Excised Tumor Specimens Research Articles

Cutting-edge insights: near-infrared imaging for surgical margin assessment in head and neck tumor resection: a systematic review and meta-analysis.

In head and neck cancer (HNC), real-time evaluation of tumor margin status following surgical excision of the tumor is of critical importance. This systematic review aimed to assess the effectiveness of near-infrared fluorescence (NIRF) imaging for the real-time delineation of tumor margins in HNC resections. Two investigators independently conducted a comprehensive search following the Preferred Reporting Items for a Systematic Review and Meta-analysis of Diagnostic Test Accuracy Studies (PRISMA-DTA) guidelines across the PubMed, Scopus, Embase, and China National Knowledge Infrastructure (CNKI) databases until August 1st, 2023. Observational studies were included, while other studies with inappropriate study design were excluded. The primary outcomes included the specificity, sensitivity, and area under the summary receiver operating characteristic (SROC) curve when using NIRF imaging to assess surgical margins. We compared fluorescence in the resection specimen margins and residual fluorescence in the surgical cavity margins as methods of utilizing NIRF to evaluate surgical margins. Diagnostic trial quality was assessed, and statistical heterogeneity was determined. The initial search yielded 1,607 articles. After reviewing the full texts, seven articles with 103 patients were included, among which five were incorporated for quantitative analysis. The selected studies had an average score of 10.1 of quality. Heterogeneity analysis revealed I2 values of 26% [95% confidence interval (CI): 0-100%] and 78% (95% CI: 52-100%) for NIRF specimen imaging with close margin considered positive or negative. Comparing NIRF imaging to the gold standard of pathology for surgical margin diagnosis, with close margin considered positive, sensitivity and specificity in excised specimens were 0.84 (95% CI: 0.39-0.98) and 0.96 (95% CI: 0.80-0.99), respectively. When a close margin was considered negative, sensitivity and specificity were 0.98 (95% CI: 0.10-1.00) and 0.96 (95% CI: 0.45-1.00), respectively. The areas under the SROC curves were 0.97 (95% CI: 0.95-0.98) and 0.99 (95% CI: 0.98-1.00), respectively. A quantitative analysis of residual fluorescence at surgical cavity margins was not performed due to an insufficient number of studies. NIRF imaging is a promising method for real-time surgical margin assessment of HNC. With its robust diagnostic capabilities in excised tumor specimens, it is also an effective technique for detecting residual tumor fluorescence in the surgical cavity for supplementary assessment. But the results should be interpreted with caution.

Oxidative Status and Lipid Metabolism Analytes in Dogs with Mast Cell Tumors: A Preliminary Study

Mast cell tumors (MCTs) are common skin neoplasms in dogs. Prognostic indicators include histologic grade, clinical stage, high Ki-67 index, elevated argyrophilic nucleolus organizer regions (AgNOR) index, c-kit mutations, and recurrence after surgery. Blood serum redox status has been shown to correlate with prognostic factors in canine lymphoma and mammary tumors. This study aimed to assess the correlation between established prognostic factors and serum redox status and lipid metabolism analytes in dogs with MCTs. Dogs with cutaneous (n = 33) or subcutaneous (n = 6) MCTs, without comorbidities, were studied. Staging was evaluated based on cytology of regional lymph nodes and ultrasound-guided liver and spleen aspiration cytology. Histologic grading and immunohistochemical staining for Ki-67 and KIT patterns were performed on excised tumor specimens. Dogs were categorized by Patnaik grading (1–3), Kiupel grading (low/high), metastatic status, Ki-67 positive nuclei per cm2 (>23 or ≤23), and KIT pattern (I, II–III). Paraoxonase-1, Butyrylcholinesterase, Cupric Reducing Antioxidant Capacity (CUPRAC), Diacron Reactive Oxygen Metabolites (d-ROMs), and oxy-adsorbent levels were measured before any therapeutic intervention. ANOVA and independent t-tests were used to detect differences in the mean values among groups. Paraoxonase-1 activity was significantly lower in Patnaik grade 3 (p = 0.003) and Kiupel high-grade (p = 0.022) MCTs. No significant differences were found in CUPRAC, d-ROMs, or oxy-adsorbent levels across different prognostic groups. This study found a significant correlation between histologic grading and Paraoxonase-1 activity, suggesting a potential role of Paraoxonase-1 as a prognostic biomarker in canine MCTs.

Expression of Heat Shock Protein 90 in Testicular Cancer: A Retrospective Cohort Study.

The HSP90 marker is believed to play a constructive role in facilitating neoplastic transformation mainly via interaction with multiple pro-survival proteins. Welldesigned studies are needed to elucidate the role of HSP90 as a diagnostic marker and therapeutic target in testicular tumors. The current study aimed to investigate the expression of HSP90 in various types of testicular cancer and highlight its expression in embryonal testicular cancer. Immunohistochemical staining for HSP90 in 84 male patients, with nonmetastatic testicular cancer, who underwent orchiectomy from 2000 to 2023, was retrospectively performed at the Laboratory Department of General Hospital of Nikaia in Greece. A total of 84 males, with a mean age of 36.2 years, who have undergone high-cord radical orchiectomy, were included in this study. Out of the included males, 28.57% had embryonal carcinoma, 23.81% had seminoma, 19.05% had yolk sac tumor, 11.9% had mature teratoma, 9.52% had immature teratoma, and 7.14% had choriocarcinoma. HSP90b was positive in all embryonal carcinoma, seminoma, and choriocarcinoma cases, while it was positive in 75% of the yolk sac tumor, 75% of mature teratoma, and 75% of immature teratoma specimens. HSP90 was found negative in all choriocarcinoma, mature teratoma, and immature teratoma specimens, while it was positive in 25% of yolk sac tumor, 8.33% of embryonal carcinoma, and 10% of seminoma cases. Concerning the expression of HSP90b, a statistically significant relationship was found between excised tumor specimens and normal parenchyma specimens, especially in sac cases (p <0.001). Regarding HSP90a expression, a statistically significant relationship (OR=21.5, p =0.021) was found between excised tumor specimens and normal parenchyma specimens, especially in embryonal carcinoma cases (p <0.001). HSP90b is highly expressed in the majority of the types of testicular tumors, both in tumor and normal parenchyma specimens, while HSP90a staining is negative in resected specimens. Further well-designed studies are needed to elucidate the role of HSP90 as a diagnostic marker and therapeutic target in testicular tumors.

Apoptotic Marker Expression of Resected Lacrimal Gland Adenoid Cystic Carcinoma Tumor Margins After Intra-arterial Chemotherapy and Globe-Sparing Excision.

Lacrimal gland adenoid cystic carcinoma (LGACC) is a rare orbital malignancy with devastating lethality. Neoadjuvant intra-arterial chemotherapy (IACC) has demonstrated cytoreductive effects on LGACC macroscopically, but limited studies have examined cellular and molecular determinants of the cytoreductive effect. This post hoc study assessed apoptotic marker expression on excised tumor specimens after neoadjuvant IACC and globe-sparing resection, emphasizing the examination of tumor margins. This retrospective study identified LGACC specimens resected in a globe-sparing technique after neoadjuvant IACC by reviewing the Florida Lions Ocular Pathology database at Bascom Palmer Eye Institute. Histopathology slides of the specimens were re-examined to confirm the diagnosis and identify the tumor margin. Immunofluorescent staining was performed for apoptotic markers, including P53, cleaved caspase-3, cleaved PARP-1, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Positive expression was determined by comparison to the negative control. Tumor specimens from 5 patients met inclusion criteria. All 5 cases were positive at the center and the margin for TUNEL, p53, and cleaved caspase-3. One case did not show positive expression of cleaved PARP-1 at the margin but was positive for the other apoptotic markers. This post hoc study demonstrated positive staining for multiple apoptotic markers in post-IACC tumor specimens at the tumor center and margin. Apoptotic marker expression along the margins of post-treatment specimens is important, as it may offer surrogate information to speculate on the state of residual cancer cells adjacent to the excision margin inadvertently remaining in the orbit.

The efficacy of dermoscopy in defining the surgical margins of cutaneous squamous cell carcinoma: a retrospective study.

To investigate the diagnostic value of dermoscopy in defining the tumor margin of cutaneous squamous cell carcinoma (cSCC) for the appropriate surgical margin. A total of 90 cSCC patients were enrolled in the study. All patients were recruited into two groups: those who preserved intact macroscopic features of neoplasms without or after incisional biopsy and those with uncertain residual tumors after excisional biopsy. A dermoscopy-defined surgical margin of 8mm outward was used according to the tumor boundaries observed with the naked eye and dermoscopy. All excised tumor specimens were divided into serial sections according to the four "3, 6, 9, 12" directions at every 4-mm interval from the dermoscopy-detected tumor margin. Pathological examination was performed at 0mm, 4mm, and 8mm margins to confirm tumor remnants. Retrospective analysis of dermatoscopic results showed inconsistent clinical and dermatoscopic borders in 43 of 90 cases (47.8%). The ability of dermoscopy to detect tumor borders showed no statistical difference between the two groups (p > 0.05). In the unbiopsy or incisional biopsy group, 66.6% of the tumors were resected with a 4-mm margin and 98.3% with an 8-mm margin, with significant differences (p = 0.047). For patients with inconspicuous clinical evidence of residual tumor after excisional biopsy, the tumor clearance rate was 53.3% at 0mm, 93.3% at 4mm, and 100.0% at 8mm. Statistically significant differences were noted between 0mm and 4mm (p = 0.017), as well as between 0mm and 8mm (p = 0.043) but did not differ between 4mm and 8mm (p>0.05). Dermoscopy defined the tumor margin of cSCC better than visual inspection alone. Direct dermoscopic-guided surgery with at least 8-mm expansion was recommended for high-risk cSCC. Dermoscopy also assisted in identifying surgical margins at the healing biopsy site, making 8mm still the recommended expansion range.

Invasive lobular carcinoma enclosed by a benign phyllodes tumor: A case report.

Introduction and importancePhyllodes tumors (PT) account for approximately 1% of all breast tumors. The coexistence of phyllodes tumor and invasive lobular carcinoma in the ipsilateral breast is extremely rare with fewer than six cases reported worldwide in the last 20 years. We hereby present the first in Taiwan.Case presentationA 43-year-old female was presented with a protruding tumor with bleeding tendency over left breast in 2016. Breast sonography revealed highly suspected malignancy (ACR BI-RADS category 5). Computed tomography scan disclosed a protruding mass occupying the left breast. Core needle biopsy showed a fibroepithelial lesion favoring fibroadenoma. Considering clinicopathological discrepancy, a nipple-sparing mastectomy was conducted. Pathology report revealed a benign PT with an incidental finding of invasive lobular carcinoma (pT1cN0) within the tumor. Due to tumor recurrence, the patient received re-operation of total mastectomy in 2017, and is under regular adjuvant hormonal therapy without cancer recurrence to date.Clinical discussionPhysicians could easily overlook carcinomas enclosed by PTs due to its occult property. However, when carcinomatous changes arise from within or along with the PT, the proposed therapeutical course may be altered. Moreover, invasive carcinoma components in PTs possess potential for lymph node metastasis. Multidisciplinary cooperation is key in detecting and managing PT with synchronous carcinomatosis.ConclusionThorough examination of the excised tumor specimen and ensuring an adequate surgical margin is necessary. Sentinel lymph node biopsy (SLNB) should be considered whenever suspicious clinical features occur in PT patients. This may aid in the detection of microscopic invasive carcinomatous change.

Artificial Intelligence-based Tumor Segmentation in Mouse Models of Lung Adenocarcinoma

BackgroundMouse models are highly effective for studying the pathophysiology of lung adenocarcinoma and evaluating new treatment strategies. Treatment efficacy is primarily determined by the total tumor burden measured on excised tumor specimens. The measurement process is time-consuming and prone to human errors. To address this issue, we developed a novel deep learning model to segment lung tumor foci on digitally scanned hematoxylin and eosin (H&E) histology slides. MethodsDigital slides of 239 mice from 9 experimental cohorts were split into training (n=137), validation (n=37), and testing cohorts (n=65). Image patches of 500×500 pixels were extracted from 5× and 10× magnifications, along with binary masks of expert annotations representing ground-truth tumor regions. Deep learning models utilizing DeepLabV3+ and UNet architectures were trained for binary segmentation of tumor foci under varying stain normalization conditions. The performance of algorithm segmentation was assessed by Dice Coefficient, and detection was evaluated by sensitivity and positive-predictive value (PPV). ResultsThe best model on patch-based validation was DeepLabV3+ using a Resnet-50 backbone, which achieved Dice 0.890 and 0.873 on validation and testing cohort, respectively. This result corresponded to 91.3 Sensitivity and 51.0 PPV in the validation cohort and 93.7 Sensitivity and 51.4 PPV in the testing cohort. False positives could be reduced 10-fold with thresholding artificial intelligence (AI) predicted output by area, without negative impact on Dice Coefficient. Evaluation at various stain normalization strategies did not demonstrate improvement from the baseline model. ConclusionsA robust AI-based algorithm for detecting and segmenting lung tumor foci in the pre-clinical mouse models was developed. The output of this algorithm is compatible with open-source software that researchers commonly use.

Translation of c-Met Targeted Image-Guided Surgery Solutions in Oral Cavity Cancer-Initial Proof of Concept Data.

Simple SummaryTranslation of tumor-specific fluorescent tracers is crucial in the realization intraoperative of tumor identification during fluorescence-guided surgery. Ex vivo assessment of surgical specimens after topical tracer application has the potential to reveal the suitability of a potential surgical target prior to in vivo use in patients. In this study, the c-Met receptor was identified as a possible candidate for fluorescence-guided surgery in oral cavity cancer. Freshly excised tumor specimens obtained from ten patients with squamous cell carcinoma of the tongue were incubated with EMI-137 and imaged with a clinical-grade Cy5 prototype fluorescence camera. In total, 9/10 tumors were fluorescently illuminated, while non-visualization could be linked to non-superficial tumor localization. Immunohistochemistry revealed c-Met expression in all ten specimens. Tumor assessment was improved via video representation of the tumor-to-background ratio.Intraoperative tumor identification (extension/margins/metastases) via receptor-specific targeting is one of the ultimate promises of fluorescence-guided surgery. The translation of fluorescent tracers that enable tumor visualization forms a critical component in the realization of this approach. Ex vivo assessment of surgical specimens after topical tracer application could help provide an intermediate step between preclinical evaluation and first-in-human trials. Here, the suitability of the c-Met receptor as a potential surgical target in oral cavity cancer was explored via topical ex vivo application of the fluorescent tracer EMI-137. Freshly excised tumor specimens obtained from ten patients with squamous cell carcinoma of the tongue were incubated with EMI-137 and imaged with a clinical-grade Cy5 prototype fluorescence camera. In-house developed image processing software allowed video-rate assessment of the tumor-to-background ratio (TBR). Fluorescence imaging results were related to standard pathological evaluation and c-MET immunohistochemistry. After incubation with EMI-137, 9/10 tumors were fluorescently illuminated. Immunohistochemistry revealed c-Met expression in all ten specimens. Non-visualization could be linked to a more deeply situated lesion. Tumor assessment was improved via video representation of the TBR (median TBR: 2.5 (range 1.8–3.1)). Ex vivo evaluation of tumor specimens suggests that c-Met is a possible candidate for fluorescence-guided surgery in oral cavity cancer.

Increased YAP1 expression is significantly associated with breast cancer progression, metastasis and poor survival.

YAP1 plays a key role as a transcriptional coactivator in the Hippo pathway. Based on conflicting reports regarding YAP1 function in cancer, this study discerned its role in breast carcinogenesis. First, a systematic review of salient breast cancer studies targeting YAP1 dysregulation was performed. Additionally, freshly excised tumor specimens of approximately200 breast cancer patients were processed for quantification of YAP1 expression at mRNA and protein levels using quantitative PCR and immunohistochemistry, respectively. YAP1 expression was ninefolds higher in tumors versus controls and significantly associated with metastasis (p <0.05) and poor survival in Pakistani breast cancer patients. These findings establish the role of YAP1 overexpression in tumorigenesis and metastasis. Hence, YAP1 inhibition may be considered a possible therapeutic strategy.

Optical palpation for tumor margin assessment in breast-conserving surgery.

Intraoperative margin assessment is needed to reduce the re-excision rate of breast-conserving surgery. One possibility is optical palpation, a tactile imaging technique that maps stress (force applied across the tissue surface) as an indicator of tissue stiffness. Images (optical palpograms) are generated by compressing a transparent silicone layer on the tissue and measuring the layer deformation using optical coherence tomography (OCT). This paper reports, for the first time, the diagnostic accuracy of optical palpation in identifying tumor within 1 mm of the excised specimen boundary using an automated classifier. Optical palpograms from 154 regions of interest (ROIs) from 71 excised tumor specimens were obtained. An automated classifier was constructed to predict the ROI margin status by first choosing a circle diameter, then searching for a location within the ROI where the circle was ≥ 75% filled with high stress (indicating a positive margin). A range of circle diameters and stress thresholds, as well as the impact of filtering out non-dense tissue regions, were tested. Sensitivity and specificity were calculated by comparing the automated classifier results with the true margin status, determined from co-registered histology. 83.3% sensitivity and 86.2% specificity were achieved, compared to 69.0% sensitivity and 79.0% specificity obtained with OCT alone on the same dataset using human readers. Representative optical palpograms show that positive margins containing a range of cancer types tend to exhibit higher stress compared to negative margins. These results demonstrate the potential of optical palpation for margin assessment.

Abstract PS4-37: A pro-tumorigenic mechanism of M2 tumor-associated macrophages (TAM) in triple-negative breast cancer

Abstract Introduction:Triple-Negative Breast Cancer (TNBC) comprises approximately 30% of all breast cancers in Indian women. Given their aggressive nature, TNBCs have high rates of systemic metastasis and mortality with only chemotherapy available for treatment. The success of immunotherapy in solid tumors has raised the hope for their utility in TNBCs as well. But only a subset of patients have a clinical response to check-point inhibitors. The cellular and molecular mechanisms that mediate the immunological response or tolerance are just beginning to be understood. Compared to ER/PR+ breast cancer, TNBC features a unique tumor microenvironment (TME) characterized by a large number of tumor-infiltrating lymphocytes (TILs) and tumor-associated macrophages (TAMs). The density of TILs in and of themselves do not accurately predict response to neoadjuvant chemotherapy or survival. M2 tumor-associated macrophages (M2-TAMs) have been reported to associate with solid tumors to facilitate epithelial to mesenchymal transition (EMT), tumor invasiveness, metastasis, and resistance to therapy. In this study, we have characterized presence of M2-TAM in the TNBC immune environment by examining expression of several biomarkers. Methods:Surgically excised tumor specimens from 88 Indian women with TNBC were accessed from the longitudinal observational series of SJNAHS tissue bank under IERB approved protocols and assayed on nCounter® PanCancer Immune Profiling Panel which comprised of 740 genes and characterises 14 different immune cell types. Transcriptome analysis using Non-negative Matrix factorization (NMF) based unsupervised clustering was done to arrive at stable subtypes characterized by different tumour microenvironments within TNBC. CIEBERSORT tool was used to analyse for distribution of cell types. Identification of macrophages was done by Immunohistochemistry (IHC) for CD68 and CD163 markers in TNBCs and a control group of ER+HER2-.Results:NMF analysis with an enriched immune gene signature of 111 genes, yielded 3 subtypes (ST) (37%, 27% and 36% with relative proportions of ST1, ST2 and ST3) within the TNBC. The three subtypes had distinct survival patterns with ST1 having the best prognosis with enriched TH1 gene signature and ST3 had poorest (log rank p=0.5). On application of this gene signature to TNBC groups in METABRIC and TCGA data, similar pattern emerged with a significant survival pattern of ST 3 having the poorest outcome (p=0.005). A closer examination of ST 3 revealed a signature enriched for M2 macrophages also known as TAM. Immunohistochemistry for Pan macrophage marker CD68 and an M2 specific marker CD163 between TNBC and ER/PR+ revealed difference in the two groups was significantly different (Mann Whitney p=0.03), with TNBC exhibiting 2.4 fold higher expression of CD163. CD 68, was enriched in ST2 and ST3. CD163 which is an M2 specific marker was highest in ST3 as compared to ST 1&amp; 2 (p=0.04). This group also correlated with signal molecules secreted by macrophages containing growth factors, cytokines and chemokines, such as TGF-β, VEGF, IL-10 and CXCL and interleukins like IL4 and IL6 suggestive of TAM recruitment and polarization. It is likely that the TAM enriched subgroup is likely to be unresponsive to PD1/PDL1 inhibitors but could be targeted by novel therapeutic strategies to directly target M2-TAM. In vitro and In vivo analysis to target M2-TAM to evaluate the response to these therapeutic compounds in cell lines as well as a mouse syngeneic model is underway.Conclusions and future directions: As a cell type within the tumor microenvironment, that promotes invasion, M2-TAMs makes an ideal therapeutic target in TNBC. In addition, this sub-type lends itself to easy identification by a simple IHC assay. Citation Format: Aruna Korlimarla, Jyothi Prabhu, Chantirika Ragulan, Gnanapriya Shivakumar, Krisha Desai, Maggie Cheang, Srinath BS, Ravi Diwakar, Sandhya Apachu, Rekha Kumar, TS Sridhar, Savitha RajaRajan, Rohini Kaluve, Annie Alexander, Anguraj Sadanandam. A pro-tumorigenic mechanism of M2 tumor-associated macrophages (TAM) in triple-negative breast cancer [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 PS4-37.

Diagnostic accuracies of intraoperative frozen section and permanent section examinations for histological grades during open biopsy of bone tumors.

A histological diagnosis obtained from an intraoperative frozen section (FS) during biopsy confirms the adequacy of tumor tissue in the specimen. However, some cases show a discrepancy among the intraoperative FS diagnosis, permanent section (PS) diagnosis of the biopsy specimen, and the final diagnosis of the excised tumor specimen. In this study, we retrospectively investigated the diagnostic accuracy of the FS and PS for different types of bone tumors. This study included 377 patients with 411 bone tumors who underwent tumor excision after an open biopsy with intraoperative FS diagnosis. FS, PS, and final diagnoses of the patients were classified into benign tumors/tumor-like lesions, intermediate malignancies, and malignant tumors. To assess diagnostic accuracy, the histological grades in FS and PS diagnoses were compared with those in the final diagnoses. The overall diagnostic accuracies of FS and PS were 93% and 97%, respectively. The accuracy of FS and PS for histological grade was 84% and 93% for chondrogenic tumors, 90% and 96% for osteogenic tumors, 97% and 98% for osteoclastic giant cell-rich tumors, 100% and 100% for tumors of undefined neoplastic nature, and 95% and 99% for other bone tumors, respectively. These data suggest that surgical planning based on PS diagnosis is recommended for chondrogenic and osteogenic tumors.

Accuracy of histological grades from intraoperative frozen-section diagnoses of soft-tissue tumors.

For excised tumor specimens, histological grades can differ between the biopsy diagnosis and the final diagnosis. We retrospectively investigated the diagnostic accuracies of histological grades for frozen-section and permanent-section diagnoses from patients with soft-tissue tumors. Frozen-section, permanent-section, and final diagnoses were classified as benign tumors/tumor-like lesions, intermediate malignancies, or malignant tumors. Diagnostic accuracies of the histological grades from the frozen and permanent sections were evaluated by comparing the final diagnoses for the resected specimens. The diagnostic accuracies of the histological grades for the frozen- and permanent-section diagnoses were 95% (387/408 lesions) and 97% (395/408 lesions), respectively. Among the tumor types, the diagnostic accuracies of the histological grades for the frozen-section and permanent-section diagnoses were 84% and 87% for adipocytic tumors, 87% and 91% for fibroblastic/myofibroblastic tumors, 99% and 100% for nerve-sheath tumors, 98% and 98% for fibrohistiocytic tumors, 90% and 98% for tumors of uncertain differentiation, 100% and 100% for vascular tumors, and 97% and 98% for other tumors, respectively. Histological grades from frozen-section diagnoses yielded low diagnostic accuracies in adipocytic and fibroblastic/myofibroblastic tumors. Treatment should be planned based on permanent-section diagnosis and radiological findings for these tumors.

Abstract P3-13-08: Usefulness of periareolar zigzag incision in oncoplastic breast-conserving surgery

Abstract Background Breast-conserving surgery (BCS) has been performed as a standard procedure because the addition of radiation therapy after breast-conserving surgery in early breast cancer patients is similar to that of mastectomy.The surgical methods for breast cancer have been developed in recent years, and the cosmetic aspect has become important.Several incisions are used to reduce the scar to the incision site during BCS. The periareolar incision is often used when the small tumor relatively close to the nipple. However, periareolar incision does not have sufficient field of sight, which can result in difficult or impossible operations if the tumor is large or far from the nipple. In plastic surgery, various methods such as zigzag incisions have been recommended to achieve satisfactory esthetic results. The periareolar zigzag incision has the advantage of a not only good surgical field but also contributed to better surgical scars. The purpose of this study was to evaluate the oncological safety of procedures by studying the status of the surgical margins of the excised tumor specimen and reduces the need for further surgery Methods From January 2016 to November 2017, we conducted a survey of female patients who underwent BCS using zigzag incision. Patients with exclusion criteria were excluded from this study if they had a Patients with bilateral breast cancer or neoadjuvant chemotherapy was included in the study. Intraoperative frozen section margin was evaluated in all patients and additional resection was performed when the positive margin was present. Final margin status was determined by examination of the permanent paraffin-embedded sections. In patients with invasive breast cancer or relatively large carcinoma in situ, axillary surgery was performed according to the presence of axillary metastasis. Patients characteristics, tumor characteristics, operative time, size of the specimen and the distance from the tumor to nipple were evaluated Results 393 patients were enrolled in the study, including 9 patients with bilateral breast cancer, and a total of 402 cases of BCS surgery were analyzed. Thirty-five patients received neoadjuvant chemotherapy. The median age of the patients was 51 (range: 25-84 years), the median time of operation was 72 minutes in patients who did not undergo axillary surgery or sentinel node biopsy only, and 83 minutes in patients who underwent axillary node dissection or supraclavicular node dissection. The median tumor size was 1.6 cm (range: 0-8.8 cm), median tumor distance from the nipple was 3.0 cm (range: 0.4-8.1 cm), mean excised specimen sized was 5.0 cm (range: 0.9-15.0 cm). Frozen biopsy of the resection margin during surgery revealed tumor-positive in 69 (17.2%) cases and re-excision was performed. All patients were discharged with no sign of infection or skin necrosis. Conclusions The periareolar zigzag incision technique has good cosmetic results and provides a sufficient surgical field, which can be useful for removing relatively large tumors or tumors far from the nipple. Citation Format: Ko B, Kim H, Chung I, Lee S, Kim J, Lee J, Son B, Ahn SH. Usefulness of periareolar zigzag incision in oncoplastic breast-conserving surgery [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-13-08.

Raman spectroscopy for rapid intra-operative margin analysis of surgically excised tumour specimens.

Raman spectroscopy, a form of vibrational spectroscopy, has the ability to provide sensitive and specific biochemical analysis of tissue. This review article provides an in-depth analysis of the suitability of different Raman spectroscopy techniques in providing intra-operative margin analysis in a range of solid tumour pathologies. Surgical excision remains the primary treatment of a number of solid organ cancers. Incomplete excision of a tumour and positive margins on histopathological analysis is associated with a worse prognosis, the need for adjuvant therapies with significant side effects and a resulting financial burden. The provision of intra-operative margin analysis of surgically excised tumour specimens would be beneficial for a number of pathologies, as there are no widely adopted and accurate methods of margin analysis, beyond histopathology. The limitations of Raman spectroscopic studies to date are discussed and future work necessary to enable translation to clinical use is identified. We conclude that, although there remain a number of challenges in translating current techniques into a clinically effective tool, studies so far demonstrate that Raman Spectroscopy has the attributes to successfully perform highly accurate intra-operative margin analysis in a clinically relevant environment.

Calibration and analysis of a multimodal micro-CT and structured light imaging system for the evaluation of excised breast tissue

A multimodal micro-computed tomography (CT) and multi-spectral structured light imaging (SLI) system is introduced and systematically analyzed to test its feasibility to aid in margin delineation during breast conserving surgery (BCS). Phantom analysis of the micro-CT yielded a signal-to-noise ratio of 34, a contrast of 1.64, and a minimum detectable resolution of 240 μm for a 1.2 min scan. The SLI system, spanning wavelengths 490 nm to 800 nm and spatial frequencies up to 1.37 , was evaluated with aqueous tissue simulating phantoms having variations in particle size distribution, scatter density, and blood volume fraction. The reduced scattering coefficient, and phase function parameter, γ, were accurately recovered over all wavelengths independent of blood volume fractions from 0% to 4%, assuming a flat sample geometry perpendicular to the imaging plane. The resolution of the optical system was tested with a step phantom, from which the modulation transfer function was calculated yielding a maximum resolution of 3.78 cycles per mm. The three dimensional spatial co-registration between the CT and optical imaging space was tested and shown to be accurate within 0.7 mm. A freshly resected breast specimen, with lobular carcinoma, fibrocystic disease, and adipose, was imaged with the system. The micro-CT provided visualization of the tumor mass and its spiculations, and SLI yielded superficial quantification of light scattering parameters for the malignant and benign tissue types. These results appear to be the first demonstration of SLI combined with standard medical tomography for imaging excised tumor specimens. While further investigations are needed to determine and test the spectral, spatial, and CT features required to classify tissue, this study demonstrates the ability of multimodal CT/SLI to quantify, visualize, and spatially navigate breast tumor specimens, which could potentially aid in the assessment of tumor margin status during BCS.

Abstract 4833: Perfusion-based bioreactor culture of primary cancer tissue maintains tumor microenvironment complexity and allow in-vitro testing of immune blockade therapy

Abstract In vitro culture of primary cancer tissue is still very limited and the generation of patient derived xenograft determine the loss of human-cancer associated stroma. In this context, the use of 3D in vitro systems based on human tissue may be an innovative system to be exploited for keeping the tumor microenvironment (TME) complexity of the tissue in vitro. Freshly excised colorectal (CRC) and breast cancer (BrCa) specimens were fragmented and cultured in 3D “sandwich-like format” between porous collagen scaffolds under perfusion flow (U-CUP, Cellec Biotek AG). The maintenance of tumor and immune-infiltrating cells, survival and phenotypic characterization were histologically assessed. In a second step cancer treatment were tested. U-CUP culture allowed the preservation, viability and expansion of tumor tissue with concomitant stromal and immune cells. Expanding cancer cells were viable after 10 and 21 days (CRC and BrCa, respectively). Administration of anti-ER treatment to Lumina A ER+ BrCa was associated with decreased expansion of cancer tissue into the scaffold after 21 days. The maintenance of immune-infiltrating cells allowed testing of immune blockade therapy. Administration of anti-PDL1 antibody, alone or in combination with anti-CTLA4, to the culture medium was associated with increased expression of markers of immune-activation (i.e. IFNγ) and decreased expression of immunosuppressive cytokine IL10. Preserving malignant, interstitial and immunocompetent cells comprised in surgically excised tumor specimens might allow a direct evaluation of the effects of various treatments on the complex TME. This engineered in vitro model could allow animal-free testing and it could be extended as a platform allowing the testing of innovative approaches for the treatment of human malignancies. Our findings shed the light on a promising system for selecting personalized treatment based on a patient’s tumor specific microenvironment. Note: This abstract was not presented at the meeting. Citation Format: Manuele Giuseppe Muraro, Simone Muenst, Celeste Manfredonia, Valentina Mele, Silvio Daester, Alexandar Tzankov, Luigi Terracciano, Walter Weber, Giulio C. Spagnoli, Giandomenica Iezzi, Ivan Martin, Savas D. Soysal. Perfusion-based bioreactor culture of primary cancer tissue maintains tumor microenvironment complexity and allow in-vitro testing of immune blockade therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4833. doi:10.1158/1538-7445.AM2017-4833

Surgical tissue handling methods to optimize ex vivo fluorescence with the activatable optical probe γ-glutamyl hydroxymethyl rhodamine green.

Optical fluorescence imaging has been developed as an aid to intraoperative diagnosis to improve surgical and endoscopic procedures. Compared with other intraoperative imaging methods, it is lower in cost, has a high safety margin, is portable and easy to use. γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) is a recently developed activatable fluorescence probe that emits strong fluorescence in the presence of the enzyme γ-glutamyl transpeptidase (GGT), which is overexpressed in many cancers, including ovarian cancer. Ex vivo testing is important for clinical approval of such probes. The diagnostic performance of gGlu-HMRG in fresh excised surgical specimens has been reported; however, details of tissue handling have not been optimized. In this study, we investigated four different tissue handling procedures to optimize imaging in excised tumor specimens. The fluorescence intensity time courses after the different tissue handling methods were compared. Additionally, the fluorescence positive areas were correlated with the presence of red fluorescent protein (RFP) in an RFP positive cell line as the standard of reference for cancer location. In the 'intact' groups, tumors yielded quick and homogeneous activation of gGlu-HMRG. In the 'rinse' and 'cut' groups, the fluorescence intensity of the tumor was a little lower than that in the intact group. In the 'pressed' groups, however, fluorescence intensity from gGlu-HMRG was lower over the entire time course, suggesting a decrease or relocation of excreted GGT. In conclusion, we demonstrate that the method of tissue handling prior to ex vivo imaging with the activatable probe gGlu-HMRG has a strong influence on the signal derived from the specimen. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Abstract LB-37: A patient-derived in vitro model of colorectal cancer: a perfusion culture system that recapitulates patient tumor composition and structure in 3-dimensions

Abstract Cancer medicine would benefit greatly from a practical technology that accurately predicts the sensitivity of individual patient tumors to drugs. Previously developed in vitro chemosensitivity models have generally failed to do this because they do not incorporate key aspects of the tumor microenvironment (TME), which clearly plays a role in drug resistance. There has been significant interest in patient-derived xenograft (PDX) chemosensitivity testing as an alternative to in vitro models, but the applicability of this approach is limited by engraftment percentages and growth rates. We have developed an in vitro technology that is designed to recapitulate the 3D tissue architecture of patient-derived colorectal tumors including accessory (endothelial and immune) cells, starting with freshly excised tumor specimens. The system incorporates: 1) low-sheer, tangential flow of nutrient medium above and below an open, synthetic 3D cell scaffold; and 2) exchange of metabolic gasses via permeable membranes which separate the culture chamber core from gassing chambers located above and below the nutrient flow compartments. This design allows establishment of gas and nutrient gradients across the cell scaffold that closely mimic conditions in vivo. Aliquots of mechanically and enzymatically dissociated, freshly excised, primary human colorectal adenocarcinomas were introduced into the 3D culture system and maintained for up to 3 weeks. Routine maintenance of the cultures involved partial medium exchanges as needed based on glucose concentrations in the circulating medium. The proportions and arrangement of key cell types (e.g., normal muscularis, tumor epithelium, stromal fibroblasts, endothelial cells, and immune cells) in the cultured explants were compared to those of the original tumor by standard histological techniques. Results revealed good agreement between the histology of the original tumor and that of the in vitro model. These results suggest that this model technology could be applied to individual patient drug selection as well as studies of colorectal cancer biology that are not feasible or practical in vivo. Citation Format: William P. Pfund, Paul A. Neeb. A patient-derived in vitro model of colorectal cancer: a perfusion culture system that recapitulates patient tumor composition and structure in 3-dimensions. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-37. doi:10.1158/1538-7445.AM2014-LB-37

Comparative Study of the Surgical Margins between Oncoplastic Breast Surgery and Quadrantectomy in Breast Conserving Surgery

Objective: To evaluate the oncological safety of oncoplastic procedures by studying the status of the surgical margins of the excised tumor specimen in comparison with standard quadrantectomies. Method: This is a study that was conducted at Queen Alia Military Hospital (QAMH), Amman, Jordan, between January 2009 until August 2011. Fifteen consecutive breast cancer patients who underwent oncoplastic surgery (group1) and 15 patients who underwent standard quadrantectomy (group 2) were studied with regard to the stage of breast cancer, the surgical procedures performed, the volume of breast tissue excised, and the histopathology of the tumor specimen, with specific details on surgical margins. Results: Patients who underwent oncoplastic surgery (group 1) were younger (mean age 48.73 years) than patients who had a classic quadrantectomy (group 2; mean age 55.76 years. The mean volume of the excised specimen in group 1 was 188.2 cm 3 , compared with 101.6 cm 3 in group 2. Surgical margins were negative in all 15 cases in group 1 and 14 out of 15 in group 2. The average width of the surgical margin was 9.2 mm in group 1 and 6.3 mm in group 2.