High-content immunofluorescence assay detecting PD-L1 expression changes in head and neck cancer patient-derived
Yuen-Keng Ng1, Stacy Magdalene Abbang2, Jishi Ye3
1Georgia Cancer Center, Medical College of Georgia, Augusta University, USA; Department of Medicine, Medical College of Georgia, Augusta University, USA.
Abstract:
Clinical uses of monoclonal antibodies against immune checkpoint molecules, such as the Program Death-Ligand 1 (PD-L1) or Program Death Protein-1 (PD-1), has transformed cancer therapy across pan-cancers. In addition to antibody therapies, there is a growing interest in identifying small molecules that could modulate PD-L1 levels in cancer cells. Yet, most current PD-L1 assays are not robust enough to be developed for drug screening purposes. Here, we report the development of a sensitive PD-L1 immunofluorescence assay that can capture PD-L1 expression heterogeneity in HNC patient tumor cultures and allows relative quantification of PD-L1 levels in cells in a streamlined and robust manner. Furthermore, this imaging-based assay can capture additional spatial or subcellular localization information of PD-L1 expression in patient cultures and has the potential to be combined with other image-based assays for future drug development purposes. Importantly, we demonstrated that this assay was robust enough to evaluate dose-dependent PD-L1-modulatory effects of drugs in patient-derived tumor cultures and demonstrated patient-to-patient variability of drug responses for PD-L1 modulation. This assay has the potential to be adopted for high-throughput drug screening for identifying small molecules modulators of PD-L1 using individual patient tumor cultures of various cancer types.
Insights
A new immunofluorescence assay accurately measures Program Death-Ligand 1 (PD-L1) expression in cancer cells. This sensitive assay aids in developing new drugs targeting PD-L1 for cancer therapy.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Monoclonal antibodies targeting immune checkpoints like Program Death-Ligand 1 (PD-L1) have revolutionized cancer treatment.
- There is a significant need for small molecules that can regulate PD-L1 levels in cancer cells.
- Existing PD-L1 assays lack the robustness required for effective drug screening.
Purpose of the Study:
- To develop a sensitive and robust immunofluorescence assay for quantifying PD-L1 expression.
- To assess the potential of this assay for drug screening and development.
- To evaluate drug-induced modulation of PD-L1 in patient-derived tumor cultures.
Main Methods:
- Development of a sensitive PD-L1 immunofluorescence assay.
- Utilizing the assay to capture PD-L1 expression heterogeneity in head and neck cancer (HNC) patient tumor cultures.
- Employing imaging-based analysis for relative quantification and spatial localization of PD-L1.
Main Results:
- The developed assay is sensitive, streamlined, and robust for quantifying PD-L1 levels.
- The assay successfully captured PD-L1 expression heterogeneity and spatial information in patient cultures.
- Demonstrated dose-dependent PD-L1 modulation by drugs and significant patient-to-patient variability in drug response.
Conclusions:
- The novel immunofluorescence assay is suitable for evaluating drug effects on PD-L1 modulation in patient-derived tumor cultures.
- This assay has the potential for high-throughput drug screening to identify small molecule modulators of PD-L1.
- The assay can be adapted for various cancer types, facilitating personalized medicine approaches.
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