DNA transposition by protein transduction of the piggyBac transposase from lentiviral Gag precursors

Abstract DNA transposon-based vectors have emerged as gene vehicles with a wide biomedical and therapeutic potential. So far, genomic insertion of such vectors has relied on the co-delivery of genetic material encoding the gene-inserting transposase protein, raising concerns related to persistent expression, insertional mutagenesis and cytotoxicity. This report describes potent DNA transposition achieved by direct delivery of transposase protein. By adapting integrase-deficient lentiviral particles (LPs) as carriers of the hyperactive piggyBac transposase protein (hyPBase), we demonstrate rates of DNA transposition that are comparable with the efficiency of a conventional plasmid-based strategy. Embedded in the Gag polypeptide, hyPBase is robustly incorporated into LPs and liberated from the viral proteins by the viral protease during particle maturation. We demonstrate lentiviral co-delivery of the transposase protein and vector RNA carrying the transposon sequence, allowing robust DNA transposition in a variety of cell types. Importantly, this novel delivery method facilitates a balanced cellular uptake of hyPBase, as shown by confocal microscopy, and allows high-efficiency production of clones harboring a single transposon insertion. Our findings establish engineered LPs as a new tool for transposase delivery. We believe that protein transduction methods will increase applicability and safety of DNA transposon-based vector technologies.

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The Old and the New: Prospects for Non-Integrating Lentiviral Vector Technology

Viruses ◽

10.3390/v12101103 ◽

2020 ◽

Vol 12 (10) ◽

pp. 1103 ◽

Cited By ~ 1

Author(s):

Luis Apolonia

Keyword(s):

Insertional Mutagenesis ◽

Lentiviral Vector ◽

Genetic Material ◽

Lentiviral Vectors ◽

Cell Types ◽

Target Cells ◽

Attachment Sites ◽

Dividing Cells ◽

Multiple Cell ◽

Episomal Dna

Lentiviral vectors have been developed and used in multiple gene and cell therapy applications. One of their main advantages over other vectors is the ability to integrate the genetic material into the genome of the host. However, this can also be a disadvantage as it may lead to insertional mutagenesis. To address this, non-integrating lentiviral vectors (NILVs) were developed. To generate NILVs, it is possible to introduce mutations in the viral enzyme integrase and/or mutations on the viral DNA recognised by integrase (the attachment sites). NILVs are able to stably express transgenes from episomal DNA in non-dividing cells or transiently if the target cells divide. It has been shown that these vectors are able to transduce multiple cell types and tissues. These characteristics make NILVs ideal vectors to use in vaccination and immunotherapies, among other applications. They also open future prospects for NILVs as tools for the delivery of CRISPR/Cas9 components, a recent revolutionary technology now widely used for gene editing and repair.

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Targeted and non-targeted effects of radiation in mammalian cells: An overview

Archives of Biotechnology and Biomedicine ◽

10.29328/journal.abb.1001023 ◽

2021 ◽

Vol 5 (1) ◽

pp. 013-019

Author(s):

Ghosh Rita ◽

Hansda Surajit

Keyword(s):

Mammalian Cells ◽

Bystander Effect ◽

Therapeutic Potential ◽

Genetic Material ◽

Direct Interaction ◽

Ultra Violet ◽

Cell Types ◽

Target Cells ◽

Effects Of Radiation ◽

Ionizing Radiations

Radiation of different wavelengths can kill living organisms, although, the mechanism of interactions differs depending on their energies. Understanding the interaction of radiation with living cells is important to assess their harmful effects and also to identify their therapeutic potential. Temporally, this interaction can be broadly divided in three stages – physical, chemical and biological. While radiation can affect all the important macromolecules of the cells, particularly important is the damage to its genetic material, the DNA. The consequences of irradiation include- DNA damage, mutation, cross-linkages with other molecules, chromosomal aberrations and DNA repair leading to altered gene expression and/or cell death. Mutations in DNA can lead to heritable changes and is important for the induction of cancer. While some of these effects are through direct interaction of radiation with the target, radiation can interact with the surrounding environment to result in its indirect actions. The effects of radiation depend not only on the total dose but also on the dose rate, LET etc. and also on the cell types. However, action of radiation on organisms is not restricted to interactions with irradiated cells, i.e. target cells alone; it also exerts non-targeted effects on neighboring unexposed cells to produce productive responses; this is known as bystander effect. The bystander effects of ionizing radiations are well documented and contribute largely to the relapse of cancer and secondary tumors after radiotherapy. Irradiation of cells with non-ionizing Ultra-Violet light also exhibits bystander responses, but such responses are very distinct from that produced by ionizing radiations.

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Therapeutic Potential of Umbilical Cord Stem Cells for Liver Regeneration

Current Stem Cell Research & Therapy ◽

10.2174/1568026620666200220122536 ◽

2020 ◽

Vol 15 (3) ◽

pp. 219-232

Author(s):

Ifrah Anwar ◽

Usman A. Ashfaq ◽

Zeeshan Shokat

Keyword(s):

Stem Cells ◽

Umbilical Cord ◽

Fas Ligand ◽

Viral Infections ◽

Therapeutic Potential ◽

Cell Types ◽

Low Risk ◽

Blood Stem Cells ◽

Umbilical Cord Stem Cells ◽

High Regeneration

The liver is a vital organ for life and the only internal organ that is capable of natural regeneration. Although the liver has high regeneration capacity, excessive hepatocyte death can lead to liver failure. Various factors can lead to liver damage including drug abuse, some natural products, alcohol, hepatitis, and autoimmunity. Some models for studying liver injury are APAP-based model, Fas ligand (FasL), D-galactosamine/endotoxin (Gal/ET), Concanavalin A, and carbon tetrachloride-based models. The regeneration of the liver can be carried out using umbilical cord blood stem cells which have various advantages over other stem cell types used in liver transplantation. UCB-derived stem cells lack tumorigenicity, have karyotype stability and high immunomodulatory, low risk of graft versus host disease (GVHD), low risk of transmitting somatic mutations or viral infections, and low immunogenicity. They are readily available and their collection is safe and painless. This review focuses on recent development and modern trends in the use of umbilical cord stem cells for the regeneration of liver fibrosis.

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NF-κB in Cancer Immunity: Friend or Foe?

Cells ◽

10.3390/cells10020355 ◽

2021 ◽

Vol 10 (2) ◽

pp. 355

Author(s):

Guilhem Lalle ◽

Julie Twardowski ◽

Yenkel Grinberg-Bleyer

Keyword(s):

Immune Responses ◽

Therapeutic Potential ◽

Cell Types ◽

Transcriptional Activators ◽

New Class ◽

Complex Interactions ◽

Cancer Immunity ◽

Distinct Cell ◽

Cancer Initiation ◽

Cancer Outcome

The emergence of immunotherapies has definitely proven the tight relationship between malignant and immune cells, its impact on cancer outcome and its therapeutic potential. In this context, it is undoubtedly critical to decipher the transcriptional regulation of these complex interactions. Following early observations demonstrating the roles of NF-κB in cancer initiation and progression, a series of studies converge to establish NF-κB as a master regulator of immune responses to cancer. Importantly, NF-κB is a family of transcriptional activators and repressors that can act at different stages of cancer immunity. In this review, we provide an overview of the selective cell-intrinsic contributions of NF-κB to the distinct cell types that compose the tumor immune environment. We also propose a new view of NF-κB targeting drugs as a new class of immunotherapies for cancer.

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Therapeutic Features and Updated Clinical Trials of Mesenchymal Stem Cell (MSC)-Derived Exosomes

Journal of Clinical Medicine ◽

10.3390/jcm10040711 ◽

2021 ◽

Vol 10 (4) ◽

pp. 711

Author(s):

Byung-Chul Lee ◽

Insung Kang ◽

Kyung-Rok Yu

Keyword(s):

Clinical Trials ◽

Therapeutic Agent ◽

Therapeutic Potential ◽

Therapeutic Option ◽

Treatment Options ◽

Genetic Material ◽

Experimental Models ◽

Attractive Alternative ◽

Cell Based Therapy ◽

Cellular Rejection

Identification of the immunomodulatory and regenerative properties of mesenchymal stem cells (MSCs) have made them an attractive alternative therapeutic option for diseases with no effective treatment options. Numerous clinical trials have followed; however, issues such as infusional toxicity and cellular rejection have been reported. To address these problems associated with cell-based therapy, MSC exosome therapy was developed and has shown promising clinical outcomes. MSC exosomes are nanosized vesicles secreted from MSCs and represent a non-cellular therapeutic agent. MSC exosomes retain therapeutic features of the cells from which they originated including genetic material, lipids, and proteins. Similar to MSCs, exosomes can induce cell differentiation, immunoregulation, angiogenesis, and tumor suppression. MSC exosomes have therefore been employed in several experimental models and clinical studies. Here, we review the therapeutic potential of MSC-derived exosomes and summarize currently ongoing clinical trials according to disease type. In addition, we propose several functional enhancement strategies for the effective clinical application of MSC exosome therapy.

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Therapeutic Attributes of Endocannabinoid System against Neuro-Inflammatory Autoimmune Disorders

Molecules ◽

10.3390/molecules26113389 ◽

2021 ◽

Vol 26 (11) ◽

pp. 3389

Author(s):

Ishtiaq Ahmed ◽

Saif Ur Rehman ◽

Shiva Shahmohamadnejad ◽

Muhammad Anjum Zia ◽

Muhammad Ahmad ◽

...

Keyword(s):

Cannabinoid Receptors ◽

Cannabinoid Receptor ◽

Therapeutic Potential ◽

Endocannabinoid System ◽

Cell Types ◽

Autoimmune Disorders ◽

Specific Receptors ◽

Different Cell Types

In humans, various sites like cannabinoid receptors (CBR) having a binding affinity with cannabinoids are distributed on the surface of different cell types, where endocannabinoids (ECs) and derivatives of fatty acid can bind. The binding of these substance(s) triggers the activation of specific receptors required for various physiological functions, including pain sensation, memory, and appetite. The ECs and CBR perform multiple functions via the cannabinoid receptor 1 (CB1); cannabinoid receptor 2 (CB2), having a key effect in restraining neurotransmitters and the arrangement of cytokines. The role of cannabinoids in the immune system is illustrated because of their immunosuppressive characteristics. These characteristics include inhibition of leucocyte proliferation, T cells apoptosis, and induction of macrophages along with reduced pro-inflammatory cytokines secretion. The review seeks to discuss the functional relationship between the endocannabinoid system (ECS) and anti-tumor characteristics of cannabinoids in various cancers. The therapeutic potential of cannabinoids for cancer—both in vivo and in vitro clinical trials—has also been highlighted and reported to be effective in mice models in arthritis for the inflammation reduction, neuropathic pain, positive effect in multiple sclerosis and type-1 diabetes mellitus, and found beneficial for treating in various cancers. In human models, such studies are limited; thereby, further research is indispensable in this field to get a conclusive outcome. Therefore, in autoimmune disorders, therapeutic cannabinoids can serve as promising immunosuppressive and anti-fibrotic agents.

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Plasma membrane integrity in health and disease: significance and therapeutic potential

Cell Discovery ◽

10.1038/s41421-020-00233-2 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Catarina Dias ◽

Jesper Nylandsted

Keyword(s):

Plasma Membrane ◽

Therapeutic Potential ◽

Calcium Influx ◽

Membrane Integrity ◽

Cell Types ◽

Physical Parameters ◽

Membrane Repair ◽

Plasma Membrane Integrity ◽

Repair Mechanisms ◽

And Function

AbstractMaintenance of plasma membrane integrity is essential for normal cell viability and function. Thus, robust membrane repair mechanisms have evolved to counteract the eminent threat of a torn plasma membrane. Different repair mechanisms and the bio-physical parameters required for efficient repair are now emerging from different research groups. However, less is known about when these mechanisms come into play. This review focuses on the existence of membrane disruptions and repair mechanisms in both physiological and pathological conditions, and across multiple cell types, albeit to different degrees. Fundamentally, irrespective of the source of membrane disruption, aberrant calcium influx is the common stimulus that activates the membrane repair response. Inadequate repair responses can tip the balance between physiology and pathology, highlighting the significance of plasma membrane integrity. For example, an over-activated repair response can promote cancer invasion, while the inability to efficiently repair membrane can drive neurodegeneration and muscular dystrophies. The interdisciplinary view explored here emphasises the widespread potential of targeting plasma membrane repair mechanisms for therapeutic purposes.

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Ferroptotic cell death triggered by conjugated linolenic acids is mediated by ACSL1

Nature Communications ◽

10.1038/s41467-021-22471-y ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Alexander Beatty ◽

Tanu Singh ◽

Yulia Y. Tyurina ◽

Vladimir A. Tyurin ◽

Svetlana Samovich ◽

...

Keyword(s):

Cell Death ◽

Therapeutic Potential ◽

Neutral Lipids ◽

Cell Types ◽

Lipid Hydroperoxides ◽

Conjugated Linolenic Acids ◽

Cellular Lipids ◽

Transcriptional Changes ◽

Acyl Chains ◽

Tumor Growth And Metastasis

AbstractFerroptosis is associated with lipid hydroperoxides generated by the oxidation of polyunsaturated acyl chains. Lipid hydroperoxides are reduced by glutathione peroxidase 4 (GPX4) and GPX4 inhibitors induce ferroptosis. However, the therapeutic potential of triggering ferroptosis in cancer cells with polyunsaturated fatty acids is unknown. Here, we identify conjugated linoleates including α-eleostearic acid (αESA) as ferroptosis inducers. αESA does not alter GPX4 activity but is incorporated into cellular lipids and promotes lipid peroxidation and cell death in diverse cancer cell types. αESA-triggered death is mediated by acyl-CoA synthetase long-chain isoform 1, which promotes αESA incorporation into neutral lipids including triacylglycerols. Interfering with triacylglycerol biosynthesis suppresses ferroptosis triggered by αESA but not by GPX4 inhibition. Oral administration of tung oil, naturally rich in αESA, to mice limits tumor growth and metastasis with transcriptional changes consistent with ferroptosis. Overall, these findings illuminate a potential approach to ferroptosis, complementary to GPX4 inhibition.

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Generation of Functional Human Cardiac Progenitor Cells by High-Efficiency Protein Transduction

Stem Cells Translational Medicine ◽

10.5966/sctm.2015-0136 ◽

2015 ◽

Vol 4 (12) ◽

pp. 1415-1424 ◽

Cited By ~ 23

Author(s):

Xiao-Hong Li ◽

Qianqian Li ◽

Lin Jiang ◽

Chunyu Deng ◽

Zaiyi Liu ◽

...

Keyword(s):

Progenitor Cells ◽

High Efficiency ◽

Protein Transduction ◽

Cardiac Progenitor Cells

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Precancerous niche (PCN), a product of fibrosis with remodeling by incessant chronic inflammation

4open ◽

10.1051/fopen/2018009 ◽

2019 ◽

Vol 2 ◽

pp. 11 ◽

Cited By ~ 4

Author(s):

Björn L.D.M. Brücher ◽

Ijaz S. Jamall

Keyword(s):

Extracellular Matrix ◽

Chronic Inflammation ◽

Genetic Material ◽

Cell Communication ◽

Cell Types ◽

Complex Signaling ◽

Different Cell Types ◽

Multiple Signaling ◽

Extracellular Environment

Fibroblasts are actively involved in the creation of the stroma and the extracellular matrix which are important for cell adhesion, cell–cell communication, and tissue metabolism. The role of fibrosis in carcinogenesis can be examined by analogy to tissues of various cancers. The orchestration of letters in the interplay of manifold components with signaling and crosstalk is incompletely understood but available evidence suggests a hitherto underappreciated role for fibrosis in carcinogenesis. Complex signaling and crosstalk by pathogenic stimuli evoke persistent subclinical inflammation, which in turn, results in a cascade of different cell types, ubiquitous proteins and their corresponding enzymes, cytokine releases, and multiple signaling pathways promoting the onset of fibrosis. There is considerable evidence that the body's attempt to resolve such a modified extracellular environment leads to further disruption of homeostasis and the genesis of the precancerous niche as part of the six-step process that describes carcinogenesis. The precancerous niche is formed and can be understood to develop as a result of (1) pathogenic stimulus, (2) chronic inflammation, and (3) fibrosis with alterations of the extracellular matrix, stromal rigidity, and mechano-transduction. This is why carcinogenesis is not just a process of aberrant cell growth with damaged genetic material but the role of the PCN in its entirety reveals how carcinogenesis can occur without invoking the need for somatic mutations.

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