoneal injection of methimazole. Seven days later, OriCell adipose-derived mesenchymal stem cells obtained from green fluorescent protein (GFP) transgenic C57BL/6 mice were nasally administered to the left nostril of these mice, and their innate odor aversion behavior to butyric acid was assessed. Mice showed significant recovery of odor aversion behavior, along with improved olfactory marker protein (OMP) expression on both sides of the upper-middle part of the nasal septal epithelium assessed by immunohistochemical staining 14 d after the treatment with ADSCs compared with vehicle control animals. Nerve growth factor (NGF) was detected in the ADSC culture supernatant, NGF was increased in the nasal epithelium of mice, and GFP-positive cells were observed on the surface of the left side nasal epithelium 24 h after left side nasal administration of ADSCs. The results of this study suggest that the regeneration of olfactory epithelium can be stimulated by nasally administered ADSCs secreting neurotrophic factors, thereby promoting the recovery of odor aversion behavior in vivo. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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19 pages, 3118 KiB

Open AccessArticle

Synovial Fluid Derived from Human Knee Osteoarthritis Increases the Viability of Human Adipose-Derived Stem Cells through Upregulation of FOSL1

by Hironori Kitajima, Takuya Sakamoto, Tetsuhiro Horie, Ayane Kuwano, Atsushi Fuku, Yasuhiko Taki, Yuka Nakamura, Ikuhiro Tanida, Hiroshi Sunami, Hiroaki Hirata, Yoshiyuki Tachi, Naoki Yamamoto, Yasuo Iida, Yasuhito Ishigaki, Sohsuke Yamada, Shigetaka Shimodaira, Yusuke Shimizu, Toru Ichiseki, Ayumi Kaneuji, Satoshi Osawa and Norio Kawahara Show full author list Hide full author list

Cells 2023, 12(2), 330; https://doi.org/10.3390/cells12020330 - 15 Jan 2023

Cited by 1 | Viewed by 3368

Abstract

Knee osteoarthritis (Knee OA) is an irreversible condition that causes bone deformity and degeneration of the articular cartilage that comprises the joints, resulting in chronic pain and movement disorders. The administration of cultured adipose-derived stem cells (ADSCs) into the knee joint cavity improves the clinical symptoms of Knee OA; however, the effect of synovial fluid (SF) filling the joint cavity on the injected ADSCs remains unclear. In this study, we investigated the effect of adding SF from Knee OA patients to cultured ADSCs prepared for therapeutic use in an environment that mimics the joint cavity. An increase in the viability of ADSCs was observed following the addition of SF. Gene expression profiling of SF-treated ADSCs using DNA microarrays revealed changes in several genes involved in cell survival. Of these genes, we focused on FOSL1, which is involved in the therapeutic effect of ADSCs and the survival and proliferation of cancer stem cells. We confirmed the upregulation of FOSL1 mRNA and protein expression using RT-PCR and western blot analysis, respectively. Next, we knocked down FOSL1 in ADSCs using siRNA and observed a decrease in cell viability, indicating the involvement of FOSL1 in the survival of ADSCs. Interestingly, in the knockdown cells, ADSC viability was also decreased by SF exposure. These results suggest that SF enhances cell viability by upregulating FOSL1 expression in ADSCs. For therapy using cultured ADSCs, the therapeutic effect of ADSCs may be further enhanced if an environment more conducive to the upregulation of FOSL1 expression in ADSCs can be established. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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14 pages, 4359 KiB

Open AccessArticle

Multipotent Mesenchymal Stromal Cells from Porcine Bone Marrow, Implanted under the Kidney Capsule, form an Ectopic Focus Containing Bone, Hematopoietic Stromal Microenvironment, and Muscles

by Nataliya Petinati, Irina Shipounova, Natalia Sats, Alena Dorofeeva, Alexandra Sadovskaya, Nikolay Kapranov, Yulia Tkachuk, Anatoliy Bondarenko, Margarita Muravskaya, Michail Kotsky, Irina Kaplanskaya, Tamara Vasilieva and Nina Drize

Cells 2023, 12(2), 268; https://doi.org/10.3390/cells12020268 - 10 Jan 2023

Cited by 1 | Viewed by 1400

Abstract

Multipotent mesenchymal stromal cells (MSCs) are an object of intense investigation due to their therapeutic potential. MSCs have been well studied in vitro, while their fate after implantation in vivo has been poorly analyzed. We studied the properties of MSCs from the bone marrow (BM-MSC) before and after implantation under the renal capsule using a mini pig model. Autologous BM-MSCs were implanted under the kidney capsule. After 2.5 months, ectopic foci containing bones, foci of ectopic hematopoiesis, bone marrow stromal cells and muscle cells formed. Small pieces of the implant were cultivated as a whole. The cells that migrated out from these implants were cultured, cloned, analyzed and were proven to meet the most of criteria for MSCs, therefore, they are designated as MSCs from the implant—IM-MSCs. The IM-MSC population demonstrated high proliferative potential, similar to BM-MSCs. IM-MSC clones did not respond to adipogenic differentiation inductors: 33% of clones did not differentiate, and 67% differentiated toward an osteogenic lineage. The BM-MSCs revealed functional heterogeneity after implantation under the renal capsule. The BM-MSC population consists of mesenchymal precursor cells of various degrees of differentiation, including stem cells. These newly discovered properties of mini pig BM-MSCs reveal new possibilities in terms of their manipulation. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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16 pages, 2964 KiB

Open AccessArticle

The Efficacy of HGF/VEGF Gene Therapy for Limb Ischemia in Mice with Impaired Glucose Tolerance: Shift from Angiogenesis to Axonal Growth and Oxidative Potential in Skeletal Muscle

by Iurii S. Stafeev I, Maria A. Boldyreva, Svetlana S. Michurina, Margarita Yu. Agareva, Arina V. Radnaeva, Mikhail Yu. Menshikov, Yu-Chen Hu, Pavel I. Makarevich and Yelena V. Parfyonova

Cells 2022, 11(23), 3824; https://doi.org/10.3390/cells11233824 - 29 Nov 2022

Cited by 2 | Viewed by 1888

Abstract

Background: Combined non-viral gene therapy (GT) of ischemia and cardiovascular disease is a promising tool for potential clinical translation. In previous studies our group has developed combined gene therapy by vascular endothelial growth factor 165 (VEGF165) + hepatocyte growth factor (HGF). Our recent works have demonstrated that a bicistronic pDNA that carries both human HGF and VEGF165 coding sequences has a potential for clinical application in peripheral artery disease (PAD). The present study aimed to test HGF/VEGF combined plasmid efficacy in ischemic skeletal muscle comorbid with predominant complications of PAD-impaired glucose tolerance and type 2 diabetes mellitus (T2DM). Methods: Male C57BL mice were housed on low-fat (LFD) or high-fat diet (HFD) for 10 weeks and metabolic parameters including FBG level, ITT, and GTT were evaluated. Hindlimb ischemia induction and plasmid administration were performed at 10 weeks with 3 weeks for post-surgical follow-up. Limb blood flow was assessed by laser Doppler scanning at 7, 14, and 21 days after ischemia induction. The necrotic area of m.tibialis anterior, macrophage infiltration, angio- and neuritogenesis were evaluated in tissue sections. The mitochondrial status of skeletal muscle (total mitochondria content, ETC proteins content) was assessed by Western blotting of muscle lysates. Results: At 10 weeks, the HFD group demonstrated impaired glucose tolerance in comparison with the LFD group. HGF/VEGF plasmid injection aggravated glucose intolerance in HFD conditions. Blood flow recovery was not changed by HGF/VEGF plasmid injection either in LFD or HFD conditions. GT in LFD, but not in HFD conditions, enlarged the necrotic area and CD68+ cells infiltration. However, HGF/VEGF plasmid enhanced neuritogenesis and enlarged NF200+ area on muscle sections. In HFD conditions, HGF/VEGF plasmid injection significantly increased mitochondria content and ETC proteins content. Conclusions: The current study demonstrated a significant role of dietary conditions in pre-clinical testing of non-viral GT drugs. HGF/VEGF combined plasmid demonstrated a novel aspect of potential participation in ischemic skeletal muscle regeneration, through regulation of innervation and bioenergetics of muscle. The obtained results made HGF/VEGF combined plasmid a very promising tool for PAD therapy in impaired glucose tolerance conditions. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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20 pages, 4836 KiB

Open AccessArticle

Supramolecular Hydrogel-Wrapped Gingival Mesenchymal Stem Cells in Cutaneous Radiation Injury

by Shasha Nie, Chunhua Ren, Xin Liang, Hui Cai, Hao Sun, Fengting Liu, Kaihua Ji, Yan Wang and Qiang Liu

Cells 2022, 11(19), 3089; https://doi.org/10.3390/cells11193089 - 30 Sep 2022

Cited by 4 | Viewed by 2076

Abstract

Radiation-induced skin wound/dermatitis is one of the common side effects of radiotherapy or interventional radiobiology. Gingiva-derived mesenchymal stem cells (GMSCs) were indicated to have therapeutic potentials in skin diseases. However, stem cells are prone to spread and difficult to stay in the skin for a long time, limiting their curative effects and application. This study investigated the therapeutic efficacy of Nap-GDFDFpDY (pY-Gel) self-assembled peptide hydrogel-encapsulated GMSCs to treat ¹³⁷Cs γ-radiation-induced skin wounds in mice. The effects were evaluated by skin damage score, hind limb extension measurement and histological and immunohistochemical analysis. In vivo studies showed that pY-Gel self-assembled peptide hydrogel-encapsulated GMSCs could effectively improve wound healing in irradiated skin tissues. In addition, it was found that GMSCs conditioned medium (CM) could promote the proliferation, migration and DNA damage repair ability of skin cells after irradiation in human keratinocyte cell line HaCaT and normal human dermal fibroblasts (HFF). Mechanistically, GMSCs-CM can promote the expression of epidermal growth factor receptor (EGFR), signal transducers and activators of transcription 3 (STAT3) and matrix metalloproteinases (MMPs), suggesting that activation of the EGFR/STAT3 signaling pathway may be involved in the repair of skin cells after exposure to radiations. In conclusion, pY-Gel self-assembled peptide hydrogel-encapsulated GMSCs have a beneficial therapeutic effect on radiation-induced cutaneous injury and may serve as a basis of novel cells therapeutic approach. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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Review

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24 pages, 875 KiB

Open AccessReview

Generation of Red Blood Cells from Human Pluripotent Stem Cells—An Update

by Shin-Jeong Lee, Cholomi Jung, Jee Eun Oh, Sangsung Kim, Sangho Lee, Ji Yoon Lee and Young-sup Yoon

Cells 2023, 12(11), 1554; https://doi.org/10.3390/cells12111554 - 05 Jun 2023

Cited by 2 | Viewed by 3981

Abstract

Red blood cell (RBC) transfusion is a lifesaving medical procedure that can treat patients with anemia and hemoglobin disorders. However, the shortage of blood supply and risks of transfusion-transmitted infection and immune incompatibility present a challenge for transfusion. The in vitro generation of RBCs or erythrocytes holds great promise for transfusion medicine and novel cell-based therapies. While hematopoietic stem cells and progenitors derived from peripheral blood, cord blood, and bone marrow can give rise to erythrocytes, the use of human pluripotent stem cells (hPSCs) has also provided an important opportunity to obtain erythrocytes. These hPSCs include both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). As hESCs carry ethical and political controversies, hiPSCs can be a more universal source for RBC generation. In this review, we first discuss the key concepts and mechanisms of erythropoiesis. Thereafter, we summarize different methodologies to differentiate hPSCs into erythrocytes with an emphasis on the key features of human definitive erythroid lineage cells. Finally, we address the current limitations and future directions of clinical applications using hiPSC-derived erythrocytes. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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22 pages, 2259 KiB

Open AccessReview

Transcriptome-Powered Pluripotent Stem Cell Differentiation for Regenerative Medicine

by Derek A. Ogi and Sha Jin

Cells 2023, 12(10), 1442; https://doi.org/10.3390/cells12101442 - 22 May 2023

Cited by 2 | Viewed by 1959

Abstract

Pluripotent stem cells are endless sources for in vitro engineering human tissues for regenerative medicine. Extensive studies have demonstrated that transcription factors are the key to stem cell lineage commitment and differentiation efficacy. As the transcription factor profile varies depending on the cell type, global transcriptome analysis through RNA sequencing (RNAseq) has been a powerful tool for measuring and characterizing the success of stem cell differentiation. RNAseq has been utilized to comprehend how gene expression changes as cells differentiate and provide a guide to inducing cellular differentiation based on promoting the expression of specific genes. It has also been utilized to determine the specific cell type. This review highlights RNAseq techniques, tools for RNAseq data interpretation, RNAseq data analytic methods and their utilities, and transcriptomics-enabled human stem cell differentiation. In addition, the review outlines the potential benefits of the transcriptomics-aided discovery of intrinsic factors influencing stem cell lineage commitment, transcriptomics applied to disease physiology studies using patients’ induced pluripotent stem cell (iPSC)-derived cells for regenerative medicine, and the future outlook on the technology and its implementation. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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41 pages, 1204 KiB

Open AccessReview

Various AAV Serotypes and Their Applications in Gene Therapy: An Overview

by Shaza S. Issa, Alisa A. Shaimardanova, Valeriya V. Solovyeva and Albert A. Rizvanov

Cells 2023, 12(5), 785; https://doi.org/10.3390/cells12050785 - 01 Mar 2023

Cited by 35 | Viewed by 12460

Abstract

Despite scientific discoveries in the field of gene and cell therapy, some diseases still have no effective treatment. Advances in genetic engineering methods have enabled the development of effective gene therapy methods for various diseases based on adeno-associated viruses (AAVs). Today, many AAV-based gene therapy medications are being investigated in preclinical and clinical trials, and new ones are appearing on the market. In this article, we present a review of AAV discovery, properties, different serotypes, and tropism, and a following detailed explanation of their uses in gene therapy for disease of different organs and systems. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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28 pages, 2021 KiB

Open AccessReview

Cell Sources for Retinal Regeneration: Implication for Data Translation in Biomedicine of the Eye

by Eleonora N. Grigoryan

Cells 2022, 11(23), 3755; https://doi.org/10.3390/cells11233755 - 24 Nov 2022

Cited by 2 | Viewed by 3626

Abstract

The main degenerative diseases of the retina include macular degeneration, proliferative vitreoretinopathy, retinitis pigmentosa, and glaucoma. Novel approaches for treating retinal diseases are based on cell replacement therapy using a variety of exogenous stem cells. An alternative and complementary approach is the potential use of retinal regeneration cell sources (RRCSs) containing retinal pigment epithelium, ciliary body, Müller glia, and retinal ciliary region. RRCSs in lower vertebrates in vivo and in mammals mostly in vitro are able to proliferate and exhibit gene expression and epigenetic characteristics typical for neural/retinal cell progenitors. Here, we review research on the factors controlling the RRCSs’ properties, such as the cell microenvironment, growth factors, cytokines, hormones, etc., that determine the regenerative responses and alterations underlying the RRCS-associated pathologies. We also discuss how the current data on molecular features and regulatory mechanisms of RRCSs could be translated in retinal biomedicine with a special focus on (1) attempts to obtain retinal neurons de novo both in vivo and in vitro to replace damaged retinal cells; and (2) investigations of the key molecular networks stimulating regenerative responses and preventing RRCS-related pathologies. Full article

(This article belongs to the Special Issue Gene and Cell Therapy in Regenerative Medicine)

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