Digital Pathology Applications for PD-L1 Scoring in Head and Neck Squamous Cell Carcinoma: A Challenging Series
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cases
- One hematoxylin–eosin (H&E)-stained section;
- One LDT PD-L1 IHC stained with Dako clone 22C3 (Dako, Carpinteria, CA, USA) or Ventana clone SP263 (Ventana, Tucson, AZ, USA) and tested on the Dako or Ventana platforms, respectively;
- One blank section for IHC that was centralized at the Pathology Department of Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy, where a pharmDx assay CDx (Dako) was performed.
2.2. Digital Pathology
2.3. PD-L1 CPS Scoring
- Presence of at least 100 viable tumor cells (adequacy criterion);
- Score range values from 0 to 100, with 100 as the maximum value;
- Evaluation of the staining at 20× magnification.
- LDTs PD-L1 (board consensus) vs. CDx PD-L1 (board consensus);
- LDTs PDL1 (board consensus) vs. LDTs PD-L1 (primary local diagnoses);
- CDx PD-L1 (board consensus) vs. LDTs PD-L1 (primary local diagnoses);
- The technical concordance of the IHC stainings for PD-L1;
- The interpretative concordance of the two PD-L1 CPS scores.
2.4. Computational Analysis
- Tissue was identified from the background using a thresholding technique based on image-specific parameters.
- Regions of interest (ROIs) for “Tumor” and “tumor + stroma” were identified and annotated manually. The stromal component was considered peritumoral when included in a microscopic field at 20× magnification, placing the tumor/peritumoral stroma interface at the center of the field.
- Automatic cell identification within the “tumor + stroma” annotations was achieved with the nuclear detector StarDist [11]. For this task, a custom script built for H&E images was used and modified to locate cell nuclei with a threshold of 0.2 and the surrounding cytoplasm based on nuclear expansion.
3. Statistical Analysis
4. Results
4.1. Inter-Assay Variability
4.2. Technical and Interpretative Evaluations
4.3. Impact of Variability on CPS Cutoffs
4.4. Computational Analysis
5. Discussion
6. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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LAB | Requirement Assigned | Sample Type | PD-L1 Clone |
---|---|---|---|
1 | Crushing artifacts | Biopsy | 22C3 |
2 | Around CPS 1 | Biopsy | 22C3 |
3 | Around CPS 20 | Surgical | 22C3 |
4 | Fragmented sample | Biopsy | 22C3 |
5 | Great inflammatory component prevalence | Surgical | 22C3 |
6 | Lymph node metastasis | Surgical (lymph node metastasis) | 22C3 |
7 | Extensive necrosis | Surgical | SP263 |
8 | Cell block 1 | Biopsy | SP263 |
9 | Cell block 2 | Biopsy | 22C3 |
10 | Tumor component prevalence | Biopsy | 22C3 |
11 | Scant material | Biopsy | SP263 |
Case ID | CPS-LDTs | CPS-LDTs | CPS-CDx | Technical Concordance | Interpretative Concordance |
---|---|---|---|---|---|
Local Evaluation | Board Evaluation | Board Evaluation | |||
1 | 25 | 20–25 | 20–25 | ||
2 | 2 | 1 | 1 | ||
3 | 30 | around 20 * | around 20 * | ||
4 | 5–9 | 10 | 15 | ||
5 | 60 | 80 | 70 | ||
6 | 10–20 | 5–10 | 5–10 | + | + |
7 | 14 | 15 | 22 | ||
8 | 50 | 50 | 50 | ||
9 | 15–20 | 25 | 15 | ||
10 | 35 | 40 | 40 | ||
11 | 2 | 1 | 2 |
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Canini, V.; Eccher, A.; d’Amati, G.; Fusco, N.; Maffini, F.; Lepanto, D.; Martini, M.; Cazzaniga, G.; Paliogiannis, P.; Lobrano, R.; et al. Digital Pathology Applications for PD-L1 Scoring in Head and Neck Squamous Cell Carcinoma: A Challenging Series. J. Clin. Med. 2024, 13, 1240. https://doi.org/10.3390/jcm13051240
Canini V, Eccher A, d’Amati G, Fusco N, Maffini F, Lepanto D, Martini M, Cazzaniga G, Paliogiannis P, Lobrano R, et al. Digital Pathology Applications for PD-L1 Scoring in Head and Neck Squamous Cell Carcinoma: A Challenging Series. Journal of Clinical Medicine. 2024; 13(5):1240. https://doi.org/10.3390/jcm13051240
Chicago/Turabian StyleCanini, Valentina, Albino Eccher, Giulia d’Amati, Nicola Fusco, Fausto Maffini, Daniela Lepanto, Maurizio Martini, Giorgio Cazzaniga, Panagiotis Paliogiannis, Renato Lobrano, and et al. 2024. "Digital Pathology Applications for PD-L1 Scoring in Head and Neck Squamous Cell Carcinoma: A Challenging Series" Journal of Clinical Medicine 13, no. 5: 1240. https://doi.org/10.3390/jcm13051240