Inflammatory Response of THP1 and U937 Cells: The RNAseq Approach.

in Cells by Layla Panahipour, Chiara Micucci, Reinhard Gruber

TLDR

  • The study compared the response of THP1 and U937 cells to LPS and IFNG, identifying a panel of highly regulated genes in both cell lines.

Abstract

THP1 and U937 are monocytic cell lines that are common bioassays to reflect monocyte and macrophage activities in inflammation research. However, THP-1 is a human monocytic leukemia cell line, and U937 originates from pleural effusion of histiocytic lymphoma; thus, even though they serve as bioassay in inflammation research, their response to agonists is not identical. Consequently, there has yet to be a consensus about the panel of strongly regulated genes in THP1 and U937 cells representing the inflammatory response to LPS and IFNG. Therefore, we have performed an RNAseq of THP1 and U937 exposed to LPS and IFNG to identify the most sensitive genes and the unique properties of each individual cell line. When applying a highly stringent threshold, we could identify 43, 8 up and 94, 103 down-regulated genes in THP1 and U937 cells, respectively. In THP1 cells, among the most strongly up-regulated genes are,,,,,, and. In U937 cells, the strongest up-regulated genes include,,,,,,,,,,, and. Even though THP1 is considerably less responsive than U937, there are genes commonly upregulated by LPS and IFNG, including the,,,,,, as well as,,,, and genes of prostaglandin synthesisand. Downregulated genes are limited toand. This head-to-head comparison revealed that THP1 is less responsive than U937 cells to LPS and IFNG and identified a panel of highly regulated genes that can be applied in bioassays in inflammation research. Our data further propose bulk RNAseq as a standard method in bioassay research.

Overview

  • The study aimed to identify the most sensitive genes in THP1 and U937 cells, human monocytic cell lines, to LPS and IFNG, and to compare their response.
  • The researchers performed an RNAseq of THP1 and U937 cells exposed to LPS and IFNG to identify the most strongly regulated genes.
  • The primary objective was to identify a panel of highly regulated genes that can be applied in bioassays in inflammation research.

Comparative Analysis & Findings

  • The study identified 43 up-regulated genes and 94 down-regulated genes in THP1 cells, and 8 up-regulated genes and 103 down-regulated genes in U937 cells.
  • The most strongly up-regulated genes in THP1 cells were,,,,,, and, while in U937 cells they were,,,,,,,,,,, and.
  • Commonly upregulated genes by LPS and IFNG in both cell lines included,,,,,, as well as,,,, and genes of prostaglandin synthesisand. Downregulated genes were limited toand.

Implications and Future Directions

  • The study revealed that THP1 cells are less responsive than U937 cells to LPS and IFNG, and identified a panel of highly regulated genes that can be applied in bioassays in inflammation research.
  • The study proposes bulk RNAseq as a standard method in bioassay research.
  • Future research can focus on further validating the identified genes and exploring their roles in inflammation processes, as well as investigating the differences in response between THP1 and U937 cells.
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