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1.
Cell ; 187(1): 95-109.e26, 2024 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-38181745

RESUMO

DddA-derived cytosine base editors (DdCBEs) and transcription activator-like effector (TALE)-linked deaminases (TALEDs) catalyze targeted base editing of mitochondrial DNA (mtDNA) in eukaryotic cells, a method useful for modeling of mitochondrial genetic disorders and developing novel therapeutic modalities. Here, we report that A-to-G-editing TALEDs but not C-to-T-editing DdCBEs induce tens of thousands of transcriptome-wide off-target edits in human cells. To avoid these unwanted RNA edits, we engineered the substrate-binding site in TadA8e, the deoxy-adenine deaminase in TALEDs, and created TALED variants with fine-tuned deaminase activity. Our engineered TALED variants not only reduced RNA off-target edits by >99% but also minimized off-target mtDNA mutations and bystander edits at a target site. Unlike wild-type versions, our TALED variants were not cytotoxic and did not cause developmental arrest of mouse embryos. As a result, we obtained mice with pathogenic mtDNA mutations, associated with Leigh syndrome, which showed reduced heart rates.


Assuntos
DNA Mitocondrial , Efetores Semelhantes a Ativadores de Transcrição , Animais , Humanos , Camundongos , Adenina , Citosina , DNA Mitocondrial/genética , Edição de Genes , RNA , Efetores Semelhantes a Ativadores de Transcrição/metabolismo , Engenharia de Proteínas
2.
Cell ; 185(10): 1764-1776.e12, 2022 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-35472302

RESUMO

Mitochondrial DNA (mtDNA) editing paves the way for disease modeling of mitochondrial genetic disorders in cell lines and animals and also for the treatment of these diseases in the future. Bacterial cytidine deaminase DddA-derived cytosine base editors (DdCBEs) enabling mtDNA editing, however, are largely limited to C-to-T conversions in the 5'-TC context (e.g., TC-to-TT conversions), suitable for generating merely 1/8 of all possible transition (purine-to-purine and pyrimidine-to-pyrimidine) mutations. Here, we present transcription-activator-like effector (TALE)-linked deaminases (TALEDs), composed of custom-designed TALE DNA-binding arrays, a catalytically impaired, full-length DddA variant or split DddA originated from Burkholderia cenocepacia, and an engineered deoxyadenosine deaminase derived from the E. coli TadA protein, which induce targeted A-to-G editing in human mitochondria. Custom-designed TALEDs were highly efficient in human cells, catalyzing A-to-G conversions at a total of 17 target sites in various mitochondrial genes with editing frequencies of up to 49%.


Assuntos
DNA Mitocondrial , Doenças Mitocondriais , Animais , Sistemas CRISPR-Cas , Citosina/metabolismo , DNA Mitocondrial/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Edição de Genes , Humanos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Doenças Mitocondriais/genética , Purinas
3.
Small ; : e2405618, 2024 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-39264000

RESUMO

Since the coronavirus pandemic, mRNA vaccines have revolutionized the field of vaccinology. Lipid nanoparticles (LNPs) are proposed to enhance mRNA delivery efficiency; however, their design is suboptimal. Here, a rational method for designing LNPs is explored, focusing on the ionizable lipid composition and structural optimization using machine learning (ML) techniques. A total of 213 LNPs are analyzed using random forest regression models trained with 314 features to predict the mRNA expression efficiency. The models, which predict mRNA expression levels post-administration of intradermal injection in mice, identify phenol as the dominant substructure affecting mRNA encapsulation and expression. The specific phospholipids used as components of the LNPs, as well as the N/P ratio and mass ratio, are found to affect the efficacy of mRNA delivery. Structural analysis highlights the impact of the carbon chain length on the encapsulation efficiency and LNP stability. This integrated approach offers a framework for designing advanced LNPs and has the potential to unlock the full potential of mRNA therapeutics.

4.
J Infect Dis ; 2023 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-37711050

RESUMO

Developing new adjuvants that can effectively induce both humoral and cellular immune responses while broadening the immune response is of great value. In this study, we aimed to develop GM-CSF- or IL-18-expressing single-stranded RNA (ssRNA) adjuvants based on the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) and tested their efficacy in combination with ovalbumin (OVA) or inactivated influenza vaccines. Notably, cytokine-expressing RNA adjuvants increased the expression of antigen-presenting cell activation markers. Specifically, GM-CSF-expressing RNA adjuvants increased CD4+T cell responses, while IL-18-expressing RNA adjuvants increased CD8+T cell responses in mice when combined with OVA. In addition, cytokine-expressing RNA adjuvants increased the frequency of polyclonal T cells in combination with the influenza vaccine and reduced the clinical illness scores and weight loss of mice after viral challenge. Collectively, our results suggest that cytokine-expressing RNA adjuvants can be applied to protein-based or inactivated vaccines to increase their efficacy.

5.
J Med Virol ; 95(12): e29309, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-38100632

RESUMO

The E6 and E7 proteins of specific subtypes of human papillomavirus (HPV), including HPV 16 and 18, are highly associated with cervical cancer as they modulate cell cycle regulation. The aim of this study was to investigate the potential antitumor effects of a messenger RNA-HPV therapeutic vaccine (mHTV) containing nononcogenic E6 and E7 proteins. To achieve this, C57BL/6j mice were injected with the vaccine via both intramuscular and subcutaneous routes, and the resulting effects were evaluated. mHTV immunization markedly induced robust T cell-mediated immune responses and significantly suppressed tumor growth in both subcutaneous and orthotopic tumor-implanted mouse model, with a significant infiltration of immune cells into tumor tissues. Tumor retransplantation at day 62 postprimary vaccination completely halted progression in all mHTV-treated mice. Furthermore, tumor expansion was significantly reduced upon TC-1 transplantation 160 days after the last immunization. Immunization of rhesus monkeys with mHTV elicited promising immune responses. The immunogenicity of mHTV in nonhuman primates provides strong evidence for clinical application against HPV-related cancers in humans. All data suggest that mHTV can be used as both a therapeutic and prophylactic vaccine.


Assuntos
Proteínas Oncogênicas Virais , Infecções por Papillomavirus , Vacinas contra Papillomavirus , Neoplasias do Colo do Útero , Humanos , Feminino , Animais , Camundongos , Papillomavirus Humano , Proteínas Oncogênicas Virais/genética , Infecções por Papillomavirus/prevenção & controle , RNA Mensageiro/genética , Proteínas E7 de Papillomavirus/genética , Camundongos Endogâmicos C57BL , Vacinação/métodos , Imunização , Neoplasias do Colo do Útero/prevenção & controle
6.
Sensors (Basel) ; 22(8)2022 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-35458902

RESUMO

In this paper, experimental validation of high precision web handling for a two-actuator-based roll-to-roll (R2R) system is presented. To achieve this, the tension control loop is utilized to regulate the tension in the unwinder module, and the velocity loop is utilized to regulate the web speed in the rewinder module owing to the limitation of the number of actuators. Moreover, the radius estimation algorithm is applied to achieve an accurate web speed and the control sequence of the web handling in the longitudinal axis is developed to manipulate the web handling for convenience. Having these, the tension control performances are validated within ±0.79, ±1.32 and ±1.58 percent tension tracking error and 1.6, 1.53 and 1.33 percent web speed error at the speeds of 0.1 m/s, 0.2 m/s, and 0.3 m/s, respectively. The tension control performance is verified within ±0.3 N tracking error in the changes of the reference tension profile at 0.1 m/s web speed. Lastly, the air floating roller is used to minimize the friction terms and the inertia of the idle roller in the tension zone so that tension control performance can be better achieved during web transportation.

7.
Small ; 16(5): e1905821, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31898870

RESUMO

Although carbon nanotubes (CNTs) are remarkable materials with many exceptional characteristics, their poor chemical functionality limits their potential applications. Herein, a strategy is proposed for functionalizing CNTs, which can be achieved with any functional group (FG) without degrading their intrinsic structure by using a deoxyribonucleic acid (DNA)-binding peptide (DBP) anchor. By employing a DBP tagged with a certain FG, such as thiol, biotin, and carboxyl acid, it is possible to introduce any FG with a controlled density on DNA-wrapped CNTs. Additionally, different types of FGs can be introduced on CNTs simultaneously, using DBPs tagged with different FGs. This method can be used to prepare CNT nanocomposites containing different types of nanoparticles (NPs), such as Au NPs, magnetic NPs, and quantum dots. The CNT nanocomposites decorated with these NPs can be used as reusable catalase-like nanocomposites with exceptional catalytic activities, owing to the synergistic effects of all the components. Additionally, the unique DBP-DNA interaction allows the on-demand detachment of the NPs attached to the CNT surface; further, it facilitates a CNT chirality-specific NP attachment and separation using the sequence-specific programmable characteristics of oligonucleotides. The proposed method provides a novel chemistry platform for constructing new functional CNTs suitable for diverse applications.


Assuntos
Nanocompostos , Nanotubos de Carbono , Peptídeos , DNA/metabolismo , Nanocompostos/química , Nanotubos de Carbono/química , Peptídeos/química , Peptídeos/metabolismo , Pontos Quânticos
8.
Analyst ; 145(12): 4079-4095, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32386402

RESUMO

DNA binding fluorescent proteins are useful probes for a broad range of biological applications. Fluorescent protein (FP)-tagging allows DNA binding proteins expressed within a living cell to be directly visualised, in real-time, to study DNA binding patterns and dynamics. Moreover, FP-tagged DNA binding proteins (FP-DBP) have allowed the imaging of single proteins bound to large elongated DNA molecules with a fluorescence microscope. Although there are numerous DNA binding proteins, only a small portion of them have been exploited to construct FP-DBPs to study molecular motion in a cell or in vitro single-molecule visualisation. Therefore, it would be informative to review FP-DBP for further development. Here, we summarise the design of FP-DBPs and their brightness, photostability, pKa, maturation rate, and binding affinity (Kd) characteristics. Then, we review the applications of FP-DBP in cells to study chromosome dynamics, DNA replication, transcription factors, DNA damage, and repair. Finally, we focus on single DNA molecule visualisation using FP-DBP.


Assuntos
Proteínas de Ligação a DNA/química , DNA/metabolismo , Corantes Fluorescentes/química , Proteínas Luminescentes/química , Animais , Linhagem Celular , Cromossomos/metabolismo , DNA/análise , Dano ao DNA/fisiologia , Reparo do DNA/fisiologia , Replicação do DNA/fisiologia , Proteínas de Ligação a DNA/metabolismo , Corantes Fluorescentes/metabolismo , Humanos , Proteínas Luminescentes/metabolismo , Microscopia/métodos , Mitose/fisiologia , Plantas , Ligação Proteica , Análise de Célula Única/métodos
9.
Nucleic Acids Res ; 46(18): e108, 2018 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-29931115

RESUMO

Fluorophore-linked, sequence-specific DNA binding reagents can visualize sequence information on a large DNA molecule. In this paper, we synthesized newly designed TAMRA-linked polypyrrole to visualize adenine and thymine base pairs. A fluorescent image of the stained DNA molecule generates an intensity profile based on A/T frequency, revealing a characteristic sequence composition pattern. Computer-aided comparison of this intensity pattern with the genome sequence allowed us to determine the DNA sequence on a visualized DNA molecule from possible intensity profile pattern candidates for a given genome. Moreover, TAMRA-polypyrrole offers robust advantages for single DNA molecule detection: no fluorophore-mediated photocleavage and no structural deformation, since it exhibits a sequence-specific pattern alone without the use of intercalating dyes such as YOYO-1. Accordingly, we were able to identify genomic DNA fragments from Escherichia coli cells by aligning them to the genomic A/T frequency map based on TAMRA-polypyrrole-generated intensity profiles. Furthermore, we showed band and interband patterns of polytene chromosomal DNA stained with TAMRA-polypyrrole because it prefers to bind AT base pairs.


Assuntos
Pareamento de Bases , DNA/química , Substâncias Intercalantes , Polímeros/química , Pirróis/química , Rodaminas/química , Coloração e Rotulagem/métodos , Adenina/química , Adenina/metabolismo , Pareamento de Bases/efeitos dos fármacos , Sequência de Bases , Benzoxazóis/química , Benzoxazóis/farmacologia , DNA/efeitos dos fármacos , Escherichia coli/genética , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/química , Corantes Fluorescentes/farmacologia , Substâncias Intercalantes/síntese química , Substâncias Intercalantes/química , Substâncias Intercalantes/farmacologia , Polímeros/farmacologia , Pirróis/farmacologia , Compostos de Quinolínio/química , Compostos de Quinolínio/farmacologia , Rodaminas/farmacologia , Imagem Individual de Molécula/métodos , Timina/química , Timina/metabolismo
10.
Analyst ; 144(3): 921-927, 2019 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-30310901

RESUMO

The recent advances in the single cell genome analysis are generating a considerable amount of novel insights into complex biological systems. However, there are still technical challenges because each cell has a single copy of DNA to be amplified in most single cell genome analytical methods. In this paper, we present a novel approach to directly visualize a genomic map on a large DNA molecule instantly stained with red and green DNA-binding fluorescent proteins without DNA amplification. For this visualization, we constructed a few types of fluorescent protein-fused DNA-binding proteins: H-NS (histone-like nucleoid-structuring protein), DNA-binding domain of BRCA1 (breast cancer 1), high mobility group-1 (HMG), and lysine tryptophan (KW) repeat motif. Because H-NS and HMG preferentially bind A/T-rich regions, we combined A/T specific binder (H-NS-mCherry and HMG-mCherry as red color) and a non-specific complementary DNA binder (BRCA1-eGFP and 2(KW)2-eGFP repeat as green color) to produce a sequence-specific two-color DNA physical map for efficient optical identification of single DNA molecules.


Assuntos
Proteínas de Ligação a DNA/metabolismo , DNA/análise , Proteínas de Fluorescência Verde/metabolismo , Processamento de Imagem Assistida por Computador/métodos , Microscopia de Fluorescência/métodos , Análise de Célula Única/métodos , DNA/química , DNA/metabolismo , Humanos
11.
Chemistry ; 24(22): 5895-5900, 2018 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-29443432

RESUMO

Bioorthogonal metabolic DNA labeling with fluorochromes is a powerful strategy to visualize DNA molecules and their functions. Here, we report the development of a new DNA metabolic labeling strategy enabled by the catalyst-free bioorthogonal ligation using vinyl thioether modified thymidine and o-quinolinone quinone methide. With the newly designed vinyl thioether-modified thymidine (VTdT), we added labeling tags on cellular DNA, which could further be linked to fluorochromes in cells. Therefore, we successfully visualized the DNA localization within cells as well as single DNA molecules without other staining reagents. In addition, we further characterized this bioorthogonal DNA metabolic labeling using DNase I digestion, MS characterization of VTdT as well as VTdT-oQQF conjugate in cell nuclei or mitochondria. This technique provides a powerful strategy to study DNA in cells, which paves the way to achieve future spatiotemporal deciphering of DNA synthesis and functions.


Assuntos
DNA/síntese química , Corantes Fluorescentes/química , Indolquinonas/química , Sulfetos/química , Timidina/química , DNA/química , Desoxirribonuclease I/metabolismo , Células HeLa , Humanos , Microscopia Confocal , Ressonância Magnética Nuclear Biomolecular , Quinolonas/química , Ribonuclease Pancreático/metabolismo
12.
Nucleic Acids Res ; 44(1): e6, 2016 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-26264666

RESUMO

Fluorescent proteins that also bind DNA molecules are useful reagents for a broad range of biological applications because they can be optically localized and tracked within cells, or provide versatile labels for in vitro experiments. We report a novel design for a fluorescent, DNA-binding protein (FP-DBP) that completely 'paints' entire DNA molecules, whereby sequence-independent DNA binding is accomplished by linking a fluorescent protein to two small peptides (KWKWKKA) using lysine for binding to the DNA phosphates, and tryptophan for intercalating between DNA bases. Importantly, this ubiquitous binding motif enables fluorescent proteins (Kd = 14.7 µM) to confluently stain DNA molecules and such binding is reversible via pH shifts. These proteins offer useful robust advantages for single DNA molecule studies: lack of fluorophore mediated photocleavage and staining that does not perturb polymer contour lengths. Accordingly, we demonstrate confluent staining of naked DNA molecules presented within microfluidic devices, or localized within live bacterial cells.


Assuntos
Proteínas de Ligação a DNA/metabolismo , DNA/metabolismo , Proteínas Luminescentes/metabolismo , Imagem Molecular , Proteínas Recombinantes de Fusão , Proteínas de Ligação a DNA/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Humanos , Proteínas Luminescentes/genética , Microscopia de Fluorescência , Imagem Molecular/métodos
13.
Small ; 13(2)2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27813273

RESUMO

Synthesis of smooth and continuous DNA nanowires, preserving the original structure of native DNA, and allowing its analysis by scanning electron microscope (SEM), is demonstrated. Gold nanoparticles densely assembled on the DNA backbone via thiol-tagged DNA binding peptides work as seeds for metallization of DNA. This method allows whole analysis of DNA molecules with entangled 3D features.


Assuntos
DNA/análise , Microscopia Eletrônica de Varredura/instrumentação , Nanofios/química , Peptídeos/metabolismo , Sequência de Aminoácidos , Ouro/química , Nanofios/ultraestrutura , Peptídeos/química , Compostos de Sulfidrila/química
14.
J Acoust Soc Am ; 139(5): 2324, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27250128

RESUMO

In this paper, the sound transmission loss (STL) of thin double plates with an air cavity between them in a rigid duct is considered using an analytical approach. The vibration motion of the plate and sound pressure field are expanded in terms of an infinite series of the modal functions. Under the plane wave condition, a low frequency solution is derived by including the first few symmetric modes. It is determined that the peak frequencies of the double plates coincide with those of each single plate. When the two plates are identical, the STL becomes zero at the natural frequencies of the single plate. However, when the two plates are not identical, the STL is always greater than zero. The location and amplitude of the dips are investigated using an approximate solution when the cavity depth is very small. It is observed that dividing the single plate into two plates with an air cavity in between degrades the STL in the low frequency range, while the equivalent surface mass density is preserved. However, when the cavity depth is not small, the STL of the single plate can be smaller than that of the double plates.

15.
Electrophoresis ; 36(17): 2057-71, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25994517

RESUMO

Long and linear DNA molecules are the mainstream single-molecule analytes for a variety of biochemical analysis within microfluidic devices, including functionalized surfaces and nanostructures. However, for biochemical analysis, large DNA molecules have to be unraveled, elongated, and visualized to obtain biochemical and genomic information. To date, elongated DNA molecules have been exploited in the development of a number of genome analysis systems as well as for the study of polymer physics due to the advantage of direct visualization of single DNA molecule. Moreover, each single DNA molecule provides individual information, which makes it useful for stochastic event analysis. Therefore, numerous studies of enzymatic random motions have been performed on a large elongated DNA molecule. In this review, we introduce mechanisms to elongate DNA molecules using microfluidics and nanostructures in the beginning. Secondly, we discuss how elongated DNA molecules have been utilized to obtain biochemical and genomic information by direct visualization of DNA molecules. Finally, we reviewed the approaches used to study the interaction of proteins and large DNA molecules. Although DNA-protein interactions have been investigated for many decades, it is noticeable that there have been significant achievements for the last five years. Therefore, we focus mainly on recent developments for monitoring enzymatic activity on large elongated DNA molecules.


Assuntos
DNA/ultraestrutura , Microscopia de Fluorescência/métodos , DNA/química , DNA/metabolismo , Metilação de DNA , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/ultraestrutura , Imagem Molecular
16.
Analyst ; 139(10): 2432-9, 2014 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-24695614

RESUMO

DNA polymerase I offers great promise for a wide range of biotechnological applications due to its capability to add labeled nucleotides into double-stranded large DNA molecules by using both polymerase and nuclease domains. Accordingly, it is crucially important to thoroughly characterize this enzyme for further developments. Although the enzyme has been thus far characterized using mainly traditional analytical instruments, here we utilized an advanced and convenient means of mass spectrometry to elucidate enzymatic functions and mechanisms by measuring DNA oligomers generated by polymerase and nuclease reactions. Our analysis revealed several novel enzymatic features, including the observation that polymerase readily dissociates from the DNA molecules containing a wide single-stranded section. From this finding, we reasoned a serious situation of DNA break because polymerase domains cannot efficiently repair the wide single-stranded section, which is susceptible to DNA breaks. Furthermore, we deduced a plausible explanation for a paradoxical question as to why two domains of polymerase and 5'-nuclease are linked by a small and flexible polypeptide in polymerase I. The polypeptide link seems to prevent a 5'-nuclease from causing DNA breaks by locating a polymerase domain closely for immediate repair reaction. Here we present experimental evidence to prove our hypothesis via a set of mass spectrometric analyses as well as single DNA molecule observation and bacterial cell growth assay. Consequently, mass spectrometric analysis for DNA polymerase I provides a meaningful biological insight that a polypeptide link can be a molecular leash to control an aggressive domain in order to prevent unmanageable damages.


Assuntos
DNA Polimerase I/química , Espectrometria de Massas/métodos , Peptídeos/química , Sequência de Bases , Dano ao DNA , Sondas de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
17.
Adv Mater ; : e2411479, 2024 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-39318072

RESUMO

Herein, a mild thermal annealing (MTA) process is presented for additive-free gelation of graphene oxide (GO) dispersions. This process transitions the GO from a nematic liquid crystal phase to a random network structure, significantly enhancing its rheological properties by order of magnitude. This transition is facilitated by the diffusion of functional groups on the GO surface, which induces hydrophobic attractions, leading to a stable network structure. Employing rheo-SAXS experiments, detailed insights are provided into the microstructural changes of GO gel under shear stress, establishing a direct correlation between its rheological behavior and microstructure. The distinctive properties of MTA-processed inks are illustrated, seamlessly integrating with 3D printing, to yield a highly porous lattice structure that demonstrates promising potential as a supercapacitor electrode. The MTA process, an additive-free approach to gelation, maintains the inherent dispersion properties of GO while offering scalability. Thus, this method brings significant economic and environmental advantages compared to conventional gelation techniques. The findings not only advance the fundamental understanding of 2D colloidal network gels but also increase the potential of GO for a wide range of applications, from gas and liquid absorbers to electrodes for energy storage and conversion, and biomedical fields.

18.
PLoS One ; 19(10): e0311726, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39388490

RESUMO

The current COVID-19 mRNA vaccines were developed and applied for pandemic-emergent conditions. These vaccines use a small piece of the virus's genetic material (mRNA) to stimulate an immune response against COVID-19. However, their potential effects on individuals with chronic inflammatory conditions and vaccination routes remain questionable. Therefore, we investigated the effects of mRNA vaccines in a mouse model of chronic inflammation, focusing on their cardiac toxicity and immunogenicity dependent on the injection route. mRNA vaccine intravenous administration with or without chronic inflammation exacerbated cardiac pericarditis and myocarditis; immunization induced mild inflammation and inflammatory cytokine IL-1beta and IL-6 production in the heart. Further, IV mRNA vaccination induced cardiac damage in LPS chronic inflammation, particularly serum troponin I (TnI), which dramatically increased. IV vaccine administration may induce more cardiotoxicity in chronic inflammation. These findings highlight the need for further research to understand the underlying mechanisms of mRNA vaccines with chronic inflammatory conditions dependent on injection routes.


Assuntos
Vacinas contra COVID-19 , COVID-19 , Miocardite , Vacinas de mRNA , Animais , Masculino , Camundongos , Administração Intravenosa , Doença Crônica , COVID-19/prevenção & controle , Vacinas contra COVID-19/administração & dosagem , Vacinas contra COVID-19/efeitos adversos , Modelos Animais de Doenças , Inflamação , Interleucina-1beta , Interleucina-6 , Camundongos Endogâmicos BALB C , Vacinas de mRNA/administração & dosagem , Vacinas de mRNA/efeitos adversos , Miocardite/induzido quimicamente , Troponina I
19.
Exp Mol Med ; 2024 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-39482535

RESUMO

Mitochondrial dysfunction induced by mitochondrial DNA (mtDNA) mutations has been implicated in various human diseases. A comprehensive analysis of mitochondrial genetic disorders requires suitable animal models for human disease studies. While gene knockout via premature stop codons is a powerful method for investigating the unique functions of target genes, achieving knockout of mtDNA has been rare. Here, we report the genotypes and phenotypes of heteroplasmic MT-ND5 gene-knockout mice. These mutant mice presented damaged mitochondrial cristae in the cerebral cortex, hippocampal atrophy, and asymmetry, leading to learning and memory abnormalities. Moreover, mutant mice are susceptible to obesity and thermogenetic disorders. We propose that these mtDNA gene-knockdown mice could serve as valuable animal models for studying the MT-ND5 gene and developing therapies for human mitochondrial disorders in the future.

20.
Bioact Mater ; 38: 486-498, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38779592

RESUMO

The rapid development of messenger RNA (mRNA) vaccines formulated with lipid nanoparticles (LNPs) has contributed to control of the COVID-19 pandemic. However, mRNA vaccines have raised concerns about their potential toxicity and clinical safety, including side effects, such as myocarditis, anaphylaxis, and pericarditis. In this study, we investigated the potential of trehalose glycolipids-containing LNP (LNP S050L) to reduce the risks associated with ionizable lipids. Trehalose glycolipids can form hydrogen bonds with polar biomolecules, allowing the formation of a stable LNP structure by replacing half of the ionizable lipids. The efficacy and safety of LNP S050L were evaluated by encapsulating the mRNA encoding the luciferase reporter gene and measuring gene expression and organ toxicity, respectively. Furthermore, mice immunized with an LNP S050L-formulated mRNA vaccine expressing influenza hemagglutinin exhibited a significant reduction in organ toxicity, including in the heart, spleen, and liver, while sustaining gene expression and immune efficiency, compared to conventional LNPs (Con-LNPs). Our findings suggest that LNP S050L, a trehalose glycolipid-based LNP, could facilitate the development of safe mRNA vaccines with improved clinical safety.

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