Salivary gland tumors are rare and morphologically diverse, posing both diagnostic and scientific challenges. This study presents the first phase of the SALV-Dataset Registry; a nationwide, expertly curated, and fully digitized clinicopathological resource, designed to support research and develop artificial intelligence (AI) tools assisting in salivary gland tumor pathology diagnostics. Salivary gland tumor resections diagnosed at the Leiden University Medical Center (1999–2024) were collected through the Dutch national network and registry for histo- and cytopathology (PALGA). In total, 685 cases were included. Hematoxylin- and eosin-stained slides were digitized and independently reviewed by three teams of head and neck pathologists, in line with the 2023 WHO Classification of Head and Neck Tumours. Discordant and ambiguous cases were resolved in consensus meetings, with access to immunohistochemistry, molecular analysis, and clinical data. Interobserver agreement among the three teams was quantified (Fleiss’ kappa), and agreement between the original and consensus diagnosis was determined (Cohen’s kappa). Of the 685 tumors, 75

PURPOSE:Manual chart review (MR) of electronic health records (EHRs) is time-consuming, error-prone, and limits the reproducibility and scalability of real-world data (RWD) research. Automation and standardization using natural language processing (NLP) could improve efficiency and scalability. CTcue is an NLP-based software platform designed to extract structured and unstructured data from EHRs. This study evaluated the accuracy and efficiency of CTcue versus MR in patients with early-stage resectable non-small cell lung cancer (NSCLC). METHODS:Included were all patients with stage I to III NSCLC who underwent lung resections between January 2018 and December 2021 at the Leiden University Medical Center, the Netherlands. Demographics, tumor characteristics, treatment, and outcomes were collected. CTcue performance was compared with MR using weighted F1-scores, accuracy, precision, and recall for categorical variables and Bland-Altman analysis for continuous variables. RESULTS:Eighty-five patients (70.2% of patients from the manual cohort) were identified by both methods and included in the comparison. CTcue achieved weighted F1-scores >0.85 for seven of 15 categorical variables, including sex, tumor location, and deceased status, although some scores were based on low number of observations in both cohorts. Lower performance was observed for variables with varying terminology in documentation, such as Eastern Cooperative Oncology Group status and pathological N-stage. Continuous variables showed negligible mean differences, indicating good agreement. Survival outcomes were identical in both data sets. CONCLUSION:CTcue performance for patient selection was lower than anticipated. However, it enables accurate, efficient extraction of structured and unstructured EHR data in early-stage NSCLC. Manual validation remains necessary for variables with varying terminology. Further development of artificial intelligence-based tools-particularly for free-text data extraction-will be crucial to enhance the accuracy and scalability of future RWD research.

IntroductionThe incidence of tumour-positive surgical resection margins (TPRMs) after breast-conserving surgery (BCS) remains high, ranging from 10 to 40%. A TPRM, defined as breast cancer cells at the edge of the resected specimen at pathological evaluation, implies residual tumour and necessitates re-resection or boost radiation. To prevent these additional treatments, intraoperative near-infrared (NIR) fluorescence imaging with the topically applied, fluorescently quenched, cathepsin-activatable imaging agent AKRO-6qcICG might be used to detect residual cancer in the surgical cavity and guide additional resection during BCS. Cathepsins are proteolytic enzymes that are upregulated by breast cancer (associated) cells and therefore are suitable targets for tumour imaging. Ex vivo validation studies have shown that topically applied AKRO-6qcICG allows for clear breast cancer visualization and the detection of TPRMs. The proposed phase I/II study in healthy volunteers and breast cancer patients will assess the local and systemic safety of a single, topical dose of AKRO-6qcICG and its feasibility for intraoperative margin assessment during BCS.Methods and analysisA total of six healthy volunteers (Part A) and 16 breast cancer patients (Part B) will be enrolled. In Part A, AKRO-6qcICG will be topically applied randomly on drawn blisters in two doses as will the vehicle compound, and one blister will be untreated functioning as a negative control. Physician and subject will remain blinded. In Part B, a single dose of AKRO-6qcICG will be topically applied in the surgical cavity. The primary objective is, with the occurrence of treatment-emergent (serious) adverse events as primary outcome measure. Secondary outcome measures include local and systemic tolerability parameters such as wound healing, numeric rating scales of pain and pruritus, vital signs, electrocardiogram parameters, clinical laboratory tests and pharmacokinetic parameters. Among the exploratory outcome measures are the diagnostic accuracy of the imaging agent to detect residual tumour in the surgical cavity and the tumour-to-background ratio of the fluorescent signal.Ethics and disseminationThis protocol has been approved by the Medical Ethical Committee Leiden-Den Haag- Delft (METC- LDD). The protocol is registered at EU Clinical Trials Register number 2025-523166-24-00. The results of this study will be reported through peer- reviewed publications and conference presentations.Clinical Trial Registrationhttps://ctis.eu/trial/2025-523166-24-00?from=search

OBJECTIVES:Identifying the primary tumor in cancer of unknown primary (CUP) remains a major clinical challenge. While WGS-based tissue-of-origin (TOO) prediction has improved diagnostic precision, its use is constrained by cost and availability. We investigated whether a low-complexity surrogate, combining SMARCA4 mutations and smoking history, can identify a lung cancer origin in CUP (CUP-Lung). METHODS:We retrospectively identified 305 provisional CUP cases from two Dutch CUP referral centers (2022-2025). Integration of whole-genome sequencing (WGS)-based TOO prediction with clinicopathological data identified 58 patients (18.9%) with CUP-Lung. Associations between SMARCA4 mutations, smoking history, and CUP-Lung were assessed using comparative and regression analyses. RESULTS:SMARCA4 mutations were present in 36.2% of CUP-Lung cases, significantly higher than in the overall CUP cohort (10.7%) and TCGA lung cancer datasets (6-8%). CUP-Lung cases had higher tumor mutational burden (median 21.5 vs. 12.6 mut/Mb, p < 0.001) and enriched smoking-related mutational signatures (p < 0.001). Smoking history was reported in 87.9% of cases. Smoking history (OR 4.9, 95% CI 2.2-10.9, p < 0.001) and SMARCA4 mutation (OR 10.3, 95% CI 4.5-23.8, p < 0.001) independently predicted CUP-Lung, together conferring a 76.0% probability of lung origin. No statistical differences were found between cases with and without detectable pulmonary involvement. CONCLUSION:Combined smoking history and SMARCA4 mutations support lung cancer classification even in the absence of detectable pulmonary involvement enabling organ-directed treatment when WGS is unavailable.

PURPOSE:In hormone receptor-positive, HER2-negative, early-stage breast cancer, cyclin-dependent kinase 4 and 6 inhibition combined with endocrine therapy could represent a less toxic alternative to neoadjuvant chemotherapy (CT). The NEOLBC trial studied whether neoadjuvant ribociclib plus letrozole (RL) results in a doubling of complete cell-cycle arrest (CCCA; Ki67 <1% on IHC) compared with CT in the surgical specimen in luminal breast cancer. PATIENTS AND METHODS:This randomized phase II trial tailored neoadjuvant therapy in postmenopausal patients with early, luminal, HER2-negative, stage II/III breast cancer based on the percentage of Ki67-positive cancer cells after 2 weeks of letrozole. Patients with a Ki67 ≥1% were randomized between RL and standard CT. Secondary endpoints included pathologic response and toxicity. RESULTS:Of 161 registered patients, 70 were randomized, and 66 started the allocated treatment. The CCCA in the surgical specimen was similar for both groups: 35.3% in the RL group and 31.3% in the CT group (P = 0.73). The response according to Miller and Payne was not significantly different between the two groups, nor was the pathologic complete response rate. Overall toxicity was observed more often in the CT group. In the RL group, eight patients discontinued treatment due to toxicity, and in the CT group, 10 patients discontinued treatment. CONCLUSIONS:Although the primary endpoint was not met, the NEOLBC trial (NCT03283384) showed a similar CCCA and pathologic response for RL and CT with less toxicity. Therefore, RL as an alternative to neoadjuvant CT merits further investigation.

To evaluate the diagnostic yield and clinical impact of a comprehensive RNA-based sequencing panel (“SalvGlandDx version 2”) in the work-up of salivary gland tumors. Between 2021 and 2025, 118 salivary gland tumors were assessed by routine histological and immunohistochemical investigation, followed by SalvGlandDx v2 RNA sequencing. This panel targets the recurrent molecular alterations in salivary gland neoplasms. Pre- and post-sequencing (differential) diagnoses were compared to determine the diagnostic contribution, categorized as diagnostic confirmation (1 A), refinement (1B/2A/2B) or no impact (3). Clinical impact was scored as none (A), altered follow-up (B), radiotherapy indication (C) and/or systemic therapy (D) when comparing pre- and post-sequencing (differential) diagnoses. RNA sequencing was successful in 114 of 118 cases (97

PURPOSE:Although the European Medicines Agency approved durvalumab post-chemoradiation (CRT) only for stage III unresectable non-small cell lung cancer (UR-NSCLC) patients with PD-L1 tumor proportion scores (TPS) ≥ 1%, the Netherlands offers reimbursement irrespective of PD-L1 status. This real-world study compares survival between durvalumab-treated patients with PD-L1 TPS < 1% vs. ≥ 1%, and also evaluates its effectiveness against a historic-cohort. PATIENTS AND METHODS:This multicenter retrospective study included 2 patient cohorts with stage III UR-NSCLC: a durvalumab cohort and a historic CRT-only cohort. The durvalumab cohort was divided into PD-L1 TPS subgroups: < 1%, ≥ 1%, and unknown. Overall survival (OS) and progression-free survival (PFS) were compared between (1) durvalumab-treated patients with PD-L1 TPS < 1% vs. ≥ 1%, and (2) durvalumab cohort (including PD-L1 subgroups) vs. historic-cohort. RESULTS:229 and 339 patients were included in the durvalumab- and historic-cohorts, respectively. Although not statistically significant, durvalumab-treated patients with PD-L1 TPS ≥ 1% experienced a modestly greater benefit in OS (2-year OS 74.3% vs. 66.5%) and PFS (2-year PFS 55.5% vs. 36.2%) compared to those with PD-L1 TPS < 1%. Survival outcomes favored durvalumab over the historic cohort across PD-L1 subgroups, though PFS improvement was not statistically significant for PD-L1 TPS < 1%. CONCLUSIONS:Given these findings, patients with PD-L1 TPS < 1% may also benefit from durvalumab treatment in stage III UR-NSCLC.

Functional loss of the intracellular peptide Transporter associated with Antigen Processing (TAP) fosters resistance to T-cell based immunotherapy. We discovered the presentation of an alternative set of shared tumor antigens on such escaped cancers and developed a LRPAP1 synthetic long peptide vaccine (TEIPP24) to stimulate T-cell immunity. In this first-in-human multicenter dose-escalation study with extension cohort, HLA-A*0201-positive patients with non-small cell lung cancer progressive after checkpoint blockade were treated with TEIPP24 (NCT05898763). Dose escalation followed an adapted 3 + 3 scheme where in each cohort six patients received the TEIPP24 peptide emulsified in Montanide ISA-51 at either 20, 40, 100 µg of peptide, subcutaneously injected three times every three weeks in alternating limbs. The extension cohort of six patients received the highest safe dose of TEIPP24 combined with the PD-1 checkpoint blocker pembrolizumab. The primary objectives of the study were safety, tolerability and immunogenicity of the TEIPP24 vaccine. Secondary objectives included the evaluation of specificity and immune modulatory effects of the vaccine, antigen and immune status of the patients, progression free (PFS) and overall survival (OS) and radiological tumor response rate and duration. A total of 26 patients were enrolled across 2 institutions. Treatment was well tolerated, and vaccine-induced LRPAP1-specific CD8+ T cells were detected in 20 of 24 evaluable patients (83%). In 13 of 21 tested cases (62%) vaccine-specific CD4+ T cells were also detected. The increase in activated polyfunctional CD8+ effector T cells was influenced by vaccine dose, number of vaccines administered, induction of a CD4+ T-cell response, and the pre-existing frequency of monocytic cells. Co-administration of pembrolizumab resulted in the ex-vivo detection of activated (HLA-DR+ , PD-1+ , ICOS+ ) LRPAP1-specific CD8+ T cells. The observation of one PR, 8 stable diseases and 2 mixed responses in 24 evaluable patients after vaccination, correlated with a stronger vaccine-induced CD8+ T-cell response to this single epitope from this new class of cancer antigens.

BACKGROUND:Diagnosis of salivary gland neoplasms is challenging, especially on cytological specimens acquired by fine-needle aspiration. The recently implemented standardized Milan system for reporting salivary gland cytopathology provides an estimated risk of malignancy (ROM); yet, for two of the categories, the diagnosis of the lesion remains unclear. However, a precise diagnosis is desirable for optimal patient management, including planning of surgery and imaging procedures. METHODS:Cytological specimens (n = 106) were subjected to molecular analysis using the SalvGlandDx panel. The risk of malignancy was calculated for each detected alteration based on the diagnosis of the resection specimen. By taking into account the molecular alterations, their associated ROM, the clinical and cytological features, and the current literature, the Milan category was evaluated. RESULTS:Of n = 63 technically valid cases, 76% revealed a molecular alteration. A total of 94% of these molecularly altered cases could be assigned to a different Milan category when additionally taking molecular results into account. In only 2% of the salivary gland neoplasms of uncertain malignant potential, in which a molecular alteration was detected, the classification remained salivary gland neoplasms of uncertain malignant potential. CONCLUSION:Molecular analysis of cytological specimens provides a benefit in classifying salivary gland neoplasms on fine-needle aspiration. It can improve the ROM estimation and thus help to assign cases of formerly unknown malignant potential to clearly benign or malignant categories.

BackgroundAn adequate diagnosis for interstitial lung disease (ILD) is important for clinical decision making and prognosis. In most patients with ILD, an accurate diagnosis can be made by clinical and radiological data assessment, but in a considerable proportion of patients, a lung biopsy is required. Surgical lung biopsy (SLB) is the most common method to obtain tissue, but it is associated with high morbidity and even mortality. More recently, transbronchial cryobiopsy has been introduced, with fewer adverse events but a lower diagnostic yield than SLB. The aim of this study is to compare two diagnostic strategies: a step-up strategy (transbronchial cryobiopsy, followed by SLB if the cryobiopsy is insufficiently informative) versus immediate SLB.MethodsThe COLD study was a multicentre, randomised controlled trial in six hospitals across the Netherlands. We included patients with ILD with an indication for lung biopsy as assessed by a multidisciplinary team discussion. Patients were randomly assigned in a 1:1 ratio to the step-up or immediate SLB strategy, with follow-up for 12 weeks from the initial procedure. Patients, clinicians, and pathologists were not masked to the study treatment. The primary endpoint was unexpected chest tube drainage, defined as requiring any chest tube after transbronchial cryobiopsy, or prolonged (>24 h) chest tube drainage after SLB. Secondary endpoints were diagnostic yield, in-hospital stay, pain, and serious adverse events. A modified intention-to-treat analysis was performed. This trial is registered with the Dutch Trial Register, NL7634, and is now closed.FindingsBetween April 8, 2019, and Oct 24, 2021, 122 patients with ILD were assessed for study participation; and 55 patients were randomly assigned to the step-up strategy (n=28) or immediate SLB (n=27); three patients from the immediate SLB group were excluded. Unexpected chest tube drainage occurred in three of 28 patients (11%; 95% CI 4–27%) in the step-up group, and the number of patients for whom the chest tube could not be removed within 24 h was 11 of 24 patients (46%; 95% CI 2–65%) in the SLB group, with an absolute risk reduction of 35% (11–56%; p=0·0058). In the step-up strategy, the multidisciplinary team diagnostic yield after transbronchial cryobiopsy alone was 82% (64–92%), which increased to 89% (73–96%) when subsequent SLB was performed after inconclusive transbronchial cryobiopsy. In the immediate surgery strategy, the multidisciplinary team diagnostic yield was 88% (69–97%). Total in-hospital stay was 1 day (IQR 1–1) in the step-up group versus 5 days (IQR 4–6) in the SLB group. One (4%) serious adverse event occurred in step-up strategy versus 12 (50%) in the immediate SLB strategy.InterpretationIn ILD diagnosis, if lung tissue assessment is required, a diagnostic strategy starting with transbronchial cryobiopsy, followed by SLB when transbronchial cryobiopsy is inconclusive, appears to result in a significant reduction of patient burden and in-hospital stay with a similar diagnostic yield versus immediate SLB.FundingNetherlands Organisation for Health Research and Development (ZonMW) and Amsterdam University Medical Centers.

Lymph node micrometastases could be one of the reasons for the high recurrence rate after complete surgical resection in stage I–IIIA non‐small cell lung cancer (NSCLC). The standard evaluation of a single haematoxylin and eosin (H&E) slide of a paraffin‐embedded section of a lymph node is insufficient for the detection of micrometastases, and there is a need for additional histopathological evaluation. The association of lymph node micrometastases with survival remains as yet unresolved. The aim of this systematic review and meta‐analysis is to investigate if lymph node micrometastases and isolated tumour cells in patients with stage I–IIIA NSCLC, detected with multiple sectioning and/or immunohistochemistry (IHC) and/or reverse transcriptase polymerase chain reaction (RT‐PCR), are associated with overall survival (OS) and disease‐free survival (DFS) after surgical resection. We performed a meta‐analysis of time‐to‐event outcomes based on 15 articles using ancillary techniques to detect micrometastases. We extracted the OS and DFS every 3–6 months after surgery, for patients with and without occult lymph node micrometastasis, from the survival curves published in each article. These data were used to reconstruct OS and DFS for ‘micrometastasis’ and ‘no micrometastasis’ groups. Based on all included studies that used IHC, serial sectioning, or RT‐PCR, we found a 5‐year OS of 55% (micrometastasis) vs. 75% (no micrometastasis), and a 5‐year DFS of 53% (micrometastasis) vs. 75% (no micrometastasis). Patients with stage I–IIIA NSCLC with lymph node micrometastases detected by ancillary histopathological and molecular techniques have a significantly poorer OS and DFS compared to patients without lymph node micrometastases.

Background: Adrenal metastases are common in lung cancer patients. Adrenalectomy is advised in case of solitary metastasis. Nowadays, a new group of patients is referred for adrenalectomy: patients with advanced metastatic lung cancer with mixed response to immunotherapy. In these patients the primary lung cancer and most metastases showed good response to immunotherapy, while the adrenal metastases did not respond or were newly developed. No literature is available on adrenalectomy in these immunotherapy patients. This study aims to get insights into whether adrenalectomy is safe and effective for these patients.

Background: Approximately 20% of invasive ductal breast malignancies are human epidermal growth factor receptor 2 (HER2)-positive. These patients receive neoadjuvant systemic therapy (NAT) including HER2-targeting therapies. Up to 65% of patients achieve a pathological complete response (pCR). These patients might not have needed surgery. However, accurate preoperative identification of a pCR remains challenging. A radiologic complete response (rCR) on MRI corresponds to a pCR in only 73% of patients. The current feasibility study investigates if HER2-targeted PET/CT-imaging using Zirconium-89 (89Zr)-radiolabeled trastuzumab can be used for more accurate NAT response evaluation. Methods: HER2-positive breast cancer patients scheduled to undergo NAT and subsequent surgery received a 89Zr-trastuzumab PET/CT both before (PET/CT-1) and after (PET/CT-2) NAT. Qualitative and quantitative response evaluation was performed. Results: Six patients were enrolled. All primary tumors could be identified on PET/CT-1. Four patients had a pCR and two a pathological partial response (pPR) in the primary tumor. Qualitative assessment of PET/CT resulted in an accuracy of 66.7%, compared to 83.3% of the standard-of-care MRI. Quantitative assessment showed a difference between the SUVR on PET/CT-1 and PET/CT-2 (ΔSUVR) in patients with a pPR and pCR of −48% and −90% (p = 0.133), respectively. The difference in tumor-to-blood ratio on PET/CT-1 and PET/CT-2 (ΔTBR) in patients with pPR and pCR was −79% and −94% (p = 0.133), respectively. Three patients had metastatic lymph nodes at diagnosis that were all identified on PET/CT-1. All three patients achieved a nodal pCR. Qualitative assessment of the lymph nodes with PET/CT resulted in an accuracy of 66.7%, compared to 50% of the MRI. Conclusions: NAT response evaluation using 89Zr-trastuzumab PET/CT is feasible. In the current study, qualitative assessment of the PET/CT images is not superior to standard-of-care MRI. Our results suggest that quantitative assessment of 89Zr-trastuzumab PET/CT has potential for a more accurate response evaluation of the primary tumor after NAT in HER2-positive breast cancer.

Objectives: Lung cancer diagnosis nowadays greatly relies on molecular analysis and assessment of PD-L1 expression. Adequacy of tissue samples obtained with EBUS-TBNA plays a crucial role in this process. The 22G (Acquire) needle with Franseen tip was developed to perform transbronchial needle biopsy (TBNB) with improved quality. The objective of the present study was to verify whether the PD-L1 assessment rate is related to the type of needle used: 22G (Acquire) TBNB needle vs regular 22G (Expect) TBNA needle. Methods: International randomized clinical trial, patients with suspected (N)SCLC and an indication for staging via EBUS were recruited in 5 institutions. Patients were randomized equally between the two needles. Samples were analyzed by two blinded pathologists. The primary outcome was PD-L1 suitability rate. Results: 154 patients randomized (n=76 (Acquire) TBNB; n=78 (Expect) TBNA). 92.9% of them had a final diagnosis of malignancy. (Acquire) needle yielded a suitability for PD-L1 analysis of 80.0% (95%CI 0.68-0.94) while it was 76.7% (95%CI 0.65-0.85) with the (Expect) needle (p=0.633). As regards specimens quality, (Acquire) TBNB needle returned better results (65.3% (95%CI 0.57-0.73) vs 49.4% (95%CI 0.41-0.57, p=0.005) and yielded more tissue cores (72.0% (95%CI 0.60-.81) vs 41.0% (95%CI 0.31-0.54, p<0.01). Tissue adequacy, sensitivity for malignancy and N2/N3 disease and adequacy for molecular analysis did not statistically differ. Conclusion: The 22G (Acquire) TBNB needle provided tissue specimens of higher quality compared to the (Expect) TBNA needle. However, suitability rate for PD-L1 analysis was not influenced by the needle type.

Objectives: The landmark ADAURA study recently demonstrated a significant disease-free survival benefit of adjuvant osimertinib in patients with resected EGFR-mutated lung adenocarcinoma. However, data on prevalence rates and stage distribution of EGFR mu-tations in non-small cell lung cancer in Western populations are limited since upfront EGFR testing in early stage lung adenocarcinoma is not common practice. Here, we present a unique, real-world, unselected cohort of lung adenocarcinoma to aid in providing a rationale for routine testing of early stage lung cancers for EGFR mutations in the West-European popu-lation.Material and methods: We performed routine unbiased testing of all cases, regardless of TNM stage, with targeted next-generation sequencing on 486 lung adenocarcinoma cases between 01-January 2014 and 01 February 2020. Clinical and pathological data, including co -mutations and morphology, were collected. EGFR-mutated cases were compared to KRAS-mutated cases to investigate EGFR-specific characteristics.Results: In total, 53 of 486 lung adenocarcinomas (11%) harboured an EGFR mutation. In early stages (stage 0-IIIA), the prevalence was 13%, versus 9% in stage IIIB-IV. Nine out of 130 (7%) stage IB-IIIA patients fit the ADAURA criteria. Early stage cases harboured more L858R mutations (p = 0.02), fewer exon 20 insertions (p = 0.048), fewer TP53 co-mutations (p = 0.007), and were more frequently never smokers (p = 0.04) compared to late stage cases with EGFR mutations. The KRAS-mutated cases were distributed more evenly across TNM stages compared to the EGFR-mutated cases. Conclusion: As (neo-)adjuvant targeted therapy regimes enter the field of lung cancer treat-ment, molecular analysis of early stage non-small cell lung cancer becomes relevant. Testing for EGFR mutations in early stage lung adenocarcinoma holds a substantial yield in our pop-ulation, as our number needed to test ratio for adjuvant osimertinib was 14.4. The observed differences between early and late stage disease warrant further analysis to work towards bet-ter prognostic stratification and more personalised treatment. 2022 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PURPOSE:BRCA-deficient breast cancers (BC) are highly sensitive to platinum-based chemotherapy and PARP inhibitors due to their deficiency in the homologous recombination (HR) pathway. However, HR deficiency (HRD) extends beyond BRCA-associated BC, highlighting the need for a sensitive method to enrich for HRD tumors in an alternative way. A promising approach is the use of functional HRD tests which evaluate the HR capability of tumor cells by measuring RAD51 protein accumulation at DNA damage sites. This study aims to evaluate the performance of a functional RAD51-based HRD test for the identification of HRD BC.METHODS:The functional HR status of 63 diagnostic formalin-fixed paraffin-embedded (FFPE) BC samples was determined by applying the RAD51-FFPE test. Samples were screened for the presence of (epi)genetic defects in HR and matching tumor samples were analyzed with the RECAP test, which requires ex vivo irradiated fresh tumor tissue on the premise that the HRD status as determined by the RECAP test faithfully represented the functional HR status.RESULTS:The RAD51-FFPE test identified 23 (37%) of the tumors as HRD, including three tumors with pathogenic variants in BRCA1/2. The RAD51-FFPE test showed a sensitivity of 88% and a specificity of 76% in determining the HR-class as defined by the RECAP test.CONCLUSION:Given its high sensitivity and compatibility with FFPE samples, the RAD51-FFPE test holds great potential to enrich for HRD tumors, including those associated with BRCA-deficiency. This potential extends to situations where DNA-based testing may be challenging or not easily accessible in routine clinical practice. This is particularly important considering the potential implications for treatment decisions and patient stratification.

INTRODUCTION:Accurate diagnosis and staging of lung cancer is crucial because it directs treatment and prognosis. Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) and endoscopic ultrasound with bronchoscope fine-needle aspiration (EUS-B-FNA) are important in this process by sampling hilar/mediastinal lymph nodes and centrally located lung tumours. With the upcoming of immunotherapy and targeted therapies, assessment of programmed death ligand 1 (PD-L1) expression and molecular profiling has become important but is often impossible in cytological samples obtained through standard 22G TBNA needles. Recently, a three-pronged cutting edge 22G needle was developed that allows for transbronchial needle biopsy (TBNB). Our objective is to determine if EBUS/EUS-B-guided nodal/lung tumour sampling with Acquire 22G TBNB needles results in an improved suitability rate for the assessment of PD-L1 expression in comparison to standard 22G TBNA needles in patients with a final diagnosis of lung cancer.METHODS AND ANALYSIS:This is an investigator-initiated, parallel group randomised clinical trial. Patients are recruited at respiratory medicine outpatient clinics of participating university and general hospitals in the Netherlands, Poland and Italy. In total 158 adult patients with (suspected) lung cancer are included if they have an indication for mediastinal/hilar lymph node or lung tumour sampling by EBUS-TBNA and/or EUS-B-FNA based on current clinical guidelines. Web-based randomisation between the two needles will be performed. Samples obtained from mediastinal/hilar lymph nodes and/or primary tumour will be processed for cytology smears and cell block analysis and reviewed by blinded reference pathologists. An intention-to-treat analysis will be applied. Patients with missing data will be excluded from analysis for that specific variable but included in the analysis of other variables. This study is financially supported by Boston Scientific.ETHICS AND DISSEMINATION:The study was approved by the local Ethics Committee (Medisch Ethische Toetsingscommissie Amsterdam Medical Center (AMC)). Dissemination will involve publication in a peer-reviewed biomedical journal.TRIAL REGISTRATION NUMBER:NL7701; Pre-results.

Introduction In hormone receptor-positive, HER2-negative early-stage breast cancer (BC), cyclin-dependent kinases 4 and 6 inhibition (CDK4/6i) in combination with endocrine therapy (ET) could represent an alternative to neoadjuvant chemotherapy (NAC). Methods NEOLBC is a randomized phase II trial that tailored neoadjuvant therapy in postmenopausal patients with early, luminal (ER >50%, PR any), HER2-negative, stage II/III BC based on the percentage of Ki67 positive cancer cells after a window of opportunity of two weeks letrozole. Patients with a Ki67 >= 1% after 2 weeks were randomized between ribociclib plus letrozole (RL) and chemotherapy (CT; AC-T regimen). The primary objective was to determine if RL gives a doubling in complete cell cycle arrest (CCCA; Ki67 < 1% on IHC) as compared to CT in the surgical specimen (70% vs. 35% of patients, respectively). Secondary endpoints included pathological response, toxicity and ER pathway activity (measured by the OncoSIGNal qPCR test). Results Out of 161 registered patients, 70 patients were randomized and 66 patients started treatment; 34 RL and 32 CT. Patient characteristics were equally distributed between the two groups, except for the PR status (RL 23.5% negative vs. 50.0% negative in the CT group). In the intention to treat analysis, the CCCA in the surgical specimen was similar for both groups: 35.3% in the RL vs. 31.3% in the CT group (p = 0.73). The pathological complete response (pCR) in the breast was not significantly different between the two groups (11.8% vs. 3.1%, p = 0.36) nor was the pCR rate in breast plus lymph nodes (8.8% vs. 3.1%, p = 0.61) for the RL vs. CT group, respectively. An explorative analysis on the difference in Ki67% (decline, no change, increase) from baseline to surgery showed a decline in 73.5% vs. 50.0%, no change in 17.6% vs. 31.3% and an increase in 8.8% vs. 18.8% of patients (p = 0.06) for the RL vs. CT group, respectively. In the RL group eight patients (23.5%) discontinued ribociclib early due to toxicity (two SAE’s were observed) vs. 10 patients (31.3%) discontinuing treatment in the CT group (one SAE was observed). Secondary endpoints, including the ER pathway activity analysis, will be presented in-depth during the meeting. Conclusion Although the primary endpoint was not met, the NEOLBC trial showed a similar CCCA and pathological response at surgery for RL vs. CT. Therefore, RL as an alternative for NAC merits further investigation in follow-up studies. ClinicalTrials.gov: NCT03283384 Citation Format: Anne Florine de Groot, Danielle Cohen, Joan B. Heijns, Caroline Mandigers, Diederick M. Keizer, Hein Putter, Elma Meershoek-Klein Kranenbarg, Marjolijn Duijm-de Carpentier, Kyra Dijkstra, Elise van Leeuwen-Stok, Gerrit-Jan Liefers, Sabine Linn, Judith Kroep. The use of ribociclib/letrozole combination as an alternative for neoadjuvant chemotherapy in selected patients with early luminal breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-07-07.