Core Needle Biopsy Specimens Research Articles

Abstract OT2-03-01: RNA disruption assay (RDA) - Breast cancer response evaluation for individualized therapy (brevity/brevity-02)

Abstract Background & Rationale Identifying non-responders to chemotherapy with reliable predictive tests is crucial to response-guided neoadjuvant therapy. BREVITY aims to support RDA as a response assessment tool in breast cancer.RDA is based on RNA disruption, a qualitative and quantitative alteration of ribosomal RNA that correlates with chemotherapy response. RDA stratifies patients into 3 zones: non-response zone 1, partial-response zone 2 and response zone 3. Survival data in MA.22 trial showed significantly superior 5-year DFS KM-curves for zone 3 patients compared to zones 1 & 2. MA.22 and other studies indicated a potentially valuable clinical utility to be pursued. Trial Design. Study type: Prospective 2-phase interventional study. Samples: 2 core needle biopsy specimens taken at 2 time points during chemotherapy. 1 Follow-up: 60 months from date of study enrolment. Incl. Criteria. •Women 18+ years old and able to provide informed consent. •Newly diagnosed clinical stage I, II or III breast cancer with complete surgical excision after neoadjuvant therapy as treatment goal. •Confirmation of invasive breast cancer of any subtype or grade, tumor size > 1cm. •Scheduled for neoadjuvant chemotherapy +/- antibodies and +/- other drugs according to SoC. •Willing to have research core needle biopsies at 2 timepoints during neoadjuvant treatment. Excl. Criteria. •Prior local or systemic therapy for current breast cancer. •Participation in another interventional clinical trial with experimental drugs during neoadjuvant therapy. •Stage IV breast cancer. •Bilateral, multifocal or multicentric breast cancer. •Prior malignant disease except curatively treated basalioma of the skin or pTis of the cervix uteri. •Pregnancy or breast feeding. Trial Objectives & Endpoints Statistical Methods. Phase 1: training set will be used to estimate and visualize operating characteristics as functions of RDI. These include NPV, PPV, sensitivity, specificity, and related concepts. Suitable cut-offs will be determined that optimize NPV of zone 1 and PPV of zone 3. Phase 1 powered to estimate 25th RDI percentile as candidate for cut-off c1 with 9.5% precision E. Phase 2: analysis by interval-estimation of performance characteristics on an independent validation set, given cut-offs selected in phase 1. Primary target of validation is NPV of zone 1 that corresponds to RDA’s main clinical utility. PPV of zone 3 to be established for selected subgroups. Sample size for validation chosen to yield 95% CI estimate for NPV with 5.25% precision. Present and Target Accrual. Present AccrualTarget Accrual(must be fully evaluable). Phase 1 (Training set):71 (target 80). Phase 2 (Validation set):454 Other Contacts. Sponsor: Rna Diagnostics Inc Biopsy Collection Time Points for RDABiopsy(2 specimens)1st core biopsy2nd core biopsyTiming35 +/- 4 daysIf no drug change:55 +/- 5 days. If drug change:2-3 weeks after start of new drugs:•3-weekly: at 16 +/- 2 days•Bi-weekly: at day of 2nd dose•Weekly: at day of 4th dose Primary ObjectivesDescriptionEndpointsi.1.Phase 1: Determine 2 RDI cut-offs to have a diagnostic test optimized in terms of predictive values2.Phase 2: Establish performance characteristics for first cut-off in terms of NPVNPVPPVSecondary Objectivesi.Assess NPV in subgroups by subtypeNPVii.Assess PPV in HER2+PPViii.Compare pCR prevalence and DFS in zones 1-3 for all patients and by subgrouppCRDFSv.Test for association btw. cancer subgroups and test resultTest result (zones 1-3)vi.Compare RDA and other biomarker results (Ki67, tumor grade) Citation Format: Maureen Trudeau, Joke Tio, Foluso Ademuyiwa, Thierry Petit, Bryan Hennessy, Marina Cazzaniga, Daniele Generali. RNA disruption assay (RDA) - Breast cancer response evaluation for individualized therapy (brevity/brevity-02) [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr OT2-03-01.

Abstract P1-03-01: Heterogeneity between core needle biopsy and primary tumor tissue in early breast cancer patients: Comparison of intrinsic subtypes after different treatment regimes

Abstract Background: All initial therapeutic decisions in early breast cancer are commonly based on the expression profiles of estrogen (ER), progesterone (PR) and the human epidermal growth factor 2 (HER2) receptors. However, breast cancer is a very heterogeneous disease, and receptor changes were manifold reported during progression. Only little data is known about receptor changes after upfront therapy. Here, we compared receptor expression profiles between core needle biopsy (CNB) tissue and primary tumor tissue after different treatment regimes. Methods: In a German single center study, we retrospectively analyzed 248 breast cancer patients during primary treatment regime between 2014 and 2020. Patients had either neoadjuvant chemotherapy, neoadjuvant endocrine therapy or no upfront therapy. Tumor material was obtained by core needle biopsy (CNB) at primary diagnosis and during primary oncological surgery of the axilla and breast. Analysis of histological subtype, grading, Ki67 index and expression profiling of ER, PR, HER2, was performed using formalin-fixed, paraffin-embedded (FFPE) specimens. Immunohistochemical examination was performed according to the ASCO/CAP guidelines using the Ventana-platform. Tumors were grouped into different intrinsic subtypes according to the international St. Gallen classification in either Luminal A, Luminal B, HER2-enriched or triple negative. 35 patients were excluded because the intrinsic subtype was not specified in the CNB and final postoperative specimen, therefore 213 patients were included in the primary analysis. Results: In the primary analysis (n=213), median age was 53 years (IQR 43 - 64). Based on the initial CNB receptor profile 105 (49%) patients had a Luminal A carcinoma, 46 (22%) patients had a Luminal B HER2 negative carcinoma, 22 (10%) patients had a Luminal B HER2 positive carcinoma, 6 (3%) patients had a HER2 enriched carcinoma and 33 (16%) patients had a triple negative carcinoma. In the primary analysis 84 (39%) patients had neoadjuvant chemotherapy, 48 (23%) patients had neoadjuvant endocrine therapy and 81 (38%) patients had no upfront therapy. Overall, 77 (36%) patients had an intrinsic subtype change between CNB and definitive surgical treatment, 139 (64 %) patients had no subtype change. There were 44 (52%) changes after neoadjuvant chemotherapy, 17 (35%) changes after neoadjuvant endocrine therapy and 16 (20%) subtype changes after no upfront therapy (p<0.0001 for the effect of neoadjuvant chemotherapy). ER receptor status changed in 5 (6%) patients after neoadjuvant chemotherapy, 6 (13%) changes were observed after neoadjuvant endocrine therapy and 2 (2%) ER changes occurred after no upfront therapy. Concerning the PR receptor there were 23 (27%) receptor changes after neoadjuvant chemotherapy, 17 (35%) changes after neoadjuvant endocrine therapy and 9 (11%) changes after no upfront therapy (p<0.0001 for the effect of neoadjuvant endocrine therapy). Regarding the HER2 receptor, there were 18 (21%) receptor status changes after neoadjuvant chemotherapy, 2 (4%) changes after neoadjuvant endocrine therapy and 2 (2%) subtype changes after no upfront therapy (p=0.0001 for the effect of neoadjuvant chemotherapy).Neoadjuvant chemotherapy led significantly more often to a decrease of HER2 expression, compared to neoadjuvant endocrine therapy or no upfront therapy. Conclusions: Our results imply the high frequency of intrinsic subtype changes after neoadjuvant therapy. Subtype changes should be taken into account for an optimal and individual treatment. Further research needs to be conducted to investigate whether an individual treatment decision based on the receptor profile after primary treatment might improve clinical outcome of breast cancer patients. Citation Format: Laura Weydandt, Anne Kreklau, Ivonne Nel, Lars-Christian Horn, Bahriye Aktas. Heterogeneity between core needle biopsy and primary tumor tissue in early breast cancer patients: Comparison of intrinsic subtypes after different treatment regimes [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-03-01.

Abstract P4-07-11: Prognostic significance of changes in tumor infiltrating lymphocytes and neutrophil-to-lymphocyte ratio after neoadjuvant chemotherapy in early breast cancer patients

Abstract Introduction. There are still 30-40% of patients with early breast cancer (BC) that relapse after neoadjuvant chemotherapy (NAC). New prognostic biomarkers are needed. Current evidence shows that both markers of antitumor local and systemic immune response, such as tumor infiltrating lymphocytes (TIL) and neutrophil-to-lymphocyte ratio (NLR) have prognostic value in early BC. Most works have evaluated baseline levels, but their variation after NAC might be related to treatment resistance and should be explored.The aim of this study was to simultaneously investigate the prognostic significance of the pre-post-NAC variation of TIL and NLR in patients with early BC.Methods. Retrospective, single-center cohort of 121 patients with early BC treated with NAC between 2001 and 2010. Pre- and post-NAC TIL (CD3+) were assessed by tumor tissue microarrays performed in both diagnostic core-needle biopsy and surgical excision specimens, respectively. Pre- and post-NAC NLR were calculated based on blood tests collected within 1 month of cancer diagnosis and at the moment of surgery, respectively. The change on TIL and NLR after NAC was estimated by calculating the absolute difference of post- and pre- values of these parameters (Δpost-pre). We examined the association between the absolute difference of TIL and NLR (both divided into deciles) and survival outcomes by Cox regression. To represent in Kaplan-Meier curves the combined effect of the change of TIL and NLR in OS, we divided patients into 4 groups, depending on the sign of Δpost-pre values of TIL and NLR: 1) no TIL nor NLR increment after NAC (ΔTIL & ΔNRLpost-pre≤0); 2) no TIL but NLR increment (ΔTILpost-pre≤0, ΔNRLpost-pre>0); 3) TIL but no NLR increment (ΔTILpost-pre>0, ΔNRLpost-pre≤0); and 4) TIL and NLR increment following NAC (ΔTIL & ΔNRLpost-pre>0).Results. 47 (40%) out of 121 patients had pre- and post-NAC values of both NLR and TIL. Patients’ baseline characteristics are shown in Table 1. The median follow-up of this group was 11.5 years (IQR: 10.5 - 12.5 years). At the final follow-up date, overall survival (OS) was 87.2 %. In the univariable analysis, the increase in TIL and NLR after NAC among deciles showed a negative prognostic value for OS (TIL: HR 1.54, 95%CI 1.05-2.26, p=0.026; HR 1.32 95%CI 0.98-1.79, p=0.070). In the multivariable analysis, adjusted by cancer staging after NAC, both the increase of TIL and NLR after NAC among deciles add independent predictive value (TIL: HR 1.53, 95%CI 1.00-2.34, p=0.049; RNL: HR 1.42 95%CI 0.95-2.12, p=0.09).The subpopulation of patients with no TIL increment after NAC (Groups 1&2, N=23) showed the best prognosis, with no deaths at 10 years. Among patients with TIL increment (Groups 3&4, N=22), those with no NLR increment (Group 3, N=5) showed an intermediate prognosis (20% deaths at 10 years), whereas patients with concurrent NLR increment (Group 4, N=17) had the lowest OS (29.4% deaths at 10 years). Differences observed between groups were statistically significant (Log Rank p=0.042).Conclusion. The integrated characterization of TIL and NLR variations after NAC identifies different prognostic subgroups in early BC patients. In our cohort, both the increment of TIL and/or NLR after NAC are associated to a worse prognosis. Future validation of these findings in large, multicenter cohorts might allow treatment optimization by means of new strategies such as immunotherapy. Table 1.Patients’ baseline characteristicsN=47Age (median, range)56 (21, 78)Menstrual statusPostmenopausal25 (53,2%)Premenopausal22 (46,8%)Histologic subtypeInvasive ductal carcinoma42 (91,5%)Other subtypes4 (8,5%)Histologic gradeGrade 12 (4,3%)Grade 218 (38,3%)Grade 323 (48,9%)Unknown4 (8,6%)cTcT221 (44,7%)cT324 (51,1%)cT4a-d2 (4,2%)cNcN018 (38,3%)cN114 (29,8%)cN26 (12,8%)cN39 (19,1%)Molecular subtypeHR+ HER2-22 (46,8%)HER2+ HR+8 (17,0%)HER2+ HR-6 (12,8%)Triple negative10 (21,3%)Breast surgeryConservative26 (55,3%)Mastectomy21 (44,7%)Axillary surgerySentinel lymph node biopsy12 (25,5%)Axillary lymph node dissection35 (74,5%)Relapse typeMetastasis7 (14,9%)Local/contralateral1 (0,83%)Deaths6 (12,8%) Citation Format: Esmeralda Garcia-Torralba, Beatriz Álvarez-Abril, Carlos Bravo-Pérez, Esther Navarro Manzano, Pilar de la Morena Barrio, Alejandra Ivars Rubio, Elisa García-Garre, Gema Marín Zafra, Francisco Ayala de la Peña, Elena García-Martínez. Prognostic significance of changes in tumor infiltrating lymphocytes and neutrophil-to-lymphocyte ratio after neoadjuvant chemotherapy in early breast cancer patients [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-07-11.

The Role of Core Needle Biopsy Pathology Combined with Molecular Tests in the Diagnosis of Lymph Node Tuberculosis

BackgroundEarly lymph node tuberculosis (LNTB) diagnosis is still difficult. The majority of LN specimens require the undertaking of invasive and unpleasant procedures.PurposeTo evaluate the diagnostic efficacy of pathology when combined with molecular tests for the diagnosis of LNTB in core needle biopsy (CNB) specimens and to compare that diagnostic efficacy with that deriving from tissue specimens’ examination alone.MethodsWe retrospectively analyzed the medical records of LNTB patients who met the inclusion criteria. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and area under the curve (AUC) of pathology, molecular tests, and parallel test (positive result for either of these two assays) were calculated to evaluate their diagnostic efficacy compared with a composite reference standard.ResultsA total of 289 patients were included in the study. The overall sensitivity, specificity, PPV, NPV, and AUC of pathology, molecular tests, and parallel test were 94.5%, 97.2%, 99.6%, 71.4%, 0.96; 73.1%, 100.0%, 100.0%, 34.6%, 0.87; and 98.4%, 97.2%, 99.6%, 89.7%, 0.98, respectively. For CNB specimens, these values for pathology, molecular tests, and parallel test were 93.3%, 96.2%, 99.4%, 69.4%, 0.95; 76.4%, 100.0%, 100.0%, 40.0%, 0.88; and 99.4%, 96.2%, 99.4%,96.2%,0.98, while those same values for the tissue were 96.6%, 100.0%, 100.0%, 76.9%, 0.98; 67.1%, 100.0%, 100.0%, 25.6%, 0.84; and 96.6%, 100.0%, 100.0%, 76.9%,0.98, respectively.ConclusionThe validity of pathology and molecular testing when using CNB specimens was similar to that of tissue specimens for relevant assessment approaches. For the LNTB diagnosis, CNB specimens were preferred for the simultaneous undertaking of pathological examination and molecular testing.

Discordance of Histological Grade between Diagnostic Core-Needle Biopsy and Resection Specimen for Retroperitoneal Leiomyosarcomas

Background: Diagnostic core-needle biopsy (DCNB) allows to identify the histological subtype of soft-tissue sarcoma in retroperitoneal sarcomas (RPS) with reasonable accuracy and safety. This allows for treatment tailored to different histologic subtypes of RPS, including neoadjuvant therapy. The discrepancy between the histological grade between DCNB and the resection specimen for retroperitoneal Leiomyosarcomas (RPLMS) has been highlighted before. The histological grade on DCNB is an important tool to select patients for preoperative therapy.

Impact of intratumoral heterogeneity on the metabolic profiling of breast cancer tissue using high-resolution magic angle spinning magnetic resonance spectroscopy.

High-resolution magic angle spinning (HR-MAS) magnetic resonance spectroscopy (MRS) is a useful metabolic profiling technique for human tissue. However, the impact of intratumoral heterogeneity on the metabolite levels of breast cancers is not yet established. The purpose of this prospective study was to investigate whether the tumor cell fraction of core needle biopsy (CNB) specimens of breast cancers affect metabolic profiles assessed with HR-MAS MRS. From June 2015 to December 2016, 46 patients with 47 breast cancers were enrolled. HR-MAS MRS was used for the metabolic profiling of 285 CNB specimens from the 47 cancers. Multiple CNB samples (range 2-8) for the HR-MAS MRS experiment were obtained from surgical specimens under ultrasound guidance following surgical removal of the tumor. Tumor cell fraction was expressed as a percentage of the tumor cell volume relative to the total tumor volume contained in each CNB sample. Metabolite quantification levels were compared according to primary tumor characteristics using the t-test. Multivariate analyses were performed including primary tumor characteristics and tumor cell percentages as variables. Correlations between tumor cell percentage and metabolite levels in the CNB specimens were assessed according to the immunohistochemical status of the primary tumor. In univariate analysis, levels of choline-containing compounds, glutamate, glutamine, glycine, serine, and taurine were correlated with primary tumor characteristics. In multivariate analysis, most metabolite levels were not affected by tumor cell percentage. Tumor cell percentage showed poor correlation with metabolite levels in hormone receptor-positive cancer and triple-negative cancer, and poor to fair correlation with metabolite levels in HER2-positive cancer. This study showed that differences in the tumor cell fraction of CNB samples do not affect predictions on the primary cancer from which the samples are obtained.

Diagnosis of giant cell-rich bone tumors on core needle biopsy: A practical approach

BackgroundThe differential diagnosis of giant cell-rich bone tumors comprises a broad spectrum of lesions with prominent reactive osteoclast-like and/or neoplastic giant cells, with substantial differences in biologic behavior and clinical management. Evaluation of giant cell-rich bone tumors on small biopsies can be challenging especially in specimens with limited representative material. An accurate diagnosis requires a high level of skill on the part of both radiologist and pathologist as correlation with clinical and radiologic characteristics is critical. The objective of the study was to assess the utility of touch preparations (TP), immunohistochemistry (IHC) for mutation-specific markers H3G34W and H3K36M, and fluorescence in situ hybridization (FISH) for USP6 rearrangements and MDM2 amplification in the diagnostic workup of core needle biopsy specimens. MethodsA total of 27 core needle biopsies with TPs from patients with primary giant cell-rich bone tumors (16 giant cell tumors of bone (GCTBs) (including 3 with lung metastasis), 3 chondroblastomas (CBs), 4 primary aneurysmal bone cysts (ABCs), 2 non-ossifying fibromas (NOFs), 1 low grade osteosarcoma (OS), and 1 conventional OS with tumor giant cells were analyzed with IHC for H3G34W and H3K36M and in select cases FISH for USP6 rearrangements and MDM2 amplification. ResultsIn all cases the core biopsies were sufficient for histologic examination and diagnostic workup. 16 of 16 GCTBs were positive for H3G34W and negative for H3K36M, and 3 of 3 CBs were positive for H3K36M and negative for H3G34W. All other cases were negative for H3G34W and H3K36M. 4 of 4 primary ABCs showed rearrangement of USP6 by FISH and the low grade OS showed amplification of MDM2 by FISH. ConclusionsOn-site adequacy assessment of TPs proved to be an accurate, simple, and fast method for obtaining sufficient material for complete diagnostic workup. The application of IHC for H3G34W and H3K36M and FISH for detection of rearrangements of USP6 and amplification of MDM2 can improve the diagnostic accuracy in core needle biopsy specimens.

Predictive value of ductal carcinoma in situ with invasive breast cancer in core needle biopsies for final pathologic size of intraductal elements

Preoperative evaluations of the size of ductal carcinoma in situ (DCIS) extension in invasive breast cancer (IBC) are problematic and markers of the actual size of DCIS remain elusive. This study aimed to quantify DCIS on core needle biopsy (CNB) and investigated its association with degree of DCIS extension on paired resection specimens, instead of with presence or absence of an extensive intraductal component or margin status as in earlier studies. This series examined 150 IBCs diagnosed from paired CNB and resection specimens. The DCIS/invasion ratio was calculated using the sum of each element size from CNB. In resection specimens, cases in which the greatest dimension of DCIS extension was longer than the greatest dimension of invasive size were defined as extended DCIS (Ext-DCIS). DCIS/invasion ratio level correlated positively with the degree of Ext-DCIS (P = 0.003). Using receiver operating characteristic curve analysis, setting cases with the subgroup of DCIS extension with greatest dimension > 2.5 times that of the invasive size in the resection specimen (Ext-DCIS > 2.5) as the positive class provided the best discrimination ability for DCIS/invasion ratio (0.375). In multivariate analysis, DCIS/invasion ratio > 0.375 was significantly associated with Ext-DCIS > 2.5 (P = 0.033). In conclusion, DCIS/invasion ratio > 0.375 in CNB was identified as a predictor of Ext-DCIS > 2.5 in resection specimens, suggesting that an approach combining DCIS/invasion ratio from CNB with preoperative staging may better predict the extent of DCIS and facilitate better surgical planning.

Managing the Lesion and Counseling Patient Post–Breast Biopsy

AbstractBreast cancer is the leading cause of cancer deaths worldwide among women. Early diagnosis and prompt treatment offers a chance for cure and long-term survival among afflicted women. Triple assessment which includes clinical examination, radiological imaging and histopathological evaluation has become a standard of care in the evaluation of breast cancer. Radiological imaging involves mammography, ultrasound, and magnetic resonance imaging while pathological evaluation can be done on fine needle, core-needle, and surgical excision biopsy specimens with core needle biopsy being the workhorse for diagnosis. While triple assessment has increased the diagnostic yield and helps in the identification of early lesions, an increasing subset of patients present with a situation where the imaging and pathological characteristics of a lesion do not correlate with each other thereby causing diagnostic and therapeutic dilemma. Imaging and pathology concordance and discordance present distinctly under differing circumstances in the care of the patient. While those with concordant findings proceed to treatment or follow up directly and hence benefit from the early treatment, those with discordant findings are beset with a delay in the diagnosis and should be subjected to further evaluations for confirming or refuting a diagnosis. This article reviews the salient features of image-guided breast biopsy and the outcomes and management options in imaging and pathology concordance and discordance. We also discuss the role of a radiologist in the counselling of the patient before and after receiving the pathological report after biopsy.

Immunocytochemical Evaluation of TTF-1, Napsin-A, and p-63 for Subtyping of Non-Small Cell Lung Carcinoma and Clinicopathological Correlation.

Carcinoma of lung is the most common cause of cancer-associated mortality worldwide. About 70% of lung cancer cases are unresectable and present in advanced stages. So, cytology and small core needle biopsy specimen are available for diagnostic as well as prognostication workup. Subtyping of non-small cell lung cancer (NSCLC) is essential for the treatment and further workup study. For this, immunocytochemistry (ICC) plays a crucial role that helps in early diagnosis. Subtyping of NSCLC from cytology samples using ICC markers like TTF-1, Napsin-A, and p63 and their clinicopathological correlation are the aims of the study. This ambispective study was conducted in the pathology department of a tertiary care hospital of eastern India for a 2-year period from 2018 to 2020. In our study, 46 cytologically diagnosed cases of NSCLC were included. Subtyping was done by cytomorphology and correlated with ICC expression, histopathology, and clinicopathological parameters. In our study, adenocarcinoma (ADC) was the most common (32.61%) cancer. Most cases of ADC showed positive expression of TTF-1 and Napsin-A, and p63 was positive in most cases of squamous cell carcinoma (SCC). Concordance with cytomorphology and ICC was 87.50% and 81.81% with ADC and SCC, respectively. Cyto-ICC-histo concordance was observed in 85.51% of ADC and 66.66% of SCC cases. Sensitivity was 100%, 93.1%, and 100% for TTF-1, Napsin-A, and p63, respectively. Specificity of both TTF-1 and Napsin-A was 88.2% and for p63 was 93.8%. In small biopsy along with cytology samples, ICC is cost-effective and plays an important role in early diagnosis along with management of NSCLC.

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.

A multicenter study of interobserver variability in pathologic diagnosis of papillary breast lesions on core needle biopsy with WHO classification

BackgroundPapillary breast lesions (PBLs) comprise diverse entities from benign and atypical lesions to malignant tumors. Although PBLs are characterized by a papillary growth pattern, it is challenging to achieve high diagnostic accuracy and reproducibility. Thus, we investigated the diagnostic reproducibility of PBLs in core needle biopsy (CNB) specimens with World Health Organization (WHO) classification.MethodsDiagnostic reproducibility was assessed using interobserver variability (kappa value, κ) and agreement rate in the pathologic diagnosis of 60 PBL cases on CNB among 20 breast pathologists affiliated with 20 medical institutions in Korea. This analysis was performed using hematoxylin and eosin (H&E) staining and immunohistochemical (IHC) staining for cytokeratin 5 (CK5) and p63. The pathologic diagnosis of PBLs was based on WHO classification, which was used to establish simple classifications (4-tier, 3-tier, and 2-tier).ResultsOn WHO classification, H&E staining exhibited ‘fair agreement’ (κ = 0.21) with a 47.0% agreement rate. Simple classifications presented improvement in interobserver variability and agreement rate. IHC staining increased the kappa value and agreement rate in all the classifications. Despite IHC staining, the encapsulated/solid papillary carcinoma (EPC/SPC) subgroup (κ = 0.16) exhibited lower agreement compared to the non-EPC/SPC subgroup (κ = 0.35) with WHO classification, which was similar to the results of any other classification systems.ConclusionsAlthough the use of IHC staining for CK5 and p63 increased the diagnostic agreement of PBLs in CNB specimens, WHO classification exhibited a higher discordance rate compared to any other classifications. Therefore, this result warrants further intensive consensus studies to improve the diagnostic reproducibility of PBLs with WHO classification.

Value of intraoperative pathological diagnosis in decision-making regarding resection of well-differentiated retropharyngeal liposarcoma: A case report.

Introduction and importancePreoperative diagnosis of well-differentiated liposarcoma (WDLS) in the retropharyngeal space is challenging because of the difficulty in obtaining a biopsy tissue specimen that will yield the microscopic findings necessary for a definitive pathological diagnosis. This report describes a case of retropharyngeal WDLS that was successfully diagnosed intraoperatively, which allowed radical resection.Case presentationThe patient was a 60-year-old man suspected of having a lipomatous tumor in the retropharyngeal space. On imaging, the tumor contained linear septum-like structures that were prominent behind the larynx. Pathological examination was performed using fine-needle aspiration cytology and core needle biopsy specimens. However, no malignant features were found. Given that partial biopsy of the retropharyngeal tumor by puncture was anatomically limited, we decided to collect appropriate tissue specimens for intraoperative pathological examination. During the operation, we biopsied the tumor, including the hard portion behind the larynx, anticipating inclusion of the septum-like structures seen on imaging. A pathological diagnosis of WDLS was successfully made and the tumor was completely excised.Clinical discussionThe fibrous septum with induration is important for intraoperative diagnosis of WDLS. The fibrous septum within the tumor was palpable as a rubbery hard portion.ConclusionWDLS in the retropharyngeal space could be successfully resected surgically by making a rapid intraoperative pathological diagnosis using appropriately selected tissue sampled from a hard portion of the tumor.

Acute lymphoblastic leukemia simulating breast carcinoma

BackgroundBreast metastasis in hematological malignancies is a rare phenomenon, and it is primarily seen in acute myeloid leukemia (AML). In patients with acute lymphoblastic leukemia (ALL), this condition is even rarer.Case presentation.We present a case of a precursor B cell ALL involving breast in a 40-year-old female and its imaging features on mammography and ultrasound. Histopathology of core needle biopsy (CNB) specimen allowed us to diagnose ALL with extramedullary metastases. The patient was referred to oncology for further management.ConclusionTo conclude, ALL infiltrating breast is rare but should be given due consideration, especially in the cases of known primary hematopoietic malignancy, particularly in patients presenting with a history of sudden lumps in the breast. A CNB can give reliable results in combination with flow cytometry and immunocytochemistry, circumventing the need for an excisional biopsy and allowing the commencement of early treatment.

Predictive and prognostic impact of ferroptosis-related genes ACSL4 and GPX4 on breast cancer treated with neoadjuvant chemotherapy.

BackgroundRecent evidence shows that inducing ferroptosis may improve efficacy of tumor therapy. However, ferroptosis-related genes have been little studied in patients with breast cancer especially in the neoadjuvant setting. ACSL4 and GPX4 have been well established as the positive and negative regulator of ferroptosis, respectively. This study aimed to explore the predictive value of ACSL4 and GPX4 for patients with breast cancer administered neoadjuvant chemotherapy.MethodsThis study included patients treated with paclitaxel-cisplatin-based neoadjuvant chemotherapy. Immunohistochemistry staining of ACSL4 and GPX4 was carried out on the core needle biopsy specimens. Logistic regression was performed to explore the predictive biomarkers of pathological complete response (pCR). Survival analyses were examined by log-rank test and Cox proportional hazard regression.FindingsA total of 199 patients were included for the analyses. Both ACSL4 expression and ACSL4/GPX4 combination status could serve as independent predictive factors for pCR. The interaction for pCR was observed between ACSL4 and clinical tumor stage. Besides, ACSL4 expression, GPX4 expression, and their combination status were independent prognostic factors for disease-free survival. Analyses of the Kaplan-Meier Plotter database suggested that higher ACSL4 expression is related to better overall survival, and higher GPX4 expression is related to better distant metastasis-free survival. Pathway analyses revealed that ACSL4 and GPX4 might function in crucial pathways including apoptosis, autophagy, cell adhesion, lipid metabolism, etc.InterpretationThis study revealed the critical value of ACSL4 and GPX4 serving as novel predictive and prognostic biomarkers for patients with breast cancer receiving neoadjuvant chemotherapy. It might be a novel strategy to induce ferroptosis to promote chemosensitivity. Future studies are required to elucidate the potential mechanisms.FundingThis work was supported by Shanghai Natural Science Foundation [grant number 19ZR1431100], Clinical Research Plan of Shanghai Hospital Development Center [grant numbers SHDC2020CR3003A, 16CR3065B, and 12016231], Shanghai “Rising Stars of Medical Talent” Youth Development Program for Youth Medical Talents - Specialist Program [grant number 2018-15], Shanghai “Rising Stars of Medical Talent” Youth Development Program for Outstanding Youth Medical Talents [grant number 2018-16], Shanghai Collaborative Innovation Center for Translational Medicine [grant number TM201908], Multidisciplinary Cross Research Foundation of Shanghai Jiao Tong University [grant numbers YG2017QN49, ZH2018QNA42, and YG2019QNA28], Nurturing Fund of Renji Hospital [grant numbers PYMDT-002, PY2018-IIC-01, PY2018-III-15, and PYIII20-09], Science and Technology Commission of Shanghai Municipality [grant numbers 20DZ2201600 and 15JC1402700], and Shanghai Municipal Key Clinical Specialty.

Comparison between core needle biopsy and excisional biopsy for breast neoplasm.

This study aimed to explore clinical significance of core needle biopsy (CNB) in pathological diagnosis of breast neoplasm.Seventy one breast neoplasm samples were obtained from Tongzhou Maternal and Child Health Hospital of Beijing between the years of 2006 and 2014. Forty five specimens were obtained via CNB and cases offering 26 of them received neoadjuvant chemotherapy. Pathology, histology, and immunohistochemistry results were compared between CNB specimens and excisional biopsy.Upward and downward tendencies could be observed in CNB specimens and excisional biopsy, respectively, in all items. Tumor proportion of CNB tissues was (33 + 2)/45 = 77.78%, when ductal carcinoma in situ detected by both CNB and excisional biopsy was 31/45 = 68.89%, with a consistency of (31 + 3)/45 = 75.56%. Tumor thrombus detected by both CNB and excisional biopsy was 2/45 = 4.44%. Among cases receiving neoadjuvant chemotherapy, CNB and excisional biopsy, in mitotic figure, cytological scoring and histological grading, showed a total change rate of >50% (50%–75%), while changes in duct and cellular heteromorphism were not distinct. Cases showing changes were up to 73.08%, with 8/26 = 30.77% for rise and 11/26 = 42.31% for descent.CNB could be used for preoperative diagnosis of breast neoplasm, and help to determine proper treatment regimen, thus elevating the rate of breast conserving. However, this method still has several limitations, particularly in immunohistochemical tests of human epidermal receptor protein-2. Neoadjuvant chemotherapy may influence the accuracy of CNB diagnosis.

Rapid non-destructive volumetric tumor yield assessment in fresh lung core needle biopsies using polarization sensitive optical coherence tomography.

Adequate tumor yield in core-needle biopsy (CNB) specimens is essential in lung cancer for accurate histological diagnosis, molecular testing for therapeutic decision-making, and tumor biobanking for research. Insufficient tumor sampling in CNB is common, primarily due to inadvertent sampling of tumor-associated fibrosis or atelectatic lung, leading to repeat procedures and delayed diagnosis. Currently, there is no method for rapid, non-destructive intraprocedural assessment of CNBs. Polarization-sensitive optical coherence tomography (PS-OCT) is a high-resolution, volumetric imaging technique that has the potential to meet this clinical need. PS-OCT detects endogenous tissue properties, including birefringence from collagen, and degree of polarization uniformity (DOPU) indicative of tissue depolarization. Here, PS-OCT birefringence and DOPU measurements were used to quantify the amount of tumor, fibrosis, and normal lung parenchyma in 42 fresh, intact lung CNB specimens. PS-OCT results were compared to and validated against matched histology in a blinded assessment. Linear regression analysis showed strong correlations between PS-OCT and matched histology for quantification of tumors, fibrosis, and normal lung parenchyma in CNBs. PS-OCT distinguished CNBs with low tumor content from those with higher tumor content with high sensitivity and specificity. This study demonstrates the potential of PS-OCT as a method for rapid, non-destructive, label-free intra-procedural tumor yield assessment.

Current Methodological Challenges of Single-Cell and Single-Nucleus RNA-Sequencing in Glomerular Diseases.

Single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA-seq (snRNA-seq) allow transcriptomic profiling of thousands of cells from a renal biopsy specimen at a single-cell resolution. Both methods are promising tools to unravel the underlying pathophysiology of glomerular diseases. This review provides an overview of the technical challenges that should be addressed when designing single-cell transcriptomics experiments that focus on glomerulopathies. The isolation of glomerular cells from core needle biopsy specimens for single-cell transcriptomics remains difficult and depends upon five major factors. First, core needle biopsies generate little tissue material, and several samples are required to identify glomerular cells. Second, both fresh and frozen tissue samples may yield glomerular cells, although every experimental pipeline has different (dis)advantages. Third, enrichment for glomerular cells in human tissue before single-cell analysis is challenging because no effective standardized pipelines are available. Fourth, the current warm cell-dissociation protocols may damage glomerular cells and induce transcriptional artifacts, which can be minimized by using cold dissociation techniques at the cost of less efficient cell dissociation. Finally, snRNA-seq methods may be superior to scRNA-seq in isolating glomerular cells; however, the efficacy of snRNA-seq on core needle biopsy specimens remains to be proven. The field of single-cell omics is rapidly evolving, and the integration of these techniques in multiomics assays will undoubtedly create new insights in the complex pathophysiology of glomerular diseases.

O14 Optimising the management of primary breast cancer in older women

Abstract Introduction The risk of breast cancer increases with age and our global population is ageing. By 2040 the number of breast cancer cases diagnosed per year worldwide will double and over 40% of these will be in patients aged 70 or over. Despite this, there are few treatment guidelines specific to breast cancer in older women and none which consider the unique biological differences of this cohort. Method Surgical and core needle biopsy (CNB) specimens were obtained from an existing series of 1,785 women over the age of 70 with primary breast cancer, treated in a single institution with long-term (37+ years) follow-up. Of this cohort, 813 had primary surgical treatment. As part of previous work, it was possible to construct good quality tissue microarrays (TMAs) in 575 surgical specimens and 693 CNB specimens. Immunohistochemical staining for 32 biomarkers has been performed in all of the available TMAs. Association between histological score for each biomarker and tumour size, grade, recurrence rate, breast cancer specific and overall survival is currently being investigated in the whole cohort. Results Results to date have revealed a unique biological cluster in older women with primary breast cancer that is not seen in a comparative younger cohort. In the future, bioinformatics analysis will determine which biomarkers and in what combination, can predict chance of recurrence/overall survival in this cohort. Conclusions This information will be used to create a prognostic tool specific to assist older women with decision making regarding primary treatment of breast cancer.

Is the 21-Gene Recurrence Score on Core Needle Biopsy Equivalent to Surgical Specimen in Early-Stage Breast Cancer? A Comparison of Gene Expression Between Paired Core Needle Biopsy and Surgical Specimens.

Molecular testing on surgical specimens predicts disease recurrence and benefit of adjuvant chemotherapy in hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2-) early-stage breast cancer (EBC). Testing on core biopsies has become common practice despite limited evidence of concordance between core/surgical samples. In this study, we compared the gene expression of the 21 genes and the recurrence score (RS) between paired core/surgical specimens. Eighty patients with HR+/HER2- EBC were evaluated from two publicly available gene expression datasets (GSE73235, GSE76728) with paired core/surgical specimens without neoadjuvant systemic therapy. The expression of the 21 genes was compared in paired samples. A microarray-based RS was calculated and a value ≥ 26 was defined as high-RS. The concordance rate and kappa statistic were used to evaluate the agreement between the RS of paired samples. Overall, there was no significant difference and a high correlation in the gene expression levels of the 21 genes between paired samples. However, CD68 and RPLP0 in GSE73235, AURKA, BAG1, and TFRC in GSE76728, and MYLBL2 and ACTB in both datasets exhibited weak to moderate correlation (r<0.5). There was a high correlation of the microarray-based RS between paired samples in GSE76728 (r=0.91, 95% confidence interval [CI] 0.81-0.96) and GSE73235 (r=0.82, 95% CI 0.71-0.89). There were no changes in RS category in GSE76728, whereas 82% of patients remained in the same RS category in GSE73235 (κ=0.64). Gene expression levels of the 21-gene RS showed a high correlation between paired specimens. Potential sampling and biological variability on a set of genes need to be considered to better estimate the RS from core needle biopsy.