bims-limsir 2024-01-28 papers

Methods Mol Biol. 2024 ;2766 145-151

RNA interference (RNAi) is a widely used technique to regulate the expression of genes and proteins with a high degree of specificity that is not easily accessed by traditional pharmacological approaches. For preclinical research on rheumatoid arthritis (RA), silencing of target genes in primary immune cells can be easily achieved by the application of small interfering RNA (siRNA) and synthetic short hairpin RNA (shRNA). Cellular and systemic administration of siRNA or shRNA has been a significant advance in preclinical research on RA. In this chapter, the basic techniques for gene silencing in human-derived peripheral T cells using liposome-dependent siRNA transfection and lentiviral-mediated shRNA delivery, aiming at gene silencing of therapeutic targets, are introduced.

Keywords: Gene silencing; Lentiviral-mediated delivery; RNA interference; T cells; Transfection

DOI: https://doi.org/10.1007/978-1-0716-3682-4_15

Curr Probl Cardiol. 2024 Jan 19. pii: S0146-2806(24)00058-6. [Epub ahead of print] 102419

Inclisiran siRNA Technology in the Management of Dyslipidemia: A Narrative Review of Clinical Trials.

Gbolahan Olatunji, Emmanuel Kokori, Ismaila Ajayi Yusuf, Olumide Akinmoju, Emmanuel Egbunu, Ifeanyichukwu Muogbo, Kaleb Lema, Sai Gutham Kanagala, Samuel Owolabi, Muili Abdulbasit, Nicholas Aderinto.

RNA interference therapies, particularly small interfering RNAs (siRNAs) like Inclisiran, have shown great potential in managing dyslipidemia, a significant risk factor for cardiovascular disease. Inclisiran targets pro-protein convertasesubtilisin/kexin type 9 (PCSK9) mRNA to reduce low-density lipoprotein cholesterol (LDL-C) levels. This review evaluates Inclisiran's efficacy, safety, and clinical applications in managing dyslipidemia. A review of clinical trials evaluating Inclisiran's efficacy and safety in dyslipidemia management was conducted. PubMed, Embase, Google Scholar and Scopus were searched for relevant trials. Inclusion criteria covered clinical trials in English, published within the last six years, involving human subjects. 12 clinical trials were included in this review, demonstrating Inclisiran's consistent efficacy in reducing LDL-C levels across diverse patient populations, even in statin intolerance or resistance cases. The efficacy was observed over various durations, with some trials extending up to 4 years. Inclisiran demonstrated a favourable safety profile, with mild adverse events reported in most trials, suggesting its potential as a well-tolerated treatment option. Inclisiran's consistent efficacy and safety profile make it a promising option for managing dyslipidemia. Future studies should confirm its long-term effects and explore its clinical implications in diverse patient populations and high-risk scenarios.

Keywords: Inclisiran; LDL cholesterol; dyslipidemia; siRNA therapy

DOI: https://doi.org/10.1016/j.cpcardiol.2024.102419

Mol Med Rep. 2024 Mar;pii: 47. [Epub ahead of print]29(3):

CAR‑T cell therapy: A breakthrough in traditional cancer treatment strategies (Review).

Dahua Sun, Xiang Shi, Sanyan Li, Xiaohua Wang, Xiao Yang, Meiping Wan.

Chimeric antigen receptor (CAR)‑T cell therapy is an innovative approach to immune cell therapy that works by modifying the T cells of a patient to express the CAR protein on their surface, and thus induce their recognition and destruction of cancer cells. CAR‑T cell therapy has shown some success in treating hematological tumors, but it still faces a number of challenges in the treatment of solid tumors, such as antigen selection, tolerability and safety. In response to these issues, studies continue to improve the design of CAR‑T cells in pursuit of improved therapeutic efficacy and safety. In the future, CAR‑T cell therapy is expected to become an important cancer treatment, and may provide new ideas and strategies for individualized immunotherapy. The present review provides a comprehensive overview of the principles, clinical applications, therapeutic efficacy and challenges of CAR‑T cell therapy.

Keywords: adverse effects; chimeric antigen receptor T cell therapy; immunotherapy; malignant tumors

DOI: https://doi.org/10.3892/mmr.2024.13171

J Immunother Cancer. 2023 Nov 20. pii: e007803. [Epub ahead of print]11(11):

CD8+ chimeric antigen receptor T cells manufactured in absence of CD4+ cells exhibit hypofunctional phenotype.

Sang Yun Lee, Dong Hoon Lee, Wei Sun, Francisco Cervantes-Contreras, Ryan S Basom, Feinan Wu, Si Liu, Richa Rai, Hamid R Mirzaei, Shyril O'Steen, Damian J Green, Mazyar Shadman, Brian G Till.

BACKGROUND: Cell culture conditions during manufacturing can impact the clinical efficacy of chimeric antigen receptor (CAR) T cell products. Production methods have not been standardized because the optimal approach remains unknown. Separate CD4+ and CD8+ cultures offer a potential advantage but complicate manufacturing and may affect cell expansion and function. In a phase 1/2 clinical trial, we observed poor expansion of separate CD8+ cell cultures and hypothesized that coculture of CD4+ cells and CD8+ cells at a defined ratio at culture initiation would enhance CD8+ cell expansion and simplify manufacturing.
METHODS: We generated CAR T cells either as separate CD4+ and CD8+ cells, or as combined cultures mixed in defined CD4:CD8 ratios at culture initiation. We assessed CAR T cell expansion, phenotype, function, gene expression, and in vivo activity of CAR T cells and compared these between separately expanded or mixed CAR T cell cultures.
RESULTS: We found that the coculture of CD8+ CAR T cells with CD4+ cells markedly improves CD8+ cell expansion, and further discovered that CD8+ cells cultured in isolation exhibit a hypofunctional phenotype and transcriptional signature compared with those in mixed cultures with CD4+ cells. Cocultured CAR T cells also confer superior antitumor activity in vivo compared with separately expanded cells. The positive impact of CD4+ cells on CD8+ cells was mediated through both cytokines and direct cell contact, including CD40L-CD40 and CD70-CD27 interactions.
CONCLUSIONS: Our data indicate that CD4+ cell help during cell culture maintains robust CD8+ CAR T cell function, with implications for clinical cell manufacturing.

Keywords: CD4-CD8 Ratio; CD8-Positive T-Lymphocytes; Cell Engineering; Immunotherapy, Adoptive; Receptors, Chimeric Antigen

DOI: https://doi.org/10.1136/jitc-2023-007803