# Using adjusted local assortativity with Molecular Pixelation unveils colocalization of membrane proteins with immunological significance

## TL;DR

This paper introduces a new method to analyze how membrane proteins cluster together in cells, revealing insights into immune responses and potential therapies.

## Contribution

The paper introduces adjusted local assortativity for analyzing MPX data, enabling detailed analysis of protein colocalization.

## Key findings

- Adjusted local assortativity detects chemokine-induced changes in T cell uropod formation.
- The method recapitulates rituximab's effect on CD20 polarity in cancerous B-cell lines.
- The approach enhances understanding of immune reactions and cell surface protein reorganization.

## Abstract

Advances in spatial proteomics and protein colocalization are a driving force in the understanding of cellular mechanisms and their influence on biological processes. New methods in the field of spatial proteomics call for the development of algorithms and open up new avenues of research. The newly introduced Molecular Pixelation (MPX) provides spatial information on surface proteins and their relationship with each other in single cells. This allows for in silico representation of neighborhoods of membrane proteins as graphs. In order to analyze this new data modality, we adapted local assortativity in networks of MPX single-cell graphs and created a method that is able to capture detailed information on the spatial relationships of proteins. The introduced method can evaluate the pairwise colocalization of proteins and access higher-order similarity to investigate the colocalization of multiple proteins at the same time. We evaluated the method using publicly available MPX datasets where T cells were treated with a chemokine to study uropod formation. We demonstrate that adjusted local assortativity detects the effects of the stimuli at both single- and multiple-marker levels, which enhances our understanding of the uropod formation. We also applied our method to treating cancerous B-cell lines using a therapeutic antibody. With the adjusted local assortativity, we recapitulated the effect of rituximab on the polarity of CD20. Our computational method together with MPX improves our understanding of not only the formation of cell polarity and protein colocalization under stimuli but also advancing the overall insight into immune reaction and reorganization of cell surface proteins, which in turn allows the design of novel therapies. We foresee its applicability to other types of biological spatial data when represented as undirected graphs.