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RPPAs for Cell Subpopulation Analysis.

Kohei Kume¹, Satoshi S Nishizuka²

¹Department of Systems Biology, City of Hope Comprehensive Cancer Center, Monrovia, CA, USA.

Advances in Experimental Medicine and Biology

|December 11, 2019

Summary

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Colony lysate array (CoLA) enables proteomic profiling of drug-tolerant persister cells, revealing distinct subpopulations and signaling pathway changes linked to drug resistance in cancer biology.

Area of Science:

Cell biology
Proteomics
Cancer biology

Background:

Cellular heterogeneity presents a significant challenge in cell biology, particularly for stem cell and cancer research.
Drug-tolerant persister cells (DTPs) are crucial for understanding chemotherapeutic relapse in cancer.

Purpose of the Study:

To describe two Colony Lysate Array (CoLA) assays for proteomic profiling of individual drug-tolerant persister cells (DTPs).
To investigate proteomic profiles of DTPs and identify signaling pathways associated with drug resistance.

Main Methods:

Developed and applied Colony Lysate Array (CoLA), a modified reverse phase protein array (RPPA) method.
Analyzed individual DTPs from various cell types under different drug treatments.
Utilized hierarchical clustering and principal component analyses for proteomic data.

Keywords:

Cell subpopulation Colony lysate array Drug-tolerant persisters Heterogeneity Reverse phase protein array

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Conducted drug dose-response analysis on gastric cancer cell lines to identify signaling pathway alterations.

Main Results:

CoLA revealed two types of DTP clonal populations expressing stem cell-associated and epithelium-associated proteins.
Proteomic profiles of DTPs clustered based on drug and dose, indicating dynamic changes.
In 5-fluorouracil-tolerant gastric cancer cells, PI3K phosphorylation increased while AKT phosphorylation decreased.

Conclusions:

CoLA provides a framework for profiling proteomic heterogeneity in small cell subpopulations under stress.
These methods can be applied to diverse heterogeneous cellular samples, aiding in understanding cellular responses and drug resistance mechanisms.