A Prospective Multicenter Study of the TuFEst-LN Model for Axillary Lymph Node Status Assessment in Breast Cancer

NCT07304934 | Recruiting DMC

• 1 other identifier: 2025-1028
• Study Type: observational
• Target: 400
• Locations: 1 country
• Sites: 1

Brief Summary

Through the research of this project, we expect to validate the clinical utility of the TuFEst-LN model in assessing axillary lymph node status in breast cancer patients. Specifically, we aim to prospectively validate its ability to identify pathologically node-negative patients among clinically and radiologically assessed cN0 patients undergoing upfront surgery ，and explore the predictive value of the TuFEst-LN model combined with preoperative MRI for ypN status assessment in initially node-positive patients following neoadjuvant therapy.

Conditions

Breast Cancer; Axillary Lymph Node Metastasis

Keywords

breast cancer; cfDNA; TuFEst model; Fragmentomics; Axillary lymph node; Sentinel lymph node biopsy; Neoadjuvant therapy; Magnetic resonance imaging; Diagnostic accuracy; Machine learning

Outcome Measures

Primary Outcomes (1)

• Pathological axillary lymph node positivity rate among TuFEst-LN-negative patients

  Defined as the proportion of patients with pathological axillary lymph node positivity (pN1mi or higher) among patients predicted as negative by the locked TuFEst-LN model in Cohort 1. FOR = FN/(TN+FN) = 1-NPV.

  up to 2 weeks

Secondary Outcomes (7)

• Negative Predictive Value (NPV) of the TuFEst-LN Model

  2 weeks

• Sensitivity and False Negative Rate (FNR) for Detecting Pathological Axillary Lymph Node Positivity

  2 weeks

• Specificity of the TuFEst-LN Model

  2 weeks

• Positive Predictive Value (PPV), Overall Accuracy, and Discrimination Performance

  2 weeks

• Proportion of Patients Classified as Negative by the TuFEst-LN Model

  2 weeks

Other Outcomes (5)

• Predictive Performance of the TuFEst-LN Model for ypN Status

  2 weeks

• Predictive Performance of Preoperative MRI for ypN Status

  2 weeks

• Performance of Combined cfDNA Fragmentomics and MRI Model

  2 weeks

Study Arms (2)

1

Invasive breast cancer patients with cT1-3N0M0 disease who have received no prior neoadjuvant therapy or radiotherapy and undergo SLNB and/or ALND for axillary evaluation. A total of 300 participants are planned to be enrolled in Cohort 1

Other: No Intervention: Observational Cohort

2

Patients with newly diagnosed invasive breast cancer and biopsy-confirmed axillary lymph node metastasis without distant metastasis, who are scheduled to receive standard neoadjuvant systemic therapy followed by breast and axillary surgery . Patients who complete or prematurely discontinue neoadjuvant therapy for clinical reasons but proceed to surgery are eligible.At least 100 participants are planned to be enrolled in Cohort 2.

Other: No Intervention: Observational Cohort

Interventions

No Intervention: Observational Cohort OTHER

No Intervention: Observational Cohort

1; 2

Eligibility Criteria

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

Study Population

Breast cancer patients receiving care across the six participating centers who fulfill the eligibility criteria for either the cN0 cohort undergoing upfront surgery or the initially cN+ cohort after neoadjuvant systemic therapy.

You may qualify if:

• Participants in Cohort 1 must meet all of the following criteria:
• Histologically confirmed invasive breast cancer.
• Clinical stage cT1-3, cN0, M0 disease at enrollment.
• No clinically palpable suspicious metastatic axillary lymph nodes identified by physical examination.
• No radiologically suspicious metastatic axillary lymph nodes identified by preoperative axillary imaging (at least ultrasound examination). Patients with suspicious lymph nodes must have negative cytological or histological findings confirmed by fine-needle aspiration or core needle biopsy.
• No prior neoadjuvant therapy, including chemotherapy, targeted therapy, immunotherapy, endocrine therapy, or breast/axillary radiotherapy before enrollment.
• Planned to undergo definitive breast surgery with sentinel lymph node biopsy (SLNB) and/or axillary lymph node dissection (ALND).
• Ability to provide qualified preoperative plasma samples for cfDNA analysis.
• Participants in Cohort 2 must meet all of the following criteria:
• Histologically confirmed invasive breast cancer.
• Ipsilateral axillary lymph node metastasis confirmed by fine-needle aspiration or core needle biopsy at initial diagnosis.
• No evidence of distant metastasis (M0), and planned to receive standard neoadjuvant systemic therapy followed by definitive breast and axillary surgery.
• Completion of the planned neoadjuvant therapy, or premature discontinuation due to clinical reasons while remaining eligible for subsequent surgery.
• Ability to provide a baseline plasma sample (T0) within 24-72 hours before initiation of the first systemic treatment (including chemotherapy, immunotherapy, or targeted therapy).
• Availability of preoperative breast and axillary magnetic resonance imaging (MRI) after completion of neoadjuvant therapy. Patients unable to undergo MRI due to contraindications or other reasons may be included in the cfDNA-only analysis set but will not be included in the complete-case analysis of the combined cfDNA-MRI model.

You may not qualify if:

• Participants meeting any of the following criteria will be excluded:
• Pregnancy or breastfeeding.
• Prior surgical removal of the primary breast lesion before enrollment, resulting in inability to obtain the required preoperative blood samples.
• Presence of confirmed distant metastasis.
• Presence of supraclavicular, internal mammary, or other lymph node lesions that cannot be adequately assessed by planned surgery and pathological evaluation.
• History of another active malignancy within the previous 5 years, except for cured non-melanoma skin cancer, cervical carcinoma in situ, or other malignancies considered by investigators unlikely to affect study outcomes.
• Receipt of whole blood, plasma, or other blood product transfusion within 30 days prior to enrollment.
• Insufficient plasma sample volume, severe hemolysis, or failure to meet cfDNA sequencing quality control requirements determined by the central laboratory.
• Absence of evaluable axillary surgical pathological results.
• Any other condition considered by the investigator to make the patient unsuitable for participation in this study.

Sponsors & Collaborators

• Second Affiliated Hospital, Zhejiang University, School of Medicine: lead
• Shandong Cancer Hospital and Institute: collaborator
• Suzhou Municipal Hospital: collaborator
• Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University: collaborator
• First People's Hospital of Hangzhou: collaborator
• Ningbo Medical Center Lihuili Hospital: collaborator

Study Sites (1)

Second affiliated hospital of Medical school, Zhejiang university

Hangzhou, Zhejiang, China

RECRUITING

Related Publications (4)

• Bruhm DC, Vulpescu NA, Foda ZH, Phallen J, Scharpf RB, Velculescu VE. Genomic and fragmentomic landscapes of cell-free DNA for early cancer detection. Nat Rev Cancer. 2025 May;25(5):341-358. doi: 10.1038/s41568-025-00795-x. Epub 2025 Mar 4.

  PMID: 40038442 BACKGROUND

• Gentilini OD, Botteri E, Sangalli C, Galimberti V, Porpiglia M, Agresti R, Luini A, Viale G, Cassano E, Peradze N, Toesca A, Massari G, Sacchini V, Munzone E, Leonardi MC, Cattadori F, Di Micco R, Esposito E, Sgarella A, Cattaneo S, Busani M, Dessena M, Bianchi A, Cretella E, Ripoll Orts F, Mueller M, Tinterri C, Chahuan Manzur BJ, Benedetto C, Veronesi P; SOUND Trial Group. Sentinel Lymph Node Biopsy vs No Axillary Surgery in Patients With Small Breast Cancer and Negative Results on Ultrasonography of Axillary Lymph Nodes: The SOUND Randomized Clinical Trial. JAMA Oncol. 2023 Nov 1;9(11):1557-1564. doi: 10.1001/jamaoncol.2023.3759.

  PMID: 37733364 BACKGROUND

• Reimer T, Stachs A, Veselinovic K, Kuhn T, Heil J, Polata S, Marme F, Muller T, Hildebrandt G, Krug D, Ataseven B, Reitsamer R, Ruth S, Denkert C, Bekes I, Zahm DM, Thill M, Golatta M, Holtschmidt J, Knauer M, Nekljudova V, Loibl S, Gerber B. Axillary Surgery in Breast Cancer - Primary Results of the INSEMA Trial. N Engl J Med. 2025 Mar 13;392(11):1051-1064. doi: 10.1056/NEJMoa2412063. Epub 2024 Dec 12.

  PMID: 39665649 BACKGROUND

• Zhu Y, Zheng S, Shao Y, Zhou J, Gu X, Shen L, Li X, Liu W, Xue W, Lu H, Zhou J, Ding J, Deng H, Chen J, Yu Z, Yao Y, Xia W, Chen W, Sun S, Wang Z, Qian T, Yu X, Liu J, Chen Y, Lin Z, Huang J, Ni C. Fragmentomic liquid biopsy enables early breast cancer detection, molecular subtyping and lymph node assessment. Nat Commun. 2026 Mar 6;17(1):2276. doi: 10.1038/s41467-026-70204-w.

  PMID: 41792156 BACKGROUND

Biospecimen

Retention: SAMPLES WITH DNA

Detect tumor-related cfDNA fragments in plasma

MeSH Terms

Conditions

Breast Neoplasms

Condition Hierarchy (Ancestors)

Neoplasms by Site; Neoplasms; Breast Diseases; Skin Diseases; Skin and Connective Tissue Diseases

Central Study Contacts

Chao Ni

CONTACT

+8613989463951; drnichao@zju.edu.cn

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Target Duration: 1 Year
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: November 19, 2025
• First Posted: December 26, 2025
• Study Start: August 1, 2025
• Primary Completion (Estimated): December 31, 2027
• Study Completion (Estimated): December 31, 2027
• Last Updated: August 10, 2026

Record last verified: 2025-10

Data Sharing

• IPD Sharing: Will not share

Locations

CN (1)