Cell-type-specific signaling networks in heterocellular organoids

Area of Science:

• Biomedical Engineering
• Cell Biology
• Systems Biology

Background:

• Organoids are valuable biomimetic tissue models, but lack methods for comprehensive cell-type-specific post-translational modification (PTM) signaling analysis.
• Understanding PTM signaling is crucial for deciphering cellular functions in health and disease.

Purpose of the Study:

• To develop and validate a multivariate single-cell analysis method for PTM signaling networks in organoids.
• To enable high-throughput comparison of signaling networks between different organoid cultures.

Main Methods:

• Mass cytometry was used to simultaneously analyze 28 PTMs in over 1 million single cells from small intestinal organoids.
• Thiol-reactive organoid barcoding in situ (TOBis) was integrated for high-throughput comparative analysis.
• Colorectal cancer organoid cocultures were analyzed for cell-type-specific PTMs.

Main Results:

• Identified cell-type- and cell-state-specific PTM signaling networks in stem, Paneth, enteroendocrine, tuft, goblet cells, and enterocytes.
• Demonstrated high-throughput comparison of signaling networks using TOBis.
• Showcased cell-autonomous mimicry of stromal-induced signaling states by cancer mutations (shApc, KrasG12D, Trp53R172H).

Conclusions:

• Standard mass cytometry workflows can be adapted for high-throughput, multivariate, cell-type-specific signaling analysis of organoids.
• This method provides novel insights into healthy and cancerous organoid signaling networks.
• The findings facilitate a deeper understanding of PTM roles in organoid biology and cancer progression.