Needle Biopsy Samples Research Articles

Endoscopic ultrasound-guided side-fenestrated needle biopsy sampling is sensitive for pancreatic neuroendocrine tumors but inadequate for tumor grading: a prospective study

Accurate pretreatment grading of pancreatic neuroendocrine tumors (PanNETs) is important to guide patient management. We aimed to evaluate endoscopic ultrasound-guided fine needle biopsy sampling (EUS-FNB) for the preoperative diagnosis and grading of PanNETs. In a tertiary-center setting, patients with suspected PanNETs were prospectively subjected to 22-gauge, reverse-bevel EUS-FNB. The EUS-FNB samples (Ki-67EUS) and corresponding surgical specimens (Ki-67SURG) were analyzed with Ki-67 indexing and thereafter tumor grading, (GRADEEUS) and (GRADESURG) respectively. In total 52 PanNET-patients [median age: 66 years; females: 25/52; surgical resection 22/52 (42%)] were included. EUS-FNB was diagnostic in 44/52 (85%). In 42 available FNB-slides, the median neoplastic cell count was 1034 (IQR: 504–3667) with 32/42 (76%), 22/42 (52%), and 14/42 (33%) cases exceeding 500, 1000, and 2000 neoplastic cells respectively. Ki-67SURG was significantly higher compared to Ki-67EUS with a moderate correlation comparing Ki-67EUS and Ki-67SURG (Pearson r = 0.60, r2 = 0.36, p = 0.011). The GRADEEUS had a weak level of agreement (κ = 0.08) compared with GRADESURG. Only 2/12 (17%) G2-tumors were correctly graded in EUS-FNB-samples. EUS-guided fine needle biopsy sampling is sensitive for preoperative diagnosis of PanNET but biopsy quality is relatively poor. Therefore, the approach seems suboptimal for pretreatment grading of PanNET.

Is Free Breathing Possible During Computed Tomography-Guided Percutaneous Transthoracic Lung Biopsy? The Clinical Experience in 585 Cases.

The aim of the study was to retrospectively evaluate the safety and accuracy of computed tomography (CT)-guided percutaneous transthoracic needle biopsy (PTNB) of lung lesions during quiet breathing. We investigated the diagnostic performance and complication rate of 585 procedures in 563 patients (357 men; mean age, 67.7 years), who underwent CT-guided PTNBs during quiet breathing, aided by a respiratory targeting technique from May 2017 to July 2019. Differences between the cases with and without respiratory targeting were analyzed. Logistic regression analyses were performed to examine the development of pneumothorax and hemoptysis. Percutaneous transthoracic needle biopsy samples were successfully obtained in 574 of 585 procedures (98.1%). Final diagnoses included: 410 malignant cases, 119 benign cases, and 45 indeterminate cases. The sensitivity, specificity, and accuracy of diagnosis were 94.4%, 100%, and 95.7%, respectively. Use of respiratory targeting was associated with younger age (P = 0.004), smaller lesion size (P < 0.001), peripheral location (P = 0.003), shorter distance from the diaphragm (P < 0.001), lower lobe location (P < 0.001), prone position (P = 0.004), and visible motion artifact (P < 0.001). Pneumothorax and hemoptysis rates were 22.9% and 7.9%, respectively. Upon multivariate analysis, emphysema (P = 0.002) was the only independent risk factor for pneumothorax, whereas distance from the pleura greater than 2 cm (P < 0.001), tissue sampling 3 times or more (P = 0.003), and a less experienced operator (P < 0.001) were risk factors for hemoptysis. Computed tomography-guided PTNB during quiet breathing with respiratory targeting yielded high diagnostic performance with a slightly higher rate of complications. Free-breathing PTNB can be applied in clinical practice, based on lesion location and risk factors for complications.

Convolutional Neural Network–Based Automatic Classification of Colorectal and Prostate Tumor Biopsies Using Multispectral Imagery: System Development Study

Background Colorectal and prostate cancers are the most common types of cancer in men worldwide. To diagnose colorectal and prostate cancer, a pathologist performs a histological analysis on needle biopsy samples. This manual process is time-consuming and error-prone, resulting in high intra- and interobserver variability, which affects diagnosis reliability. Objective This study aims to develop an automatic computerized system for diagnosing colorectal and prostate tumors by using images of biopsy samples to reduce time and diagnosis error rates associated with human analysis. Methods In this study, we proposed a convolutional neural network (CNN) model for classifying colorectal and prostate tumors from multispectral images of biopsy samples. The key idea was to remove the last block of the convolutional layers and halve the number of filters per layer. Results Our results showed excellent performance, with an average test accuracy of 99.8% and 99.5% for the prostate and colorectal data sets, respectively. The system showed excellent performance when compared with pretrained CNNs and other classification methods, as it avoids the preprocessing phase while using a single CNN model for the whole classification task. Overall, the proposed CNN architecture was globally the best-performing system for classifying colorectal and prostate tumor images. Conclusions The proposed CNN architecture was detailed and compared with previously trained network models used as feature extractors. These CNNs were also compared with other classification techniques. As opposed to pretrained CNNs and other classification approaches, the proposed CNN yielded excellent results. The computational complexity of the CNNs was also investigated, and it was shown that the proposed CNN is better at classifying images than pretrained networks because it does not require preprocessing. Thus, the overall analysis was that the proposed CNN architecture was globally the best-performing system for classifying colorectal and prostate tumor images.

Ultrasound-guided lymph node biopsy sampling to study the immunopathogenesis of rheumatoid arthritis: a well-tolerated valuable research tool

BackgroundAnalyses of lymphoid organs are required to further elucidate the pathogenesis of inflammatory diseases like rheumatoid arthritis (RA). Yet, invasive tissue collection methods are scarcely applied, because they are often considered burdensome, although patients do not always consider invasive methods as a high burden. We aimed to investigate the perspectives of study participants undergoing ultrasound-guided inguinal lymph node (LN) needle biopsy sampling and determine the molecular and cellular quantity and quality of LN biopsies.MethodsTogether with patient research partners, questionnaires were developed to evaluate the motives, expectations, and experiences of participants undergoing a LN biopsy. Healthy controls and RA(-risk) patients were asked to complete these questionnaires before and after the procedure. RNA and lymphocyte yields from obtained LN biopsies were also calculated.ResultsWe included 50 individuals, of which 43 (86%) reported their pre- and post-procedure experiences. The median reported pain on a 5-point Likert scale (1 not to 5 very painful) was 1. Interestingly, almost all (n = 32; 74%) study participants would undergo a second procedure and more than half (n = 23; 54%) would encourage others to take part in the LN biopsy study. Motives for current and future participation were mostly altruistic. Inguinal hematoma occurred frequently, but no other significant or unexpected complications ensued. The LN biopsies yielded sufficient and high-quality RNA and lymphocyte numbers.ConclusionsUltrasound-guided inguinal LN biopsy sampling is well-tolerated, safe, and provides sufficient material for further molecular and cellular analyses. Our participants’ positive experiences endorse the application of this research tool to further elucidate the pathogenesis of RA and other inflammatory diseases.

Expression of Alpha Methyl Acyl-CoaRacemase (AMACR) Immunohistochemistry in Carcinoma Prostate in Tertiary Care Hospital

Objective: Prostate needle biopsy samples from cancer patients at a tertiary care hospital were analysed for the presence of the potential screening indicator alpha-methylacyl-COA RACEMES. Material and Methods: This was a Cross-sectional study carried out at Department of Histopathology, Lady Reading Hospital Peshawar from June, 2021 to December, 2021.Regardless of the patient's age, tumour grade, or histological type, all adenocarcinoma prostate specimens diagnosed at the Lady Reading Hospital Peshawar on the basis of immunohistochemistry and routine histology were examined. Standard and mean deviation were computed for quantitative factors of patient like age and frequencies, percentages were calculated for qualitative factors like AMACR expressions. Results: Outof 90 cases, AMACR positivity was found in 68 (85%), while negativity was seen in 22 (15%). Nine (11.3%) of the negative cases had one staining (weak, non-circumferential) and three cases (3%) had no stainings (No cytoplasmic staining). Conclusion: Positive AMACR staining on prostate needle core necropsies may be utilised to support a cancer diagnosis when the focus at issue is 1 mm in highest diameter. Basic hematoxylin and eosin standards for malice require analysis of cell-specific markers like P63 or 34E12. AMACR labelling must be interpreted in the context of other labels that can be used to find out more. Keywords: Adenocarcinoma of the prostate, Alphamethylacyl-CoA racemase (AMACR), Immunohistochemistry.

Minimally Invasive Tissue Sampling Findings in 12 Patients With Coronavirus Disease 2019

BackgroundMinimally invasive tissue sampling (MITS), a postmortem procedure that uses core needle biopsy samples and does not require opening the body, may be a valid alternative to complete autopsy (CA) in highly infectious diseases such as coronavirus disease-19 (COVID-19). This study aimed to (1) compare the performance of MITS and CA in a series of COVID-19 deaths and (2) evaluate the safety of the procedure.MethodsFrom October 2020 to February 2021, MITS was conducted in 12 adults who tested positive before death for COVID-19, in a standard, well-ventilated autopsy room, where personnel used reinforced personal protective equipment. In 9 cases, a CA was performed after MITS. A thorough histological evaluation was conducted, and the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was evaluated by real-time reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry.ResultsThe diagnoses provided by MITS and CA matched almost perfectly. In 9 patients, COVID-19 was in the chain of events leading to death, being responsible for diffuse alveolar damage and mononuclear T-cell inflammatory response in the lungs. No specific COVID-19 features were identified. Three deaths were not related to COVID-19. All personnel involved in MITS repeatedly tested negative for COVID-19. SARS-CoV-2 was identified by RT-PCR and immunohistochemistry in the MITS samples, particularly in the lungs.ConclusionsMITS is useful for evaluating COVID-19–related deaths in settings where a CA is not feasible. The results of this simplified and safer technique are comparable to those of CA.

A Rare Case of Primary Hepatic Adenosquamous Carcinoma Mimicking Liver Abscess

A 61-year-old woman with a 3-week history of chills, fevers to 39°C, anorexia, weight loss, and fatigue was referred to our hospital due to the progressive discomforts over the last week. Her documented medical history was notable for well-controlled type 2 diabetes. Physical examination was unremarkable. Notable laboratory tests of the blood revealed a hemoglobin level of 90 g/L, a white blood cell count of 11.7×109/L with neutrophil 90.7%, albumin 31.6 g/L, gamma glutamyl transferase 331 IU/L, alkaline phosphatase 748 IU/L. Direct microbiologic detection and cultures were negative for hepatitis B and C virus, bacterial and fungal pathogens, and human immunodeficiency virus. A computed tomography scan of the chest was unremarkable. Abdominal contrast-enhanced computed tomography discovered an ill-defined and sub-circular lowdensity region measuring 7.6×6.1cm with patchy enhancement in the right lobe of liver (Figure A), which implied a liver abscess. Empiric antibiotics were changed to meropenem combined with metronidazole on day three of hospitalisation. However, the fever did not alleviate with poorly therapeutic effect. Ultrasound-guided biopsy was performed and purulent fluid was collected to make a definitive diagnosis of the liver mass. Surprisingly, The histopathological detection of a needle biopsy sample confirmed neoplastic necrosis with negative bacterial and fungal culture of the purulent fluid. The patient then underwent right hemihepatectomy and histopathologic examination further revealed that the lesion included a mixture of a poorly differentiated adenocarcinoma with mucin production and squamous cell carcinoma with keratinization, as well as large patchy necrosis (Figure B). On immunostaining, the lesion stained strongly with CK5/6, P63, and P40, and was also partially positive for CK7, CK8/18, and Cam5.2 with negative staining for hepatocyte and alpha-fetoprotein, which were consistent with adenosquamous carcinoma of the liver. Primary hepatic adenosquamous carcinoma is a rare variant of intrahepatic cholangiocarcinoma with approximately 10 cases reported within 5 years.1,2 The pathogenesis of adenosquamous carcinoma of the liver is largely unknown owing to limited data in the literature. Fever, jaundice, weight loss, anorexia, and abdominal pain are the most frequently reported clinical manifestations.3 Adenosquamous carcinoma of the liver usually presents atypical imaging characteristics, which is difficult to distinguish it from liver abscess, hepatocellular carcinoma, and liver metastasis. Adenosquamous carcinoma of the liver is a highly aggressive neoplasm and the median survival time is 6 months after surgery.4 Therefore, preoperative diagnosis is of vital importance to predict a poor prognosis. Despite postoperative chemoradiation, the patient passed away 7 months after surgery.

Morphological and p40 immunohistochemical analysis of squamous differentiation in endoscopic ultrasound guided fine needle biopsies of pancreatic ductal adenocarcinoma

The basal-like molecular subtype of pancreatic ductal adenocarcinoma (PDAC) is associated with poor prognosis and upregulation in TP63ΔN (p40) network. Adenosquamous histology can be observed. This study assessed immunohistochemical p40 expression in fine needle biopsy (FNB) samples with PDAC and association with cytomorphological features of squamous differentiation and clinical data. 106 EUS FNBs with PDAC were assessed for eight cytomorphological features of squamous differentiation. P40 H-score (intensity 0–3 × percentage positive nuclei) was analysed for association with morphological features, patient age, gender, operability, chemotherapy and survival. P40 H-score in 14 paired FNBs and resections was compared. P40 h-score was 1–3 in 31%, 4–30 in 16% and > 30 in 13% of FNBs. It was significantly associated with intercellular bridges, elongated cell shape, sharp cell borders, angular nuclei with homogenous chromatin (p < 0.001) and dense cytoplasm (p = 0.002). Keratinisation was not seen. Inoperable patients (n = 81) had a shorter median survival for h-score > 30 (n = 9, 1.8 months) than for h-score ≤ 30 (n = 66, 6.7 months) not quite reaching statistical significance (p = 0.08). P40 was significantly associated with squamous morphology in FNBs with PDAC. P40 H-score > 30 showed a trend towards shorter survival in inoperable patients. Squamous differentiation may be a treatment target in PDAC.

Concordance between core needle biopsy and surgical excision specimens for Ki-67 in breast cancer - a systematic review of the literature.

The biomarkers oestrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) are routinely measured in patients with breast cancer with international consensus on how they should be interpreted. There is evidence to support use of other biomarkers to give more detailed predictive and prognostic information. Ki-67 is one example, and measures the proliferative activity of cancer cells. It is important that this can be performed at diagnosis of breast cancer for patients who do not have initial surgical treatment (mainly older women) and those receiving neoadjuvant therapies. A systematic review was performed to assess concordance of measurement of Ki-67 between core needle biopsy (CNB) samples and surgical excision (SE) samples in patients with invasive breast cancer. MEDLINE and Embase databases were searched. Studies were eligible if performed within the last 10 years; included quantitative measurement of Ki-67 in both CNB and SE samples with no prior breast cancer treatment; measured concordance between two samples; and had full text available. A total of 22 studies, including 5982 paired CNB and SE samples on which Ki-67 was measured, were appraised. Overall, there appeared to be concordance; however, reliability was unclear. Where given, the Cohen's kappa coefficient (κ) of correlation between samples ranged from 0.261 to 0.712. The concordance rate between CNB and SE where measured as a percentage had a range from 70.3 to 92.7% CONCLUSIONS: Assessment of level of concordance of Ki-67 between CNB and SE samples is hampered by different methodologies. International consensus on Ki-67 measurement is urgently needed.

S1016 Comparison of Tissue Acquisition Techniques and Needle Design for EUS-Guided Fine Needle Biopsy Sampling of Pancreatic Lesions: A Randomized Controlled Pilot Clinical Trial

S1016 Comparison of Tissue Acquisition Techniques and Needle Design for EUS-Guided Fine Needle Biopsy Sampling of Pancreatic Lesions: A Randomized Controlled Pilot Clinical Trial

Profiling of 3D Genome Organization in Nasopharyngeal Cancer Needle Biopsy Patient Samples by a Modified Hi-C Approach.

Nasopharyngeal cancer (NPC), a cancer derived from epithelial cells in the nasopharynx, is a cancer common in China, Southeast Asia, and Africa. The three-dimensional (3D) genome organization of nasopharyngeal cancer is poorly understood. A major challenge in understanding the 3D genome organization of cancer samples is the lack of a method for the characterization of chromatin interactions in solid cancer needle biopsy samples. Here, we developed Biop-C, a modified in situ Hi-C method using solid cancer needle biopsy samples. We applied Biop-C to characterize three nasopharyngeal cancer solid cancer needle biopsy patient samples. We identified topologically associated domains (TADs), chromatin interaction loops, and frequently interacting regions (FIREs) at key oncogenes in nasopharyngeal cancer from the Biop-C heatmaps. We observed that the genomic features are shared at some important oncogenes, but the patients also display extensive heterogeneity at certain genomic loci. On analyzing the super enhancer landscape in nasopharyngeal cancer cell lines, we found that the super enhancers are associated with FIREs and can be linked to distal genes via chromatin loops in NPC. Taken together, our results demonstrate the utility of our Biop-C method in investigating 3D genome organization in solid cancers.

MP60-14 POST-BIOPSY CELL-FREE DNA IN BLOOD AS A TOOL FOR MOLECULAR TESTS IN LOCALIZED PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Markers (MP60)1 Sep 2021MP60-14 POST-BIOPSY CELL-FREE DNA IN BLOOD AS A TOOL FOR MOLECULAR TESTS IN LOCALIZED PROSTATE CANCER Marinella Corbetta, Chiara Chiereghin, Ilaria De Simone, Giulia Soldà, Monica Zuradelli, Michele Giunta, Davide Maffei, Giovanni Lughezzani, Nicolò Maria Buffi, Rodolfo Hurle, Alberto Rosario Saita, Paolo Casale, Rosanna Asselta, Massimo Lazzeri, Giorgio Ferruccio Guazzoni, and Stefano Duga Marinella CorbettaMarinella Corbetta More articles by this author , Chiara ChiereghinChiara Chiereghin More articles by this author , Ilaria De SimoneIlaria De Simone More articles by this author , Giulia SoldàGiulia Soldà More articles by this author , Monica ZuradelliMonica Zuradelli More articles by this author , Michele GiuntaMichele Giunta More articles by this author , Davide MaffeiDavide Maffei More articles by this author , Giovanni LughezzaniGiovanni Lughezzani More articles by this author , Nicolò Maria BuffiNicolò Maria Buffi More articles by this author , Rodolfo HurleRodolfo Hurle More articles by this author , Alberto Rosario SaitaAlberto Rosario Saita More articles by this author , Paolo CasalePaolo Casale More articles by this author , Rosanna AsseltaRosanna Asselta More articles by this author , Massimo LazzeriMassimo Lazzeri More articles by this author , Giorgio Ferruccio GuazzoniGiorgio Ferruccio Guazzoni More articles by this author , and Stefano DugaStefano Duga More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002095.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The analysis of circulating cell-free DNA (ccfDNA) and its tumoral fraction, the circulating tumor DNA (ctDNA), represents an innovative strategy for cancer management. It can provide information on the molecular profile of patients, useful for early cancer diagnosis and monitoring. However, considering localized prostate cancer (PCa), the presence of a pseudo-capsule surrounding the organ limits the release of ccfDNA in the circulation and its tumoral fraction is under the threshold for detection. We hypothesized that during the biopsy the needle punctures performed on the organ are likely to cause a release of prostate-derived ccfDNA, suitable for further molecular tests. METHODS: We collected needle biopsies and a blood sample (3 mL) before the start of the biopsy procedure from 38 patients undergoing transperineal prostate biopsy. A second blood sample was collected at the end of the procedure, at different sampling times: for 13 patients after 60 minutes, for 19 after 120 minutes and for 6 patients 4 post-biopsy blood samples were taken (after 10, 30, 60 and 120 minutes). ccfDNA was extracted from 1 mL of plasma using the Maxwell RSC ccfDNA Plasma Kit (Promega) and quantified with a Qubit fluorometer (Thermo Fisher). The size distribution analysis was performed using a TapeStation (Agilent). Patients’ specific somatic mutations were identified with the TruSight RNA PanCancer panel (Illumina) on bioptic material and were searched in ccfDNA of the corresponding patient using a targeted sequencing (NEBNext Ultra II DNA Library Prep kit, NEB) of selected amplicons. RESULTS: We demonstrated a significant increase in the amount of ccfDNA after the biopsy procedure, both after 60 and 120 minutes (P≤0.0024). The size distribution analysis revealed the presence of longer DNA molecules, up to 1 kb, in the post-biopsy samples compared to the pre-biopsy condition. Longer molecules are probably prostate-derived and freshly released after the end of the biopsy, not yet being degraded by nucleases in the circulation. Five patient-specific somatic variants were detected with an experiment of targeted RNA-sequencing performed on needle biopsy samples. The NGS analysis of amplicons targeting the selected variants in pre- and post-biopsy ccfDNA in the corresponding patients revealed an enrichment in ctDNA (from 2.2 to 164 fold) in the post-biopsy sample CONCLUSIONS: This study opens the possibility of exploiting the enrichment of ctDNA in the post-biopsy condition for the analysis of somatic mutations/epigenetic modifications in ccfDNA for the first time in the context of localized PCa. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1047-e1047 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Marinella Corbetta More articles by this author Chiara Chiereghin More articles by this author Ilaria De Simone More articles by this author Giulia Soldà More articles by this author Monica Zuradelli More articles by this author Michele Giunta More articles by this author Davide Maffei More articles by this author Giovanni Lughezzani More articles by this author Nicolò Maria Buffi More articles by this author Rodolfo Hurle More articles by this author Alberto Rosario Saita More articles by this author Paolo Casale More articles by this author Rosanna Asselta More articles by this author Massimo Lazzeri More articles by this author Giorgio Ferruccio Guazzoni More articles by this author Stefano Duga More articles by this author Expand All Advertisement Loading ...

194P Stromal tumor-infiltrating lymphocyte scores in core needle biopsy and its correlation to complete pathological response to neoadjuvant systemic therapy

Neoadjuvant systemic therapy (NAST) is increasingly being used to treat locally advanced breast cancer with the goal of less extensive surgery of the breast and axilla. The best possible outcome of NAST is complete pathological response (pCR) in both sites. A high density of tumor-infiltrating lymphocytes (TIL) is prognostic for pCR to NAST in some histologic subtypes. The aim of our study was to prospectively assess TIL and pCR correlation on core needle biopsy (CNB) samples in patients before NAST.

Analytical Validation and Performance of a 7-Gene Next-Generation Sequencing Panel in Uveal Melanoma

Introduction: Gene expression profiling (GEP) is widely used for prognostication in patients with uveal melanoma (UM). Because biopsy tissue is limited, it is critical to obtain as much genomic information as possible from each sample. Combined application of both GEP and next-generation sequencing (NGS) allows for analysis of RNA and DNA from a single biopsy sample, offers additional prognostic information, and can potentially inform therapy selection. This study evaluated the analytical performance of a targeted custom NGS panel for mutational profiling of 7 genes commonly mutated in UM. Methods: One hundred five primary UM tumors were analyzed, including 37 formalin-fixed paraffin-embedded (FFPE) and 68 fine-needle aspiration biopsy specimens. Sequencing was performed on the Ion GeneStudio S5 platform to an average read depth of >500X per region of interest. Results: The 7-gene panel achieved a positive percent agreement of 100% for detection of both single-nucleotide variants and insertions/deletions, with a technical positive predictive value of 98.8% and 100%, respectively. Intra-assay and inter-assay concordance studies confirmed the assay’s reproducibility and repeatability. Discussion/Conclusion: The 7-gene panel is a robust, highly accurate NGS test that can be successfully performed, along with GEP, from a single small-gauge needle biopsy sample or FFPE specimen.

Pathology Quality Control for Multiplex Immunofluorescence and Image Analysis Assessment in Longitudinal Studies.

Immune profiling of formalin-fixed, paraffin-embedded tissues using multiplex immunofluorescence (mIF) staining and image analysis methodology allows for the study of several biomarkers on a single slide. The pathology quality control (PQC) for tumor tissue immune profiling using digital image analysis of core needle biopsies is an important step in any laboratory to avoid wasting time and materials. Although there are currently no established inclusion and exclusion criteria for samples used in this type of assay, a PQC is necessary to achieve accurate and reproducible data. We retrospectively reviewed PQC data from hematoxylin and eosin (H&E) slides and from mIF image analysis samples obtained during 2019. We reviewed a total of 931 reports from core needle biopsy samples; 123 (13.21%) were excluded during the mIF PQC. The most common causes of exclusion were the absence of malignant cells or fewer than 100 malignant cells in the entire section (n = 42, 34.15%), tissue size smaller than 4 × 1 mm (n = 16, 13.01%), fibrotic tissue without inflammatory cells (n = 12, 9.76%), and necrotic tissue (n = 11, 8.94%). Baseline excluded samples had more fibrosis (90 vs 10%) and less necrosis (5 vs 90%) compared with post-treatment excluded samples. The most common excluded organ site of the biopsy was the liver (n = 19, 15.45%), followed by soft tissue (n = 17, 13.82%) and the abdominal region (n = 15, 12.20%). We showed that the PQC is an important step for image analysis and that the absence of malignant cells is the most limiting sample characteristic for mIF image analysis. We also discuss other challenges that pathologists need to consider to report reliable and reproducible image analysis data.

Abstract 3163: Progressive prostate cancer LNCaP subclones partially retain regressive communications with patient-derived fibroblasts

Abstract The tumor microenvironment of prostate cancer (PCa) plays an important role in disease progression, responsiveness to therapy, or inversely restraint of tumor growth. PCa cell evolution including loss of androgen receptor dependency is associated with intratumor heterogeneity, generating progressive PCa cell subclones. The tumor stroma of PCa contains multiple functionally different populations of fibroblasts that regulate PCa cell progression, e.g., cancer-promoting or cancer-restraining. Fibroblasts secret a number of growth factors, cytokines, the ECM molecules, and miRNAs in a cellular characteristic-specific manner. Thus, we hypothesize that diversity of fibroblasts may induce heterogenous PCa cell progression. To investigate the heterotypical communications between PCa cells and fibroblasts, we used 3 subclones of androgen-sensitive LNCaP cells and 4 patient-derived fibroblasts. Androgen-low-sensitive E9 and F10 cells were obtained from the parental androgen-sensitive LNCaP cell population through use of a limiting dilution method in regular culture condition. In contrast, androgen-insensitive AIDL cells were established from LNCaP cells by continuous passaging under hormone-depleted condition. Commercially available normal human prostate stromal cells (PrSC) was purchased from Lonza Group Ltd. Three original fibroblasts pcPrFs (-M5, -M28, -M31) were isolated from needle biopsy samples of patients with advanced PCa. To examine the effects of fibroblasts on cancer-related gene expression in PCa cells, in vitro co-culture experiment was performed using cell culture insert. In all PCa cells, LNCaP and its subclones, expressions of NFKB1, SRC, and MYC mRNA were not dramatically changed by co-culturing with 3 PCa patient-derived fibroblasts. Only in LNCaP cells, however, they were uniformly down-regulated by co-culturing with PrSC. Only in LNCaP cells, expression of CDH2 mRNA was up-regulated by co-culturing with pcPrF-M5 and pcPrF-M28. In all PCa cells, expression of NKX3-1 mRNA was uniformly up-regulated by co-culturing with pcPrF-M28 and pcPrF-M31. In LNCaP, E9, and F10 cells but not in AIDL cells, expression of GSTP1 mRNA was uniformly up-regulated by co-culturing with pcPrF-M31. Cell growth of LNCaP cells was not remarkably affected by co-culturing with pcPrF-M28 and pcPrF-M31. The heterogeneous stromal compartment in PCa tissues contains multiple functionally different populations of fibroblasts that are also associated with intratumor heterogeneity. Our data showed that up-regulations of NKX3-1 and GSTP1 mRNA expression by co-culturing with pcPrF-M28 and pcPrF-M31 were commonly observed in LNCaP subclones differing in androgen sensitivity. This result suggests that certain fibroblasts may restrain PCa cell progression by up-regulations of tumor suppressor gene expression even in progressive poorly differentiated PCa cells. Citation Format: Kenichiro Ishii, Chise Matsuda, Yasuhisa Nakagawa, Taku Shirai, Yoshifumi Hirokawa, Takeshi Sasaki, Kazuhiro Iguchi, Masatoshi Watanabe. Progressive prostate cancer LNCaP subclones partially retain regressive communications with patient-derived fibroblasts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3163.

Intra- and inter-observer variability in tumor infiltrating lymphocyte scoring in breast cancer core needle biopsy.

e12626 Background: Morphological evaluation of tumor lymphocyte infiltration (TIL) in breast cancer (BC) is gaining importance in the clinical setting, as it provides good prognostic information. Most institutions adhere to TIL working group guidelines for evaluating TIL, while having to settle on an acceptable error margin due to its subjective nature, which leads to intra and inter-observer scoring discordance. We aimed to analyze both at our institution, using experienced and inexperienced examiners, with a continuous variable scoring system. Methods: 209 BC core needle biopsy (CNB) samples were stained using hematoxylin-eosin. The percentage of stromal TIL was scored using a numerical variable ranging from 1 to 100. The examination group consisted of two experienced pathologists (pathologist A and B) and one inexperienced examiner - a medical oncology resident with a learning process containing study of the TIL working group analysis criteria and about 100 samples analysed together with an experienced pathologist Pathologist A and the resident analyzed the study samples twice, pathologist B once. Intraclass correlation coefficient (ICC) was used to measure overall intra and inter-observer agreement. Statistical analysis was performed using Google Sheets and Python. Results: 203 CNB samples were analysed (6 were excluded due to inadequate quality or an inconclusive diagnosis). Patients were aged 26 to 79 years (median 49). Sample size ranged from 1 to 16 mm (median 8). The proportion of BC subtypes was: luminal A-like 18%, luminal B-like 39%, HER2+ 10%, luminal B-like HER2+ 12%, TNBC 18%, not defined 2%. The highest proportion of high stromal TIL (≥50%) was seen in HER2+ (30%) and TNBC (22%) subtypes, as observed by pathologist A. We found good intra and inter-observer ICC (Table). Conclusions: Acceptable intra and inter-observer variability was observed in experienced pathologists, suggesting that the proposed methodology could reliably be used in clinical practice, research and trials. Interestingly, variability analysis of scores from a trained non-pathologist has also produced good results. However, it is important to note that inexperienced scoring could be prone to errors, for example counting non-lymphocyte cells as such, not recognizing regions of necrosis or sample damage, or not distinguishing between the tumor and peripheral stroma. Intraclass correlation: two-way random, single score (ICC2); pathologist A and resident (R) first and second analysis (A1, A2, R1, R2); pathologist B analysis (B1). Interpretation of ICC: &lt;0.50 poor; 0.50-0.75 moderate; 0.75-0.90 good (0.75 = minimal acceptable value for a reliable clinical measurement); &gt;0.90 excellent. Clinical trial information: 2018-000547-11. [Table: see text]

Comparison of breast cancer surrogate subtyping using a closed-system RT-qPCR breast cancer assay and immunohistochemistry on 100 core needle biopsies with matching surgical specimens

BackgroundRoutine clinical management of breast cancer (BC) currently depends on surrogate subtypes according to estrogen- (ER) and progesterone (PR) receptor, Ki-67, and HER2-status. However, there has been growing demand for reduced immunohistochemistry (IHC) turnaround times. The Xpert® Breast Cancer STRAT4* Assay (STRAT4)*, a standardized test for ESR1/PGR/MKi67/ERBB2 mRNA biomarker assessment, takes less than 2 hours. Here, we compared the concordance between the STRAT4 and IHC/SISH, thereby evaluating the effect of method choice on surrogate subtype assessment and adjuvant treatment decisions.MethodsIn total, 100 formalin-fixed paraffin-embedded core needle biopsy (CNB) samples and matching surgical specimens for 98 patients with primary invasive BC were evaluated using the STRAT4 assay. The concordance between STRAT4 and IHC was calculated for individual markers for the CNB and surgical specimens. In addition, we investigated whether changes in surrogate BC subtyping based on the STRAT4 results would change adjuvant treatment recommendations.ResultsThe overall percent agreement (OPA) between STRAT4 and IHC/SISH ranged between 76 and 99% for the different biomarkers. Concordance for all four biomarkers in the surgical specimens and CNBs was only 66 and 57%, respectively. In total, 74% of surgical specimens were concordant for subtype, regardless of the method used. IHC- and STRAT4-based subtyping for the surgical specimen were shown to be discordant for 25/98 patients and 18/25 patients would theoretically have been recommended a different adjuvant treatment, primarily receiving more chemotherapy and trastuzumab.ConclusionsA comparison of data from IHC/in situ hybridization and STRAT4 demonstrated that subsequent changes in surrogate subtyping for the surgical specimen may theoretically result in more adjuvant treatment given, primarily with chemotherapy and trastuzumab.

New Insight in the Using of Core Needle Biopsy As First Line in Preoperative Diagnosis of Thyroid Nodule

Background: The diagnosis of thyroid nodule by core needle biopsy is different from post resection samplesbecause of difficulties in applying histological criteria for both samples due to limitation in core needlebiopsy sample .Objective: To assess the utility of core needle biopsy in diagnosis of thyroid nodule.Material and Method: We use core needle biopsy as first line method instead of fine needle aspirationand to reach final diagnosis after resection sample . Core needle biopsy results were divided to six groups)unsatisfactory, benign, indeterminate, follicular neoplasm, suspicious for malignancy and malignant)according to The Bethesda system for reporting thyroid cytopathology (1,2,3) .Result : A total of 45 thyroid nodules taken from 45 patients (30 female and 15 male) . Malignant changeswere seen in 19 nodules after surgery and 26 after core needle biopsy. Benign nodules were diagnosed in 11cases after surgery and in 12 cases after core needle biopsy . Final diagnoses were concluded for 34 nodules(76%). Twenty one nodules were diagnosed as malignant nodules and 13 nodules were diagnosed as benignnodules .Conclusions: Core needle biopsy is reliable and valuable tool for the diagnosis of thyroid nodule and canbeen used as an alternative to fine needle aspirate especially in patients with inconclusive results .

A not so worthless attempt to develop primary culture from breast FNAC/CNB samples!

Breast cancer is the second leading death-causing cancer in women. Since a longtime, continuous breast cancer cell lines have been used for drug discovery based research. However, these cell lines may not represent the characteristics of original as they are believed to undergo changes due to repeated passaging. Primary cultures directly obtained from the tumor are gaining more attention worldwide as these are believed to mimic the disease process more accurately than that of continuous cell lines. The major problem with the usage of primary culture is to get the sufficient number of cells for the experiments. Here, in this study we attempted to develop primary culture both from FNAC and core needle biopsy (CNB) samples of the breast cancer patients. Overall, we believe that it is not so difficult to establish primary cultures of breast cancer cells from clinical FNAC/CNB samples. However, the starting population in FNAC samples is less which can be a hindrance in getting an adequate number of cells to reach a monolayer. CNB samples may be better quantitatively and stepwise troubleshooting of problems faced can help in the successful establishment of monolayer and primary cultures that can be propagated. We believe, our experience might be helpful for those who are struggling with the development of the short-term primary culture of breast cancer from the small-sized tissue samples.