Iranian Biomedical Journal
Volume:28 Issue: 5, Nov 2024

Chimeric antigen receptor (CAR) T-cell therapy is a groundbreaking approach for treating certain hematologic malignancies and solid tumors. However, its application is limited by severe toxicities, particularly cytokine release syndrome (CRS) and cell-associated neurotoxicity syndrome (ICANS), dramatically limit its broader application. IL-1 plays a crucial role in both enhancing CAR T-cell efficacy and driving these toxic effects. This review systematically examines the dual functions of IL-1, highlighting its role in promoting CAR T-cell activation and persistence while contributing to CRS and ICANS pathogenesis. Strategies to mitigate IL-1-driven toxicities, including IL-1 receptor antagonists, monoclonal antibodies, IL-1 trapping, and interference with IL-1 production, show promise in reducing adverse effects without compromising therapeutic efficacy. Understanding the complex role of IL-1 in CAR T-cell therapy may lead to optimized treatment strategies, improving safety and expanding clinical applicability. Further research is essential to refine IL-1-targeted interventions and enhance the therapeutic potential of CAR T-cell therapy.

One of the primary drivers of liver fibrosis is the excessive accumulation of extracellular matrix (ECM), primarily caused by the over-proliferation of hepatic star-shaped cells (HSCs). The activation of HSCs by transforming growth factor beta (TGF-β) has a critical role in initiating fibrosis. Recent studies have suggested that miRNA-133a significantly regulates the fibrogenesis process, which its downregulation is associated with the fibrosis progression. Understanding the role of miRNA-133a provides potential therapeutic insights for targeting TGF-β signaling and mitigating liver fibrosis. We investigated whether exosomes could attenuate liver fibrosis by enhancing the antifibrotic effects of miR-133a.

The LX-2 cell line was treated with TGF-β for 24 hours, followed by an additional 24 hours of treatment with exosomes. After this treatment period, we assessed the mRNA expression levels of α-SMA, collagen 1, and miR-133a, as well as the protein levels of p-Smad3.

TGF-β exposure significantly increased the expression level of α-SMA and collagen 1 genes and elevated the levels of p-Smad3 protein. Additionally, it resulted in a significant downregulation of miR-133a compared to the control group. Exosome administration effectively reduced the TGF-β-induced upregulation of p-Smad3, α-SMA, and collagen 1 genes, but increased miR-133a expression levels.

Our findings indicate that by partially mitigating the downregulation of miR-133a, exosomes can effectively inhibit the persistent activation of HSCs. Furthermore, in the context of in vitro liver fibrosis, exosomes can suppress the TGF-β/Smad3 pathway, reducing the accumulation of ECM.

Developing miRNA-mediated cell engineering introduces a novel technology for cell reprogramming and generating patient-specific tissues for therapeutic use, facilitating basic research on human adult stem cells. Furthermore, optimizing a reprogramming method without transduction minimizes the risk of tumorigenesis, especially for reprogrammed cells. This study aimed to explore the use of liposomes as vehicles for delivering miRNAs to cells, focusing on their role in regulating gene networks and facilitating nuclear reprogramming.

This study utilized cationic liposomal nanoparticles preserved under different conditions to introduce miRNAs into hMSCs. Using qPCR, the effective induction of pluripotency factors (OCT4, SOX2, and NANOG) was examined.

Results indicated that miR-302a and miR-34a regulate pluripotency by interacting with key transcription factors, including OCT4, SOX2, and NANOG. Notably, the expression pattern of OCT4 showed that lipoplexes containing miR-302a increased the expression of this gene, while in the case of miR-34a, it decreased. Additionally, the study found that pluripotency precursors can be induced by delivering liposomal microRNA (LP-miRs).

LP-miRs, as small-molecule therapeutics, can influence reprogramming/engineering and the conversion of cells into other lineages. These findings have significant implications for our understanding of the mechanisms underlying the regulation of pluripotency and may have potential applications in regenerative medicine.

Leishmaniasis is a vector-borne disease prevalent in 98 countries worldwide. The current treatment has shortcomings, including drug resistance and adverse effects, highlighting the need for novel medications and treatment strategies. This study aimed to investigate the anti-leishmanial effect of thymoquinone (TQ) during the regulation of autophagy in the macrophage cell line (RAW 264.7).

After culturing the macrophage cell line, an MTT assay was performed to assess the cytotoxicity effects of the agents at different concentrations of TQ, HCQ (hydroxychloroquine), MET (metformin), and GLU (glucantime). The study groups included PBS, GLU, TQ, TQ + MET, GLU + MET, TQ + HCQ, GLU + HCQ, HCQ, and MET.  The cells were then infected with L. major and treated with TQ, while autophagy was regulated using HCQ and MET. Subsequently, the infection index, the number of amastigote loads, and the fold change in the expression of specific autophagy-related genes (LC3, P62, and Beclin) in the treatment groups were evaluated.

There was a significant decrease in the percentage of the infected macrophages treated with TQ and also the autophagy inhibitor HCQ compared to the control group. Macrophages treated with HCQ + TQ showed a significant reduction in the infection index and amastigote load compared to the TQ-treated group. Additionally, using HCQ as an autophagy inhibitor, along with TQ or GLU, enhanced the clearance of parasites and reduced the infection index of macrophages.

Downregulating autophagy could be a promising approach for Leishmania therapy, by which the leishmanicidal effect of TQ and GLU will be enhanced.

Tuberculosis (TB) is among the top ten causes of death in South Africa and worldwide and remains a high priority of the World Health Organization. South Africa has a rich tradition for using medicinal plants to treat various diseases, including TB; however, the safety and efficacy of these plants require thorough investigation and confirmation. This study examined the anti-inflammatory and anti-TB activities of three indigenous medicinal plants against two strains of TB.

Water and methanol root extracts of E. elephantina (Burch.), L. lanceolata, and T. burchellianum were tested in vitro for their activity against Latin American Mediterranean (LAM) and East African Indian (EAI) lineages. Additionally, the phytochemical screening, anti-inflammatory properties, and cytotoxicity activities of these extracts were evaluated.

Aqueous extracts of E. elephantina and T. burchellianum exhibited anti-inflammatory activity at a concentration of 200 μg/ml, while
L. lanceolata demonstrated no activity on macrophage cells. The
E. elephantina extracts showed no cytotoxicity against the Vero cells at high concentration (200 μg/ml); however, both L. lanceolata and T. burchellianum aqueous extracts were found to be cytotoxic at 200 μg/ml. Unfortunately, none of the three plant extracts showed anti-TB activity against LAM and EAI stains.

While the tested extracts lacked direct anti-TB effects, their anti-inflammatory properties and safety profile highlight their potential for adjunctive therapy in TB management or other inflammatory conditions. Further studies are needed to explore the mechanisms and potential clinical applications of these findings.

Injectable products, particularly human vaccines, must be free from fever-inducing agents and thoroughly tested for pyrogens as part of a quality control. Consequently, manufacturing facilities are required to conduct appropriate pyrogen tests per pharmacopoeial standards. This study aimed to evaluate the reliability of the monocyte activation test (MAT) in quantifying pyrogenic content in the recombinant hepatitis B vaccine.

We assessed pyrogen activity in the active pharmaceutical ingredient (API), formulated vaccine, and aluminum hydroxide by comparing the limulus amebocyte lysate (LAL), rabbit pyrogen test (RPT), and MAT, measuring activity in relative pyrogen unit (RPU) as per the European Pharmacopoeia. Monocytes from healthy donors were isolated and identified via flow cytometry to measure the CD14+ marker frequency.

The study found that the pyrogenic concentration of lipoteichoic acid (LTA) in the MAT was 50,000 ng/mL (5.19 EEU/mL). In contrast, the same concentration in the rabbit pyrogen test (RPT) was deemed non-pyrogenic based on rectal temperature assessments. The MAT showed sensitivity to the API and adjuvant, with a detection limit of 2.5 EU/mL for IL-6, outperforming the RPT, which had a detection limit of 5 EU/mL.

A strong IL-6 response to both lipopolysaccharide (LPS) and LTA stimulation was observed, indicating that IL-6 could serve as a valuable marker for pyrogen testing. The MAT appears to be an effective alternative to the RPT for assessing pyrogenicity, demonstrating commendable consistency and accuracy across various testing systems allowed by the Ph. Eur. General MAT Chapter, especially given the RPT's limitations in controlling pyrogenicity in injectable products.

In the present study, we attempted to adapt an adherent and serum-dependent Chinese hamster ovary DG44 cell line to a serum-free suspension culture and optimize the culture condition to achieve a higher yield of recombinant human follicle stimulating hormone (r-hFSH) with acceptable quality. This approach helps to mitigate the risks associated with blood-borne pathogens, reduces lot-to-lot variability, and lowers costs, making it suitable for industrial processing and scale-up.

The cell adaptation was performed using different chemically defined SFM. This process was followed by optimization through statistical experimental design, focusing on selected physicochemical parameters, including chemical supplementation of the medium and temperature shift. Both small- and large-scale cultures were conducted to test the reproducibility of the optimized condition. The expressed protein was evaluated for comparability with the standard molecule according to the Pharmacopeia guidelines.

response surface methodology (RSM) analysis indicated that supplementation of the culture medium with galactose and sodium butyrate (NaBu), along with a temperature downshift, were the main parameters leading to increased cell viability (10%), r-hFSH level (96%), and more importantly, the glycosylation content (49%) of r-hFSH compared to the control condition.

As r-hFSH isoforms generated during in vivo post-translational modifications typically exhibit different serum/plasma half-lives and bioactivity due to their incorporated sialic acid content/glycosylation, further optimizations of r-hFSH production are necessary to enhance its biological activity. In this study, following a primary screening of the studied parameters, optimization of culture conditions based on selected parameters resulted in enhanced quality and quantity of the produced r-hFSH. However, further examination is necessary before transitioning to industrial production.

Muscular dystrophy is an inherited disease with clinical and genetic heterogeneity. Muscle weakness is the primary symptom of these disorders that often leads to disability and death. The overall prevalence for all types of muscular dystrophies worldwide is 19.8-25.1 per 100.000 population. Autosomal recessive types of muscular dystrophies are more common in Iran, likely due to the high rate of consanguineous marriage. We aimed at deciphering molecular defects in three unrelated families with muscular dystrophies not related to Duchene muscular dystrophy (MD) or limb girdle muscular dystrophies. We are reporting families having affected children with MD owing to the mutations in three genes related to the COL6A (collagen type VI, alpha subunit) gene family.

Three unrelated families, who had at least one member affected with MD and for whom a definite molecular diagnosis was not provided by routine methods, were investigated by WES and confirmed by Sanger sequencing.

In the first family, a homozygous variant was found in the COL6A3 gene (NM_004369.4:c.4390C>T:p.Arg1464Ter), which explains the clinical symptoms observed in this family. In the second family, two homozygote missense variants with possible relevance to the patient’s phenotype were identified in COL6A1 and COL6A2 genes (NM_001848.2:c.803A>G: p.Glu268Gly and NM_001849.3:c.2489G>A:p.Arg830Gln). Also, a heterozygous pathogenic variant in the COL6A2 gene (NM_001849.3: c.1053+1G>T) was detected in the third family.

WES can serve as an effective method for detecting the causative mutations in families with unresolved cases of MD. The data provided herein broadens the spectrum of mutations causing MD in Iran.