Sentinel Lymph Node Analysis Research Articles

Frozen Section Analysis of Sentinel Nodes in Patients With Oral Squamous Cell Carcinoma

Frozen section (FS) analysis of sentinel nodes offers potential on-table diagnosis and treatment for occult metastasis in oral squamous cell cancer. Systematic analysis of FS during sentinel node biopsy has not been illuminated in the literature. To systematically review pooled data from studies using FS analysis in evaluating sentinel nodes in patients with cT1-T2 N0 oral squamous cell cancer. An academic librarian led the search of CENTRAL, CINAHL, Cochrane Database of Systematic Reviews, Embase, and MEDLINE for studies published in English between January 2000 and January 2023. Two authors independently screened cohort studies, case series, and randomized clinical trials, in which FS analysis was used to evaluate sentinel nodes in patients with cT1-T2 N0 oral squamous cell cancer. Data were extracted by 2 reviewers. Reporting quality was estimated using the Diagnostic Precision Study Quality Assessment Tool. Data analysis was performed between April and July 2023, and the meta-analysis was completed using the bivariate random-effects model. The primary outcome was the pooled sensitivity of FS sentinel node analysis. Secondary outcomes included evaluation of the FS technique, rate of occult metastasis, false-negative rate, and survival. Seventeen articles with 878 patients met the eligibility criteria. Although protocols varied, confirmatory serial step sectioning was performed in all studies. Occult metastasis was found in 263 of 878 patients (30%), and FS analysis identified 173 cases (65.8%). Following serial sectioning, an additional 90 positive results were identified, leading to 47 patients undergoing staged completion neck dissection. The pooled sensitivity of FS was 0.71 (95% CI, 0.60-0.80), the diagnostic odds ratio was 110, and the false-negative rate was 34.2%. The Cochrane Q value was 15.62 (df = 16; P = .48) and τ2 = 0.36. In this systematic review and meta-analysis, evaluated studies showed various techniques, in which pooled sensitivity reached 0.71, providing a benchmark for comparison to other 1-stop approaches. Due to the high false-negative rate of approximately one-third of patients, intraoperative FS must always be supplemented by serial sectioning. On-table diagnosis remains a key objective for sentinel node biopsy, and FS detection may be improved by standardizing protocols.

The importance of sentinel lymph node intraoperative frozen analysis for indication of lateral neck dissection in patients with medullary thyroid cancer.

Our research seeks to evaluate the utility of intraoperative frozen analysis of sentinel lymph nodes (SLNs) in the lateral cervical compartment (LCC) as a tool to inform decision-making regarding therapeutic neck dissection in patients with medullary thyroid carcinoma (MTC). This is particularly relevant due to the variability observed in guidelines regarding the indication for lateral neck dissection in this patient population. The study comprised 64 patients (25 males, 39 females) aged between 29 and 81 years, with a median age of 59, who underwent surgery for MTC at stage T1-3N0-1M0 between January 1, 2012, and December 31, 2020. A standardized surgical approach involving total thyroidectomy with central neck dissection was adopted. LCC dissection was reserved for patients with clinically apparent nodal metastases. In patients lacking clinical evidence of nodal involvement, SLNs were identified using patent blue dye, excised, and subjected to intraoperative frozen analysis. If metastasis was confirmed, LCC dissection was subsequently performed. Among the study participants, 14 individuals (21.9%) underwent therapeutic LCC dissection due to clinical lymph node (LN) metastases. This intervention resulted in clinical remission for 9 patients, while disease progression was observed in 5 cases, leading to 2 fatalities. In the remaining cohort of 50 patients clinically negative for nodal involvement, SLNs were successfully identified and examined in 38 cases, revealing metastases in 6 patients (15.8%). Among both subsets of patients with analyzed SLNs, irrespective of metastatic status, one patient each required repeat surgery due to disease recurrence; however, all patients eventually achieved clinical remission. Lymphatic mapping in the LCC plays a pivotal role in detecting early metastases, thereby aiding in the avoidance of unnecessary repeat neck surgeries, and ultimately improving the prognosis in patients with MTC.

Feasibility and Diagnostic Accuracy of Ultrastaging in the Detection of Micrometastases in Sentinel and Non-sentinel Lymph Nodes in Cervical Cancer: A Single-Center Retrospective Study With a Five-Year Follow-Up Period.

Cervical cancer is the fourth most common cause of malignant tumor-related deaths among women in developing nations. Cervical cancer has been estimated to cause 527.600 new cases and 265.700 deaths globally per year. This study aimed to evaluate patients with cervical cancer by ultrastaging all the lymph nodes (LN), sentinel LN (SLN) and non-SLN, to increase the sensitivity of the detection of LN metastases and the diagnostic accuracy in cervical cancer with a five-year follow-up. Thisis a retrospective study of 14 cervical cancer cases from 2017 to 2019 at the Municipal Emergency Clinical Hospital of Timisoara, Romania. The cases were selected based on their high risk of LN involvement but negative intraoperative pathologic LN. After re-evaluating all paraffin block biopsy samples from 29 cases, 14 cases were included in the study, which met all criteria for ultrastaging on surgical biopsy samples. Patients' ages included in the study ranged from 43 to 70 years (median: 57.14 years). According to the International Federation of Gynecology and Obstetrics (FIGO) staging, the majority of the patients were in stage IB: seven cases (50%). The study revealed a positive correlation between patient age and FIGO staging, with Pearson's correlation coefficient of 0.707 and a p-value of less than 0.05, indicating that older patients were more likely to be diagnosed with a higher FIGO stage. The mean follow-up was 34.5 months, and the median follow-up was 36 months (range: 6-60 months). We obtained 167 nodes, with a mean of 11.92 nodes/case. Twenty-one LN were found to be positive with the ultrastaging method. We detected 11 LN with macrometastases (MAC) (52.38%), seven with micrometastasis (MIC) (33.3%), and three with tumor cell islets (14.4%). That would be 13% of newly diagnosed ultrastaging cases as positive nodes.This ultrastaging method detected nodal MIC in eight (57.1%) out of the 14 patients, who initially tested negative for LN involvement using the routine Hematoxylin and Eosin (HE) method. The detection of micrometastases in these patients underscored the superior sensitivity of ultrastaging, which was further highlighted by the subsequent relapse of four (28.57%) out of these eight patients. The study also found no correlation between the FIGO standardization and the number of MIC found in these patients. Predicting cervical LN metastasis (LNM) is crucial for improving survival rates and reducing recurrence. Very few small cohort studies used an ultrastaging method to assess non-SLNs; most of them only assessed SLNs. We showed in our study that the ultrastaging method, both in the case of SLN and non-SLN, is superior compared with H&E analysis, with a 13% rate of new positive nodule diagnosis. Metastatic involvement of non-SLN was found in over 50% of all cases (8/14) according to the ultrastaging method. Additionally, our study confirms that the sensitivity of SLN ultrastaging is high for the presence of both MIC and MAC in SLN pelvic LN. As a result, we feel that ultrastaging is the most effective method for SLN analysis in patients with early-stage cervical cancer, and bilateral detection is preferable, significantly reducing false-negative results. The routine use of SLN along with ultrastaging would lead to more accurate surgical staging and better oncological follow-up of cases.

Abstract 1503: Melanoma-derived lymphatic extracellular vesicles suppress dendritic cell differentiation and maturation in the sentinel lymph node

Abstract Spread to regional lymph nodes is the most common site of metastasis from melanoma and almost all solid tumors. Lymphatic metastases begin in the first lymph nodes draining from the primary tumor, i.e., the sentinel lymph nodes (SLN), preceded by the development of the premetastatic niche (PMN) initiated by primary cutaneous melanoma. Melanoma mediators are known to play a critical role in developing the PMN through the immune subversion in SLN. The DCs are a vital component of LN immunity that directly relates to eliciting T-cell functions. Our multiplex immunofluorescence (MxIF) analysis of SLNs indicates significantly decreased levels of DCs and cytotoxic-T cells in both core and interface areas of tumors in recurred patients treated with immunotherapy compared to non-recurred melanoma patients. The goal of this study is to determine whether the lymphatic extracellular vesicles (L-EVs) of melanoma patients regulate DC tolerization in SLN in order to stimulate the development of PMN. To identify whether L-EVs stimulate DC suppression, we extracted L-EVs from the lymphatic fluid of patients with melanoma downstream of primary cutaneous melanomas, compared it with controls (non-malignant post-operative fluid collected from the lymph node dissection field, and evaluated their effect on DC maturation by challenging these L-EVs in monocytic (CD14+) and hematopoietic stem cells (CD34+) cells. We found that L-EVs derived from melanoma patients suppress differentiation of CD11c+HLA-DR+ DCs and their maturation (defined by CD83 and CD86) compared to control L-EVs in CD14+ based DC maturation assay. We also found a reduction of conventional type 1 DC (cDC1) in CD34+-based DC maturation assay those challenged with patients’ L-EVs compared to control. To understand the mechanism of DC suppression in patients' L-EVs, we also conducted a total RNA sequencing analysis of subsets of DCs from maturation experiments to evaluate differently regulated transcriptomic profiles. The gene ontology analysis indicates that patient-derived L-EVs interfere with tRNA processing, oncostatin M & IL-10 signaling, and other immunoregulatory pathways to alter the hematopoietic cell lineage pathways in the DC maturation. Furthermore, the proteomic analysis of L-EVs also indicated several differentially regulated cargo-identified proteins in patient L-EVs, which are associated with immunomodulation, including regulation of DC function compared to control L-EVs. In conclusion, our study revealed that the patient's L-EVs carry differential proteomic cargoes secreted by cutaneous melanoma, which may be involved in regulating DC differentiation and maturation to diminish their functions and support the developing PMN in SLN. Citation Format: Shankar Suman, Wendy K. Nevala, James W. Jakub, Ray Guo, Alexey A. Leontovich, Raymond M. Moore, Chathu L. Atherton, Liyi Geng, Jeffrey E. Johnson, Sarah A. McLaughlin, Svetomir N. Markovic. Melanoma-derived lymphatic extracellular vesicles suppress dendritic cell differentiation and maturation in the sentinel lymph node [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1503.

Intraoperative Frozen Section Analysis and an Audit of its Diagnostic Accuracy: A Cross-sectional Study from Maharashtra, India

Introduction: Frozen Section (FS) is a diagnostic technique performed intraoperatively to obtain relevant information about the primary diagnosis, margin status, or sentinel lymph nodes, which helps guide the course of surgery. With the prevalent use of FS, it is imperative to recognise and scrutinise its diagnostic pitfalls and make efforts for rectification. Aim: To compare diagnostic results obtained on FS with final paraffin-embedded histopathology and calculate the Diagnostic Accuracy (DA) of FS. Materials and Methods: This cross-sectional study was conducted in the Department of Pathology at a tertiary care hospital, DVVPFs Medical College in Ahmednagar district, Maharashtra, India over a period of two years from January 1, 2020, to December 31, 2021. The diagnosis provided on specimens received for intraoperative FS was compared with the final histopathological diagnosis, considered as the gold standard for the same specimen. The results were categorised as concordant, discordant, and deferred cases. The diagnosis on FS and the final histopathological diagnosis, along with relevant clinical data, were entered into an excel sheet. Further, the DA of FS was calculated. All the discordant cases were analysed. Statistical analysis utilised simple percentage calculations from the excel sheet. Results: A total of 130 cases were analysed using FS, with the most common indication being sentinel lymph node analysis to detect metastatic deposits in 72 (55.4%) cases. Out of the total 130 cases, 119 (91.5%) were concordant, 10 (7.7%) were discordant, and 1 (0.8%) was deferred due to an insufficient specimen. Therefore, the overall DA rate was found to be 91.5% with an error rate of 8.5%. Upon analysis of the 10 discordant cases, the cause of inaccuracy was technical error in 7 (70%) cases and interpretation error in 3 (30%) cases. Conclusion: The DA obtained in the present study was somewhat lower than expected due to technical errors during FS sectioning, leading to artifacts, especially during lymph node processing. Analysis of the discordant cases unveiled this deficit. Therefore, such assessment studies should be performed periodically as they assist in highlighting the shortfalls and provide a plan to boost DA.

Distribution pattern and pathologic analysis of metastatic sentinel and non-sentinel lymph nodes in lymphatic basin dissection for clinical T2/T3 oral cancer with clinical N0 status

Distribution pattern and pathologic analysis of metastatic sentinel and non-sentinel lymph nodes in lymphatic basin dissection for clinical T2/T3 oral cancer with clinical N0 status

The Evaluation of Sentinel Lymph Node Biopsy Using Radiocolloid in First Stage Endometrial Cancer.

Detection of a sentinel lymph node (SLN) in patients with endometrial cancer (EC) reduces the rate of unnecessary systemic lymph dissection. The aim of this study was to assess the SLN detection rate, accuracy of the method using Tc-99m-SENTI-SCINT and the rate of metastatic nodal involvement in patients with preoperative first stage EC. A prospective study of SLN biopsy of 41 patients with stage I EC was conducted after cervical application of 4mCi Tc-99m-SENTI-SCINT. Planar lymphoscintigraphy and single-photon emission computed tomography/computed tomography (SPECT/CT) of the pelvis were performed, followed by site-specific lymphadenectomy in intermediate-risk patients if no SLN was detected per hemipelvis and pelvic lymphadenectomy in all high-risk patients. Pre-operative detection rate of planar lymphoscintigraphy was 80.49 [95% confidence interval (CI): 68.36-92.62] and of SPECT/CT 95.12 (95% CI: 88.52-101.7). The total intraoperative SLN detection rate was 95.12 (95% CI: 88.52-101.7) per patient and 26.83 (95% CI: 19.91-33.75) bilaterally. The average number of SLNs removed was 1.6±0.8. The most common anatomical location of SLN was the right external iliac region. The SLN metastatic rate was 17%. Both sensitivity and negative predictive value regarding metastatic involvement were 100%. The SLN detection rate, sensitivity and negative predictive value using Tc-99m-SENTI-SCINT in patients with EC in our study were high. The application of ultra-staging in the histopathological analysis of SLN increases the detection of nodal metastases and improves the staging in these patients.

Indocyanine green in diagnostics and reconstructive surgery for breast cancer

To analyze the efficacy of sentinel lymph node detection using indocyanine green (ICG) in breast cancer. The study included 153 breast cancer patients (stage 0-I-II-III, cN0 or cN1-ycN0) between October 2022 and April 2023. All patients underwent sentinel lymph node biopsy using ICG. Sentinel lymph node was successfully detected in 150 cases out of 153 ICG injections. Optimal period between ICG injection and its accumulation in regional lymph nodes was 10-12 min. Sensitivity of sentinel lymph node detection was 98.0% that exceeds the approximate value of radioisotope method adopted as a standard (96.9-97.2%), as well as sensitivity of proprietary blue dye (90.6-95.0%). Specificity of this method was 100%. The authors' experience regarding application of indocyanine green is consistent with numerous world statistics and proves the possibility of its effective use for sentinel lymph node biopsy. It is necessary to introduce this diagnostic method as a leading or alternative method for analysis of sentinel lymph node in national clinical guidelines.

Lack of Clinical Value for Immunohistochemistry for Sentinel Lymph Node Assessment in Invasive Lobular Carcinoma.

The distinct histologic appearance of invasive lobular carcinoma (ILC) may pose diagnostic challenges for sentinel lymph node (SLN) analysis. We evaluated the impact of cytokeratin immunohistochemistry (IHC) on SLN assessment in ILC and its contribution to pathologic nodal upstaging. We identified ILC patients treated with SLN surgery at our institution between September 2008 and August 2021. IHC for SLN assessment was employed at the discretion of the pathologist. Differences between groups evaluated with and without IHC were compared using Chi-square tests. Overall, 608 cases of ILC were identified in patients who underwent SLN surgery. IHC was used in 301 cases (49.5%)and was not associated with cT category, pT category, or tumor grade. Use of IHC increased detection of SLN+ disease when isolated tumor cells (ITCs) were included in the analysis (35.9% with IHC vs. 21.2% without IHC; p<0.001). There was no effect on nodal upstaging to micrometastatic disease (pN1mi) or greater (21.9% with IHC vs. 21.2% without IHC; p=0.82). IHC did not increase the number of positive SLNs detected (median 1 with and without IHC) nor did it increase axillary lymph node dissection (ALND) rates (11.6% with IHC vs. 15.3% without IHC; p=0.18). IHC improved detection of pN0(i+) disease among ILC patients undergoing SLN surgery. IHC did not increase upstaging to pN1mi or higher categories of nodal disease, detection of a greater number of positive SLNs, or ALND rates. Our data suggest routine use of IHC for SLN assessment in ILC patients does not add clinical utility.

A nomogram for predicting three or more axillary lymph node involvement before breast cancer surgery

Based on the American College of Surgeons Oncology Group (ACOSOG)-Z0011, a useful nomogram has been constructed to identify patients who do not require intraoperative frozen sections to evaluate sentinel lymph nodes in the previous study. This study investigated the developed nomogram by ultrasonography (US) and positron emission tomography (PET)/computed tomography (CT) as a modality. In the training set, 89/1030 (8.6%) patients had three or more positive nodes. Larger tumor size, higher grade ultrasonographic ALN classification, and findings suspicious of positive ALN on PET/CT were associated in multivariate analysis. The areas under the receiver operating characteristic curve (AUC) of the nomogram were 0.856 [95% CI 0.815–0.897] in the training set. The AUC in the validation set was 0.866 [95% CI 0.799–0.934]. Application of the nomogram to 1067 patients who met the inclusion criteria of ACOSOG-Z0011 showed that 90 (8.4%) patients had scores above the cut-off and a false-negative result was 37 (3.8%) patients. And the specificity was 93.8%, and the negative predictive value was 96.4%. The upgraded nomogram improved the predictive accuracy, using only US and PET/CT. This nomogram is useful for identifying patients who do not require intraoperative analysis of sentinel lymph nodes and considering candidates for identifying neoadjuvant chemotherapy. The patients consisted of clinical T1-2 and node-negative invasive breast cancer. The training and validation set consisted of 1030 and 781 patients, respectively. A nomogram was constructed by analyzing factors related to three or more axillary lymph node metastases. The patients who matched the ACOSOG-Z0011 criteria were selected and applied to the new nomogram.

Evaluation of Intraoperative Frozen Section Analysis with Final Histopathology Results for Sentinel Lymph Node Biopsy: Z0011 Criteria Eligible Versus Ineligible Breast Cancer Patients.

Background Intraoperative frozen section analysis (FSA) of sentinel lymph nodes (SLNs) declined in the post American College of Surgeons Oncology Group Z0011 (ACOSOG Z0011) trial era. However, for those patients who do not meet the ACOSOG Z0011 criteria, FSA continues to be a valuable tool in intraoperative decision-making for axillary lymph node dissection (ALND). The aim of this study was therefore to retrospectively evaluate the benefit and accuracy of FSA of Z0011 criteria eligible versus ineligible patients and identify possible predictive factors for false negative results. Methods Intraoperative FSA was performed on SLNs of 522 cT1–T3 breast cancer patients between 2008 and 2013. Clinicopathologic characteristics were retrospectively assessed by chart review. Results Overall FSA sensitivity and specificity was 67.8% and 100%. Sensitivity was generally higher for macrometastasis than for micrometastasis. The Z0011 eligible group showed a sensitivity and specificity of 72.7% and 100% versus 62.1% and 100% in the Z0011 ineligible group. Importantly, subgroup analysis of ≤ 2 versus > 2 positive SLNs of the Z0011 eligible group demonstrated both a 100% specificity and sensitivity. Several clinicopathologic factors were associated with a higher rate of false negative results in the Z0011 ineligible patient group. FSA was beneficial for 22.2% of Z0011 ineligible patients and for only 0.6% of Z0011 eligible patients regarding intraoperative decision-making for ALND. Conclusions FSA continues to be especially beneficial in the intraoperative assessment of SLNs in the Z0011 ineligible group to prevent second stage ALND. Despite an overall lower FSA sensitivity in the Z0011 eligible patient group, FSA offers in both groups a comparable high sensitivity and diagnostic accuracy for macrometastasis.

Intraoperative sentinel lymph node analysis after neoadjuvant chemotherapy in breast cancer patient - A 5 year single centre cohort study

Introduction: Intraoperative analysis (IOA) of sentinel lymph nodes is effective, and reliable for assessing node in breast cancer allowing for immediate clearance. Metasin® is a quantitative PCR Assay that detects CK 19 and mammaglobin. ABS guidance does not advocate IOA methods after neo-adjuvant chemotherapy (NACT) due to concern regarding detection of residual disease. We completed a retrospective comparative review of all consecutive patients who had IOA during SNB following NACT.

A prediction model for early systemic recurrence in breast cancer using a molecular diagnostic analysis of sentinel lymph nodes: A large-scale, multicenter cohort study.

BackgroundThe one‐step nucleic acid amplification (OSNA) assay can quantify the cytokeratin 19 messenger RNA copy number as a proxy for sentinel lymph node (SN) metastasis in breast cancer. A large‐scale, multicenter cohort study was performed to determine the prognostic value of the SN tumor burden based on a molecular readout and to establish a model for the prediction of early systemic recurrence in patients using the OSNA assay.MethodsSN biopsies from 4757 patients with breast cancer were analyzed with the OSNA assay. The patients were randomly assigned to the training or validation cohort at a ratio of 2:1. On the basis of the training cohort, the threshold SN tumor burden value for stratifying distant recurrence was determined with Youden's index; predictors of distant recurrence were investigated via multivariable analyses. Based on the selected predictors, a model for estimating 5‐year distant recurrence–free survival was constructed, and predictive performance was measured with the validation cohort.ResultsThe prognostic cutoff value for the SN tumor burden was 1100 copies/μL. The following variables were significantly associated with distant recurrence and were used to construct the prediction model: SN tumor burden, age, pT classification, grade, progesterone receptor, adjuvant cytotoxic chemotherapy, and adjuvant anti–human epidermal growth factor receptor 2 therapy. The values for the area under the curve, sensitivity, specificity, and accuracy of the prediction model were 0.83, 63.4%, 81.7%, and 81.1%, respectively.ConclusionsUsing the OSNA assay, the molecular readout–based SN tumor burden is an independent prognostic factor for early breast cancer. This model accurately predicts early systemic recurrence and may facilitate decision‐making related to treatment.

Intraoperative prediction of non-sentinel lymph node metastases in breast cancer using cytokeratin 19 mRNA copy number: A retrospective analysis.

One-step nucleic acid amplification (OSNA) is a molecular procedure used intraoperatively for the detection of sentinel lymph node (SLN) metastases. The aim of the present study was to define a cut-off of cytokeratin (CK)19 mRNA copy number predictive of positive completion axillary lymph node dissection (ALND). The OSNA procedure was employed for SLN analysis in 812 patients with T1-T2 N0 breast cancer. A total of 197 patients with SLN metastases were retrospectively analyzed. A total of 40 patients (20%) had non-SLN metastases. Receiver operating characteristics curve analysis established a cut-off of 5,000 CK19 mRNA copy number with 75% sensitivity and 72% specificity. The positive and negative predictive values were 40.5 and 92%, respectively. Multivariate analysis showed that this cut-off and tumor localization in the outer or lower-outer quadrant of the breast were significantly associated with non-SNL involvement (P<0.001 and P=0.025, respectively). The findings of the present study support the conventional cut-off of 5,000 copies for intraoperative decision to perform ALND, whereas ALND can safely be avoided in patients with tumor located outside the outer or lower-outer quadrant of the breast if the CK19 mRNA copy number is <5,000.

Distribution pattern and pathologic analysis of metastatic sentinel and non-sentinel lymph nodes in lymphatic basin dissection for clinical T2/T3 oral cancer with clinical N0 status

ObjectiveThe localization pattern of metastatic sentinel lymph node (SN) and non-SNs and pathologic analysis of metastatic lymph nodes in SN lymphatic basin dissection (SLBD) were investigated in patients with cT2/T3cN0 oral squamous cell carcinoma (OSCC). MethodsThis prospective multicenter trial involved 10 institutions nationwide in Japan. A total of 57 patients were enrolled. The lateral neck was divided into 5 lymphatic basins. The lymphatic basin containing SNs was defined as the SN lymphatic basin.All patients underwent SLBD with backup selective neck dissection (I-III) combined with primary tumor removal. When SNs were found outside of levels I-III, including in the contralateral neck, SLBD was performed by removing the compartments containing SNs separately. SN metastasis was classified as isolated tumor cells (ITCs), micrometastasis, or macrometastasis. ITCs are defined as a lesion no larger than 0.2 mm in largest dimension and are classified as pN0. ResultsSN metastasis was observed in 22 cases. All metastatic lymph nodes, including false-negative cases, were detected in the SN lymphatic basin. Isolated tumor cells in the SNs did not affect prognosis, whereas micrometastasis tended to have poor prognosis.After adjusting for other risk factors, a positive SN remained a significant predictor of poor 5-year overall survival in pT2-4 OSCC. ConclusionSLBD for intraoperative SN biopsy is a sufficient therapeutic procedure and is valuable for determining pathologic nodal stage in OSCC. SN positivity was demonstrated to be an independent predictor of poor prognosis in patients with pT2-4 disease undergoing SLBD with backup selective neck dissection (I-III).

High-resolution three-dimensional imaging for precise staging in melanoma

IntroductionMany cancer guidelines include sentinel lymph node (SLN) staging to identify microscopic metastatic disease. Current SLN analysis of melanoma patients is effective but has the substantial drawback that only a small representative portion of the node is sampled, whereas most of the tissue is discarded. This might explain the high clinical false-negative rate of current SLN diagnosis in melanoma. Furthermore, the quantitative assessment of metastatic load and microanatomical localisation might yield prognosis with higher precision. Thus, methods to analyse entire SLNs with cellular resolution apart from tedious sequential physical sectioning are required. Patients and methodsEleven melanoma patients eligible to undergo SLN biopsy were included in this prospective study. SLNs were fixed, optically cleared, whole-mount stained and imaged using light sheet fluorescence microscopy (LSFM). Subsequently, compatible and unbiased gold standard histopathological assessment allowed regular patient staging. This enabled intrasample comparison of LSFM and histological findings. In addition, the development of an algorithm, RAYhance, enabled easy-to-handle display of LSFM data in a browsable histologic slide-like fashion. ResultsWe comprehensively quantify total tumour volume while simultaneously visualising cellular and anatomical hallmarks of the associated SLN architecture. In a first-in-human study of 21 SLN of melanoma patients, LSFM not only confirmed all metastases identified by routine histopathological assessment but also additionally revealed metastases not detected by routine histology alone. This already led to additional therapeutic options for one patient. ConclusionOur three-dimensional digital pathology approach can increase sensitivity and accuracy of SLN metastasis detection and potentially alleviate the need for conventional histopathological assessment in the future. German clinical trials register(DRKS00015737).

Dibenzoylmethane derivative inhibits melanoma cancer in vitro and in vivo through induction of intrinsic and extrinsic apoptotic pathways

Malignant melanoma has a low incidence, but is the most lethal type of skin cancer. Studies have shown that dibenzoylmethanes (DBMs) have interesting biological activities, including antineoplastic properties. These findings led us to investigate whether news DBM derivatives exert antitumor effects against skin cancers. In a previous study, we found that 1,3-diphenyl-2-benzyl-1,3-propanedione (DPBP) has high in vitro antineoplastic activity against murine B16F10 melanoma cells, with an IC50 of 6.25 μg/mL. In the current study, we used transdermal and topical formulations of DPBP to evaluate its activity and molecular mechanism of action in a murine model of melanoma. The compound induces tumor cell death with high selectivity (selectivity index of 41.94) by triggering apoptosis through intrinsic and extrinsic pathways. DPBP treatment reduced tumor volume as well as serum VEGF-A and uric acid levels. Hepatomegaly and nephrotoxicity were not observed at the tested doses. Histopathological analysis of sentinel lymph nodes revealed no evidence of metastases. According to the observed data, the DPBP compound was effective for the topical treatment of melanoma cancer, suggesting that it acts as a chemotherapeutic or chemopreventive agent.

In-depth proteomics analysis of sentinel lymph nodes from individuals with endometrial cancer

SummaryEndometrial cancer (EC) is one of the most common gynecological cancers worldwide. Sentinel lymph node (SLN) status could be a major prognostic factor in evaluation of EC, but several prospective studies need to be performed. Here we report an in-depth proteomics analysis showing significant variations in the SLN protein landscape in EC. We show that SLNs are correlated to each tumor grade, which strengthens evidence of SLN involvement in EC. A few proteins are overexpressed specifically at each EC tumor grade and in the corresponding SLN. These proteins, which are significantly variable in both locations, should be considered potential markers of overall survival. Five major proteins for EC and SLN (PRSS3, PTX3, ASS1, ALDH2, and ANXA1) were identified in large-scale proteomics and validated by immunohistochemistry. This study improves stratification and diagnosis of individuals with EC as a result of proteomics profiling of SLNs.

Sentinel lymph node processing in gynecological cancer histopathology and molecular biology.

As sentinel lymph nodes (SLNs) are the first nodes receiving drainage from primary tumors, they provide important prognostic information about the nodal status of a tumor. SLN biopsy has modified the lymph node assessment by pathologists. This review highlights the different ways of histopathological and molecular SLN assessment according to the different gynecological cancers. Other than in breast cancer and melanoma, frozen section (FS) analysis of SLN in gynecological malignancies is still considered an important diagnostic tool. Intraoperative evaluation of the SLN allows to determine the need of completing lymph node dissection in case of metastasis. Intraoperative FS has a high negative predictive value (NPV) and is more sensitive than imprint cytology (IC) alone. If on intraoperative examination on FS the SLN is negative, subsequent analysis of the entire lymph node and histological ultrastadification has the potential to detect occult low volume metastases or to ascertain that a SLN is really negative. This reduces the morbidity compared to systematic pelvic and paraaortic lymph node dissection. Inclusion of the entire lymph node tissue in paraffin blocks after cutting it in 2 mm thick slices and histopathological ultrastaging with serial sections provides important prognostic information about the need of adjuvant treatment. Three sections at 200-250 µm seem to identify the majority of micrometastases. This review discusses different histopathological protocols and molecular [qRT-PCR and one-step nucleic acid amplification (OSNA®)] aspects of SLN evaluation in gynecological cancer.

Abstract PS1-51: A nomogram for predicting three or more axillary lymph node involvement before breast cancer surgery

Abstract Introduction Focus has moved to de-escalate of axillary surgery in the management of early breast cancer. Based on the American College of Surgeons Oncology Group-Z0011, our team created the nomogram to be useful for identifying patients who do not require intraoperative analysis of the sentinel lymph node. The nomogram could identify as patients likely to three or more positive axillary lymph nodes by preoperative imaging. But the nomogram had several limitations. This study investigated a developed nomogram by excluding the chest computed tomography (CT) and adding 18F-Fluorodeoxyglucose (FDG) positron emission tomography (PET)/CT as a modality. Material and Methods Patients underwent preoperative ultrasonography (US) and PET/CT. The training set consisted of 1030 patients with clinical T1-2 and node-negative invasive breast cancer. Factors associated with ≥ 3 involved axillary lymph nodes (ALNs) were evaluated by logistic regression analysis. The validation set consisted of 781 independent patients. The nomogram was applied to 1,067 patients who met the selection criteria of the ACOSOG-Z0011. Results Of the 1,030 patients, 89 (8.6%) had ≥ 3 positive nodes. Multivariate analysis according to be used for making a new nomogram to predict the probability of involvement ≥ 3 nodes, there was significantly associated with larger tumor size, higher grade ultrasonographic ALN classification, and findings suspicious of positive ALN on PET/CT. The areas under the receiver operating characteristic curve of the nomogram were 0.856 (95% confidence interval [CI], 0.815 to 0.897) for the training set and 0.866 (95% CI, 0.799 to 0.934) for the validation set. Application of the nomogram to the patients who met the ACOSOG-Z0011 showed that 90/1067 (8.4%) had scores above the cut-off and the false-negative rate was 37/977 (3.8%). And the specificity was 93.8%, and the negative predictive value was 96.4%. Conclusion The upgraded nomogram improved predictive accuracy, and that was possible using only US and PET/CT. It reduced operation time and cost, with a very low re-operation rate. This nomogram is useful for identifying patients who do not require intraoperative analysis of sentinel lymph nodes. Citation Format: Young-Joon Kang. A nomogram for predicting three or more axillary lymph node involvement before breast cancer surgery [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS1-51.