Immunophenotyping with (phospho)protein profiling and fluorescent cell barcoding for single-cell signaling analysis...

Area of Science:

• Immunology
• Oncology
• Biotechnology

Background:

• The tumor microenvironment in hematologic cancers impacts disease progression and treatment resistance.
• Understanding immune cell dynamics is crucial for developing effective cancer therapies.
• Peripheral blood is ideal for single-cell analysis due to its liquid nature.

Purpose of the Study:

• To develop an advanced flow cytometry protocol for single-cell analysis of hematologic cancers.
• To combine multi-parameter immunophenotyping with single-cell (phospho)protein profiling.
• To provide a method for studying immune cell composition and signaling in cancer.

Main Methods:

• Developed a flow cytometry protocol integrating immunophenotyping and (phospho)protein profiling.
• Utilized fluorescent cell barcoding to pool multiple samples for simultaneous analysis.
• Enabled high-throughput, cost-effective, and reduced-variability sample processing.

Main Results:

• Demonstrated the protocol's suitability for pre-clinical studies by correlating (phospho)protein profiles with drug sensitivity.
• Showcased the ability to analyze immunophenotype and cell signaling at single-cell resolution.
• Highlighted reduced experimental variability and increased throughput via sample multiplexing.

Conclusions:

• The protocol offers a robust method for dissecting immune cell function in hematologic malignancies.
• Findings can provide insights into cancer pathogenesis and identify novel drug targets.
• The approach has potential as a biomarker to guide clinical decisions in cancer treatment.