Set Of Biopsies Research Articles

Abstract 1196: BALF liquid biopsy provides a novel platform for speedy and accurate EGFR mutation testing in advanced NSCLC patients

Abstract Background EGFR mutation testing is an essential step for the therapeutic decision in newly diagnosed advanced NSCLC patients and usually performed by using DNA extracted from biopsied tumor tissue with a turnaround time(TAT) of 2-3 weeks. Plasma liquid biopsy using ctDNA is minimally invasive and less time-consuming, but shows limited usefulness due to low sensitivity. We conducted a prospective study to demonstrate clinical usefulness of bronchoalveolar lavage fluid (BALF) liquid biopsy using extracellular vesicle(EV)-derived DNA in advanced NSCLC patients by comparing the sensitivity and TAT with conventional tissue biopsy and plasma liquid biopsy Methods The analysis set for liquid EGFR genotyping is 224 BALF samples and 110 plasma samples obtained from newly diagnosed 224 advanced non-squamous NSCLC patients. EGFR genotyping was done through peptide nucleic acid(PNA)-mediated real-time PCR after extracting BALF EV DNA and plasma cfDNA separately. The sensitivity, specificity, and concordance rate of BALF liquid biopsy were calculated in comparison to matched tissue genotyping and plasma liquid biopsy. In addition, TAT was compared between BALF liquid biopsy and tissue genotyping. Results Tissue genotyping revealed 93 EGFR mutant cases (41.5%) and 131 wild-type cases (58.5%). The sensitivity, specificity, positive predictive value, negative predictive value, and concordance rate of BALF liquid biopsy to tissue genotyping were 97.8%, 96.9%, 96.8%, 98.4%, and 97.7%, respectively. BALF liquid biopsy showed almost identical performance with standard tissue genotyping, while only 5 cases were mismatched each other. On the other hand, plasma liquid biopsy using cfDNA (n=110) revealed 48.5% sensitivity, 86.3% specificity, and 63.6% concordance rate in comparison to tissue genotyping. The mean turnaround times of BALF liquid biopsy was significantly shortened to 2.6±2.0 days in contrast to 13.9±12.4 days of tissue genotyping. (p<0.001). Conclusion BALF liquid biopsy can be developed as a novel platform for EGFR mutation testing in advanced NSCLC with almost identical performance with conventional tissue genotyping and significantly shorten TAT to 1-2 days. Citation Format: In Ae Kim, Jae Young Hur, Hee Joung Kim, Wan Seop Kim, Kye Young Lee. BALF liquid biopsy provides a novel platform for speedy and accurate EGFR mutation testing in advanced NSCLC patients [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 1196.

Intravascular Ultrasound-Guided Transvenous Biopsy of Abdominal and Pelvic Targets Difficult to Access by Percutaneous Needle Biopsy: Technique and Initial Clinical Experience

PurposeTo report initial clinical experience with intravascular ultrasound (US)-guided transvenous biopsy (TVB) for perivascular target lesions in the abdomen and pelvis using side-viewing phased-array intracardiac echocardiography catheters. Materials and MethodsIn this single-institution, retrospective study, 48 patients underwent 50 intravascular US-guided TVB procedures for targets close to the inferior vena cava or iliac veins deemed difficult to access by conventional percutaneous needle biopsy (PNB). In all procedures, side-viewing phased-array intracardiac echocardiography intravascular US catheters and transjugular liver biopsy sets were inserted through separate jugular or femoral vein access sheaths, and 18-gauge core needle biopsy specimens were obtained under real-time intravascular US guidance. Diagnostic yield, diagnostic accuracy, and complications were analyzed. ResultsIntravascular US-guided TVB was diagnostic of malignancy in 40 of 50 procedures for a diagnostic yield of 80%. There were 5 procedures in which biopsy was correctly negative for malignancy, with a per-procedure diagnostic accuracy of 90% (45/50). Among the 5 false negatives, 2 patients underwent repeat intravascular US-guided TVB, which was diagnostic of malignancy for a per-patient diagnostic accuracy of 94% (45/48). There were 1 (2%) mild, 2 (4%) moderate, and 1 (2%) severe adverse events, with 1 moderate severity adverse event (venous thrombosis) directly attributable to the intravascular US-guided TVB technique. ConclusionsIntravascular US-guided TVB performed on difficult-to-approach perivascular targets in the abdomen and pelvis resulted in a high diagnostic accuracy, similar to accepted thresholds for PNB. Complication rates may be slightly higher but should be weighed relative to the risks of difficult PNB, surgical biopsy, or clinical management without biopsy.

Extent of High-Grade Prostatic Adenocarcinoma in Multiparametric Magnetic Resonance Imaging-Targeted Biopsy Enhances Prediction of Pathologic Stage.

Multiparametric magnetic resonance imaging (mpMRI) of prostate with targeted biopsy has enhanced detection of high-grade prostatic adenocarcinoma (HG PCa). However, utility of amount of HG PCa (Gleason pattern 4/5) in mpMRI-targeted biopsies versus standard 12-core biopsies in predicting adverse outcomes on radical prostatectomy (RP) is unknown. To examine the utility of amount of HG PCa in mpMRI-targeted biopsies versus standard 12-core biopsies in predicting adverse RP outcomes. We performed a retrospective review of prostate biopsies that had corresponding RP, 1 or more mpMRI-targeted biopsy, and Grade Group 2 disease or higher. For the 169 cases identified, total millimeters of carcinoma and HG PCa and longest length HG PCa in a single core were recorded for 12-core biopsies and each set of mpMRI-targeted biopsies. For RP specimens, Gleason grade, extraprostatic extension, seminal vesicle involvement, and lymph node metastasis were recorded. The main outcome studied was prostate-confined disease at RP. A logistic regression model was used to test which pre-RP variables related to this outcome. Univariate analysis showed significant associations with adverse RP outcomes in 5 of 8 quantifiable variables; longest millimeter HG PCa in a single 12-core biopsy, highest Grade Group in any core, and total millimeter HG in mpMRI-targeted biopsies showed no statistical association (P = .54, P = .13, and P = .55, respectively). In multivariate analysis, total millimeter carcinoma in all cores, highest Grade Group in any core, and longest millimeter HG PCa in a single mpMRI-targeted core provided additional predictive value (P < .001, P = .004, and P = .03, respectively). Quantitation of HG PCa in mpMRI-targeted biopsies provides additional value over 12-core biopsies alone in predicting nonorgan confined prostate cancer at RP. Linear millimeters of HG PCa in mpMRI-targeted biopsies is a significant parameter associated with higher pathologic stage and could be of value in risk models.

Diagnosis of coeliac disease by flow cytometry of intraepithelial lymphocytes: a new ‘gold’ standard?

ObjectiveThe analysis of intraepithelial lymphocytes (IELs) by flow cytometry of duodenal biopsies—the ‘IEL’ lymphogram—has been proposed as a diagnostic test for coeliac disease. However, its clinical applicability has been limited...

For men enrolled in active surveillance, pre-biopsy biparametric magnetic resonance imaging significantly reduces the risk of reclassification and disease progression after 1 year

Aims To assess the level of disease progression at confirmatory staging biopsies after 1 year of active surveillance (AS) and compare the detection rate of significant prostate cancers (PCas) in patients who underwent pre-biopsy biparametric magnetic resonance imaging (bpMRI) before the first set of diagnostic transrectal ultrasonography-guided biopsies (TRUS-bx) with the detection rate in patients who did not undergo pre-biopsy bpMRI. Materials and methods Comparison of two patient groups enrolled in AS. Patients in Group A (n = 127) underwent pre-biopsy bpMRI followed by TRUS-bx ± targeted biopsies. Patients in Group B (n = 127) were enrolled in AS based on biopsy results from TRUS-bx only. Results Overall, 6% of the patients in Group A and 20% of the patients in Group B had an upgrade in Gleason grade from insignificant to significant PCa at confirmatory staging biopsies (odds ratio [OR], 3.5; p = .002; 95% confidence interval [CI], 1.6–7.9). Conclusions Patients who underwent pre-biopsy bpMRI before the first set of diagnostic biopsies had a reduced risk of reclassification and disease progression after 1 year of AS. Thus, pre-biopsy bpMRI improves the selection of men who should be enrolled in AS.

Use of Apixaban in Heart Transplant Patients Receiving Biopsies: A Case Series

Purpose The need for routine biopsies poses a challenge in the management of heart transplant recipients with an indication for anticoagulation. Per local institutional protocol, all patients receive at least 7 endomyocardial biopsies within 90 days of transplant to assess for rejection. Apixaban is a direct acting anticoagulant with a short half-life allowing for minimal time off anticoagulation to accommodate a biopsy. To our knowledge, apixaban has never been evaluated in the setting of routine biopsies. The objective of this study was to determine the safety and efficacy of apixaban in heart transplant patients undergoing biopsies. Methods This retrospective case series evaluated patients who received a heart transplant and were treated with apixaban within 90 days post-transplant. This research was approved by the Eastern Virginia Medical School IRB. Per institutional protocol, apixaban was held for 1-2 doses prior to routine biopsy and restarted the night following biopsy. The primary outcome was the occurrence of a bleeding or thrombotic event. Results A total of 13 patients received apixaban within 90 days following heart transplant. The median age was 54 years (IQR 37-59) with a median weight of 96 kg (IQR 72-101). None of the patients had either a bleeding or thromboembolic event while on apixaban. During therapy, patients underwent an average of 8 biopsies. The median time from transplant to initiation of apixaban was 39.5 ± 26.1 days (range 9-77). The number of patients on apixaban for their biopsies during post-op weeks 3-6 are as such: 3, 6, 6, 7. No patients required reversal therapy at any point and patients remained on therapy for a median of 276 (IQR 45-245) days. Indication for apixaban was stroke prevention with atrial fibrillation (15%), acute DVT (70%), and both (15%). Of the DVTs 30% were femoral while 70% were upper extremity. Compliance with the apixaban dosing protocol was excellent. Dosing was adjusted according to indication and renal function. Conclusion Based on the data from this case series, apixaban is safe to use for anticoagulation of heart transplant recipients undergoing routine biopsies in the setting of an established institutional protocol. Using apixaban allows for a short duration off anticoagulation to accommodate a biopsy without any increased risk of bleeding.

A135 ASSESSING THE HISTOLOGICAL QUALITY OF ENDOSCOPIC BIOPSY SAMPLES OBTAINED USING NOVEL MULTIBITE FORCEPS FROM A PORCINE GASTRIC SPECIMEN

Abstract Background Tissue sampling is often limited to acquisition of one to two biopsy samples during a single pass. The ability to obtain more than two biopsies during a single pass can improve diagnostic yield however is potentially limited by poor specimen quality and loss of specimens. The multibite forceps used in this study have a unique geometry with the ability to store up to six biopsy samples taken consecutively with easy removal of the samples when shaken in solution. If multiple biopsies can be taken during a single pass with preserved specimen quality then we can reduce procedure time, improve efficiency and sensitivity of biopsies. Aims To evaluate the histological quality of the first biopsy sample compared to the last (sixth) biopsy sample acquired consecutively with the multibite forcep during a single act. Methods A porcine stomach was coloured with surgical dye to create six separate segments. An experienced endoscopist used single use disposable MultiCROC multi-sampling biopsy forceps to acquire six consecutive biopsies. Biopsies were manually separated into the order of which they were acquired (biopsy one through six) and each sample was placed in formalin solution. A total of 35 sets of 6 biopsies were obtained producing a total of 210 samples. Samples were randomized and two independent pathologists who were blinded to the biopsy order assessed the histological quality of specimens. Specimens were evaluated for presence of full thickness mucosa, absence of fragmentation, crush artifact and diagnostic utility. Each pathologist then scored each specimen and the mean scores were used to compare the histological quality of the first biopsy vs. the sixth biopsy for each set. Results Our preliminary results include 12 of the 35 sets of biopsies. Using a paired sample t test, there was no significant difference between the mean score given to biopsy one and biopsy six for all twelve pairs [3.62 (SD 1.46) vs. 3.67 (SD 1.15), correlation factor=0.498 and p=.10). There was no significant difference between the first and sixth biopsy when comparing the presence of full thickness mucosa [0.59 (SD 0.49) vs. 0.59 (SD 0.44), p=.086], absence of fragmentation [0.50 (SD 0.50) vs. 0.73 (SD 0.34), p=0.06], absence of crush artifact [0.96 (SD 0.15) vs. 0.91 (SD 0.30), p=0.77), and specimen size [1.64 (SD 0.92) vs. 1.70 (SD 0.65), p=0.56). Conclusions No significant differences were found between the histological quality of the first biopsy and the sixth biopsy. Additional parameters such as specimen size, full thickness mucosa, absence of fragmentation and absence of crush artifact revealed no significant differences between the first and sixth biopsy. This preliminary data thus far shows that there is no difference between the histological quality when multiple biopsies are retrieved consecutively. Funding Agencies NoneNone

Multiparametric MRI may Help to Identify Patients With Prostate Cancer in a Contemporary Cohort of Patients With Clinical Bladder Outlet Obstruction Scheduled for Holmium Laser Enucleation of the Prostate (HoLEP).

Objective: To investigate the value of standard [digital rectal examination (DRE), PSA] and advanced (mpMRI, prostate biopsy) clinical evaluation for prostate cancer (PCa) detection in contemporary patients with clinical bladder outlet obstruction (BOO) scheduled for Holmium laser enucleation of the prostate (HoLEP).Material and Methods: We retrospectively analyzed 397 patients, who were referred to our tertiary care laser center for HoLEP due to BOO between 11/2017 and 07/2020. Of those, 83 (20.7%) underwent further advanced clinical PCa evaluation with mpMRI and/or prostate biopsy due to elevated PSA and/or lowered PSA ratio and/or suspicious DRE. Logistic regression and binary regression tree models were applied to identify PCa in BOO patients.Results: An mpMRI was conducted in 56 (66%) of 83 patients and revealed PIRADS 4/5 lesions in 14 (25%) patients. Subsequently, a combined systematic randomized and MRI-fusion biopsy was performed in 19 (23%) patients and revealed in PCa detection in four patients (5%). A randomized prostate biopsy was performed in 31 (37%) patients and revealed in PCa detection in three patients (4%). All seven patients (9%) with PCa detection underwent radical prostatectomy with 29% exhibiting non-organ confined disease. Incidental PCa after HoLEP (n = 76) was found in nine patients (12%) with advanced clinical PCa evaluation preoperatively. In univariable logistic regression analyses, PSA, fPSA ratio, and PSA density failed to identify patients with PCa detection. Conversely, patients with a lower International Prostate Symptom Score (IPSS) and PIRADs 4/5 lesion in mpMRI were at higher risk for PCa detection. In multivariable adjusted analyses, PIRADS 4/5 lesions were confirmed as an independent risk factor (OR 9.91, p = 0.04), while IPSS did not reach significance (p = 0.052).Conclusion: In advanced clinical PCa evaluation mpMRI should be considered in patients with elevated total PSA or low fPSA ratio scheduled for BOO treatment with HoLEP. Patients with low IPSS or PIRADS 4/5 lesions in mpMRI are at highest risk for PCa detection. In patients with a history of two or more sets of negative prostate biopsies, advanced clinical PCa evaluation might be omitted.

Abstract PS4-09: Pathologic features of the inter-regimen biopsy predict response to neoadjuvant therapy in the I-SPY 2 TRIAL

Abstract Background: I-SPY 2 is a neoadjuvant platform trial open to patients with locally advanced, molecular high-risk breast cancer. In a concerted pursuit of mid-therapy response biomarkers, we evaluated inter-regimen biopsies, to identify patients who may be candidates for treatment de-escalation. In a pilot study, we observed that absence of carcinoma in an inter-regimen biopsy may predict pathologic complete response (pCR). In this expanded study of 100 participants, we sought to confirm that finding and assess pathologic features of the inter-regimen biopsy as predictors of tumor response to neoadjuvant therapy. Methods: Digital H&amp;E images of 100 inter-regimen (12 week) image-guided breast biopsies +/- ancillary immunohistochemistry (p63 and/or cytokeratin) were reviewed by 9 I-SPY affiliated pathologists to record 1) tumor bed and 2) presence/absence of residual invasive carcinoma (IC) (with tumor cellularity scored as 0-100%). The data set included 393 cores (mean 3.9 (2-4) cores/biopsy). Fisher’s exact t-test was used for association of presence/absence of IC with pCR, and tumoral hormone receptor (HR) and HER2 status. Association between biopsy tumor cellularity and residual cancer burden (RCB) indices used Pearson’s correlation. Results: In the biopsy set, 84 (84%) had ≥80% inter-observer diagnostic agreement on both 1) presence of tumor bed and 2) presence/absence of IC (53 IC+ /31 IC-). IC+/IC- biopsies had equal numbers of evaluable tissue cores. The primary tumors were 63% HR+/37% HR-. The presence of IC in the biopsy correlated with tumoral HR/HER2 status (p=0.0014: 74%: HR+HER2-; 62%: TN; 60%: HR+HER2+; 10%: HR-HER2+). Of 31 patients with IC- biopsies, 25 (80%) went on to pCR, whereas only 7/53 (13%) of patients with IC+ biopsies had pCR, conferring an odds ratio for pCR of 26, Fisher p=7.5E-10. Overall, IC- biopsies had a positive predictive value (PPV) for pCR of 81%, with a PPV for HR- tumors of 94% vs. 67% for HR+ tumors (Table 1). In the 6 IC- biopsies from patients with non-pCR (“false-negatives”), most were HR+ (5/6, Table 1), and tumor bed size in the resection specimen was smaller than for IC+ biopsies with non-pCR: 276 mm2 (0.4-1000 mm2) vs. 1166 mm2 (1-11960 mm2). In contrast, the 46/53 IC+ biopsies in patients with non-pCR had a PPV for predicting non-pCR of 86%, (PPV for HR+ tumors: 94% vs. PPV for HR- tumors: 66%. Tumor cellularity in the biopsy (mean 37%, [2.5-93%]) did not correlate with RCB index (p=0.57) or RCB breast-only index (p = 0.17) at resection. Conclusion: In this 100 biopsy set from the I-SPY2 trial, the absence of residual carcinoma in inter-regimen biopsies was highly predictive of pCR, particularly for HR- tumors. The “false-negative” biopsies (IC-/non-pCR) were predominantly HR+ tumors with small residual tumor beds at resection. Conversely, the presence of carcinoma in inter-regimen biopsies was highly predictive of non-pCR, particularly for HR+ tumors. These data demonstrate the utility, and the limitations, of the inter-regimen biopsy as one tool to identify patients who may benefit from therapeutic de-escalation. Table 1: PPV for pCR/non-PCR by Inter-regimen Biopsy StatusInter-regimen biopsy with or without Invasive carcinoma (IC+/-)pCRnon-pCRPPV (Sensitivity) for pCR(IC- Biopsies)PPV (Sensitivity) for non-pCR(IC+ biopsies)IC- biopsiesAll25681% (78%)-HR+10567% (83%)-HR-15194% (75%)-IC+ biopsiesAll746-86% (88%)HR+236-94% (88%)HR-510-66% (91%) Citation Format: Jodi M Carter, Molly E Klein, Sara J Venters, Kimmie Rabe, I Tolgay Ocal, Kamaljeet Singh, Denise M Wolf, Sunati Sahoo, Shuko Harada, Laila Khazai, Malini Harigopal, Alexander D Borowsky, Gregor Krings, Ronald Balassanian, Yunn-Yi Chen, Kimberley Cole, Sonal Shad, Amy Delson, Lamorna Brown-Swigart, I-SPY 2 TRIAL Consortium, Laura Esserman, Laura van ‘t Veer, W Fraser Symmans. Pathologic features of the inter-regimen biopsy predict response to neoadjuvant therapy in the I-SPY 2 TRIAL [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 PS4-09.

Abstract PS18-10: Intratumoural heterogeneity in PgR expression: Molecular and prognostic significance

Abstract Background: PgR is only expressed significantly in ER+ breast cancers and is generally considered a classical estrogen-sensitive protein, however, while ER+ cells are almost always expressed in a non-clustered pattern across the tumour bed, PgR+ cells form clusters or clumps with surrounding areas of PgR- cells when assessed by IHC in about 20-25% of PgR+ tumours. This is an ill-described and unexplained feature of intratumoural heterogeneity of a widely measured biomarker. The prognostic significance of the PgR clumpiness is also unknown. Aims: to determine the (i) biologic and (ii) prognostic significance of PgR clumpiness in ER+ primary breast cancer. Methods: (i) Biologic study: 40 primary ER+PgR+ tumours were identified with distinct PgR clumpiness by IHC. Areas of PgR+ and PgR- cells (PgR-rich and PgR-poor areas, respectively) were needle microdissected from unstained sections by juxtaposing them with PgR-stained sections from the same tumour. Areas were also dissected from 8 homogeneous PgR+ and 8 PgR- tumours for reference. NanoString gene expression analysis using a 50-gene code set (45 oestrogen-responsive or proliferation genes + 5 housekeeping genes) was performed on RNA extracts from the samples. (ii) Prognostic study: we developed a PgR heterogeneity score (PgR Het-score, range 0-18) as the product of (A) the number of PgR+ clumps/cm2 of tumour area (range 0 to 6) and (B) the level of overall PgR-positivity (0-4% or 96-100%, score 0; 5-19% or 80-95%, score 1; 20-39 or 60-79%, score 2; 40-59%, score 3). PgR Het-scores were derived from images (Hamamatsu NanoZoomer-XR) of PgR-stained sections from two sets of tumour biopsies taken at diagnosis after which all patients received adjuvant endocrine therapy: 322 tumours from the Royal Marsden and from Southern Sweden collected for a case:control study of risk of recurrence (recurrence:no recurrence, 1:1) and 591 patients from the TransATAC cohort study with time to recurrence as the end-point. Results: (i) Biologic study: In unsupervised hierarchical clustering of RNA expression from all samples, 33/40 pairs of PgR-rich and PgR-poor areas from a single tumour paired together and all 8 positive and negative control samples were grouped separately. Eight genes were differentially expressed between the PgR-rich and -poor areas (FDR&amp;lt;0.05: PGR, SERPINA3, AURKA, MSMB, PDZK1, IGSF1, FKBP5, SLC2A3). PGR had a 35-fold difference indicating that transcriptional differences explained the PgR IHC differences. The progesterone-regulated genes FKBP3 and SERPINA3 both showed significantly higher expression in PgR-rich areas consistent with the PgR differences having functional impact. AURKA was also more highly expressed in the PgR-rich areas indicating that the PgR-rich areas may also be more proliferative. ER and most of known estrogen-regulated genes were not differentially expressed and therefore show an unexpected divergence from PgR expression and regulation downstream of ER. (ii) Prognostic study: The PgR Het-score showed no consistent association with clinical factors including tumour size, nodal status, grade, age. Each of the individual components as well as the composite PgR Het score were significantly associated with risk of recurrence in POLAR: (A) p=0.02; (B) p=0.002; (PgR Het score) p=0.008. However, there was no association of the PgR Het score with risk of recurrence in TransATAC. Conclusion: Major, functionally significant differences in PgR expression occur within some ER+ tumours that are not explained by differences in ER expression and are not associated with differences in expression of most other estrogen-dependent genes. Further work is on-going to provide a mechanistic explanation for the PgR heterogeneity. Given the discordance in results from the two clinical studies its prognostic significance is uncertain. This work was funded by BCRF. Citation Format: Lila Zabaglo, Richard Buus, Eugene Schuster, Belinda Yeo, Marie Klintman, Ivana Sestak, Jack Cuzick, Ian Smith, Mitch Dowsett. Intratumoural heterogeneity in PgR expression: Molecular and prognostic significance [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 PS18-10.

Differential Diagnosis of New-Onset Pulmonary Ground Glass Changes During the COVID-19 Pandemic: a Case Report

The paper describes a clinical case of hypersensitivity pneumonitis in a young patient during the COVID-19 pandemic when the patient having complaints of dyspnea and interstitial lung changes as ground glass opacity was initially unambiguously diagnosed with viral COVID-related bilateral pneumonia. The chief computed tomography manifestations in the lung were infiltrative inflammatory changes as numerous areas of predominantly interstitial infiltration by the type of ground glass, with consolidation areas and reticular changes, varying in extent, peripheral localization, mainly in the lower lobes and with the 25–50% involvement of the right and left lung parenchyma. The diagnosis of hypersensitivity pneumonitis was confirmed by cytological and histological examinations of the biopsy material obtained during bronchoscopy using a set of biopsies and endobronchial cryobiopsy.This clinical case demonstrates that the chest computed tomography detection of pulmonary ground glasstype changes is a reason for further patient examinations and requires morphological verification of the diagnosis, since the cost of a diagnostic error is the life of a patient.

UNIQUE GENE EXPRESSION IN GUT-DERIVED PHAGOCYTIC IMMUNE CELLS FROM IBD PATIENTS

Abstract Introduction/Aim Inflammatory bowel diseases (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), have a complex immunopathogenesis involving chronic, dysregulated inflammation of the gut in response to commensal microbes. Current therapies, such as Anti-TNF drugs, target this immune dysregulation, but many patients exhibit recurrent inflammation despite treatment (refractoriness). Research shows that IBD patient phagocytic immune cells have altered cytokine production and bacterial clearing defects. We thus set out to analyze the transcriptome of phagocytic cells (CD11b+) isolated from patient gut biopsies. To shed light on potential causes of refractoriness, we examined cells from both not inflamed (successfully treated) and inflamed (refractory) patients on Anti-TNF therapy. Methods We used magnetic sorting to isolate CD11b+ cells from the lamina propria of a diverse set of IBD patient biopsies and performed RNA sequencing (n = 61). We used differential expression analysis to compare combinations of four patient biopsy variables: location (ileum or colon), condition (inflamed or uninflamed), IBD type (CD or UC), and patient treatment (Anti-TNF therapy or other). We also performed pathway analysis on genes significantly upregulated or downregulated (p &amp;lt; 0.05) between these variables. Results The highest number of differentially expressed genes was found in the colon (5578 genes upregulated) vs ileum (5582). Genes upregulated in colon mapped to innate immune pathways, while ileum pathways were mainly metabolic. Only 29 genes were differentially expressed in UC vs CD, regardless of inflammation status or location. 26 genes were differentially expressed between inflamed and uninflamed biopsies on Anti-TNF therapy (n = 16). Genes upregulated in inflamed biopsies were associated with neutrophils, while genes in uninflamed biopsies were associated with adaptive immunity, short chain fatty acid synthesis, transcription and translation, and protein secretion. Conclusion CD11b+ cells of the colon are more immunologically active, likely due to their interaction with the dense microbiome of the colon. In contrast, the metabolic pathways upregulated in ileum may point to a role for phagocytes in digestion and metabolism. These results suggest that intestinal CD11b+ gene expression is highly influenced by regional microenvironment. Fewer differences were seen comparing CD11b+ cells from CD vs UC, suggesting that this cell population is not solely responsible for the clinical differences between these diseases. The small number of differentially expressed genes in Anti-TNF refractoriness shows a clear difference between innate and adaptive immune pathways, as well as increased general cell activity in Anti-TNF responders. These genes should be further investigated to determine their precise role in refractoriness.

Specimen Radiography for MRI-Guided Biopsies

Background: Specimen radiography is important for the biopsy of breast microcalcifications, and MRI is limited in the detection of microcalcifications. It is unknown whether or not the presence of microcalcifications on MRI-guided biopsies is significant. Purpose: To determine whether specimen radiography of MRI-guided biopsy samples provides any added benefits in tissue assessment. Materials and Methods: This is an IRB-approved, HIPPA-compliant retrospective review of MRI-guided biopsy reports whose tissue underwent specimen radiography from 2010 to 2017. Pathology reports were queried to compare samples with and without calcium and reviewed to determine if calcifications correlated with the lesion of interest. If there was a correlation, the original MRI was reviewed. Final pathology reports were also reviewed if excision was performed. Results: A total of 889 patients ages 22 - 85 were included with 140 (15.7%, 140/889) containing calcifications. Of 140 specimens, 119 (85.0%, 119/140) cases separated the calcifications. A total of 41 (34.5%, 41/119) were malignant or high-risk lesions/atypia of which 15 (36.6%, 15/41) showed a higher-grade lesion in the specimen containing calcium. Out of these 15, 4 (26.7%, 4/15) were pathologically associated with calcium; however, pathologic diagnosis was not dependent on the presence of calcifications. All 4 were high-risk lesions and none were malignancies. MRI in these cases showed three enhancing masses and one non-mass enhancement. None were upgraded at excision. Conclusion: The presence of microcalcifications on MRI-guided biopsies does not aid in tissue assessment and does not impact pathologic diagnosis. Specimen radiography provides no added benefits in the setting of MRI-guided biopsies.

Terminal Deoxynucleotidyl Transferase Commonly Expresses in Germ Cell Tumors: Evaluation on a Large Series from Multiple Centers.

AimThe concrete features of expression of terminal deoxynucleotidyl transferase (TdT) are needed to be revealed in male and female germ cell tumors (GCTs).MethodsTdT immunostaining was performed in 195 GCTs, and the tumor and/or tumorous components included seminomas, germ cell neoplasias in situ (GCNISs), dysgerminomas, embryonal carcinomas (ECs), extragonadal germinomas, yolk sac tumors (YSTs), teratomas, and spermatocytic tumors. Twenty-one sex cord-stromal tumors were also added. Expression of the classical germ cell tumor markers (PLAP, OCT4, SALL4, CD117, and D2-40) was compared to that of TDT.ResultsNearly all (tumors or tumorous components) seminomas (99%, 107/108), GCNISs (98%, 51/52), dysgerminomas (94%, 17/18), ECs (100%, 15/15), and extragonadal germinomas (100%, 11/11) were positive for TdT. None of the cells in YSTs (0/38), teratomas (0/19), spermatocytic tumors (0/1), or sex cord-stromal tumors (0/21) were immunoreactive for TdT staining. The normal testicular and ovarian gonadal tissues were also negative for TdT. However, TdT presented with significant loss of antigen immunoreactivity in the paraffin-embedded tissues older than 3 years, giving rise to weak or moderate staining in a subset of cases. The expressions of TdT showed no significances with PLAP, OCT4, SALL4, CD117, and D2-40 during the diagnosis of the most GCTs (P>0.05), except for with PLAP, SALL4, or CD117 in YST (P=0.000 each), and D117 (P=0.000) or D2-40 (P=0.006) in ECs.ConclusionOur findings further verify that TdT can serve as a new GCT marker for seminomas, GCNISs, dysgerminomas, ECs, and extragonadal germinomas, with a highly positive rate. Awareness of TdT positivity in GCTs contributes to the prevention of erroneous diagnoses, particularly in the setting of core needle biopsies. To determine the properties where TdT staining may not be apparent in some old archived paraffin-embedded tissues, one could circumvent the potential misinterpretations of false-negative immunohistochemistry results.

A single-center experience: Liver biopsy results during a year.

Background and Aim:Liver biopsy is the gold standard method for the diagnosis and treatment of liver diseases. In this study, we aimed to evaluate the results of liver biopsies performed in a year in our clinic. In addition, we also aimed if these liver biopsies could reveal the etiology of liver disease in patients with elevations of transaminases or/and alkaline phosphatase levels or liver masses.Materials and Methods:Patients who had liver biopsies for persistently elevated transaminases or/and alkaline phosphatase levels, protocol biopsies after liver transplantation, or liver masses in our hepatology clinic between 2011 and 2012 were included in the study. Liver biopsy decisions were made by experts during the hepatology council. Liver biopsies were previously performed using classical percutaneous liver biopsy or ultrasonography-guided Sonocan® liver biopsy sets. The pathology results of liver biopsies and clinical data of the matching patients were obtained from the liver biopsy record archives and patient files, respectively.Results:Totally, 479 liver biopsy results (male=252, 52.6%, mean age 49±14.5 years) were evaluated in the study. Of these patients, 432 (male=228) underwent percutaneous liver biopsy and 47 (male=24) underwent Sonocan® needle biopsy. The most common histopathologic diagnoses in the percutaneous liver biopsy group were chronic hepatitis B (n=127, 29.4%), normal histopathological findings (n=50, 11.6% and 32 of them were protocol biopsies after liver transplantation), and nonalcoholic steatohepatitis (NASH, n=41, 9.5%). The most common histopathologic diagnoses in the Sonocan® group were 25 liver metastasis out of 29 liver tumors (n=25, 53.2% of all) chronic hepatitis B (n=5, 10.6%), and NASH (n=3, 6.4%).Conclusion:In this study, diversity in liver biopsy results indicates the importance of histopathological evaluation. The most prevalent pathology in the liver biopsies was chronic hepatitis B, which is the most common chronic liver disease in Turkey. The metastatic liver tumor was the most common among the liver masses.

Prognostic significance of morphological patterns of Gleason grade 5 prostatic adenocarcinoma diagnosed on needle biopsy

Grading is one of the best prognostic indicators of prostate cancer with Gleason grade 5 having the worst outcome. The prognostic influence of grade 5 patterns remains uncertain. A total of 646 prostate needle biopsy sets with Gleason score (GS) 9-10 prostatic adenocarcinoma were prospectively analysed. Patterns of grade 5 were correlated with radical prostatectomy (RP) adverse findings of high tumour volume (TV), extra-prostatic extension (EPE), seminal vesicle invasion (SVI) and lymph node involvement (LNI) in 472 and biochemical recurrence (BCR) in 338 patients after RP. Mean age and serum PSA were 69 years (range 37-91) and 26.1 ng/mL (range 1.4-1800), respectively. Gleason scores were 4+5=9 in 539 (83%), 5+4=9 in 94 (15%) and 5+5=10 in 13 cases (2%). Clusters/cords, single cells, sheets and comedocarcinoma were found in 86%, 69%, 26% and 18% with a pure pattern in 25% of cases. Comparing cases with and without sheets, there were no significant differences with RP high TV (p=0.8577), EPE (p=0.5372), SVI (p=0.5183) and LNI (p=0.4323). However, the presence of sheets predicted a significantly higher BCR rate (p=0.0033), while for tumours with single cells, the interval to BCR was significantly shorter (p<0.0001). Comparing cases with and without the other patterns, two other significant differences were found. Comedocarcinoma predicted high TV (p=0.0230) and single cells predicted EPE (p=0.0101). This study shows that all patterns currently used to assign a Gleason grade 5, including sheets, comedocarcinoma, single cells and clusters/cords, are associated with aggressive outcomes validating their inclusion in grade 5.

An Unexpected Esophageal Inflammatory Lesion

Question: An 81-year-old man, operated 5 years earlier for a squamous cell carcinoma of the lower third of the esophagus (pT1), had a surveillance endoscopy which showed a circumferential erosive lesion (grade 3 esophagitis), located above the gastro-esophageal anastomosis (Figure A). A suspicion of squamous high-grade dysplasia was raised based on the first set of biopsies, which were difficult to interpret given the prominent associated inflammation. Therefore, additional biopsies were taken and sent to our department.

Tumor and microenvironment response but no cytotoxic T-cell activation in classic Hodgkin lymphoma treated with anti-PD1

AbstractClassic Hodgkin lymphoma (cHL) is the cancer type most susceptible to antibodies targeting programmed cell death protein 1 (PD1) and is characterized by scarce Hodgkin and Reed-Sternberg cells (HRSCs), perpetuating a unique tumor microenvironment (TME). Although anti-PD1 effects appear to be largely mediated by cytotoxic CD8+ T cells in solid tumors, HRSCs frequently lack major histocompatibility complex expression, and the mechanism of anti-PD1 efficacy in cHL is unclear. Rapid clinical responses and high interim complete response rates to anti-PD1 based first-line treatment were recently reported for patients with early-stage unfavorable cHL treated in the German Hodgkin Study Group phase 2 NIVAHL trial. To investigate the mechanisms underlying this very early response to anti-PD1 treatment, we analyzed paired biopsies and blood samples obtained from NIVAHL patients before and during the first days of nivolumab first-line cHL therapy. Mirroring the rapid clinical response, HRSCs had disappeared from the tissue within days after the first nivolumab application. The TME already shows a reduction in type 1 regulatory T cells and PD-L1+ tumor-associated macrophages at this early time point of treatment. Interestingly, a cytotoxic immune response and a clonal T-cell expansion were not observed in the tumors or peripheral blood. These early changes in the TME were distinct from alterations found in a separate set of cHL biopsies at relapse during anti-PD1 therapy. We identify a unique very early histologic response pattern to anti-PD1 therapy in cHL that is suggestive of withdrawal of prosurvival factors, rather than induction of an adaptive antitumor immune response, as the main mechanism of action.

Variability in the Distribution of Histological Disease Activity in the Colon of Patients with Ulcerative Colitis.

Histological activity scores have been developed and validated. However, data on the distribution of histological inflammation within one segment in patients with ulcerative colitis [UC] are lacking. This impacts on the reliability of histological activity scores. The aim of this study was to assess the variability in histological activity within one endoscopic segment in patients with UC. Biopsies were taken in sequential patients with UC in three adjacent contiguous regions within a macroscopically homogeneous colonic segment. Biopsies were scored for Geboes score [GS], Robarts histological index [RHI] and Nancy histological index [NHI]. Variability was assessed by Kappa statistics for categorical outcomes and intraclass correlation coefficient [ICC] for continuous outcomes. A total of 161 biopsy sets from 55 endoscopic segments of 21 patients were analysed. Endoscopically active disease was present in 45% of segments. The continuous histological scores showed excellent agreement between the different regions. The ICC for RHI in all segments was 0.974 (95% confidence interval [CI] 0.958-0.984; p < 0.0001) and 0.98 [95% CI: 0.968-0.988; p < 0.0001] for the numerically converted GS. The categorical NHI showed higher variability: κ = 0.574 [95% CI: 0.571-0.577; p < 0.0001]. In all segments the highest variability was seen in samples with NHI = 2. When dichotomizing based on histological remission, substantial agreement was seen for all scores, with κ > 0.734 for all cut-offs. The homogeneity in the distribution of histological disease activity was comparable between colonic segments. The distribution of histological disease activity in UC follows a homogeneous pattern in different locations of one segment.

Interobserver Variability in Assessment of Renal Mass Biopsies.

The main goal of this study was to assess the histopathological efficacy of renal mass biopsy and to check the concordance between pathological results and biopsy of the final specimen, as well as interobserver variability in the assessment of biopsy cores. A hundred sets of core biopsies of postoperative specimens (renal masses) have been performed. Three core biopsies of the intact specimen had been performed once the kidney with the tumor, or the tumor alone were resected. The urologist aimed to obtain two cores from the peripheral sides of the tumor and one core from its center. The surgical specimen was evaluated by a single pathologist, whereas biopsy samples were referred to three independent pathologists who were blinded to the final results of the renal mass biopsy. Nondiagnostic biopsy rates ranged from 13% to 22%. Sensitivity and specificity ranged 83-97% and 97-99% by excluding nondiagnostic results. The concordance between assessment of surgical specimen and biopsy in the Fuhrman grading system ranged 36.5-77.0%, respectively. Interobserver agreement between the three pathologists was substantial or moderate, depending on the tumor subtype. The Krippendorff's alpha coefficient, calculated by excluding the nondiagnostic results, was 0.28 (moderate agreement) for the Fuhrman grading system. The agreementregarding grading of biopsies between threepathologists ranged from moderate to substantial.Therefore, a team ofdedicated uropathologists should be engaged in final diagnosis of renal mass biopsy rather than single one before implementing the proper treatment.