Neutrophil Biomarker Test for Predicting Clinical Benefit From Immunotherapy Based on Flow Cytometry Analysis

NCT07246759 | Not Yet Recruiting

• 1 other identifier: NEUTROFLOW
• Study Type: observational
• Target: 600
• Locations: 3 countries
• Sites: 3

Brief Summary

The NeutroFlow study is a multi-center clinical trial designed to develop a computational model that converts flow cytometry results into a prediction of clinical benefit. The study analyzes Ly6Ehi neutrophils in biological samples from patients treated with immune checkpoint inhibitors to evaluate their likelihood of benefiting from treatment. Blood samples are collected prior to treatment and used to support the ongoing development of the algorithm.

Conditions

Non Small Cell Lung Cancer (NSCLC); Melanoma (Skin Cancer); Triple Negative Breast Cancer (TNBC); RCC, Renal Cell Cancer; HNSCC

Keywords

Immunotherapy; Biomarker; Neutrophils; Flow Cytometry; Immune checkpoint inhibitor

Outcome Measures

Primary Outcomes (2)

• Quantification of Ly6E high (Ly6Ehi) neutrophils in blood using the NeutroFlow flow cytometry assay

  Peripheral blood samples will be collected from patients up to 1 month prior to treatment initiation. Ly6Ehi neutrophil populations will be quantified using the NeutroFlow multiparametric flow cytometry assay

  Baseline (up to 1 month before treatment initiation)

• Prediction of clinical benefit rate using baseline Ly6Ehi neutrophils levels

  The patients clinical benefit (CB) will be defined according to RECIST 1.1 criteria to one of the following categories: complete response (CR), partial response (PR), or stable disease (SD). Baseline Ly6Ehi neutrophil levels will be used as input in a computational predictive model to estimate the likelihood of CB for each patient. The model will be trained and validated with independent patient cohorts, using cross-validation and ROC AUC metrics to assess predictive performance. Sensitivity, specificity, positive predictive value, and negative predictive value will also be calculated. Predictions will be assessed at multiple timepoints: 6, 12, 18, and 24 months post-initiation of anti-PD-(L)1 therapy. Subgroup analyses will include age, sex, cancer type, disease stage, and treatment line.

  Baseline (blood draw) to 6, 12, 18, and 24 months post treatment initiation

Secondary Outcomes (4)

• Clinical benefit rate across individual cancer types

  Baseline to 6, 12, 18, and 24 months post treatment initiation (per indication)

• Clinical benefit rate by PD-(L)1 treatment regimen

  Baseline to 6, 12, 18, and 24 months post treatment initiation

• Correlation between Ly6Ehi neutrophil levels and PD-L1 status (TPS/CPS)

  Baseline (PD-L1 and neutrophils assessed prior to treatment)

• Correlation between Ly6Ehi neutrophil levels and other clinical response-associated parameters

  Baseline (clinical parameters collected prior to treatment)

Study Arms (5)

Patients with stage IV or stage III unresectable non-small cell lung cancer (NSCLC)

Patients with stage IV or stage III unresectable non-small cell lung cancer (NSCLC) treated with immune checkpoint inhibitors as a first-line treatment

Other: Blood sample collection

Patients with stage IV or stage IIIb-d malignant melanoma

Patients with stage IV or stage IIIb-d malignant melanoma treated with immune checkpoint inhibitors as a first-line treatment

Other: Blood sample collection

Patients with stage IV head and neck squamous cell carcinoma (HNSCC)

Patients with stage IV head and neck squamous cell carcinoma (HNSCC) treated with immune checkpoint inhibitors as a first-line treatment

Other: Blood sample collection

Patients with stage IV renal cell carcinoma (RCC)

Patients with stage IV renal cell carcinoma (RCC) treated with immune checkpoint inhibitors as a first-line treatment

Other: Blood sample collection

Patients with stage IV triple-negative breast cancer (TNBC)

Patients with stage IV triple-negative breast cancer (TNBC) treated with immune checkpoint inhibitors as a first-line treatment

Other: Blood sample collection

Interventions

Blood sample collection OTHER

Blood sample collection prior treatment initiation

Patients with stage IV head and neck squamous cell carcinoma (HNSCC); Patients with stage IV or stage III unresectable non-small cell lung cancer (NSCLC); Patients with stage IV or stage IIIb-d malignant melanoma; Patients with stage IV renal cell carcinoma (RCC); Patients with stage IV triple-negative breast cancer (TNBC)

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

At least 600 treatment-naïve patients due to be treated with a first line regimen that includes PD- 1/PD-L1 inhibitors with the following diagnoses are planned to be enrolled in the study over a period of 3 years or as needed: 1. Stage IV or Stage III-unresectable NSCLC 2. Stage IV or Stage IIIb-d malignant melanoma 3. Stage IV HNSCC 4. Stage IV RCC 5. Stage IV TNBC

You may qualify if:

• Patients newly diagnosed with advanced-stage/metastatic NSCLC, melanoma, HNSCC, RCC, or TNBC, who are due to receive first-line treatment with a PD-(L)1 inhibitor, either as monotherapy or in combination with other agents, according to current standard-of-care regimens, including (but not limited to) the following approved options: NSCLC. Monotherapy: Pembrolizumab, Atezolizumab, Cemiplimab. Combination: Pembrolizumab + chemotherapy; Nivolumab + Ipilimumab; Cemiplimab + chemotherapy; Atezolizumab + chemotherapy + Bevacizumab.
• Melanoma. Monotherapy: Nivolumab, Pembrolizumab. Combination: Nivolumab + Ipilimumab; Nivolumab + Relatlimab.
• HNSCC. Monotherapy: Pembrolizumab, Cemiplimab. Combination: Pembrolizumab + chemotherapy.
• RCC. Combination only: Nivolumab + Ipilimumab; Nivolumab + Cabozantinib; Pembrolizumab + Lenvatinib or Axitinib; Avelumab + Axitinib.
• TNBC. Combination only: Pembrolizumab + chemotherapy.
• Male or female aged at least 18 years
• ECOG PS: 0/1-2
• Normal hematologic, renal and liver function:
• Absolute neutrophil count \> 1500/mm³ Platelets \> 100,000/mm³ Hemoglobin \> 9 g/dL Creatinine concentration ≤ 1.4 mg/dL, or creatinine clearance \> 40 mL/min, Total bilirubin \< 1.5 mg/dL ALT + AST levels ≤ 3 times above the upper normal limit

You may not qualify if:

• Any concurrent and/or other active malignancy that has required systemic treatment within 2 years of the first dose of treatment.
• For NSCLC: presence of activating EGFR, ALK, ROS1, RET, NTRK alterations linked to an approved first-line targeted drug.

Sponsors & Collaborators

• OncoHost Ltd.: lead
• European Institute of Oncology: collaborator
• University Hospital Heidelberg: collaborator
• Hospital Universitario Virgen Macarena: collaborator

Study Sites (3)

Heidelberg University Hospital

Heidelberg, 69120, Germany

Location

Intituto Europeo di Oncologia SRL

Milan, 20121, Italy

Location

Virgen Macarena University Hospital - Servicio Andaluz de Salud

Seville, 41071, Spain

Location

Related Publications (8)

• Alturki NA. Review of the Immune Checkpoint Inhibitors in the Context of Cancer Treatment. J Clin Med. 2023 Jun 27;12(13):4301. doi: 10.3390/jcm12134301.

  PMID: 37445336 BACKGROUND

• Man J, Millican J, Mulvey A, Gebski V, Hui R. Response Rate and Survival at Key Timepoints With PD-1 Blockade vs Chemotherapy in PD-L1 Subgroups: Meta-Analysis of Metastatic NSCLC Trials. JNCI Cancer Spectr. 2021 Jan 27;5(3):pkab012. doi: 10.1093/jncics/pkab012. eCollection 2021 Jun.

  PMID: 34084999 BACKGROUND

• Martins F, Sofiya L, Sykiotis GP, Lamine F, Maillard M, Fraga M, Shabafrouz K, Ribi C, Cairoli A, Guex-Crosier Y, Kuntzer T, Michielin O, Peters S, Coukos G, Spertini F, Thompson JA, Obeid M. Adverse effects of immune-checkpoint inhibitors: epidemiology, management and surveillance. Nat Rev Clin Oncol. 2019 Sep;16(9):563-580. doi: 10.1038/s41571-019-0218-0.

  PMID: 31092901 BACKGROUND

• Hurwitz JT, Vaffis S, Grizzle AJ, Nielsen S, Dodson A, Parry S. Cost-Effectiveness of PD-L1 Testing in Non-Small Cell Lung Cancer (NSCLC) Using In Vitro Diagnostic (IVD) Versus Laboratory-Developed Test (LDT). Oncol Ther. 2022 Dec;10(2):391-409. doi: 10.1007/s40487-022-00197-1. Epub 2022 May 13.

  PMID: 35556235 BACKGROUND

• Lei Y, Li X, Huang Q, Zheng X, Liu M. Progress and Challenges of Predictive Biomarkers for Immune Checkpoint Blockade. Front Oncol. 2021 Mar 11;11:617335. doi: 10.3389/fonc.2021.617335. eCollection 2021.

  PMID: 33777757 BACKGROUND

• Arora S, Velichinskii R, Lesh RW, Ali U, Kubiak M, Bansal P, Borghaei H, Edelman MJ, Boumber Y. Existing and Emerging Biomarkers for Immune Checkpoint Immunotherapy in Solid Tumors. Adv Ther. 2019 Oct;36(10):2638-2678. doi: 10.1007/s12325-019-01051-z. Epub 2019 Aug 13.

  PMID: 31410780 BACKGROUND

• McKean WB, Moser JC, Rimm D, Hu-Lieskovan S. Biomarkers in Precision Cancer Immunotherapy: Promise and Challenges. Am Soc Clin Oncol Educ Book. 2020 May;40:e275-e291. doi: 10.1200/EDBK_280571.

  PMID: 32453632 BACKGROUND

• Benguigui M, Cooper TJ, Kalkar P, Schif-Zuck S, Halaban R, Bacchiocchi A, Kamer I, Deo A, Manobla B, Menachem R, Haj-Shomaly J, Vorontsova A, Raviv Z, Buxbaum C, Christopoulos P, Bar J, Lotem M, Sznol M, Ariel A, Shen-Orr SS, Shaked Y. Interferon-stimulated neutrophils as a predictor of immunotherapy response. Cancer Cell. 2024 Feb 12;42(2):253-265.e12. doi: 10.1016/j.ccell.2023.12.005. Epub 2024 Jan 4.

  PMID: 38181798 BACKGROUND

Biospecimen

Retention: SAMPLES WITH DNA

Blood withdrawal at baseline before treatment initiation

MeSH Terms

Conditions

Carcinoma, Non-Small-Cell Lung; Melanoma; Triple Negative Breast Neoplasms; Carcinoma, Renal Cell; Squamous Cell Carcinoma of Head and Neck

Condition Hierarchy (Ancestors)

Carcinoma, Bronchogenic; Bronchial Neoplasms; Lung Neoplasms; Respiratory Tract Neoplasms; Thoracic Neoplasms; Neoplasms by Site; Neoplasms; Lung Diseases; Respiratory Tract Diseases; Neuroendocrine Tumors; Neuroectodermal Tumors; Neoplasms, Germ Cell and Embryonal; Neoplasms by Histologic Type; Neoplasms, Nerve Tissue; Nevi and Melanomas; Skin Neoplasms; Skin Diseases; Skin and Connective Tissue Diseases; Breast Neoplasms; Breast Diseases; Adenocarcinoma; Carcinoma; Neoplasms, Glandular and Epithelial; Kidney Neoplasms; Urologic Neoplasms; Urogenital Neoplasms; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Urogenital Diseases; Kidney Diseases; Urologic Diseases; Male Urogenital Diseases; Carcinoma, Squamous Cell; Head and Neck Neoplasms

Central Study Contacts

Michal Harel VP of Translational Medicine, PhD

CONTACT

97248537558; Michal@oncohost.com

Shani Raveh Shoval VP of Clinical Affairs, PhD

CONTACT

shani@oncohost.com

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Sponsor Type: INDUSTRY
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: November 16, 2025
• First Posted: November 24, 2025
• Study Start: November 15, 2025
• Primary Completion (Estimated): September 30, 2027
• Study Completion (Estimated): May 30, 2028
• Last Updated: November 24, 2025

Record last verified: 2025-11

Locations

DE (1); IT (1); ES (1)