{
  "abstract": "Background Chemoimmunotherapy offers clinical benefit for patients with advanced non-small cell lung cancer (NSCLC) and a PD-L1 tumor proportion score (TPS) <1%. 1–4 However, the absence of robust predictive biomarkers in this population limits the implementation of precision treatment strategies.5–18 We hypothesized that direct quantification of PD-1/PD-L1 engagement may provide a more accurate predictor of response to anti-PD-1/PD-L1 therapies in PD-L1-negative NSCLC.Methods PD-1/PD-L1 interactions were quantified in formalin-fixed paraffin-embedded (FFPE) tumor samples from patients with PD-L1-negative NSCLC using QF-Pro®, a high-throughput, automated imaging platform based on quantitative functional proteomics through an enhanced and amplified Förster resonance energy transfer (FRET) approach. FRET efficiency was measured as a surrogate for PD-1/PD-L1 complex formation. Patients were stratified by FRET efficiency (highest 40% vs. lower 60%). The primary endpoint was progression-free survival (PFS). Secondary endpoints included overall survival (OS) and objective response rate (ORR).Results Twenty-seven patients with PD-L1-negative NSCLC treated with chemoimmunotherapy were included. At a median follow-up of 16.8 months (95% confidence interval [CI], 11.4–22.2), the median PFS for the cohort was 9.2 months (95% CI, 4.0–15.2). Patients with high FRET efficiency demonstrated a longer median PFS compared to those with low FRET efficiency (14.6 vs. 5.4 months; hazard ratio [HR], 0.42; 95% CI, 0.15–1.14; p = 0.083) (figure 1A). Median OS was also longer in the high FRET group (14.8 vs. 7.1 months; HR, 0.52; 95% CI, 0.18–1.59; p = 0.259), although not statistically significant (figure 1B). Notably, the ORR was significantly higher in the high FRET group (37.1% vs. 14.7%; p = 0.007) (figure 2).Conclusions Quantitative assessment of PD-1/PD-L1 complex formation via FRET efficiency shows promise as a functional biomarker for predicting immunotherapy benefit in patients with PD-L1-negative NSCLC. These findings support further investigation into PD-1/PD-L1 engagement as a predictive tool in this biomarker-limited population.References Han Y, Liu D, Li L. PD-1/PD-L1 pathway: current researches in cancer. Am J Cancer Res. 2020 Mar 1;10(3):727–742.Liu J, Chen Z, Li Y, Zhao W, Wu J, Zhang Z. PD-1/PD-L1 Checkpoint Inhibitors in Tumor Immunotherapy. Front. Pharmacol. 2021;12:731798.Imani M, Mohajeri N, Rastegar M, Zarghami N. Recent advances in FRET-Based biosensors for biomedical applications. Anal Biochem. 2021;630:114323.Clinical N, Guidelines P, Guidelines N. Non-Small Cell Lung. 2024.Niu M, Yi M, Li N, Luo S, Wu K. Predictive biomarkers of anti-PD-1/PD-L1 therapy in NSCLC. Exp Hematol Oncol [Internet]. 2021;10(1):1–13.Sánchez-Magraner L, Gumuzio J, Miles J, Quimi N, Martínez Del Prado P, Abad-Villar MT, et al. Functional engagement of the PD-1/PD-L1 complex but not PD-L1 expression is highly predictive of patient response to immunotherapy in non-small-cell lung cancer. Journal of Clinical Oncology. 2023;41(14):2561–70.Mamdani H, Matosevic S, Khalid AB, Durm G, Jalal SI. Immunotherapy in lung cancer: current landscape and future directions. Front Immunol. 2022;13(February):1–12.Lahiri A, Maji A, Potdar PD, Singh N, Parikh P, Bisht B, et al. Lung cancer immunotherapy: progress, pitfalls, and promises. Mol Cancer [Internet]. 2023;22(1):1–37. Available from: https://doi.org/10.1186/s12943-023-01740-y Sanchez-Magraner L, Miles J, Baker CL, Applebee CJ, Lee DJ, Elsheikh S, et al. High PD-1/PD-L1 checkpoint interaction infers tumor selection and therapeutic sensitivity to anti-PD-1/PD-L1 treatment. Cancer Res. 2020;80(19):4244–57.Reck M, Rodríguez-Abreu D, Robinson AG, Hui R, Csőszi T, Fülöp A, et al. Pembrolizumab versus chemotherapy for PD-L1-positive non-small-cell lung cancer. New England Journal of Medicine. 2016;375(19):1823–33.Garon EB, Rizvi NA, Hui R, Leighl N, Balmanoukian AS, Eder JP, et al. Pembrolizumab for the treatment of non-small-cell lung cancer. New England Journal of Medicine. 2015;372(21):2018–28.Herbst RS, Baas P, Kim DW, Felip E, Pérez-Gracia JL, Han JY, et al. Pembrolizumab versus docetaxel for previously treated, PD-L1-positive, advanced non-small-cell lung cancer (KEYNOTE-010): a randomised controlled trial. The Lancet. 2016;387(10027):1540–50.Nosaki K, Saka H, Hosomi Y, Baas P, de Castro G, Reck M, et al. Safety and efficacy of pembrolizumab monotherapy in elderly patients with PD-L1-positive advanced non-small-cell lung cancer: pooled analysis from the KEYNOTE-010, KEYNOTE-024, and KEYNOTE-042 studies. Lung Cancer [Internet]. 2019;135(July):188–95. Available from: https://doi.org/10.1016/j.lungcan.2019.07.004 Gandhi L, Rodríguez-Abreu D, Gadgeel S, Esteban E, Felip E, De Angelis F, et al. Pembrolizumab plus chemotherapy in metastatic non-small-cell lung cancer. New England Journal of Medicine. 2018;378(22):2078–92.Rittmeyer A, Barlesi F, Waterkamp D, Park K, Ciardiello F, von Pawel J, et al. Atezolizumab versus docetaxel in patients with previously treated non-small-cell lung cancer (OAK): a phase 3, open-label, multicentre randomised controlled trial. The Lancet. 2017;389(10066):255–65.Brahmer J, Reckamp KL, Baas P, Crinò L, Eberhardt WEE, Poddubskaya E, et al. Nivolumab versus docetaxel in advanced squamous-cell non-small-cell lung cancer. New England Journal of Medicine. 2015;373(2):123–35.Borghaei H, Paz-Ares L, Horn L, Spigel DR, Steins M, Ready NE, et al. Nivolumab versus docetaxel in advanced nonsquamous non-small-cell lung cancer. New England Journal of Medicine. 2015;373(17):1627–39.Carbone DP, Reck M, Paz-Ares L, Creelan B, Horn L, Steins M, et al. First-line nivolumab in stage IV or recurrent non-small-cell lung cancer. New England Journal of Medicine. 2017;376(25):2415–26.Ethics Approval The study was conducted using archived formalin-fixed paraffin-embedded (FFPE) tissue blocks obtained from the Pathology Department of the Instituto Nacional de Cancerología. Five 5 μm sections per sample were processed for hematoxylin and eosin staining to identify immune-infiltrated regions, and subsequently for FRET analysis using donor-only (anti-PD-1-ATTO488) and donor-acceptor (anti-PD-L1-ALEXA594) staining. This protocol was reviewed and approved by the Institutional Ethics Committee of the Instituto Nacional de Cancerología. As this research involves the use of anonymized archival material and poses no more than minimal risk, informed consent was waived in accordance with applicable national regulations.Consent Written informed consent was obtained from the patient for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.Abstract 1304 Figure 1Kaplan-meier curves of progression free survival (PFS) (A) and overall survival (OS) (B) stratifying patients based on their level of PD-1/PD-L1 engagement. Red curves represent high PD-1/PD-L1 patients, blue line represents low PD-1/PD-L1 patientsAbstract 1304 Figure 2Objective response rate (A) and Suboptimal response (B) of analysed patients related to their PD-1/PD-L1 engagement, measured as mean FRET efficiency.",
  "authors": [
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Enrique Caballe-Perez"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Luis Antonio Cabrera-Miranda"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "David Dávila-Dupont"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Graciela Cruz-Rico"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Norma-Hernández Pedro"
    },
    {
      "affiliations": [
        "HAWK Biosystems, Derio, Bizkaia, Spain"
      ],
      "name": "Juan Gumuzio"
    },
    {
      "affiliations": [
        "HAWK Biosystems, Derio, Bizkaia, Spain"
      ],
      "name": "Markel Rementeria"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Jose María Lucio-Lozada"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Diana Flores-Estrada"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Cesar Saith Castillo-Ruiz"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Elí Emmanuel Santana-Aguilar"
    },
    {
      "affiliations": [
        "Instituto Nacional de Cancerología (INCan), Ciudad de México, EM, Mexico"
      ],
      "name": "Oscar Arrieta"
    }
  ],
  "title": "1304 Preliminary results on PD-1/PD-L1 axis engagement among PD-L1-negative NSCLC patients receiving chemoimmunotherapy",
  "uid": "fa9e90ff-d563-54e2-883f-5b7fa04ef7e3"
}