Cancer Biopsies Research Articles

Semen sEV tRF-Based Models Increase Non-Invasive Prediction Accuracy of Clinically Significant Prostate Cancer among Patients with Moderately Altered PSA Levels.

PSA screening has led to an over-diagnosis of prostate cancer (PCa) and unnecessary biopsies of benign conditions due to its low cancer specificity. Consequently, more accurate, preferentially non-invasive, tests are needed. We aim to evaluate the potential of semen sEV (small extracellular vesicles) tsRNAs (tRNA-derived small RNAs) as PCa indicators. Initially, following a literature review in the OncotRF database and high-throughput small RNA-sequencing studies in PCa tissue together with the sncRNA profile in semen sEVs, we selected four candidate 5'tRF tsRNAs for validation as PCa biomarkers. RT-qPCR analysis in semen sEVs from men with moderately elevated serum PSA levels successfully shows that the differential expression of the four tRFs between PCa and healthy control groups can be detected in a non-invasive manner. The combined model incorporating PSA and specific tRFs (5'-tRNA-Glu-TTC-9-1_L30 and 5'-tRNA-Val-CAC-3-1_L30) achieved high predictive accuracy in identifying samples with a Gleason score ≥ 7 and staging disease beyond IIA, supporting that the 5'tRF fingerprint in semen sEV can improve the PSA predictive value to discriminate between malignant and indolent prostate conditions. The in silico study allowed us to map target genes for the four 5'tRFs possibly involved in PCa. Our findings highlight the synergistic use of multiple biomarkers as an efficient approach to improve PCa screening and prognosis.

Upgrading of Grade Group 1 Prostate Cancer at Prostatectomy: Germline Risk Factors in a Prospective Cohort.

Localized prostate tumors show significant spatial heterogeneity, with regions of high-grade disease adjacent to lower-grade disease. Consequently, prostate cancer biopsies are prone to sampling bias, potentially leading to underestimation of tumor grade. To study the clinical, epidemiologic and molecular hallmarks of this phenomenon, we conducted a prospective study of grade upgrading: differences in detected prostate cancer grade between biopsy and surgery. We established a prospective, multi-institutional cohort of men with Grade Group 1 (GG1) prostate cancer on biopsy who underwent radical prostatectomy. Upgrading was defined as detection of GG2+ in the resected tumor. Germline DNA from 192 subjects was subjected to whole-genome sequencing to quantify ancestry, pathogenic variants in DNA damage response genes and polygenic risk. Of 285 men, 67% upgraded at surgery. PSA density and percent of cancer in pre-prostatectomy positive biopsy cores were significantly associated with upgrading. No assessed genetic risk factor was predictive of upgrading, including polygenic risk scores for prostate cancer diagnosis. In a cohort of low-grade prostate cancer patients, a majority upgraded at radical prostatectomy. PSA density and percent of cancer in pre-prostatectomy positive biopsy cores portended the presence of higher-grade disease, while germline genetics was not informative in this setting. Patients with low-risk prostate cancer, but elevated PSA density or percent cancer in positive biopsy cores, may benefit from repeat biopsy, additional imaging or other approaches to complement active surveillance. Further risk stratification of patients with low-risk prostate cancer may provide useful context for active surveillance decision-making.

Results of a Prospective Randomized Multicenter Study Comparing Indocyanine Green (ICG) Fluorescence Combined with a Standard Tracer Versus ICG Alone for Sentinel Lymph Node Biopsy in Early Breast Cancer: The INFLUENCE Trial.

For clinically node-negative early breast cancer patients, sentinel lymph node biopsy (SLNB) using dual localization with blue dye and radioisotope (RI) is currently standard of care. Documented disadvantages with these tracers have prompted exploration of alternative agents such as fluorescent indocyanine green (ICG), which demonstrates high detection rates combined with other tracers. Results of a randomized study evaluating ICG as a single tracer for SLN identification are presented. Overall, 100 patients with unilateral, clinically node-negative, biopsy-proven invasive breast cancer (≤5cm) scheduled for SLNB were recruited in two separate randomized cohorts, with 50 patients receiving ICG alone. Cohort 1 received ICG alone (n=25) or combined with RI [Technetium99] (n=25), while Cohort 2 received ICG alone (n=25) or combined with blue dye (n=25). The primary outcome was sensitivity for SLN identification. Among evaluable patients (n=97), the overall SLN identification rate was 96.9% (ICG alone = 97.9%; ICG + RI = 100%; ICG + blue dye = 92%). Node positivity rates were 14.9% for ICG alone, 16% for ICG combined with RI, and 20% for ICG combined with blue dye. There were no significant differences (p<0.05) in performance parameters, with ICG alone being non-inferior to tracer combinations for procedural node positivity rates when adjusted for specific factors. These results support potential use of ICG as a sole tracer agent for routine SLNB, thereby avoiding disadvantages of RI and/or blue dye. The latter can be safely withheld as a co-tracer without compromising detection of positive nodes in primary surgical patients.

Liquid biopsy in prostate cancer: A novel dual biomarker analysis approach integrating circulating tumor cells and circulating tumor DNA data.

This study aimed to explore the potential of liquid biopsy as a diagnostic tool by integrating two key biomarkers, Circulating Tumor Cells (CTCs) and Circulating Tumor DNA (ctDNA), and to enhance the detection fidelity of prostate cancer. A dual biomarker analysis approach was employed to synergize the sensitivities of CTCs and ctDNA. Various genetic mutations of ctDNA and tissues were scrutinized, investigating their prevalence, co-existence, and mutual exclusivity. The findings uncovered a more intricate mutation landscape than previously anticipated, indicating a complex interplay between cellular and genetic aberrations in prostate cancer. Through harnessing the combined power of CTCs and ctDNA, our dual biomarker approach provides a more comprehensive understanding of prostate cancer genetics. This has the potential to revolutionize early detection and guide personalized therapeutic interventions.

Concordance of PD-L1 Positivity Between Fine-Needle Aspiration Biopsy and Histologic Specimens in Metastatic Triple Negative Breast Cancer

Concordance of PD-L1 Positivity Between Fine-Needle Aspiration Biopsy and Histologic Specimens in Metastatic Triple Negative Breast Cancer

Application of Lightweight Deep Learning Model-Based Shear Wave Elastic Imaging with Abdominal Probe to Diagnose Prostate Cancer: A Biomedical Approach

We aimed to investigate the diagnostic value of lightweight convolutional neural network (CNN) model abdominal probe shear wave elastic imaging (SWE) in the perineal diagnosis and guided puncture biopsy of prostate cancer (PCa), and to provide reference for the clinical diagnosis of PCa. 100 PCa patients were assigned to group I (malignant) and group II (benign), with 50 cases in each. Ultrasonic elastic imaging based on lightweight convolutional neural network denoising model was adopted for detection. In both systolic and diastolic blood pressure (SBP/DBP), there was not a significant intergroup difference (P &gt; 0.05). The levels of prostate specific antigen (PSA) and its free variant (fPSA) in group II were markedly lower (P &lt; 0.05). Patients in group II had obviously more cystic components and fewer solid components. Patients with hyperechogenicity was more in group II. Patients had clearly fewer irregular margins and outward margin spread in group II. Patients without focal hyperechogenicity and punctate hyperechogenicity was more in group II, and the number of calcifications in group II was less. Patients with type 0 and type I was more and patients with type IIa and type IIb was less in group II. The Emean level of patients in group II was clearly higher, and the Emax level and Esd level of patients in group II were clearly lower. The SI level of patients was clearly lower in group II TTP was higher in group II (P &lt; 0.05). Multivariate logistic regression analysis of abdominal probe SWE for transperineal diagnosis of PCa and guided puncture biopsy showed that internal echoes had the greatest OR and were associated with the occurrence of PCa. Ultrasonic elastic imaging index based on the lightweight convolutional neural network denoising model can be used for the benign and malignant diagnosis of PCa patients.

Clinical and pathological features of renal tumours among women previously treated for breast carcinomas – the CanSeRe study

IntroductionRenal cell carcinoma (RCC) and breast carcinoma (BC) are frequent tumours, yet their co-occurrence in the same patient is a unique scenario. Few studies explored the characteristics of such patients without specific focus on pathological data. In this retrospective study, we aimed to describe the clinico-pathological features of RCC patients with a history of BC and compare them to a control cohort of RCC women free of previous BC. MethodsAll adult women treated for BC at a high-volume cancer institution between 2007 and 2020 and who subsequently developed a RCC were retrospectively included. Their clinical and pathological characteristics were compared to an independent cohort of consecutive women undergoing percutaneous kidney tumour biopsy for localized kidney cancer in a second high-volume cancer institution. ResultsA total of 113 patients were identified from 2 different institutions. We observed a lower rate of clear cell RCC in the Kidney-breast (KB) group compared to the Kidney-only (KO) group, suggesting a potential association between breast cancer and non-ccRCC. The KB group had a higher proportion of locally advanced tumours and high-grade lesions. Although recurrence-free survival favored the KO cohort, no significant difference was found in cancer-specific survival and overall survival rates between the groups. Noteworthy, patients in the KB group had a higher prevalence of family history of cancer. ConclusionOur findings highlight the need for further research to elucidate the underlying mechanisms and clinical implications of RCC coexisting with breast carcinoma. Understanding the characteristics of this unique population can guide clinical strategies and improve patient outcomes. Level of evidence4.

402.1: Minimizing risks in hepatocellular cancer biopsy: Coaxial needle technique with tract cauterization.

402.1: Minimizing risks in hepatocellular cancer biopsy: Coaxial needle technique with tract cauterization.

The probability of implantation metastasis after peripheral lung cancer biopsy.

At present, it is known that there is a possibility of pleural cavity and needle tract implantation metastasis after lung cancer puncture biopsy, but clinicians have not paid attention to this phenomenon, and the probability of occurrence is unknown. In this study, we aimed to study the probability of implantation metastasis after peripheral lung cancer biopsy. The intraoperative isolated completely collapsed fresh intact human lung lobes of 30 patients with peripheral lung cancer were taken, and the tumor body was punctured. The pleural pinholes and puncture needle tips were repeatedly rinsed with normal saline respectively. The flushing solution was prepared as inoculum, and then the inoculum was smeared for microscopic examination to find cancer cells. The inoculum was inoculated subcutaneously into nude mice, and then the probability of pleural cavity and needle tract implantation metastasis after lung biopsy was indirectly obtained by obtaining the nude mice tumorigenesis rate. The tumorigenesis rate of nude mice in the pleural pinholes group was 3.3%, and the tumorigenesis rate of nude mice in the puncture needle tip group was 3.3%. Patients with lung nodules suspected of being early stage cancer and who are fit for surgical resection may benefit from forgoing lung biopsies to avoid pleural cavity or needle tract seeding with tumor cells.

Surgical and irradiated case of early breast cancer in a patient with Ehlers–Danlos syndrome

BackgroundEhlers–Danlos syndrome (EDS) is a rare inherited connective tissue disease characterized by hyperextensibility of the skin and joints and tissue fragility of the skin and blood vessels, Vascular EDS is the most severe form of EDS, with abnormal arterial fragility. There have been no reports of breast cancer occurring in patients with vascular EDS. Here, we report here a very rare case of breast cancer in a patient with vascular EDS.Case presentationA 46-year-old woman with vascular EDS underwent partial left mastectomy and sentinel lymph node biopsy for left breast cancer (cStage 0) detected by medical examination. The final pathological diagnosis was invasive ductal carcinoma of the breast (pStage IA) [hormone receptor-positive, HER2 score 2 equivocal (FISH-positive), Ki-67LI 18%, luminal-HER2 type]. BluePrint was submitted as an aid in determining the postoperative treatment strategy, BluePrint Molecular Subtype HER2-type. However, the 10-year breast cancer mortality risk using Predict was low (5%). After consultation with the patient, the decision was made to administer postoperative radiation to the preserved breast along with hormone therapy only. There was no delay in postoperative wound healing, and the patient was free of metastatic recurrence for 9 months after surgery.ConclusionWe performed surgery, postoperative radiotherapy, and hormonal therapy in a breast cancer patient with vascular EDS without major complications.

Unexpected Expression and Function of FcεRI in Immortalized Breast Cancer Cells: A Cautionary Null Study.

The high-affinity IgE receptor, FcεRI, is typically associated with type 2 effectors such as mast cells (MC). The relatively unique expression profile of FcεRI and accumulating evidence from pre-clinical and clinical settings, such as MC interactions with tumors, have led us to study MCs as a potential therapeutic target in breast cancer. Our work identified MCs interacting with tumor cells at primary sites using the 4T1 (BALB/c) adenocarcinoma model in vivo. However, this analysis was complicated by a surprising finding that the tumor cells intrinsically and strongly expressed FcεRI. We further studied the expression and function of FcεRI in breast cancer cells in vitro. The 4T1 cells expressed FcεRI to a level similar to mouse bone marrow-derived MC (BMMC). Additionally, two established breast cancer cultures derived from human T-47D cells, one estrogen-dependent (E3) and the other estrogen-withdrawn (EWD8), also expressed FcεRI with EWD8 cells showing the greatest abundance. Functional analyses indicated that IgE-mediated antigen stimulation did not elicit classic Ca2+ flux in breast cancer cells as seen in the respective species' MCs; however, FcεRI crosslinking could stimulate IL-6 production from the T-47D derivatives. Preliminary analysis of primary breast cancer biopsy datasets using R2: Genomics Analysis and Visualization Platform was discordant with our in vivo model and in vitro observations. Indeed, FcεRI mRNA abundance declined in metastatic breast cancers compared to non-cancerous breast tissue. Altogether, we report a previously unidentified and immunologically substantive difference between breast cancer models and human primary tumors. Investigators pursuing FcεRI-relevant therapeutics in this context should be aware of this translational barrier.

A Green Lantern for the Surgeon: A Review on the Use of Indocyanine Green (ICG) in Minimally Invasive Surgery.

Indocyanine green (ICG) fluorescence imaging has revolutionized surgical practice across various medical and surgical specialties. This article reviews the clinical applications of ICG in abdominal, urological, thoracic, and gynecological surgery. ICG fluorescence imaging has been widely adopted in general surgery for various applications, including perfusion assessment, intraoperative visualization of the ureter, and tumor localization. It is particularly valuable in evaluating anastomotic leaks and aiding in precise tumor resection during minimally invasive surgeries. Studies have shown mixed results on its effectiveness in reducing anastomotic leak rates, highlighting the need for further research. In thoracic surgery, ICG facilitates the identification and resection of pulmonary bullae, as well as the precise localization of pulmonary nodules during video-assisted surgery. In urology, ICG aids in localizing renal tumors and guiding selective arterial occlusion during partial nephrectomy. Its role in identifying the lymphatic pathway in prostate cancer and sentinel lymph node biopsy in gynecological cancer is also discussed. Despite its benefits, the use of ICG fluorescence faces challenges such as limited tissue penetration, the potential for false results, a lack of standardized protocols, and high equipment costs. Nonetheless, it remains a powerful tool that could improve surgical outcomes.

Diagnostic properties of miR-146a-5p from liquid biopsies in prostate cancer: A meta-analysis

BackgroundSeveral studies on biomarker properties of microRNAs from liquid biopsy in prostate cancer (PCa) identified miR-146a-5p as a potential novel diagnostic marker. However, other studies with the same or similar topic failed to confirm the supposed discriminatory ability of miR-146a-5p, for which reason we aimed at elucidating the potential biomarker role of circulatory/urinary miR-146a-5p in PCa by conducting a qualitative and quantitative data synthesis. MethodsEligible articles were identified by searching PubMed, Scopus and Web of Science databases. Open MetaAnalyst software was used for pooling data on sensitivity, specificity, likelihood ratio and diagnostic odds ratio (OR) of miR-146a-5p. ResultsA total of 15 articles were eligible for qualitative data synthesis, while the results from 13 studies with 2080 participants were included in the meta-analysis. The established between-study heterogeneity was high, while the expression of hsa-miR-146a was associated with a diagnostic OR of 3.544 (P < 0.001; 95 %CI 2.186–5.747). Pooled sensitivity was found to be lower than 70 % (0.655, 95 %CI 0.573–0.729, P < 0.001), while the obtained value for specificity was 65 % (95 %CI 0.583–0.709, P < 0.001). Segregating studies according to ethnicity, sample type or the type of controls did not result in significantly higher sensitivity and specificity in subgroups, compared to the overall pooled data. ConclusionsThe resulting pooled sensitivity, specificity and diagnostic OR do not qualify miR-146a-5p for a reliable diagnostic biomarker of PCa.

Circulating small extracellular vesicle RNA profiling for the detection of T1a stage colorectal cancer and precancerous advanced adenoma.

It takes more than 20 years for normal colorectal mucosa to develop into metastatic carcinoma. The long time window provides a golden opportunity for early detection to terminate the malignant progression. Here, we aim to enable liquid biopsy of T1a stage colorectal cancer (CRC) and precancerous advanced adenoma (AA) by profiling circulating small extracellular vesicle (sEV)-derived RNAs. We exhibited a full RNA landscape for the circulating sEVs isolated from 60 participants. A total of 58,333 annotated RNAs were detected from plasma sEVs, among which 1,615 and 888 sEV-RNAs were found differentially expressed in plasma from T1a stage CRC and AA compared to normal controls (NC). Then we further categorized these sEV-RNAs into six modules by a weighted gene coexpression network analysis and constructed a 60-gene t-SNE model consisting of the top 10 RNAs of each module that could well distinguish T1a stage CRC/AA from NC samples. Some sEV-RNAs were also identified as indicators of specific endoscopic and morphological features of different colorectal lesions. The top-ranked biomarkers were further verified by RT-qPCR, proving that these candidate sEV-RNAs successfully identified T1a stage CRC/AA from NC in another cohort of 124 participants. Finally, we adopted different algorithms to improve the performance of RT-qPCR-based models and successfully constructed an optimized classifier with 79.3% specificity and 99.0% sensitivity. In conclusion, circulating sEVs of T1a stage CRC and AA patients have distinct RNA profiles, which successfully enable the detection of both T1a stage CRC and AA via liquid biopsy.

CCL2/CCR2 Expression in Locally Advanced Prostate Cancer and Patient Long-Term Outcome: 10-Year Results from the TROG 03.04 RADAR Trial.

This study investigated the prognostic value of the chemokine C-C motif ligand 2 (CCL2) and its receptor C-C motif chemokine receptor 2 (CCR2) expression in locally advanced prostate cancer treated with radiotherapy and androgen deprivation using the 10-year outcome data from the TROG 03.04 RADAR clinical trial. CCL2 and CCR2 protein expression in prostate cancer biopsies at the time of diagnosis were quantified by immunohistochemistry and digital quantification. CCR2 protein expression was detected in prostate cancer cells and was associated with prostate-specific antigen serum concentration (p = 0.045). However, neither CCL2 nor CCR2 tissue expression could predict prostate cancer progression, or other clinicopathological parameters including perineural invasion and patient outcome. In serum samples, CCL2 concentration at the time of diagnosis, as assayed by enzyme-linked immunosorbent assay, was significantly higher in patients with prostate cancer compared with benign prostatic hyperplasia (median difference 0.22 ng/mL, 95% CI, 0.17-0.30) (p < 0.0001) and normal controls (median difference 0.13 ng/mL, 95% CI, 0.13-0.17) (p < 0.0001). However, circulating CCL2 was not statistically significant as a predictor of disease progression and patient outcome. In conclusion, this study shows that although CCL2 and CCR2 are expressed in prostate cancer, with an increased level of CCL2 in the serum, neither CCL2 nor CCR2 expression has a clinical prognostic value in locally advanced prostate cancer.

Examining breast cancer screening recommendations in Canada: The projected resource impact of screening among women aged 40-49.

To quantify the resource use of revising breast cancer screening guidelines to include average-risk women aged 40-49 years across Canada from 2024 to 2043 using a validated microsimulation model. OncoSim-Breast microsimulation platform was used to simulate the entire Canadian population in 2015-2051. We compared resource use between current screening guidelines (biennial screening ages 50-74) and alternate screening scenarios, which included annual and biennial screening for ages 40-49 and ages 45-49, followed by biennial screening ages 50-74. We estimated absolute and relative differences in number of screens, abnormal screening recalls without cancer, total and negative biopsies, screen-detected cancers, stage of diagnosis, and breast cancer deaths averted. Compared with current guidelines in Canada, the most intensive screening scenario (annual screening ages 40-49) would result in 13.3% increases in the number of screens and abnormal screening recalls without cancer whereas the least intensive scenario (biennial screening ages 45-49) would result in a 3.4% increase in number of screens and 3.8% increase in number of abnormal screening recalls without cancer. More intensive screening would be associated with fewer stage II, III, and IV diagnoses, and more breast cancer deaths averted. Revising breast cancer screening in Canada to include average-risk women aged 40-49 would detect cancers earlier leading to fewer breast cancer deaths. To realize this potential clinical benefit, a considerable increase in screening resources would be required in terms of number of screens and screen follow-ups. Further economic analyses are required to fully understand cost and budget implications.

Comparison of state-of-the-art biopsy systems for ultrasound-guided breast biopsy using a chicken breast phantom.

To compare different biopsy systems with different-sized needles by determining the weight of the tissue cores, which is one of the important factors for precise pathological diagnoses, and to provide a rationale for choosing the appropriate breast biopsy system with the appropriate needle for breast cancer biopsy. Six different vacuum-assisted biopsy (VAB) systems and one core needle biopsy (CNB) system with different-sized needles in different modes were compared, representing 15 total combinations. Tissue cores were obtained from a chicken breast phantom, which is a common substitute for human breast tissue. Five cores were taken for each combination and weighed. The CNB combination provided significantly lighter tissue cores compared with the VAB combinations with the same-size (14-G) needle (P < 0.01). The combinations using the thickest needle obtained the heaviest among all systems (P < 0.02). The untethered battery-free VAB system yielded the lightest specimen among the VAB systems with the same-sized (12-G) needle (P < 0.04). The percent coefficient of variation (%CV) of the core weights obtained using VAB without a basket was significantly smaller compared with the core weights obtained using VAB with a basket (P < 0.01). VAB systems can yield larger tissue cores compared with CNB systems. The size of the tissue cores varies even with the same-sized needle among different VAB systems. When performing a breast tissue biopsy, it is important to consider not only CNB versus VAB but also what specific device to use with which needle size.

High early mortality after percutaneous liver biopsy in metastatic cancer: national analysis

ObjectiveThe study aimed to assess outcomes in patients undergoing liver biopsy for metastatic cancer, focusing on mortality rates and chemotherapy following their biopsy.MethodsHospital Episode Statistics data from 2010 to 2019...

The clinical value of carbon nanoparticles in sentinel lymph node biopsy for early vulvar cancer

ObjectiveCarbon nanoparticle (CNP)-guided sentinel lymph node biopsy (SLNB) has been extensively adopted as a cost-effective and highly efficient method for tracing malignant tumors except for those associated with vulvar cancer. The current study aimed to validate the feasibility and efficacy of CNPs in tracking sentinel lymph nodes (SLNs) in patients with early vulvar cancer. MethodsWe retrospectively reviewed patients with vulvar cancer at our institution from January 2016 to April 2022 who were pathologically diagnosed and underwent SLNB or inguinofemoral lymphadenectomy (IFLND). CNPs were the only lymphatic tracer used in SLNB. Patient demographics, perioperative outcomes and follow-up results, including overall survival (OS) and progression-free survival (PFS), were compared between the SLNB and IFLND groups. ResultsData from 52 patients were collected and investigated. Forty groins of 22 patients who underwent SLNB with CNP tracing were included. Black-stained SLNs were detected in 32 groins of 19 patients, and the rates of CNP detection by patient and by groin were 86.4 % and 80 %, respectively. Patients who underwent SLNB had better perioperative outcomes than those who underwent IFLND in certain aspects (groin drainage rate: 41.2 % and 80 %, respectively, p < 0.05; daily drainage volume (ml): 12.49 and 36.4, respectively, p < 0.05; and inguinal wound healing rate: 100 % and 80 %, respectively, p < 0.05). The results of survival analysis indicated similar prognoses for node-negative patients who underwent CNP-guided SLNB or IFLND. ConclusionsSentinel lymph node mapping with CNPs in vulvar cancer is feasible and demonstrates considerable biosecurity. With a satisfactory SLN detection rate achieved expediently, CNPs are a promising lymphatic tracer worthy of further utilization in vulvar cancer and could be an alternative option to canonical tracers.

The surgical technique and protocol for dynamic sentinel node biopsy for penile cancer at a Southeast Asian regional hospital.

Lymph node status is a key prognostic factor in penile cancer. The European Association of Urology (EAU) recommends intermediate-risk (pT1a, Grade 2) or high-risk (pT1b or greater) penile cancer patients with clinically non-palpable inguinal lymph node (cN0) to undergo either an invasive bilateral modified inguinal lymph node dissection (ILND) or dynamic sentinel node biopsy (DSNB). DSNB has been reported to have acceptable false negative rates, and lower rates of long-term morbidity compared to ILND. We developed a protocol for DSNB at a regional hospital in Singapore that was adopted from St James's University Hospital, Leeds Teaching Hospitals Trust. Four patients with cN0 penile cancer underwent DSNB between November 2021 and October 2022 according to this protocol. Our surgical technique and protocol are described. The patients' oncological characteristics and their outcomes were evaluated. In this small case series, there was no complication attributable to the performance of DSNB, and there was no groin that was documented to be false negative over a median follow up of 15.5 months (range, 12 to 22 months). Using our protocol, 5 of 8 groins (62.5%) were able to avoid ILND in the cN0 setting. We recommend the adoption of DSNB for the surgical staging of inguinal lymph nodes for patients with intermediate to high-risk penile cancer and non-palpable inguinal nodes due to its significantly lower risks of long-term morbidity compared to ILND. Appropriate specialist training and a multi-disciplinary team is vital to ensure the success of the procedure.