Pancreatic ductal adenocarcinoma remains one of the most lethal malignancies with only an 11% 5-year survival rate. Oncogenic microRNAs (miRNA), particularly miR-21, miR-155, and miR-18a, drive tumor progression by silencing tumor suppressors and promoting chemoresistance. Single-target miRNA inhibition has shown limited clinical efficacy because of complex network redundancy and compensatory pathway activation, necessitating multitarget therapeutic approaches. We designed and validated a trispecific miRNA sponge construct containing high-affinity target sites for all 3 oncomiRs, demonstrated in silico through miRNAsong analysis with an approximately 11-kcal/mol thermodynamic specificity gap relative to off-targets. The sponge was functionally tested in the human pancreatic ductal adenocarcinoma cell lines PANC-1 and AsPC-1 using comprehensive assays including dual-luciferase reporter, quantitative reverse transcription polymerase chain reaction, flow cytometry-based apoptosis analysis, scratch-wound migration, and gemcitabine chemosensitization studies. In AsPC-1 cells, the trispecific sponge achieved 99.0%-99.9% silencing of target miRNAs and 349-fold reporter reduction, inducing a 6.1-fold increase in apoptosis and approximately 44% reduction in wound closure at 72 hours compared with nontargeting controls. PANC-1 cells showed moderate but significant responses with 65%-98% miRNA silencing, a 2.2-fold increase in apoptosis, and approximately 51% reduction in wound closure at 72 hours under identical assay conditions. Mechanistically, simultaneous miRNA inhibition synergistically reactivated tumor suppressor genes PDCD4, ESR1, and NOTCH2 (6.3-7.7-fold upregulation) and sensitized chemoresistant cells to gemcitabine by 1.5- to 1.8-fold. Across the evaluated functional endpoints, the trispecific sponge conferred approximately 1.2- to 95-fold changes relative to the nontargeting control and, in many instances, elicited equal or greater effects than single-target constructs, consistent with a broader network-level impact rather than uniform superiority in every assay. This platform represents a promising strategy for pancreatic ductal adenocarcinoma therapy warranting preclinical development and clinical translation. SIGNIFICANCE STATEMENT: Pancreatic cancer chemoresistance drives poor survival. The trispecific microRNA sponge simultaneously targets miR-21, miR-155, and miR-18a, synergistically reactivating tumor suppressors and enhancing gemcitabine efficacy more than single-target approaches. This multitarget microRNA strategy represents a novel therapeutic platform for overcoming chemoresistance in pancreatic cancer.

As the COVID-19 pandemic continues to challenge global health systems, the reliability of diagnostic tests remains a critical concern. The most accurate way to identify SARS-CoV-2 infection is nucleic acid amplification tests (NAATs), especially real-time PCR (RT-PCR) assays. However, changes in SARS-CoV-2 primer and probe binding sites might compromise the accuracy of these diagnostic tests and increase false-negative rates. Real-time PCR serves as the gold standard for SARS-CoV-2 detection but shows 2–29% false-negative rates. The present study analyzed ~ 26,000 SARS-CoV-2 genomic sequences from the Global Initiative on Sharing All Influenza Data (GISAID) database to shed light on genetic variants that affected the performance of ongoing setup RT-PCR primer and probe set. This study assesses 12 primer sets for detecting SARS-CoV-2 variants from late 2019 to early 2023 across four frameworks: chronological, geographical, variant-wise, and diagnostic metrics. We validated computational predictions using clinical specimens and Sanger sequencing. Our findings indicate a correlation between amplification failures and single-point mutations or other genetic alterations in the primer and probe binding sites, leading to false-negative results in RT-PCR testing. Our findings provide crucial data for RT-PCR assay design and enhancement. Specifically, our analysis provided quantitative mismatch rates (0.15–77.15%), identified critical binding site mutations causing RT-PCR failures, and established temporal performance patterns tracking variant-driven primer degradation. These results enable evidence-based primer selection and highlight the need for continuous surveillance in viral pandemics. These findings recommend implementing multiplex RT-PCR assays and continuous primer surveillance for reliable COVID-19 diagnosis.

Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal cancers worldwide, and there is an urgent demand for novel therapeutic approaches. MicroRNAs (miRNAs) are promising molecules to regulate gene expression in PDAC. This study investigates the role of miR-142 in cell proliferation and apoptosis in two cell lines, AsPC-1 and PANC-1, with a focus on gene expression changes of selected genes induced by overexpression of miR-142-5p. The molecular features in PDAC were evaluated using KEGG, and significant genes in cell cycle regulation, apoptosis, and PI3k/AKT pathway were selected (CDK6, MCL1, PIK3CA). DIANA-microT-CDS and TargetScan version 7.1 were used to select miR-142 based on prediction score. Selected genes did not contain any predicted seed site matches for the 3p arm of miR-142. miR-142 was overexpressed by transfecting a vector carrying the pre-miRNA-142 sequence into two pancreatic cancer cell lines. Overexpression of miR-142 and its effect on downregulating target genes was confirmed by quantitative real-time PCR (RT-qPCR). Predicted target genes (CDK6 and MCL1) were evaluated by the Luciferase assay. Apoptosis and cell cycle arrest were performed to analyze cell proliferation and cell viability under the effect of miR-142. Our results showed that miR-142-5p reduced the expression of its target genes. Cell cycle arrest was increased in both cell lines, while apoptosis increased in PANC-1. Overall, miR-142 may function as a tumor suppressor in PDAC by downregulating the overexpressed cancer-related genes and suppressing proliferation in AsPC-1 and PANC1.

Despite current standard treatments such as chemotherapy, radiotherapy, and hormone therapy, the five-year survival rate for patients with metastatic castration-resistant prostate cancer (mCRPC) remains about 50%. While p53 gene therapy has been widely investigated in cancer research, its clinical efficacy is limited by tumor evasion mechanisms, including the upregulation of p53 negative regulators, such as MDM2. This study aims to evaluate the impact of miR-340-mediated downregulation of MDM2 on the therapeutic efficacy of p53 gene therapy in p53 non-expressing PC3 cells. Lentiviral vectors, produced using HEK293T cells, were employed to generate scramble, miR-340-overexpressing, p53-overexpressing, and co-overexpressing PC3 cells. Quantitative polymerase chain reaction (qPCR) was used to determine the expression of p53, MDM2, p21, VEGF, and miR-340. Cell viability, apoptosis, necrosis, cell cycle alterations, cellular migration, proliferation in 2D/3D culture, and HUVECs’ angiogenic potential were assessed in vitro. Additionally, the efficacy of radiotherapy and docetaxel chemotherapy was evaluated through MTT, flow cytometry, and colony formation assay. miR-340 reduced MDM2 mRNA expression by 66% compared to the scramble control and by 45% compared to p53-overexpressing PC3 cells. miR-340 also decreased MDM2 protein expression by 30% compared to both the scramble control and p53-overexpressing PC3 cells. In PC3 cells co-expressing miR-340 and p53, miR-340 induced a 21-fold increase in p21 mRNA levels and a 58.8% increase in p53 protein expression relative to p53-overexpressing cells. Moreover, miR-340/p53 gene therapy effectively inhibited cell proliferation, migration, and angiogenic potential of PC3 cells, without markedly affecting apoptosis rates. Interestingly, this combined gene therapy substantially enhanced the therapeutic response to radiotherapy and low-dose docetaxel chemotherapy. In conclusion, miR-340 can enhance the efficacy of p53 gene therapy and holds promise for the management of mCRPC.

Lentiviral vectors (LVs) have revolutionized gene therapy by enabling stable gene integration into dividing and non-dividing cells, addressing critical challenges in treating genetic disorders. The transition from second to third-generation LVs has increased biosafety by minimizing the risk of replication-competent lentiviruses and expanded their clinical applicability. Despite significant advancements, producing high-titer functional LVs, particularly at an industrial scale, remains a considerable challenge due to the need for enhanced scalability, cost-efficiency, and effectiveness. This Review delves into cutting-edge innovations in LV production, from optimized transient transfection in various cell lines to the development of stable producer cell lines. Stable producer cell lines offer unparalleled scalability but face challenges related to viral protein cytotoxicity. Inducible systems have emerged as pivotal tools for addressing these problems, allowing for precise gene expression and controlled production. Additionally, advancements in bioprocess engineering, ranging from optimized culture conditions, including pH and media composition, to novel bioreactor technologies like structured fixed-bed systems, continue to redefine industrial-scale LV production. These breakthroughs, coupled with the analysis of costs and efficiencies of various methodologies, can further illustrate the potential for large-scale LV production and facilitate widespread therapeutic applications.

Background: This study focuses on the findings related to Latency- Associated Transcript (LAT)- derived miRNAs and their interactions with ephrin family genes, especially EFNA3. It contextualizes, these results within the broader Glioblastoma multiform (GBM) research landscape. Method: The differential expression of the Ephrin (EFN) family in GBM was analyzed using TCGA and GEO databases, alongside survival data from the Kaplan-Meier Plotter. Bioinformatics predicted LAT-derived miRNAs targeting EFN genes, which were validated in vitro. Luciferase assays confirmed miR-H2 and miR-H3's targeting of EFNA3, while qRT-PCR and Western blotting assessed their effects on mRNA and protein levels. Results: EphrinA3 is significantly overexpressed in GBM tissues, and its expression level is correlated with the prognosis of GBM patients. Both miR-H2-3p and miR-H3-3p effectively target EphrinA3, resulting in approximately a twofold reduction in luciferase activity when assessed individually. Notably, this suppressive effect is enhanced fourfold in cells expressing LAT transcript. Furthermore, both miRNAs significantly downregulate EFNA3 expression at both the mRNA and protein levels. Loss-of-function experiments indicate that LAT-derived miRNAs play a critical regulatory role in modulating EFNA3 expression. Conclusion: The study highlights the potential of targeting EphrinA3 through HSV-1-derived miRNAs, particularly miR-H2 and miR-H3, as a promising therapeutic strategy. These findings suggest that modulating EphrinA3 expression could enhance treatment efficacy while minimizing off-target effects, paving the way for innovative approaches to combat GBM. Further research is warranted to explore the clinical implications of these insights in developing effective therapies for this aggressive cancer.

[This corrects the article DOI: 10.1016/j.heliyon.2024.e26971.].

Glioblastoma Multiforme (GBM) is the most frequent and invasive primary malignant brain tumor. One approach to improve the effectiveness of GBM treatment is the combination of miRNA-targeted therapy with TMZ. This study aimed to assess the effect of miR-124 overexpression on TMZ resistance in GBM cell lines. Additionally, we examined how miR-124 overexpression affects the expression of genes involved in DNA repair processes. We conducted a bioinformatics prediction for target genes of miR‑124‑3p and then overexpressed miR-124 in U-87 and U-251 cell lines through lentiviral transduction. Sixty genes were identified as potential targets of miR-124-3p, which revealed overlap among 504 target mRNAs and upregulated genes across four GEO datasets. PRRX1, ETS, VIM, and PTBP1 genes were selected based on their contributions to DNA repair and related processes such as autophagy including Beclin-1 and Atg-5. The MTT assay results showed that only the U87 cell line overexpressing miR-124 exhibited significantly greater sensitivity to TMZ treatment. The qRT-PCR analysis revealed a significant reduction in mRNA levels of all DNA repair-related genes and two autophagy-related genes in both cell lines. The results might indicate that after TMZ-induced genomic damage, cells activate the DNA repair pathways, ultimately leading to the development of resistance. In the context of TMZ treatment, autophagy is considered a protective process for cancer cells, and definitive proof of its association with the anti-cancer activity of miR-124 requires further supplementary tests. So, modulating DNA repair pathways with miR-124 could enhance the chemosensitivity of Glioma cells to TMZ.

BACKGROUND:Various forms of decellularized extracellular matrix (dECM), including patches, powders, and hydrogels, have been applied to tissue engineering. Due to a broad need for alternatives to dECM, mostly derived from animal sources, human amniotic membrane (AM) and umbilical cord (UC) as disposable birthing materials can be suitable candidates. The present study developed hydrogels from AM and UC hydrogels and compared their physicochemical and biological properties. MATERIALS AND METHODS:The decellularized and powdered AM and UC tissues were solubilized with pepsin to form pre-gel solutions. The developed hydrogels underwent biological and physicochemical assessments using techniques such as western blot, scanning electron microscopy, immunohistochemistry, and histopathology. RESULTS:UC hydrogel demonstrated a higher elastic modulus and shorter gelation time. Although the western blot results did not show significant differences in concentration of the main ECM components, specific staining showed a higher content of mucopolysaccharides in UC hydrogel as well as collagen fibers in AM hydrogel. Both hydrogels induced a fibroblast-like morphology in the cytoplasm of mesenchymal stromal cells (MSCs). Both hydrogels are suitable for 3D culture systems and support in vivo myogenic differentiation of MSCs. Finally, the hydrogels were found to be biocompatible in vivo and showed infiltration and colonization by host cells in mice. CONCLUSION:This study highlights significant bio-physicochemical variations between human UC and AM hydrogels, emphasizing the need for careful consideration in their application for tissue reconstruction, in vitro culture systems, and cell-delivery techniques.

Latency-associated transcript (LAT)-derived miRNAs are presumably the primary anti-apoptotic impellers and the other herpes simplex virus-1 (HSV-1) genes. However, the mechanisms by which this group of miRNAs negatively regulates apoptosis have not been fully elucidated. This study evaluated the hypothesis that LAT-derived miRNAs, specifically miR-H2 and miR-H3, serve as key regulatory effectors of LAT-mediated neuronal protection by targeting MAPT and SNCA, thereby interfering with caspase activation and apoptosis pathways. In silico analyses, along with in vitro and in vivo experiments, were conducted to identify key target genes of LAT-derived miR-H3 and miR-H2 and to evaluate their regulatory effects on MAPT and SNCA in sub-stable SH-SY5Y cells and latently HSV-1-infected mice, using quantitative real-time PCR (qRT-PCR) and Western blotting. The individual roles of miR-H2 and miR-H3 were investigated in vitro using luciferase reporter assays and a loss-of-function approach involving miRNA sponges. Additionally, the neuroprotective effects of LAT-derived miR-H2 and miR-H3 were evaluated through MTT and LDH assays, flow cytometry, and a quantitative analysis of apoptosis gene expression. It has been demonstrated that miR-H2 and miR-H3 overexpression directly target and suppress the expression of MAPT and SNCA genes, reducing their protein levels in the stable SH-SY5Y cells and during latency in TGs and Hip samples of mice. In addition, conditional miRNA sponge and luciferase assay approaches indicated the potential capacity of miR-H2 and miR-H3 to regulate MAPT and SNCA expression. Cells expressing miR-H2 and miR-H3 demonstrated an 80 % increase in viability and over 40 % enhancement in cell cycle progression, linked to the downregulation of MAPT and SNCA in SH-SY5Y cells. Significant alterations in CASP3 and BCL2 expression were observed in both cellular and mouse models, underscoring the anti-apoptotic effects associated with the negative regulation of caspase cascade inducers, Tau and α-syn, by miR-H2 and miR-H3. Considering the results, the neural survival prompted by LAT-derived miRNAs suggests a positive correlation between the disruption of MAPT and SNCA expression, affecting both anti-apoptotic and pro-apoptotic processes. Future studies investigating the neuroprotective properties of HSV-1 miRNAs, particularly their impact on multi-structural peptide deficiencies, will offer valuable insights into HSV-1-host co-evolution and mechanisms for counteracting neuronal apoptosis. In addition to endogenous miRNAs, LAT-derived miRNAs should also be regarded as promising candidates with significant therapeutic potential for managing the molecular mechanisms underlying disorders that lead to the onset and progression of neurodegeneration.

Abstract Background Lung cancer is a globally pervasive and deadly disease, claiming more than 1 million lives annually. Therefore, the identification of mutations in crucial cancer-related genes is paramount for guiding optimal chemotherapy strategies. The distribution of EGFR, KRAS, ALK, and ROS1 mutations varies across diverse ethnic populations. Nonetheless, there is limited data available on the prevalence of these mutations and their correlation with PD-L1 expression among Iranian lung cancer patients. Aim This study involved an analysis of EGFR, KRAS, ALK, and ROS1 gene mutations in lung cancer patients, followed by an assessment of the correlation between PD-L1 expression and clinicopathological variables. Methods Mutational profiling was conducted by examining EGFR (exons 18–21) and KRAS (exon 2) through pyrosequencing. Detection of ALK and ROS1 rearrangements, alongside PD-L1 expression, was carried out using immunohistochemistry techniques. Results EGFR mutations were identified in 23.4% of cases, exhibiting a notably higher occurrence in females (p = 0.001). KRAS mutations were present in 7.1% of cases, with no significant association found between KRAS mutations and sex (p = 0.229). ALK rearrangements were found in 4.9% of cases, while ROS1 rearrangements were present in 0.6% of patients. The overall prevalence of PD-L1 protein expression was 36.85%. Notably, PD-L1 expression was detected in 24.8% of cases with EGFR mutations, 20% of cases with KRAS mutations, 64.7% of cases with ALK rearrangements, and in 100% of cases with ROS1 rearrangements. Conclusion Although no correlation was found between PD-L1 expression and EGFR, KRAS mutations, and ROS1 rearrangements, a noteworthy association was identified between ALK rearrangements and elevated PD-L1 expression.

Four significant influenza outbreaks have occurred over the past 100 years, and the 1918 influenza pandemic is the most severe. Since influenza viruses undergo antigenic evolution, they are the pathogens most likely to trigger a new pandemic shortly. Intranasal vaccination offers a promising strategy for preventing diseases triggered by respiratory viruses by eliciting an immunoglobulin A (IgA) response, limiting virus replication and transmission from the respiratory tract more efficiently than intramuscular vaccines. Combining intranasal administration and mRNA-lipid nanoparticles can be an ideal strategy for limiting the extent of the next flu pandemic. This study explored the immunogenicity of intranasally delivered mRNA encapsulated in mannose-histidine-conjugated chitosan lipid nanoparticles (MHCS-LNPs) as a vaccine against influenza A (H1N1) in BALB/c mice. Intranasal administration of mRNA-MHCS-LNPs resulted in the generation of influenza A (H1N1) hemagglutinin-specific neutralizing antibodies in vaccinated animals. The enzyme-linked immunosorbent assay (ELISA) results indicated a notable increase in the quantity of immunoglobulin G (IgG) and IgA antibodies in serum and the bronchoalveolar lavage fluid (BALF), respectively, and exhibited influenza A-specific IFN-γ secretion in vaccinated mice, as well as a noticeable alteration in IL-5 production. Overall, this study demonstrated an effective immunogenic response against respiratory viral infections through intranasal delivery of an mRNA-MHCS-LNP vaccine.

The SARS-CoV-2 pandemic, first reported as an acute respiratory disease in December 2019 in Wuhan, China, has profoundly impacted global communities, with approximately seven million deaths reported by the World Health Organization to date. Coronaviruses exhibit a high recombination rate due to RNA-dependent RNA polymerase transcription errors, resulting in mutations that are selectively conserved under evolutionary pressures and transmitted to subsequent generations. In this study, we investigated the role of host microRNAs (miRs) in exerting evolutionary pressures on SARS-CoV-2. We identified miRNAs binding to the Wuhan strain genome and compared their binding regions with mutated strains. Our bioinformatics analysis revealed a significant number of conserved mutations within the seed regions of microRNA binding sites in the later variants. For functional validation, we focused on hsa-miR-6512, targeting the Wuhan strain’s surface glycoprotein gene (S). This miRNA’s binding site is lost in later variants, including Alpha, and this mutation was conserved across Beta, Gamma, Mu, and Omicron. Expression of hsa-miR-6512 was assessed, and its interaction with the S gene was compared between Wuhan and Omicron strains. Our findings suggest that miRNA–genome interactions may influence the evolutionary trajectory of SARS-CoV-2 and propose miRNAs as promising targets for therapeutic strategies.

Rapid, cost-effective, and sensitive diagnostic tools are essential for managing viral outbreaks such as COVID-19. In this study, we developed a label-free localized surface plasmon resonance (LSPR) aptasensor based on polyvalent G12-aptamer-conjugated gold nanoparticles (apt@AuNPs) synthesized via a one-pot method for the detection of heat-inactivated SARS-CoV-2 in throat samples. The aptasensor exploits the specific binding of the G12 aptamer to spike proteins present either on intact viral particles or in fragmented/soluble forms resulting from heat inactivation, inducing a measurable redshift in the optical spectrum within minutes upon interaction with viral particles. Our platform achieves a detection limit comparable to PCR, enabling rapid and sensitive virus identification without the need for complex sample preparation or labeling. The colorimetric response is readily observable, facilitating point-of-care applications. Notably, the sensor achieves reliable specific detection at Ct values < 37 ( 103 copies/mL), aligning with clinical diagnostic standards for SARS-CoV-2 positivity. Furthermore, the modular design of this biosensor enables adaptation to detect other viral pathogens by substituting target-specific aptamers, providing a versatile and scalable solution for the early diagnosis and monitoring of infectious diseases. This approach holds significant promise for enhancing pandemic preparedness and response through accessible, rapid, and accurate viral detection.

Glioblastoma (GBM), a grade IV brain tumor, presents a severe challenge in treatment and eradication due to its high genetic variability and the existence of stem-like cells with self-renewal potential. Conventional therapies fall short of preventing recurrence and fail to extend the median survival of patients significantly. However, the emergence of gene therapy, which has recently obtained significant clinical outcomes, brings hope. It has the potential to be a suitable strategy for the treatment of GBM. Notably, microRNAs (miRNAs) have been noticed as critical players in the development and progress of GBM. The combined usage of hsa-miR-34a and Cytosine Deaminase (CD) suicide gene and 5-fluorocytosine (5FC) prodrug caused cytotoxicity against U87MG Glioma cells in vitro. The apoptosis and cell cycle arrest rates were measured by flow cytometry. The lentiviral vector generated overexpression of CD/miR-34a in the presence of 5FC significantly promoted apoptosis and caused cell cycle arrest in U87MG cells. The expression level of the BCL2, SOX2, and P53 genes, target genes of hsa-miR-34a, was examined by quantitative real-time PCR. The treatment led to a substantial downregulation of Bcl2 and SOX2 genes while elevating the expression levels of Caspase7 and P53 genes compared to the scrambled control. The hsa-miR-34a hindered the proliferation of GBM cancer cells and elevated apoptosis through the P53-miR-34a-Bcl2 axis. The CD suicide gene with 5FC treatment demonstrated similar results to miR-34a in the apoptosis, cell cycle, and real-time assays. The combination of CD and miR-34a produced a synergistic effect. In vivo, anti-GBM efficacy evaluation in rats bearing intracranial C6 Glioma cells revealed a remarkable induction of apoptosis and a significant inhibition of tumor growth compared with the scrambled control. The simultaneous use of CD/miR-34a with 5FC almost entirely suppressed tumor growth in rat models. The combined application of hsa-miR-34a and CD suicide gene against GBM tumors led to significant induction of apoptosis in U87MG cells and a considerable reduction in tumor growth in vivo.

Glioblastoma multiforme (GBM) patients have a high recurrence rate of 90%, and the 5-year survival rate is only about 5%. Cytosine deaminase (CDA)/5-fluorocytosine (5-FC) gene therapy is a promising glioma treatment as 5FC can cross the blood-brain barrier (BBB), while 5-fluorouracil (5-FU) cannot. Furthermore, 5-FU can assist reversing the immunological status of cold solid tumors. This study developed mesenchymal stem cells (MSCs) co-expressing yeast CDA and the secretory IL18-FC superkine to prevent recurrent tumor progression by simultaneously exerting cytotoxic effects and enhancing immune responses. IL18 was fused with Igk and IgG2a FC domains to enhance its secretion and serum half-life. The study confirmed the expression and activity of the CDA enzyme, as well as the expression, secretion, and activity of secretory IL18 and IL18-FC superkine, which were expressed by lentiviruses transduced-MSCs. In the transwell tumor-tropism assay, it was observed that the genetically modified MSCs retained their selective tumor-tropism ability following transduction. CDA-expressing MSCs, in the presence of 5-FC (200 mu g/ml), induced cell cycle arrest and apoptosis in glioma cells through bystander effects in an indirect transwell co-culture system. They reduced the viability of the direct co-culture system when they constituted only 12.5 % of the cell population. The effectiveness of engineered MSCs in suppressing tumor progression was assessed by intracerebral administration of a lethal dose of GL261 cells combined in a ratio of 1:1 with MSCs expressing CDA, or CDA and sIL18, or CDA and sIL18-FC, into C57BL/6 mice. PET scan showed no conspicuous tumor mass in the MSC-CDA-sIL18-FC group that received 5-FC treatment. The pathological analysis showed that tumor progression suppressed in this group until 20th day after cell inoculation. Cytokine assessment showed that both interferon- gamma (IFN-gamma) and interleukin-4 (IL-4) increased in the serum of MSC-CDA-sIL18 and MSC-CDA-sIL18-FC, treated with normal saline (NS) compared to those of the control group. The MSC-CDA-sIL18-FC group that received 5-FC treatment showed reduced serum levels of IL-6 and a considerably improved survival rate compared to the control group. Therefore, MSCs co-expressing yeast CDA and secretory IL18-FC, with tumor tropism capability, may serve as a

This study investigated the efficacy of using chitosan/alginate nanoparticles loaded with recombinant human bone morphogenetic-2 (rhBMP-2) and SMAD4 encoding plasmid to enhance the chondrogenesis of human bone marrow mesenchymal stem cells (hBM-MSCs) seeded on an extracellular matrix (ECM). The research treatments included the stem cells treated with the biological cocktail (BC), negative control (NC), hBM-MSCs with chondrogenic medium (MCM), hBM-MSCs with naked rhBMP-2 and chondrogenic medium (NB/C), and hBM-MSCs with naked rhBMP-2 and chondrogenic medium plus SMAD4 encoding plasmid transfected with polyethyleneimine (PEI) (NB/C/S/P). The cartilage differentiation was performed with real-time quantitative PCR analysis and alizarin blue staining. The data indicated that the biological cocktail (BC) exhibited significantly higher expression of cartilage-related genes compared to significant differences with MCM and negative control (NC) on chondrogenesis. In the (NB/C/S/P), the expression levels of SOX9 and COLX were lower than those in the BC group. The expression pattern of the ACAN gene was similar to COL2A1 changes suggesting that it holds promising potential for cartilage regeneration.

The survival rate of lung cancer is low due to the high frequency of drug resistance in patients with mutations in the driver genes. Overexpression of anti-apoptotic genes is one of the most prominent features of tumor drug resistance. EGFR signaling induces the expression of anti-apoptotic genes. Also, microRNAs (miRNAs) have a critical role in regulating biological functions such as apoptosis; a process mostly eluded in cancer progression. The mutation screening was performed on one thousand non-small cell lung carcinoma patients to enroll clinical samples in this study. Bioinformatics analysis predicted that miRNAs ( miR-29a , miR-143 ) might regulate MCL-1 and cIAP-2 expression. We investigated the expression of MCL-1 , cIAP-2 , miR-29a , and miR-143 encoding genes in adenocarcinoma patients with or without EGFR mutations before treatment. The potential role of miR-29a and miR-143 on gene expression was evaluated by overexpression and luciferase assays in HEK-293T cells. EGFR mutations were found in 262 patients (26.2%) with a greater incidence in females (36.23% vs. 20.37%, P = 0.001). The expression levels of MCL-1 and cIAP-2 genes in patients with mutated EGFR were higher than those of wild-type EGFR . In contrast, compared to those of patients with wild-type EGFR , the expression levels of miR-29a and miR-143 were lower in the patients carrying EGFR mutations. In cell culture, overexpression of miR-29a and miR-143 significantly downregulated the expression of MCL-1 and cIAP-2 . Dual-luciferase reporter experiments confirmed that miR-29a and miR-143 target MCL-1 and cIAP-2 mRNAs, respectively. Our results suggest that upregulation of EGFR signaling in lung cancer cells may increase anti-apoptotic MCL-1 and cIAP-2 gene expression, possibly through downregulation of miR-29a-3p and miR-143-3p .

mRNA-based therapeutics have revolutionized medicine and the pharmaceutical industry. The recent progress in the optimization and formulation of mRNAs has led to the development of a new therapeutic platform with a broad range of applications. With a growing body of evidence supporting the use of mRNA-based drugs for precision medicine and personalized treatments, including cancer immunotherapy, genetic disorders, and autoimmune diseases, this emerging technology offers a rapidly expanding category of therapeutic options. Furthermore, the development and deployment of mRNA vaccines have facilitated a prompt and flexible response to medical emergencies, exemplified by the COVID-19 outbreak. The establishment of stable and safe mRNA molecules carried by efficient delivery systems is now available through recent advances in molecular biology and nanotechnology. This review aims to elucidate the advancements in the clinical applications of mRNAs for addressing significant health-related challenges such as cancer, autoimmune diseases, genetic disorders, and infections and provide insights into the efficacy and safety of mRNA therapeutics in recent clinical trials.