The Impact of 3D Nichoids and Matrix Stiffness on Primary Malignant Mesothelioma Cells

Our plain-English summary of The Impact of 3D Nichoids and Matrix Stiffness on Primary Malignant Mesothelioma Cells, by S. Oliveto, P. Ritter, G. Deroma, et al (Genes, 2024).

The study

• Authors: S. Oliveto, P. Ritter, G. Deroma, A. Miluzio, C. Cordiglieri, M.R. Benvenuti, L. Mutti, M.T. Raimondi, S. Biffo
• Journal: Genes
• Published: 2024;15(2)
• Cited by: 3 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Malignant mesothelioma is a type of cancer that affects the mesothelium. It is an aggressive and deadly form of cancer that is often caused by exposure to asbestos. At the molecular level, it is characterized by a low number of genetic mutations and high heterogeneity among patients. In this work, we analyzed the plasticity of gene expression of primary mesothelial cancer cells by comparing their properties on 2D versus 3D surfaces. First, we derived from primary human samples four independent primary cancer cells. Then, we used Nichoids, which are micro-engineered 3D substrates, as three-dimensional structures. Nichoids limit the dimension of adhering cells during expansion by counteracting cell migration between adjacent units of a substrate with their microarchitecture. Tumor cells grow effectively on Nichoids, where they show enhanced proliferation. We performed RNAseq analyses on all the samples and compared the gene expression pattern of Nichoid-grown tumor cells to that of cells grown in a 2D culture. The PCA analysis showed that 3D samples were more transcriptionally similar compared to the 2D ones. The 3D Nichoids induced a transcriptional remodeling that affected mainly genes involved in extracellular matrix assembly. Among these genes responsible for collagen formation, COL1A1 and COL5A1 exhibited elevated expression, suggesting changes in matrix stiffness. Overall, our data show that primary mesothelioma cells can be effectively expanded in Nichoids and that 3D growth affects the cells' tensegrity or the mechanical stability of their structure.

In plain English

This was a laboratory study of cancer cells, not a study of people. The authors took malignant pleural mesothelioma cells from surgically removed tumors of male patients aged 70 to 75 with a history of past asbestos exposure. They grew the cells on micro-engineered scaffolds called Nichoids, comparing a layered three-dimensional (3D) Nichoid with a flat, one-floor (2D) Nichoid made of the same material. Cells on the flat scaffold declined in number after day 3, while cells on the 3D scaffold remained constant. Gene-activity sequencing of cells from four patients showed that the 3D-grown samples were more alike one another than the 2D-grown samples. On the 3D scaffold, 143 genes were more active and 84 less active. These changes centered on genes for the extracellular matrix, the material surrounding cells, and on cell adhesion. The collagen genes COL1A1 and COL5A1 rose, and a second test on three samples confirmed increases in COL1A1 and THBS2. The authors concluded that 3D Nichoids induce specific gene changes that remodel extracellular matrix proteins. They noted that the physiological relevance of these changes is yet to be defined.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: laboratory (in vitro) study comparing primary tumor cells grown on 3D versus flat 2D Nichoid scaffolds, with RNA sequencing and RT-qPCR
• Who was studied: Primary malignant pleural mesothelioma cells, epithelioid subtype, derived from freshly resected surgical specimens of male patients aged 70 to 75 with a history of past asbestos exposure and no prior chemotherapy or radiotherapy. RNA was sequenced from four primary cell samples (T1, T3, T38, T39). Ethics approval was from the Ethics Committee of Bari on 11 November 2021.
• Exposure: The condition studied was the culture surface: a multi-tiered 3D Nichoid versus a flat, one-floor 2D Nichoid of the same grid and material, used as the control. The patients' past asbestos exposure is mentioned, but no fiber type or exposure details are given.
• Main results:
  • Mesothelioma cells on the flat 2D Nichoid declined in number after day 3, while cells on the 3D Nichoid remained constant (monitoring up to day 7).
  • Principal component analysis showed that the 3D-grown samples were more transcriptionally similar to one another than the 2D-grown samples.
  • Comparing genes shared by all cell samples, 1254 genes were expressed only in 3D Nichoids, 1390 only in 2D Nichoids, and 5348 in both.
  • On 3D compared with 2D Nichoids, 143 genes were upregulated and 84 downregulated (absolute log2 fold change greater than 1, adjusted p value of 0.05 or less).
  • Gene Ontology analysis of the differentially expressed genes showed enrichment for extracellular matrix organization and cell adhesion.
  • On 3D Nichoids, the collagen genes COL1A1 (log2FC = 2.361) and COL5A1 (log2FC = 2.404) showed elevated expression.
  • Other genes upregulated on 3D Nichoids included KRT18 (log2FC = 8.875), LAMC2 (log2FC = 5.908) and THBS2 (log2FC = 2.602).
  • RT-qPCR on three independent primary mesothelioma cell samples grown for 4 days confirmed upregulation of COL1A1 and THBS2 in 3D Nichoids.
  • In iRegulon analysis, NFATC1 ranked first for regulated targets and FOXN1 had the highest enrichment signal.
• The authors' conclusion: The authors concluded that 3D Nichoids can induce specific transcriptional changes leading to remodeling of extracellular matrix proteins, suggesting crosstalk between 3D shape and gene expression, with cell viability uncompromised.
• Limitations the authors state:
  • The authors state that the physiological relevance of the gene changes is yet to be defined.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Mesothelioma, Mesothelium, Collagen.

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• Migration of mesothelioma cells correlates with histotype-specific synthesis of extracellular matrix — S. Scarpa, A. Giuffrida, et al. (1999)
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